Analysis of Products Marketed as Henna for Hair: Part Two

Table of Contents

Introduction

This article is the second in a series comparing products sold as henna for hair. The purpose of this series is to describe and compare henna for hair products commonly found online on sites such as Amazon or eBay as well as in ethnic grocery stores, health food stores, and import stores in the USA.

Part One introduced objectives and methodology. It compared products that are marketed as either “pure henna,” or which are described as henna blended with other natural herbs for a red hair color result. To read part one, click here.

This article, Part Two, will continue using the same format as Part One. Part Two examines “henna for hair” products claiming to color hair brown or brunette. First, product labels are analyzed. Second, the plant powder is described in terms of texture (sift), color, and odor. After mixing, the qualities of the resulting pastes are also described. Finally, each past sample is tested using paper chromatography to determine the presence of added dyes. The products are compared to Ancient Sunrise pure henna and indigo powders.

Many products labeled and sold as henna contain additional ingredients, either plant-based or synthetic. Products whose labels claim to be “100% natural” or who use similar terms often fail to disclose all ingredients. Many products are imported from countries whose regulations are less stringent or loosely enforced. Additionally, products vary in quality. Poor sift results in powders containing larger pieces of plant particulates as well as sand. Products with poor packaging will become stale more quickly, causing the dye to be less effective. In many cases, products marketed as natural and safe in all actuality contain added dyes such as azo-dyes, metallic salts, and parapheneylene-diamine (PPD). To learn more about the difference between BAQ henna, compound henna, and other products claiming to be henna, read this article.

The FDA has a standard for henna and products entering the United States labeled as a “henna” hair coloring product. These guidelines do not appear to be regularly enforced as are regulations for products labeled as henna for use on skin. The result is that many products labeled as “henna for hair” which do not meet the guidelines make their way into the United States with relative ease.

The FDA forbids the sale of and is empowered to confiscate of any henna product labeled for skin use, or products showing images of henna used on the skin. Customs and border protection is empowered to search the importing company’s website to determine if henna is intended for use on skin, and may seize and destroy henna that appears to be imported for use on skin.

“Black henna,” or products labeled as henna containing PPD, have been known to cause skin reactions and sensitization. For more information about black henna, read the articles “Henna is Not Black,” “What You Need to Know About PPD,” and Chapter One of the Ancient Sunrise E-book.

The purpose of these studies is to determine the quality of those products in comparison to Ancient Sunrise® Henna for Hair products and to test for the existence of dye additives. This series investigates the following categories:

  • Part One: “Pure” henna, herbal henna mixes, and red result henna for hair products
  • Part Two (this article): Brunette result henna for hair products
  • Part Three: Black result henna for hair products

Premixed “Henna for Hair” Products

All of the products analyzed in this section claim to contain a blend of multiple plant dye powders and in some cases additional synthetic ingredients. All claim to contain henna. Most claim to contain indigo. Because henna and indigo require separate mixing and dye release processes, Ancient Sunrise packages plant powders separately. The most effective method for mixing henna and indigo involves mixing henna first with an acidic component and allowing time for dye release, normally 8-12 hours at room temperature. Indigo does not require an acidic component nor dye-release time because the dye releases and demises rapidly. It is mixed with only water and then combined with dye released henna just before applying to the hair. To learn more about the proper way to mix henna and indigo for brunette results, read this article.

Premixed “henna for hair” products may provide a simpler mixing process but in turn sacrifice full coverage and color permanence. Most of the products in this article recommend mixing the powder with water to form a paste and applying immediately. This is most likely due to indigo dye’s rapid demise. Without proper dye-release, the lawsone and indigo dye molecules will not bind to the hair as effectively. The result may be lighter than desired and will fade over time.

Finally, many of the following products report a number of “ayurvedic herbs” in their ingredients in addition to plant dye powders. Most of these herbs do not affect the color result but claim to condition the hair and scalp and/or promote hair growth. Please note that this article is not meant to support or deny the effectiveness of these claims. Descriptions of the herbs should not be interpreted as recommendations.

Sample Selection and Label Analysis

Eight “henna for hair” products were selected for this study based on the following criteria:

  • 1: The product is marketed as a “henna for hair” product or a plant-based product containing henna which is meant for coloring hair.
  • 2. The product claims to color hair a shade of brown or brunette.

The traditional method for achieving a brunette result using plant dye powders involves a combination of henna and indigo. Many of the products selected for this article include both henna and indigo in their ingredients disclosures. These products will be compared with Ancient Sunrise Rajasthani Twilight Henna and Ancient Sunrise Zekhara Indigo plant dye powders.

Below are the images of the labels for the selected samples. In the future, each sample will be referred to by its assigned number.

#1

This product is made in the United States and is widely available at health stores and online. The labeling and packaging are thorough in comparison to many “henna for hair” products and are reminiscent of the labeling and packaging of box hair dye. The color on the label is “chestnut.”

The ingredients listed are “henna, indigo, centaurea, rhubarb, and beetroot.” Centaurea, also known as cornflower, can be used as a natural blue dye for fabric. Whether or not it is an effective dye for hair is unknown. Most natural fabric dyeing processes involve boiling and the use of a mordant. Similarly, rhubarb and beetroot have been used to dye fabrics yellow and red respectively. To learn more about what ingredients do or do not effectively color hair, read “Does it Dye Hair?” Rhubarb is acidic and may aid in dye-release, but the instructions do not include a dye-release period.

The internal pamphlet offers detailed instructions and information regarding application, patch testing, and a warning against use near the eyes. The instructions are to mix the powder with water, apply, and leave for one hour.

#2

This product is made in India. It claims to be for dark hair and gives hair a “rich brown color and shine.” The ingredients listed are as follows: “Mehndi (Lawsonia alba), Aam Beej (Mangifera indica), Neem (Azadirachta indica), Arjun (Terminalia arjuna), Gambhari (Gmelina arborea), Daruhaldi (Berberis aristata), Kikkar gaund (Acacia arabica).” While henna is listed as the first ingredient, indigo is not mentioned at all.

The product is described as “ayurvedic,” which is a term relating to an alternative system of medicine often involving a number of herbs native to South Asia. Aam beej is mango seed. This is not a dye but is meant to condition hair. Ancient Sunrise supplies mango seed butter here. Neem is another popular ayurvedic herb used for hair conditioning in either a powder or oil form. Neem does not dye hair. Arjun is an ayurvedic herb. It appears to have potential dyeing properties on fabric but needs a mordant. Its ability to dye hair is unknown. Gambhari, also known as English Beechwood, is used in ayurvedic medicine and claims to condition hair and stimulate growth. It does not dye hair. Daruhaldi is also known as Indian Barberry. The plant is used to dye fabrics and tan leather due to its high level of tannins. Its effectiveness in coloring hair is unknown. Kikkar gaund, also known as Gum Arabic, is both eaten and used in topicals. Because it contains a high amount of mucilage, it can act as a thickener or binder when mixed with water. It is a common ingredient in both cosmetics and food. It does not have dye properties.

Like sample #1, this product’s instructions recommend mixing the powder with water. It recommends leaving the paste in the hair for half an hour for conditioning, and one hour or longer for “maximum color highlights.” In one Amazon review, a customer said that the product made her hair “really red.” It is likely that the absence of indigo and the ineffectiveness of the other herbs meant for coloring caused the result to be red rather than brunette.

#3

 

This product is from India. The color result advertised on the label is “dark brown.” The ingredients are as follows: “Indigofera tinctoria, Lawsonia Inermis, Embelica officianalis, Eclipta alba, Azadiracta indica, Bacopa monnieri, Vetiveria zizaniodes. The first three ingredients are indigo, henna, and amla powder respectively. A dark brunette mix should have a higher proportion of indigo to henna. Amla powder functions as an acid and aids henna/indigo mixes in binding to the hair effectively for a darker result. Eclipta alba, also known as bhringraj, is an ayurvedic herb known for its potential hair growth properties. Azadiracta indica, as mentioned before in sample #2, is also known as neem. Bacopa monnieri, also known as brahmi, is an ayurvedic herb that claims to promote hair growth. Finally, vetiveria zizaniodes is also called vetiver, which is another ayurvedic herb meant to condition hair and stimulate growth. Vetiver has a pleasant scent. Ancient Sunrise carries a soap bar which contains cardamom and vetiver. Except for the initial three ingredients, the remaining reported ingredients are ayurvedic herbs without dyeing properties.

The package includes gloves, a plastic cap, and an instructional pamphlet. It includes warnings about sensitivity and instructions for patch testing. The product warns about potential headaches and itching. This is most likely in reference to the reaction to indigo powder, which causes some people to have discomfort. To learn more about plant dye powder allergies, read “Plant Dye Powders and Seasonal Allergies.”

The product recommends using coconut oils or other oils to relieve itching or headaches. Itching after using plant dye powders is often due to acidity or failure to fully rinse out all residue. Because this product contains ingredients other than henna and indigo, it is difficult to determine if something else may cause such a reaction. To learn more about dryness and itching after using plant dyes, read “Why Hair Feels Dry After Henna and How to Fix It.”

The instructions recommend mixing the powder with warm water and a teaspoon of salt. Ancient Sunrise recommends adding salt to indigo paste to help the dye bind more effectively to the hair. Because this product is a premixed powder, it is not possible to mix salt with indigo paste separately before adding it to the henna paste. This product recommends leaving the paste in the hair for 2-3 hours. It also recommends using a hairdryer after rinsing. Heat can help deepen the resulting color. However, as with the other samples, this product does not recommend a dye-release period.

#4

This product is from India. The ingredients are listed as follows: “Mehindi, Harred, Berhera, Amla, Shikakai, Coffee, and other Herbal…”

Mehindi is another word for henna powder. Harred, also known a harad or haritaki, is an ayurvedic herb with a variety of reported health benefits. Its scientific name is Terminalia chebula. It does not dye hair. Berhera (Terminalia bellerica) goes by many names including behara and belleric. It appears that the combination of haritaki, belleric, and amla powders is often known as “triphala,” a popular ayurvedic herbal remedy. Amla has been mentioned before and is an effective dye-release agent as well as a popular ayurvedic herb. Shikakai is an herb used for cleansing and conditioning hair. The addition of coffee is most likely for the purpose of color, but coffee is not an effective hair dye. Katha (Acacia catechu), also known as catechu, is an herb used both in medicines and in food as a spice. It is used as a dye for wool, silk, and cotton. Its effectiveness as a hair dye is unknown.

The ingredients list does not include indigo powder. Overall, the product appears to be henna along with a number of ayurvedic herbs, with coffee and katha potentially affecting the color result.

This product recommends mixing the liquid with “light hot water” (warm water?) in an iron bowl and letting it sit for 2-3 hours. The use of an iron bowl when mixing henna is an old tradition and is meant to affect the dye-release in a way that causes a darker result. It is now known that using iron is not necessary. There are no additional instructions regarding preparing the hair, applying, processing and rinsing. There is no inner pamphlet, standard warnings or patch test instructions.

The product claims to contain no chemicals or dyes and claims to cause zero side effects. Most likely it means no synthetic dyes. Caffeine is transdermal. The addition of coffee in this product may cause jitters and headaches.

#5

This product labeled as henna with herbal conditioner for a dark brown result. The country of origin is India.

The ingredients are listed as follows: “Henna, Amla, Kali Harar, Tulsi, Bahera, Lodh, Jamun, Chandan, Kattha, Shikakai, Bhringraj.” Amla, shikakai, bahera, and bhringraj have been described previously in this article.

“Kali Harar” seems to be a misprint of “Kali Harad,” also known as harad , harade, or harred. This is the same herb as haritaki, which has been previously described. Tulsi (Ocimum sanctum) is also known as holy basil. It is an ayurvedic herb used for many purposes. Lodh (Symplocos racemosa) is a tree used for ayurvedic medicine and whose bark is used for yellow dye. Its effectiveness as a hair dye is unknown. A brief account of the plant being used for dyeing hair yellow can be found here. Jamun (Eugenia jambolana) is also known as black plum. The bark and fruit are used in avurveda. While jamun does not appear to have dyeing properties, it does contain anthocyanins which may aid in cooling or neutralizing the brighter tones from henna. Ancient Sunrise Nightfall Rose fruit acid powder is made from the purple aronia fruit also contains a high amount of anthocyanins. Ancient Sunrise Nightfall Rose Powder can be found here. Chandan is also known as sandalwood. It can be used to dye wool or fabrics red or brown but requires boiling and a mordant. Its effectiveness as a hair dye is known. Kattha is another name for katha, or catechu, which is mentioned in sample # 4.

The instructions for this product recommend mixing the powder with water and letting it sit overnight. This product does not appear to contain indigo. Because amla and other fruit powders are included with henna, the powder may be acidic enough to be able to dye release overnight when mixed with water. Like other samples, the product recommends using an iron container, which is not necessary. The instructions recommend leaving the paste in the hair for ½ an hour to 1 hour or longer.

#6

 

This product is assembled and sold by an American company. The origins of the plant dye powders is not listed. This brand is commonly found in health stores in the US. The outer packaging reports that the ingredients are certified organic and contain no ammonia, peroxide, metallic salts, or PPD.

The ingredients are as follows: “Indigofera tinctoria (indigo) leaf powder, Cassia auriculata (senna) powder, Lawsonia inermis (henna) powder, Phyllanthus emblica (amla) fruit powder.” All reported ingredients save for amla powder are plant dye powders. Amla functions as an acidic component for dye-release and also assists in neutralizing warmer tones from henna and cassia.

The labeling and insert include warnings and patch testing instructions required from the FDA. Unlike some previous products in this article, this product recommends against using any metallic container or mixing implements. Metal such as stainless steel is fine to use when mixing henna. It does not cause any issues.

The instructions recommend mixing the pre-mixed powder with boiling water and applying it to the hair once it is cool enough. It claims that its “finer mesh botanicals do not need to cure.” That implies that only coarser sifts require dye-release time, which is untrue. The process of dye release involves the lawsone precursor molecules migrating from the dry powdered plant leaves into an acidic liquid environment where the molecules remain stable until the user is ready to apply. The boiling-water method will force henna to dye-release rapidly but the resulting paste will contain fewer stable aglycones and will therefore be weaker. It will often result in lighter, brassier, impermanent results.

Additional recommendations involve replacing some of the boiling water with egg or yogurt (“for extra conditioning,”), lemon juice or chamomile tea (“to bring out golden highlights”), hibiscus tea (to “intensify reds”) and coffee (“to enhance brown tones”). We know that these ingredients have little to no effect on the hair color result. Lemon juice can in fact cause the resulting hair color to oxidize much darker over time. Egg and yogurt will coat the hair and inhibit dye uptake. Coffee will not permanently color hair brown and will cause jitters and headaches. To learn more about what not to add to henna, read “Don’t Put Food on Your Head” and “Does It Dye Hair?”

#7

 

 

This product is from India. It is labeled as “brown henna.” A single box contains six thin, flat packets. Each packet further contains a small foil pouch containing 10 grams of product and an instructional paper. Each pouch is meant to be enough for one application.

The instructions recommend mixing a packet of powder with 30-40 ml of water (roughly two to three tablespoons). The paste is then to be applied and left in the hair for 30-40 minutes.

There is no ingredient disclosure in any part of the packaging. The small amount of product per application in combination with the short processing time suggests the presence of PPD or other synthetic dyes.

#8

This final product is made in India. I can be bought online as well as in ethnic stores. “Marron” is another word for brown. The ingredients are as follows: “Natural Henna, Aritha, Amla, Shikakai, 2 Nitro, PPD & Other Natural Herbs.” While henna is the first ingredient listed, we can see at the end of the list, “2 Nitro, PPO, & Other Natural Herbs.” It is impossible to know what the other “natural herbs” are.

PPD is clearly listed. 2 Nitro is also known as 2-Nitro-P-phenylenediamine. It is a derivative of PPD. Aritha is also known as soapnut, which is used for cleaning. It contains natural saponins. It does not dye hair. Whether or not it is ideal to use a saponin in a henna mix is unknown. The natural soap may or may not affect dye uptake.

It is likely that the majority of the color effect in this product comes from PPD and its derivative. Indigo is not listed as an ingredient, yet the product claims to color hair brown. The instructions recommend mixing the powder with water and letting it sit for 30 minutes before applying, then leaving it in the hair for 2-3 hours. This is a very long time for a product containing PPD. Most box hair colors that contain PPD recommend processing for no more than half an hour.

Powder and Paste Qualities

5 grams of each product was measured into respective containers. Color, scent, and texture was noted for each sample. Each sample was mixed with 15 ml of room temperature distilled water. Prior to stirring, one can often see larger plant particulates which rise the surface of the water. Color, scent, and texture of the resulting pastes were noted.

While plant dye powders can vary in color, it is not always indicative of quality or freshness. A common misconception is that fresh henna powder has a bright green color. In some cases, green dye is added to the powder to make the appearance more appealing. Pure henna powder can be pale brownish green to bright green in color. Indigo powder tends to be very bright green. When mixed with water, indigo will turn a deep green-blue. Additional herbal plant powders may affect the color of the powder and paste. What is not ordinary is for powders and pastes to be vivid orange or red, or very dark brown or black.

#1

The powder is a light olive green color. It has an earthy, plant scent without any strong synthetic, floral, or herbal scents. The sift appeared average to the naked eye. When water was added, the liquid turned a dark green color almost instantly. Some larger plant particulates (pieces of leaf, vein, or petiole) floated to the surface of the liquid. After stirring, the resulting paste was dark green, slightly gritty, and had a faint herbal scent.

#2

The color of this powder was a light, yellow-olive color. The scent was plant-like and neutral. The powder included some soft lumps that broke with a light touch. When mixed with water, the resulting paste was a medium green. Some larger plant particulates were visible floating on the surface of the liquid prior to stirring, but not as many as in sample #1.

#3

The powder was chalky sage green. The sift appeared normal to the naked eye. No strong scent was noted, but the classic “frozen peas” scent of indigo could be detected. Upon adding water, the paste turned a medium to deep green color. A fair amount of particulates were floating on the surface of the water. The paste had a stretchy, mucous texture which is common with hennas from certain regions.

#4

This powder was a dusty, light brown color. The scent of amla and other herbs was predominant. A considerable amount of larger plant particulates could be seen both in the powder and floating on the surface of the added water. The paste was a gray-brown color with a lumpy, gritty, and mucous texture.

#5

The powder was a dusty, light brown color, somewhat lighter than that of sample #4. No chunks or large plant particulates were visible. The powder had a notable “herbal henna blend” scent. When mixed with water, the paste frothed. The paste had a faint metallic and chlorine scent along with the herbal scent.

#6

The powder was a bright, light green color. There were not large plant particulates visible in the powder. The classic indigo scent was present. There was no noticeable herbal or synthetic fragrance scent. When water was added, the powder turned a bright green color. After mixing, the paste was thick and dense like wet clay. The paste was a similar color to cooked spinach.

#7

The powder was a green-gray color with some soft lumps that broke apart under light pressure. No discernible scent was noted. The paste frothed when mixed with water. The color of the paste was a chalky sage green.

#8

Immediately upon opening the package, this product had a strange, strong scent that was difficult to describe or identify. The color of the powder was terracotta orange. When water was added, some plant particulates could be seen floating on the surface. The paste was a bright orange-brown and had a thick consistency like icing.

Ancient Sunrise Rajasthani Twilight Henna (ASRT)

This powder had a pale, yellow-green color and scent like dried grass or straw. After adding water, there were very few plant particulates noticeable on the surface. The paste had a mucous-like and lumpy consistency similar to cottage cheese upon first stirring. The paste became smoother and stringy after stirring. The color of the paste was a muddy greenish brown.

Ancient Sunrise Zekhara Indigo (ASZI)

This powder is fine and fluffy with a bright green color like matcha tea powder. It has a distinct scent that is slightly metallic and reminiscent of frozen peas. When mixed with water, the paste is dense and smooth like wet clay. The paste turns a deep, blue-green.

2:1 Ratio of ASZI and ASRT

For the purposes of this article, a mixture of two parts indigo and one part henna was created to mimic a premixed product intended to color hair dark brunette. Normally, henna and indigo pastes are mixed separately before being combined. This resultant powder naturally showed characteristics of both ASRT and ASZI. The color was a lighter green than indigo powder alone, and mixed to a dense and slightly slippery paste.

Chromatography

Paper chromatography is a simple method used to separate dyes and/or to determine the existence of dyes in a substance. A small sample of the substance is placed on a strip of paper which is lowered into a glass chamber where the end of the paper strip wicks a liquid solvent. As the solvent moves up the strip and through the sample, the dye or dyes are pulled up along the paper at varying rates. By comparing chromatography results of different “henna for hair” product samples, one can see speculate on the dyes contained in each sample. Part One of this series discusses the chromatography method in further detail.

Here is a time-lapse video of a paper chromatography test from Part One.

 

This group of samples was tested under the same conditions as the set from Part One. Samples were tested in

1) A solvent of 99% isopropyl alcohol for 20 minutes,

2) A solvent of equal parts distilled water and 99% alcohol for 15 minutes, and

3) A solvent of equal parts distilled water and 100% acetone for 15 minutes.

It was necessary to test samples under multiple conditions because the target dyes were unknown. As will be seen in the results, some dyes will react differently to each solvent. Unlike the samples in Part One, which were mixed with a leon juice dilution and allowed time for dye release, these samples were mixed with distilled water only. The resulting pastes were applied to the paper strips and tested immediately. This is because a number of the samples in this group contain indigo which releases rapidly and requires a neutral or alkaline environment. In addition, most of the products’ instructions recommended mixing with water only. In order to maintain consistency across samples, room temperature distilled water was used with each sample rather than following individual product mixing instructions.

Results

99% Isopropyl Alcohol for 20 Minutes

Each sample was tested on five individual paper chromatography strips. Most results were consistent; occasional inconsistent results were disregarded and attributed to variation in paper strip composition or other unforeseeable factors.

Below is a comparison of all samples including Ancient Sunrise Rajasthani Twilight Henna (ASRT), Ancient Sunrise Zekhara Indigo (ASZI), and a 2:1 ratio of ASZI and ASRT. One strip of each sample was selected to create this image. Note that the solvent front (the highest point reached by the solvent) is consistently lower in samples 1-5. This is due to inconsistency in the paper strip manufacturing rather than a difference in the product samples. However, it cannot be ruled out that the change in paper density may have affected the way we visually interpret the results.

Additionally, this image was taken after all samples had been tested and the paper strips had fully dried for several hours. Some samples lost color while others deepened in color. This will be discussed in the Limitations and Considerations section. Following images will show samples immediately following testing.

99% isopropyl alcohol is a virtually anhydrous solution. In other words, there is very little water. While a very small amount of water exists in the sample paste and in the solvent, it may not be enough to cause oxidation. Therefore, this condition should not reveal much lawsone or indigo dye. We see significant differences in the colors of samples 7, 8, and 2:1.

Sample#1

The result had a very pale, yellow-green tone. While the stain appears mostly even from the point of application to the highest point reached by the solvent (also known as the solvent front), the deeper part of the stain appears to end about two thirds of the way up. This sample reported henna, indigo, centaurea, and beetroot as its ingredients.

Sample#2

Sample #2 had results which were paler in color. This was an herbal blend which contained henna along with many other herbs which may or may not have dyes. While Sample #1 contained some amount of indigo, this sample does not.

The result also showed some very slight green-blue vertical stripes which may suggest that dye was added to affect the color of the powder. This was seen in a sample in Part One of this series. Because it has been believed that a green henna powder indicates freshness, some manufacturers add dye to change the powder’s appearance. It is possible that this is what happened here. Part One discusses the practice in more detail.

Sample #3

The result was a yellow-green tone similar to the result for sample #1. Like #1, this sample also reported indigo in its ingredient list. The dyes involved in this sample did not move with the solvent as much as the previous two; much of the coloration remained lower near the point of application rather than rising all the way to the solvent front.

Sample #4

This sample’s results was closer to that of Sample #2. Sample#4 and #2 are both herbal blends which do not list indigo in the ingredients. Thus, the result is very pale because whatever henna may be included in the mix is diluted with herbal ingredients which contain no dye. However, this sample shows some dark red-brown banding that appears just above the point of application and does not move further up the strip. This is inconsistent with henna’s lawsone dye. It is unclear what may cause such results.

Sample #5

This was another henna/avurvedic herbal blend which did not include indigo in the ingredients list. Similar to Sample #4, the result was mostly pale with some darker red-brown banding just above the point of application.

Sample #6

It should be noted that while the solvent front for this sample is much higher than the previous five samples, this is most likely due to a change in the manufacturing of the paper strip rather than a factor of the sample itself. This sample showed a gray-green color which was darkest about halfway between the point of application and the solvent front. One can see some banding about a centimeter above the point of application. This sample reported indigo as its first ingredient, followed by henna, cassia, and amla. The result is visually similar to the results of samples #1 and #3.

Sample #7

This sample differed from previous samples. There was a light red-brown band just above the point of application as well as some green vertical striping, most noticeable in the paper strip second from the right. As the sample dried and oxidized, the overall color deepened further and the green tone was overtaken. This effect was unlike that of most other samples which darkened very little. This product did not include a list of ingredients. The green coloration is very likely due to the same kind cause as in sample #2: an added synthetic dye rather than indigo.

Sample #8

This sample showed results unlike any of the previous seven samples. The overall stain was deep orange with a very noticeable red-brown band above the point of application. While the product reported two forms of PPD in addition to henna and some ayurvedic herbs, this does not explain the vivid orange color. There is very likely another dye or number of other dyes which were not listed.

ASRT

The Ancient Sunrise Rajasthani Twilight henna sample showed a pale stain that was similar to samples #1-6 but had a warmer orange tone. This would make sense as this is a pure henna whereas the other samples in this collection are blends of henna and other plant powders.

ASZI

This sample was pure Ancient Sunrise Zekhara Indigo. The result was a deep olive green color with some hints of banding about halfway between then application point and the solvent front. As the sample dried, the color oxidized to a pale, gray blue.

2:1 ASZI/ASRT

This sample was a blend of two parts Ancient Sunrise Zekhara Indigo and one part Ancient Sunrise Rajasthani Twilight henna. This mix is what is recommended for dark brunette results. The color was much like the results for pure indigo powder, but with a slightly warmer tone due to the addition of henna. When the paper strips dried, the color oxidized to a cooler brown tone.

Of the eight samples, sample #6 came closest in appearance to the 2:1 indigo/henna mix. Sample #6 reports indigo as its first ingredient, followed by cassia auriculata, henna, and amla.

Sample #3 also reports indigo as its first ingredient, then followed by henna, amla, and a number of herbs that do not dye. The results from sample #3 also have a similar, gray-green tone, but much lighter. It is likely that the strength of the plant dyes was diluted with the addition of other plan powders.

1:1 Dilutions for 15 Minutes

The results from an anhydrous solvent such as isopropyl alcohol will differ from that of a solvent containing water. In prior tests, using pure acetone as a solvent yielded results that were not useful; therefore, pure acetone was not used as a solvent condition in this series at all. In the case of the lawsone and indigo dye molecules, they require at least some water in order to oxidize. While the samples in the 99% isopropyl alcohol condition above were mixed with a few milliliters of water, it was not quite enough to allow the colors of the dye molecules to show.

This section reports results of the using A) a 1:1 mixture of isopropyl alcohol and distilled water and B) a 1:1 mixture of acetone and distilled water. Overall, the stains on the samples were much more visible. Results from both conditions were relatively similar to one another. Samples #7 and #8 were especially unique.

Below are results for both conditions. The top image shows the results for the 1:1 alcohol/water solvent condition. The bottom image shows results for the 1:1 acetone/water solvent condition.

Sample #1

Results from this sample were a pale orange tone which was darkest below the halfway point between the point of application and the solvent front, suggesting a wide dye band. As the paper strip dried, the color appeared lighter. The acetone/water solvent condition yielded a darker result and seemed to show a muted, red-brown section from the point of application to roughly halfway up, and a lighter orange tone closer to the solvent front.

Sample #2

Sample #2 appeared very similar to sample #1 but lighter in color. In the 1:1 alcohol/water solvent condition, the stain more even from the point of application to the solvent front. In the 1:1 acetone/water condition, a similar red-brown tone is visible from the point of application to about 2/3rds of the way up. While there was a light green streak in the results for the 99% isopropyl alcohol condition, a green tinge can be seen in the 1:1 alcohol/water condition. The green dye moved all the way up to the solvent front, where it created a thin line. The green tone was no longer visible under the 1:1 acetone/water condition. This suggests that whatever caused the green color is a dye that interacts differently with acetone and alcohol.

A faint green line is perceptible at the very edge of the solvent front in sample #2.

Sample #3

Sample #3 was, like in the first condition, very similar to samples #1 and 2. No distinct banding is noticeable, but the stain gradually fades as it approaches the solvent front. In the 1:1 acetone/water condition, the solvent moved much further up the paper strip. This is most likely due to a variation in the paper itself. As the samples from the 1:1 acetone/water solvent condition dried, the color oxidized moderately and showed a soft brown tone.

Sample #4

Sample #4 was similar to the previous three samples. There is no distinct banding and the color appears relatively even from the point of application to the solvent front, but fades somewhat as it approaches the solvent front. There is a muted red-brown tone from the point of application to about halfway up to the solvent front. Above the halfway point, the color is lighter and brighter. The color of the results deepened slightly after the paper strip dried.

Sample #5

The paper strip in the center of the 1:1 alcohol/water solvent should be disregarded as an inconsistent result most likely caused by a variation in the paper strip. In the 1:1 acetone/water solvent condition, the height of the solvent fronts matched. Sample #5 showed an even, light orange stain and no distinct banding.

Sample #6

This sample was very similar to samples #1-3. The result was pale and even with no distinct banding. On some strips in the 1:1 alcohol/water condition, the color appears darker neat the point of application and gradually fades toward the solvent front. In the 1:1 acetone/water condition, the color appears to be more concentrated near the solvent front in a brighter orange band.

Sample #7

The darker color is much more prominent in this condition in comparison to the result from using 99% isopropyl alcohol solvent. As the paper strip dried, the color oxidized further. This sample did not include an ingredients list in its packaging. The deep, almost purple-gray tone suggests that this sample may contain PPD. A previous test of black hair dye containing PPD yielded results which had a similar color, but much darker.

Sample #8

The bright orange color detected in the first condition is even more prominent in this second condition. While pure henna can color hair and skin a deep orange color similar to this, it is clear that this vivid orange tone is cause by some other type of dye. The lawsone dye molecule requires a slow and steady dye release period in an acidic environment. Given the right conditions, a good lawsone stain will oxidize to a deep orange to reddish-brown on light hair. This product, on the other hand, was tested immediately after being mixed with distilled water. It yielded a vivid orange result immediately.

ASRT

Both conditions resulted in a light, orange tone which slightly after the paper strips dried. In comparison to the 99% isopropyl alcohol condition, the dye is more visible. This is likely because the lawsone dye molecule was unable to release and oxidize as well in an anyhydrous condition; in other words, some amount of water is necessary for lawsone to create a visible stain.

ASZI

While the initial result of the 1:1 alcohol/water solvent condition was paler, some potential banding can be seen in the 1:1 acetone/water condition. The result shows some muted blue-green tone upon removal. After the paper strips dried and oxidized, the color of the results in both conditions deepened to a blue-violet tone. This is in line with the normal oxidation process of the indigo molecule.

2:1 ASZI/ASRT

In both conditions, the results of the 2:1 mixture of ASZI and ASRT appeared initially pale orange in tone. This is likely because the lawsone dye from the henna was more visually prominent at first. However, as the paper strips dried and the dyes oxidized, more of the cool tones from the indigo molecule emerged, turning the overall color a muted brown tone. Based on this result, one would expect that other samples which include more indigo than henna should show a similar result. However, with the exception of samples #7 and 8, the products tested in this article all yielded pale orange brown results that look more similar to the ASRT sample than the 2:1 mixture.

Discussion

Samples # 1, 3, and 6 reported indigo in their ingredients disclosures. One would expect for the chromatography results to appear similar to that of the 2:1 mixture of Ancient Sunrise Zekhara Indigo and Ancient Sunrise Rajasthani Twilight henna, and different from the sample of Ancient Sunrise Rajasthani Henna alone.

In the 1:1 solvent/water dilution conditions, the sample of Ancient Sunrise Zekhara Indigo alone yielded results which were green at first, and which oxidized to a pale violet tone. This tone deepened the result of the 2:1 mixture. This is especially noticeable in the 1:1 alcohol/water condition. After drying, the result was a light, cool brown. One would expect to see a similar tone in samples #1, 3, and 6. Samples #1 and 3 show some slightly cooler tones in comparison to the ARST sample and other samples which did not include indigo. Sample #6 did produce a greenish color in the 99% isopropyl alcohol condition. After the paper strip dried, that color oxidized to a cooler brown tone that was more prominent that the previous five samples of the same condition. In the 1:1 acetone/water condition, the result of sample #6 was much lighter, with a golden band appearing just below the solvent front.

Assuming that the product tested did include henna and (in the case of #1, 3, and 6) indigo, it is likely that the addition of other plant powders diluted the appearance of the lawsone and indigo dyes. The quality of the plant dye powders could have also affected the results.

The most salient results came from samples #7 and 8. Sample #7 did not include any ingredients disclosure on its packaging whatsoever. Nor was any information about ingredients available online. The fact that Sample #7 provided extremely small packets of powder, each of which is meant as one application, suggests a high likelihood of the product containing oxidative dyes. The product is reminiscent of many small, concentrated powdered hair dyes which contain PPD which are sometimes used to create “black henna.” These powders have little to no scent. A true henna product requires at least 100 grams of powder to color collar-length hair of average thickness. It is meant to be made into a thick paste, rather than a thin, paint-like liquid.

This is an example of a powdered PPD-based hair dye. Very little powder is necessary for a complete application because the dye is concentrated. 

Sample #8 reported PPD and 2 Nitro, a derivative of PPD, on its ingredients list. However, the results of the chromatography tests yielded deep orange results. PPD alone results in a deep, violet-black stain depending on the concentration. This product appears to have a lower concentration of PPD (this does not mean it is safe!) and another dye which is not reported. The ingredients list includes the phrase “other natural herbs.” It is possible that one of the “other natural herbs” is responsible for the orange color. However, it is more likely that the product includes a synthetic dye, possibly an azo dye or a concentrate food coloring, to bring about this result.

Sample #2 showed some faint, green-blue vertical streaks in the 99% alcohol condition. In the 1:1 alcohol/water condition, the same color can be seen at the solvent front. This result different from the blue-green tones of indigo. Unlike indigo, it did not oxidize to a deeper blue-violet tone over time. Additionally, the dye appeared in streaks rather than as a consistent stain. Sample #2 did not report indigo in its ingredients list. It is likely that this sample included a synthetic dye meant to alter the color of the powder itself. A similar effect appeared in Part One of this series.

In all, samples #1-6 showed visually similar results with some variation. They were visually comparable to the ASRT sample and 2:1 ARZI/ASRT samples. Sample #2 showed some blue-green streaks which suggest an added synthetic dye. Sample #7 yielded a very dark brown result which suggests the presence of PPD. Sample #8 yielded a vivid orange result which is very likely due to an added synthetic dye.

Limitations and Considerations

The purpose of this article was to compare a number of products labeled and sold as “henna for hair” products which claim to color hair brunette or brown. It is important to note that the results of the paper chromatography tests are in no way meant to indicate the color result of using any of these products on hair. Paper and hair do not dye the same way, nor was the purpose of this article to demonstrate hair color results.

The hope was that the results of the paper chromatography tests would show distinct color bands which could be compared to Ancient Sunrise henna and indigo results to determine the presence of lawsone and indigo dyes. However, very few results showed distinct dye bands. Future tests should consider new solvent conditions which may show clearer results. Tests using more advanced chromatography methods would be able to separate and identify dyes more successfully.

As noted earlier, the paper strips used for the chromatography tests varied in density. This caused the solvent to move more quickly and higher up the strip in some tests, and more slowly in others. This was an unforeseen factor caused by inconsistent manufacturing. Future tests should make sure to use high quality paper strips which are reliably consistent.

The results of the chromatography tests changed in color as the paper strips dried and as the dyes oxidized. Some dyes oxidized more than others while some colors faded rather than darkened. Because there was no way to test all samples at the exact same time, each paper strip was at a different stage of oxidation when all samples were completed. While photos of each sample was taken just after removal from the processing tank, variation in lighting and camera settings made it difficult to visually compare results. Future studies should take into consideration camera and lighting control.

The final step in this exploration of premixed “henna for hair” products would be to investigate a number of products marketed for coloring hair black. No doubt many products labeled as henna in this category will contain PPD. In some cases, “black henna” sold for both hair and skin is simply a highly concentrated PPD product with little to no henna content. Future investigations should follow the same or similar procedures as Part One and Two of this series to investigate such “black henna” products.

Achieving a Darker Result Using Ancient Sunrise® Henna: Dos and Don’ts

The dye from the henna (lawsonia inermis) plant is called lawsone. If extracted and isolated from the plant, lawsone is a bright orange color. When henna leaves are harvested, dried, and made into powder, and that powder is subsquently mixed with an acidic liquid to form a paste, the lawsone precursor molecules which exist in the henna is released as an intermediate molecule called an aglycone. This aglycone molecule can attach to keratin—such as that which forms the outer layers of hair stands and skin– and then oxidize to its final, stable form. The result on light hair is anywhere between a bright, fiery copper to a deep auburn.

There is no such thing as “brown henna,” or “black henna.” Products with such labels most likely contain some henna along with additional plant dye powders, or even synthetic dyes such as para-phenylenediamine (PPD). This goes for both products marketed for hair use as well as for use on the skin. The truth is, pure henna will only color keratin a variation of orange to red-orange.

In order to achieve a darker result when using henna, something must be added to the henna mix, or the hair/skin must be exposed to heat during or after processing.

This article will explain what can be done to safely and effectively deepen henna results on hair*, as well as what should not be done.

*Note: The same kinds of rules do not always apply in the same way to henna used on the skin. For more information on henna as body art, read “Henna for Body Art 101: How to Achieve a Dark, Long-Lasting Stain” in BecomingMoonlight.Blog. Please also note that if you live in the United States, the FDA does not allow the use of henna for body art purposes (i.e., coloring the skin). Here are the US FDA regulations for the use of henna for the purpose of body art. These regulations have the force of law: https://www.fda.gov/cosmetics/productsingredients/products/ucm108569.htm

Do: Mix your henna powder with an acidic fruit juice

For a rich, vibrant result, it is important that the henna paste properly dye-releases. Water alone is not enough and will cause light, brassy results. A mildly acidic liquid allows the maximum release of aglycone molecules by keeping them in a hydrogen-rich environment. Water releases some dye molecules, but cannot keep them in their intermediate state as well. The result from a water-only mix is lighter and often less permanent because dye molecules either have not released from the plant material or have released and oxidized to a final state which cannot bond to keratin; therefore, fewer aglycones are available to color the hair. The dye molecules bond to keratin by way of a Michael Addition, which requires the extra hydrogen ions that exist in an acidic solution.


The sample on the left was dyed with henna mixed with an acidic liquid. The sample on the right was dyed with henna mixed with water.

Leaving an acidic paste at room temperature allows for a slower, and better-controlled release of the maximum amount of aglycones. More dye molecules become available in the paste over time, while the acidity prevents rapid oxidation of those molecules (demise). At room temperature, an acidic henna mix is ready after eight to twelve hours. To learn more about proper dye release, read Chapter Six of the Ancient Sunrise Henna for Hair E-Book and “Henna 101: How to Dye-Release Henna” in this blog.

The liquid does not have to be overly acidic. A pH of 5.5 is sufficient. Lemon juice, with a pH of 2-3, is very acidic. Lemon juice can be diluted with 1-3 parts water for an effective mixing liquid. Undiluted lemon juice should be used with care and avoided by those with sensitive skin. To read more about using lemon juice in henna mixes, read The article titled “Should You Be Using Lemon Juice In Your Henna Mix?”

Other fruit juices such as orange, apple, and cranberry are effective for mixing with henna. However, cranberry is often recommended for keeping results lighter and brighter, as the antioxidant content in cranberry juice may prevent darkening.

Ancient Sunrise® also offers fruit acid powders which can be used with distilled water to create a mildly acidic solution. The fruit acid powder called Malluma Kristalovino is gentle on sensitive skin and can help make results deeper. Nightfall Rose fruit acid powder adds subtle ash tones to henna. Amla fruit acid powder can help a henna/indigo mix bind more effectively to hair for deeper, cooler brunette shades.

Don’t: Mix your henna with coffee

Mixing coffee with henna has been recommended by other sources as a way to deepen resulting colors. This has been proven to be ineffective. Not only will adding coffee do very little to the color, but the trans-dermal nature of caffeine will leave a person with jitters or a bad headache. Henna paste needs to be left on for at least three hours. During that time, caffeine would be entering the bloodstream through the skin at a rather rapid rate. Additionally, the paste would smell quite unpleasant.

Do: Apply heat during processing and/or after rinsing

Heat causes the outer cuticle layers of hair strands to open up, thus allowing better dye penetration. Once you have applied the henna paste to your hair and have wrapped it up, keep your head warm by covering it with a thick, knitted cap or a towel. You may also choose to aim a hairdryer at your head for intervals of a few minutes at a time, or sit somewhere warm and sunny. Heat can both speed up processing time and ensure a more saturated result.

After the henna paste has been rinsed out, you may choose to use heat again to speed up the oxidation process. Hennaed hair is naturally lighter and brighter first upon rinsing, and will take several days to a week to settle into its final color. Using a hair dryer or heat styling tools can cause oxidation to occur more quickly. Continual use will darken hennaed hair more and more over time. This darkening is permanent, and can only be reversed with the use of lightening products. Those who wish to avoid causing their hennaed hair to darken should avoid excessive use of heat styling.

This sample has been dyed with henna. The right side was heated with an iron.

Don’t: Mix henna paste with hot or boiling liquid

Many henna for hair products instruct users to mix the powder with hot or boiling water. This technique leads to a rapid release, and subsequent demise, of the dye molecule. As stated above, an acidic liquid allows for more aglycones to be available in the paste at the time of application. The boiling-water method of mixing henna causes lighter, brassier results. With henna, as many other good things in life, patience is key.

Do: Apply henna to clean hair

The sebum, dirt, minerals, and product buildup in unwashed hair prevent dye uptake. For the best results, apply henna to hair that has been treated with Ancient Sunrise Rainwash mineral treatment followed by a clarifying shampoo. At the very least, shampoo your hair very well. Skip the conditioner. Particularly oily or resistant hair can be washed with a few drops of dish-washing detergent to ensure it is ready for dyeing.

If you are a no-poo or low-poo person, this does mean you will have to break your regimen just once if you want the best results. There is just no way around it. Baking soda and vinegar, clay, natural herbs, or any other washing methods will not remove sebum, dirt, and mineral buildup effectively enough for the purposes of coloring hair with henna.

Don’t: Add oils to a henna for hair mix

Just as oils on the hair will prevent effective dye uptake, so will oils added to a henna mix. While some might believe that adding oils or even other ingredients such as milk or yogurt to a henna mix may help, they do not. To read more about what not to add to a henna mix, read Don’t Put Food On Your Head.

Certain types of essential oils, called “terps” (short for monoterpene alcohols) are added to henna pastes made for body art. When used on skin, “terped” henna results in deep burgundy to near-black stains. However, essential oils should not be used in henna for hair. They can cause the resulting color to be muddy (not darker in a desirable way). In addition, leaving a paste containing essential oils on the head for an extended period of time will lead to headaches and scalp irritation.

Do: Add indigo for brunette results

As discussed earlier, the lawsone molecule from henna can provide orange to auburn results when henna plant powder is used alone. In order to achieve brunette tones, another plant dye powder must be added. Indigo plant dye powder contains dye of the same name. This is the dye that was originally used to color denim, and is still used today in many textile traditions. The type of indigo powder used in henna mixes is called vashma indigo. This is made from indigo leaves that have been partially fermented before being dried and powdered. If used on its own on light hair, indigo may color hair grey-blue, sometimes violet, and sometimes a blueish green. The effect is difficult to control and not as permanent as henna.

When used in the right ratios, henna and indigo together will color light hair virtually any natural brunette shade from medium brown to warm black. Unlike henna, indigo does not need acid and time for dye release. It must be mixed with only water just prior to application. To add indigo to henna, mix the powder with distilled water until it is a similar consistency as the henna paste, then combine it thoroughly with dye-released henna paste and apply immediately.

To learn more about indigo, read Chapter Five of the Ancient Sunrise Henna for Hair E-book. To learn what henna/indigo ratio is best for your desired outcome, see “Henna for Hair 101: Choosing Your Mix.”

The sample on the left was dyed with henna only. The sample on the right was dyed with indigo only. Those in the middle were dyed with various ratios of the two.

Don’t: Add black walnut powder, anything claiming to be “Buxus” or “Katam,” or synthetic dyes

Black walnut powder is sometimes mentioned in natural hair care communities for the use of dyeing hair brown. The effect is not as permanent as henna. Additionally, black walnut is known to cause allergic reaction for many people. It is therefore best avoided.

Buxus dioica, also called katam, is a plant that works similarly to indigo when used with henna. The result is shades of brunette. However, buxus was only grown in and exported from Yemen. The conflict occurring within the country has ceased exports and production of buxus and other goods. There are a few vendors which claim to carry buxus. At best, those products are in all actuality indigo powder labeled as buxus. At worst, they contain dangerous or ineffective ingredients.

Do not add synthetic (store-bought or salon) hair dyes to your henna mix. They are not compatible and are not meant to be used in the same mixture. Oxidative dyes color the hair through a very different chemical process than henna. Do not try to add other types of dyes, such as fabric dye or food coloring.

You can, however, safely use semi/demi-permanent or oxidative dyes over hair that has been colored with Ancient Sunrise® henna for hair products (and no other henna product), as the plant powders sold by Ancient Sunrise® have been tested in an independent lab to ensure they do not contain mineral salts or other adulterants which may react with synthetic hair products.

Also Don’t: Use premixed “henna for hair” products

Some “natural” hair coloring products which promise a brunette or black result declare a combination of henna, indigo, and/or other plant ingredients. Because henna and indigo must be prepared separately, any product which blends the plant dye powders together is likely to produce inferior results.

Some products labeled as henna for hair may also contain azo dyes (such as Red 33) or oxidative dyes (such as PPD). The requirement for ingredients declarations varies from country to country so that some products manufactured outside of the US do not report all of the ingredients which they contain. While such products are not allowed to enter the United States, all too often they slip by. It is best to stick with purchasing pure plant dye powders in separate packages and mixing them yourself. While a pre-mixed product may seem tempting, opting for Ancient Sunrise® products and methods allows you to keep your peace of mind.

Do: Repeated applications of henna

While henna does not “coat” the hair, repeated applications will cause your hair to be more saturated with dye each time. We often recommend to only color new hair growth after a person has achieved their desired color. This is because repeated applications will cause darkening over time. However, if you are looking for a deeper, richer color, feel free to reapply henna to the entire length of your hair until you get it to where you like. Unlike with conventional dyes, repeated applications will not damage the hair; in fact, continuing to use henna will condition and strengthen the hair.

Leaving the paste in your hair longer can also contribute to a deeper result. However, only do this if you are not using indigo. Indigo’s dyeing power begins to slow after about three hours, after which the henna part of a henna/indigo mix will continue to color the hair. The result of leaving a henna/indigo mix on the hair longer than three hours may be redder than desired.

Don’t: Re-henna too hastily

Keep in mind that henna’s color naturally deepens over the course of the week following application. If you rinse your henna out and immediately feel that it is just a couple of shades too bright, wait at least a few days before reassessing. You may find that a little bit of time is all you needed to reach your desired shade. Reapplying too quickly may cause you to overshoot, and end up with a final color that is much darker than intended.

Do: Comment on this article or contact Ancient Sunrise® Customer Service if you have any additional questions about deepening your hennaed hair results.

Analysis of Products Marketed as Henna for Hair: Part One

Table of Contents

Introduction

There is a wide variety of products available for coloring hair which claim to be pure henna, or which claim to contain pure henna powder along with additional natural plant ingredients. There is no international standard for what can be marketed as henna, and it is often difficult to tell which products are what they claim to be. Many henna for hair products lack an ingredient declaration. A package may show only some ingredients or have no ingredients list at all.

Additionally, products which claim to contain all-natural ingredients vary in quality as there are no internationally agreed upon standards for “all natural”. Powders may be poorly sifted, containing sand or larger plant particles which make for difficult application and removal. Stale or low dye-content henna products may contain additives such as additional dyes or metallic salts to compensate for poor quality of materials. Green dye may be added to henna powder to make it look fresher.

The FDA has a standard for henna and products entering the United States labeled as a “henna” hair coloring product. These guidelines are do not appear to be regularly enforced as are regulations for products labeled as henna for use on skin.

The FDA forbids the sale of and is empowered to confiscate of any henna product labeled for skin use, or products showing images of henna used on skin. Customs and border protection is empowered to search the importing company’s website to determine if henna is intended for use on skin, and may seize and destroy henna that appears to be imported for use on skin.

“Black henna,” or products labeled as henna containing PPD, have been known to cause skin reactions and sensitization. For more information about black henna, read the articles “Henna is not Black,” “What You Need to Know About PPD,” and Chapter One of the Ancient Sunrise E-book.

This article is the first of a series that will test and compare various “henna for hair” products which have been found online on sites such as Amazon and Ebay, in import stores in the USA, and in ‘health food stores’. The purpose of these studies will be to determine the quality of those products in comparison to Ancient Sunrise® Henna for Hair products, and to test for the existence of dye additives. This series will feature articles investigating products within the following categories:

  • 1.“Pure” henna, herbal henna mixes, and red result henna for hair products
  • 2. Brunette result henna for hair products
  • 3. Black result henna for hair products

This article will cover the first category by investigating nine samples of retail henna hair dye products which are labeled and/or advertised as 100% pure henna, a 100% natural mix of henna and additional “herbs”, or as henna-based hair products which claim to color the hair red. Subsequent articles will report on henna for hair products claiming to color hair brunette and black.

The first part of this article will report on product labeling, visual and textural qualities of the products as dry powders, and visual and textural qualities of products when mixed with liquid to form a paste. The second part of this article will report the results of paper chromatography tests designed to determine the existence of dyes, lawsone or otherwise, in each sample.

Sample Selection and Label Analysis

Nine “henna” hair dye products were selected based on the following criteria:

1) The product claims to contain only 100% natural henna.

OR

2) The product claims to be 100% natural, containing henna along with additional “herbs.”

AND

3) The product either claims a red result or does NOT claim to dye the hair brunette or black.*

*Products that claim to contain henna and other plant powders for brunette and black hair results will be tested and reported in future articles.

Not all products that are marketed as 100% pure henna state explicitly state a hair color result, as some are marketed for both hair and skin, as well as for mixing with other plant dye powders such as indigo. Henna has been used for hair, skin, and home remedies for centuries. It can be assumed that if a product claims to be 100% henna, it should provide a reddish result on lighter hair..

These products are a random collection of henna products purchased in shops and online, easily accessible to the retail consumer. In fact, very few brands, if any, submit their plant dye powders to the same testing standards as Ancient Sunrise®. Ancient Sunrise® products are sent to an independent laboratory for multiple tests prior to sale to ensure they do not contain adulterants, accidental or deliberate.

Below are images of the labels for the nine samples. Beyond this section, each sample will be referred to as its corresponding number. They will be tested alongside Ancient Sunrise® Rajasthani Twilight henna powder, which will be referred to as “AS”.

Sample #1

The product claims to be 100% natural. The ingredients are reported as “100% Natural Henna Powder.” The product also claims to be organic.

This product fits into criterion 1. There is no information suggesting there are other ingredients other than henna. The label would lead the everyday consumer to assume that the contents of the packaging are pure, natural henna powder.

However, descriptions on sites selling this product claim a “Beautiful golden brown color” which is inconsistent with results expected from pure henna. This would suggest either 1) The product is not what it claims to be or 2) Poor quality product and/or poor instructions lead to a lighter result.

Instructions are included on the back of the packaging. On the side of the box, it states that this is a product of India.

Sample #2

This product fits both criteria 1 and 3, as it claims to be 100% natural henna powder, and also reports a red result in addition to showing multiple images of red hair on the packaging.

No instructions were provided. The label includes an address of an import company. The website listed does not exist. Some additional searching found that the company has a contract with a manufacturing and export company in Pakistan, and that this is one type of product they export, along with clothing and gift items.

Sample #3

While the front of this package claims it is 100% natural henna, and states that the shade is copper, the back is unclear. It says “Copper: for dark blond, light brown hair,” which most likely means that dark blonde or light brown hair may be colored to a copper color. It also mentions “neutral henna” which suggests that it contains cassia obovata. However, on the website, the only ingredient listed for this product is “Lawsonia inermis (henna) leaf extract.” Based on the reported ingredients, this product fits criterion 1 as a product that claims to be pure henna.

A pamphlet included in the box provides instructions which recommend mixing the paste with hot water and applying once the mixture has cooled.

While the packaging says that the product is made in France, there is no indication of the origin of the henna powder. If it does contain henna, it is most likely mixed and/or assembled in France with ingredients imported from elsewhere.

Sample #4

This product claims to contain henna under the insert’s first section, “Composition.” It claims to provide a deep red result. However, the description is confusing, as it states that lawsone and “mannitte” are responsible for coloring. If mannite is to be understood as the sugar alcohol, mannitol, it would not affect color.

The product claims to be “Henna with natural herbs,” but these additional ingredients are not reported anywhere. In the chromatography section, it will become clear that this product clearly contains more than just henna powder.

This product includes instructions on the insert, and is a product of Egypt.

Sample #5

This product’s label includes something close to an ingredients list, as it reports henna powder along with additional herbs which are meant to affect the color result as well as to condition the hair.

This product fits criterion 2. There is no indication of color result, other than stating that the blend of herbs and henna “give the dark color”. In fact, the label states “It does not contain any colour or dye.” This may be intended to mean that the product does not contain any synthetic dyes. If the product does contain henna, it would color light hair a red tone. None of the additional herbs listed are ones which produce a dye, but some plant powders can affect the tone of hennaed hair. This is most likely what the packaging means to suggest.

The instructions recommend mixing the product with water in an “iron vessel”* and letting it sit for to hours. If the product does contain amla powder, it may be enough to create an acidic mix.

The product was manufactured in India.

*An iron dye pot can be effective at decreasing the vibrancy of dye colors in a boiling pot to dye yarn or cloth, but will not change the result of henna on hair. The iron pot may cause the dye released paste surface to look browner, but there is no significant effect on the hair.

Sample #6

This product, like #5, is labeled as a henna and herbal mix for conditioning hair, but does not explicitly state a color result. On the product’s site, it is described as providing a “rich burgundy shade.”

The front of the packaging includes the words “100% natural” and “Mehendi,” another term for henna. It is unclear whether this should be read as two phrases or one, as the from also clearly states that there are nine additional herbs. The back of the package describes supposed benefits of the added herbs. The phrase “Rajasthani Mehendi” means that the henna in this product is from the Rajasthan region of India, which is where much of the world’s henna, including Ancient Sunrise® henna, is grown.

The ingredients list states that henna powder is the first ingredient, followed by powder forms of the following: aloe, neem, brahmi, bhringraj, amla, hibiscus flower, shikakai, jatamansi, and methi. All of these ingredients are commonly known and sold in South Asian stores as health and beauty herbs.

This product has instructions on the back of the packaging. It recommends mixing the powder with water and letting it sit for 2-3 hours. Like sample #5, it is possible that some of the added herbs are acidic enough that water would be enough. It also suggests adding oil or curd for “extra softness.” Doing so would affect the dye uptake, leading to lighter results. To learn more about what not to add to henna mixes, read Henna for Hair 101: Don’t Put Food On Your Head.

It is from India, and is a very popular “henna for hair” product both in South Asia and in the States, and is widely available online and in international grocery stores.

Sample #7

This product claims to contain henna. The brand’s website describes it as a 100% natural product. The site also sells “henna neutral” (cassia powder) and “henna black/basma” (indigo powder), which would suggest that their “henna red” is their henna (lawsonia inermis) powder. Therefore, this product seems to fit criterion 1, a product is labeled/advertised as a 100% pure henna powder product, but it is not entirely clear due to a lack of ingredient declaration.

However, the table on the back seems inconsistent with true henna results, as they suggest that, one would see some variation of brunette or copper results. Like sample #1, it is possible that this is either due to the product containing additional ingredients, or ineffective mixing/application methods.

I will still include this product in within this group of samples because it is both labeled as henna, includes the word “red,” and the website claims it is a 100% natural product.

There is a US address on the back, but no indication of where the henna was grown.

Sample #8

This product fits criterion 1 as it claims to be 100% natural henna. It includes instructions on the back of the packaging. The brand’s website appears to sell additional hair care products such as shampoos, conditioners, and oils. The product has an Indian address on its packaging.

Sample #9

This product qualifies for criterion 1 as a product that claims to be pure henna powder. The information made available on this packaging is minimal. It states that it contains natural “hina” (henna) powder with no added chemicals, and that henna has been used as a dye for centuries. There are no instructions. The address listed is in London, UK, but the source of the henna is not listed. An internet search found that this brand, like sample #2, is part of an international exporting company that sells a variety of items such as health product, gifts, and books. It specializes in Islamic items.

Powder and Paste Qualities

There was a wide variation in powder sift, texture, and scent among the samples. When mixed with liquid, the resulting pastes also varied in qualities.

Ancient Sunrise® henna is grown in the Rajasthan region of India and is finely sifted. Many of the Rajasthani hennas have a slimy, stringy texture when made into a paste. This is because henna plants will have more or less natural mucilage depending on the cultivar (variety of plant). Not all hennas have this texture, and some are naturally more creamy. Pakistani henna tends to have less mucilage and a greater coefficient of expansion. The higher coeffient of expansion and lower mucilage leads to paste cracking when henna is used as body art; it does not make any difference in hair application.

It could be assumed that hennas manufactured in a country were also grown there, unless the address is outside of where henna is naturally grown, such as those products with company addresses in Europe or the United States. In the case of the latter, it not possible to determine the origin of the henna. More information on this textural quality of henna pastes can be found on page 23 and 24 of Chapter Four: Henna Science and Microscopy from the Ancient Sunrise free E-book.

Quality henna powder should not be gritty, chunky, or contain visible pieces of plant leaf and/or stem. Henna should not bubble or turn frothy when mixed with liquid. Some other plant powders which contain saponins, such as Zizyphus, do froth. Bubbles may also indicate some reaction between the acidic liquid and an unknown ingredient in the powder.

Pure henna has a neutral, plant-like odor similar to dried straw or hay. It does not smell floral or spicy; it smells like leaves. Pure henna powder does not have a perfumed, pine, camphor, or eucalyptus scent. Henna powders can vary in color from light green to olive green. While color can be indicative of the powder’s freshness and quality, it is not always the case. Powders can be made greener by adding dyes.

For products labeled as henna for use on hair, the FDA has the following specifications:

“It shall not contain more than 10 percent of plant material from Lawsonia alba Lam. (Lawsonia inermis L.) other than the leaf and petiole, and shall be free from admixture with material from any other species of plant.

Moisture, not more than 10 percent.

Total ash, not more than 15 percent.

Acid-insoluble ash, not more than 5 percent.

Lead (as Pb), not more than 20 parts per million.

Arsenic (as As), not more than 3 parts per million.”

In other words, a product labeled as henna needs to be made from the leaf or petiole (leaf stalk) of the plant rather than the bark, roots, or any other part. It should be a dry good. It should contain no more than 15 percent inorganic filler, or ash. This can include substances such as sand and metallic salts. “Free from admixture with material from any other species of plant” means that products containing henna mixed with other herbs or plant materials are illegal to enter the United States. Several of the products selected for this article are labeled as herb mixes. Others, while labeled as pure henna, smell like additional herbs were included but not reported on the labeling or ingredients.

Please note that even if a henna product conforms to FDA standards, this does not necessarily make it a quality product. This list is simply the minimum for what the FDA considers an acceptably safe product for sale and use within the United States. A product labeled as henna for hair could be roughly ground henna leaves with 15% sand and still be legal. It is also important to mention that this article does not aim to determine the legality or safety of any of the products tested. This article serves only to report on observable physical qualities and on the presence of dyes in each product.

Below are the observations on each sample.

#1. This product was labeled as 100% natural, organic henna powder. No other ingredients were reported. The color was not of any concern. However, there was a faint herbal scent that suggested additional plant powders were added, most likely one or more of those commonly found in henna herbal mixes, such as shikakai, neem and so forth. The sift was fine. When mixed into a paste, the consistency was acceptable; its a slimy, gel-like texture was similar to Rajasthani hennas, such as the AS sample. This makes sense, given that product was from India.

#2. This product also claimed to be pure henna. However, there was a very noticeable herbal scent. The color of the powder was much deeper than a henna powder should be, almost like the color of nutmeg powder. Larger particles in the powder suggested a low sift. When mixed into a paste, the product frothed slightly and had a gritty consistency. The paste also showed a darker color than would be normal for a henna paste that had just been mixed. Shikakai powder has a red tone and herbal scent, lathers slightly when mixed. This may explain the qualities in found with this product, but would mean that the product is definitely not pure henna.

#3 The product claimed to be pure henna. There was nothing out of the ordinary about the color or texture of this product. Some of the powder formed soft lumps which could be broken with light pressure. There was no discernible scent beyond a normal plant-like scent. The color was acceptable for henna powder. The paste consistency was thinner and runnier than the others, despite all powders being mixed with the same amount of liquid.

#4. The product was labeled as “henna with natural herbs.” There was a very strong herbal scent of something other than henna. The powder was a vivid, brick red, and did smell like the other “herbal hennas” and like samples #1 and #2. The powder had large plant particulate matter and long, thin pieces of plant fiber. When liquid was added, the liquid itself immediately turned blood red even prior to mixing. The paste, after mixing, was blood red. This suggests that the product contains a water-soluble red dye. The paste texture was much grittier than a normal henna paste should be. Shikakai would not be a sufficient explanation for the color of this powder because while it has a red color, shikakai does not produce a dye.

#5. This product was labeled as henna with added herbs. The scent matched this. The powder showed large stem and/or leaf fiber particles which were easily visible to the naked eye. The paste consistency reflected this lower level of sift. There was a slight sliminess to the paste; if better sifted, the product may have produced a paste similar to a Rajasthani henna. The light green-brown color of the powder and paste was similar to how a henna product should appear.

#6. This product, like #5, was labeled as an herbal henna. Both had similar added herbs reported on their packaging. The product was better sifted than #5, but some plant particulates were visible. The powder had an herbal scent. The paste showed more of the shiny, slimy mucilage texture than #5, but not as much as can be seen in the AS sample. This would match the packaging’s statement that the henna was from the Rajasthan region of India. The powder and paste colors were a light green-brown consistent with natural henna products.

#7. This product was the “red henna” product of three sold by the company: Red (henna), Neutral (cassia), and Black (indigo). One might assume that the products are sold for the purpose of mixing, much like Ancient Sunrise plant dye powders, but it is not clear. The powder was of a light green color with pieces of plant fiber visible. The powder had an odor consistent with henna, but the product frothed when mixed with liquid. The paste was quite gritty.

#8. This product was labeled as 100% natural henna. The color was of a light greenish straw tone, along the lines of a standard henna powder color. In color, it was the sample most similar to the AS sample. When mixed with liquid, the resulting paste was noticeable gritty, but with some mucilage. There was no herbal scent.

#9. The product was also labeled as 100% natural henna. The color of the powder was deeper than what might be accepted from a henna powder, almost a muddy green. The powder appeared fairly well sifted, with some very thin, long plant fibers visible. The paste was fairly smooth but not slimy, and a mid-brown color. There was no herbal odor.

AS. This powder is finely sifted, soft to the touch, and a pale green color. No large leaf or stem pieces or fibers as visible. There is a faint dried-hay scent. The paste was a clay-green color had a smooth, slimy texture. In fact, in comparison the other samples, this sample took more focus to apply the paste to paper chromatography strips because of its mucous-like texture.

Chromatography

Paper chromatography is a simple method used to separate dyes in order to determine the presence of one or more dyes. The method involves placing a small sample of a substance onto a thin strip of absorbent paper, then hanging the strip so the end of it is just below the surface of a liquid solvent.

As the solvent travels up the strip, dyes move along with it. Dyes which differ in chemical structure will move at different rates before stopping. The result is that each individual dye will create its own mark at a certain place between the point of application and the solvent front (the highest point that the solvent reaches). Multiple dyes will show as multiple marks.

Below is the result of a paper chromatography test done on store-bought food coloring. One can see that the green dye is a combination of yellow and blue dyes, which were separated through the chromatography process.

One drawback of the simpler paper chromatography method is that it alone cannot identify the dye in each sample unless the target dye is already known. Because Ancient Sunrise henna powders are tested by an independent lab, it can be assumed that the only dye present is lawsone, which will show a dye band distinct from other dyes’ banding. If a product results in one or more bands that appear different from lawsone, it can be assumed that other dyes are present.

It was necessary to test each product under multiple conditions to achieve as full a picture as possible. Because the target dyes involved were unknown, it was not possible to test for specific dyes. Instead, each sample was tested multiple times. Depending on the nature of the solvent used, different dye bands were more or less visible on the results.

Chromatography Process

Unlike chromatography tests in which the target dye is known, here we are looking for the presence of dyes, whatever they may be. While we want to see the presence of lawsone, we also want to see what else may be there. “Henna” products may contain a wide variety of unknown dyes, some of which react to some solvents but not others. While a dye may show up on one type of test, it may not at all in another. Therefore, multiple solvents and solvent strengths should be used.

Pre-trial tests were conducted to determine which solvents and solvent strengths yielded the most useful results. Solvent type and strength also affected how long the sample strips were left in the tank before being removed. Leaving samples in the tank too long can risk dyes being “bleached out” by the solvents so that results became unclear. Pre-trial tests helped to determine the best processing times.

The most conclusive results came from sampling the pastes with the following solvents, strengths, and times:

  • 1. 99% isopropyl alcohol for 20 minutes
  • 2. 1:1 mixture of isopropyl alcohol and distilled water for 15 minutes
  • 3. 1:1 mixture of acetone and distilled water for 15 minutes

It was found that 100% acetone moved so quickly up the paper strips that no useful results were obtained. Therefore, samples on 100% acetone will not be reported in this article.

First, each product was made into a paste following classic henna dye-release methods, which will be described in more detail below. In addition, to account for the possibility of time and acidity breaking down any non-lawsone dyes, each product was tested again by mixing each powder with only distilled water and testing immediately.

Below are the methods and results of each set.

Set 1: Dye Released Paste Samples

The Lawsone Molecule in Henna

The molecule responsible for the orange/red color achieved by henna is called lawsone. In the henna plant, lawsone precursors exist. When the plant powder is mixed with a mildly acidic mix, the precursors are released as intermediary aglycone molecules in the process that is commonly known as dye release. Aglycones can release in a water-only mixture as well but will oxidize to their final stable state very quickly. The additional hydrogen atoms present in acidic liquid allow the aglycones to stay in their intermediate state longer so that the dye can release more fully before it is applied to the hair. Lawsone in its final state is oxidized and unable to bond to keratin.

The process of oxidation is what gives henna its final color. This is why hair is often lighter and brighter initially after rinsing henna. The color deepens over the course of subsequent days. When the dye molecules in henna paste oxidize before bonding to keratin, this is called demise. This is why pure lawsone on its own is useless as a hair dye.


In a mildly acidic liquid, the lawsone precursor (center) is maintained so that it can attach to keratin, after which it oxidizes to its final state (right).

Mixing and Dye Release

Each powder sample was mixed with a prepared solution of distilled water and lemon juice. Lemon juice was added to distilled water until the solution showed a pH of roughly 5.5, as indicated on a litmus strip. The resulting solution was 10 parts distilled water to 1 part lemon juice, or 100ml distilled water and 10ml lemon juice.

Five grams of each powder sample was mixed with the acidic solution to form a paste. The acidic solution was added first to the AS sample to determine an appropriate ratio for creating a paste similar to one that would be used for coloring hair. Thus, the result was 15ml acidic solution for each 5g powder sample*. All prepared pastes were kept in air-tight dark glass jars to prevent excess exposure to oxygen and light. Starting from the moment the final paste was mixed and sealed, all samples were left at 65 degrees Fahrenheit for 12.5 hours for dye release.

*Due to each powder sample varying in particle sift, some pastes were thinner or thicker than others. The variation in paste viscosity may have been a slight factor in test results, but not in any way that could lead to false results.

Set 1 Process

After dye release, a small amount of each sample was applied to paper chromatography strips 0.5 inches from the base of each strip. When not being used, all samples were kept refrigerated at approximately 37-40 degrees Fahrenheit. All samples were tested within 48 hours of dye release. Strips were hung in a glass chromatography tank containing a selected solvent so that the bottom edge of the strips sat just under the surface of the solvent. A glass plate was placed over the top of the tank to reduce airflow. After the determined time, the strips were removed and analyzed.

Below is a short time-lapse video showing how the sample traveled with the solvent. This particular set shows one paper strip for each sample and five strips at a time. One minute of real time is translated to one second of time in the video.

Results

These images show results comparing all paste samples across solvents and times. Samples are all tested numerous times, but for the sake of visual reporting, one or two paper strips of each sample for each test were chosen based on which were the most representative of the group. Outliers were those whose results appeared least like the others, most likely due to an odd variation in the paper strip causing inconsistent solvent wicking. These were disregarded. The remainders showed consistent results with little variation.

99% Isopropyl Alcohol for 20 Minutes

One can see that all of these strips show mostly pale dyes that range from green to orange to light brown, and that few show clear dye bands below the solvent front. In the world of chromatography, most of these would not be usable results. However, for our purposes, they are interesting. This is because we know that lawsone does not show much color in its intermediate aglycone state. The presence of water can cause oxidation of the aglycone to its stable final form. So, when using an anhydrous solvent, it would be expected that the dye does not turn up bright orange. If it does, it might suggest that final-state (oxidized) lawsone was present in the product, or that an orange dye other than lawsone was added to the product.

If you are familiar with the process of coloring your hair with henna, you will know that the final color result takes a few days to mature. The lawsone molecule exists in a precursor state called an aglycone when it is mixed in an acidic paste and allowed to dye release. As long as the paste is not exposed to oxygen, and if it is not left to demise, most of these molecules remain aglycones. It is through the process of bonding to the keratin in your hair, and to the oxygen molecules in the air that they become stable lawsone molecules, and you see the deeper color. This is also why henna paste applied to the skin is left for several hours, and the design deepens over the following days.

BUT, if a lower quality henna powder is “fixed” with added lawsone, that lawsone would be the stable-molecule type. This is sometimes done by henna sellers to make the product appear to have a higher lawsone content, or as an attempt to make the product more effective. Manufacturers and distributors who do not have a solid understanding of the chemistry of henna do not realize that adding lawsone will do nothing to the efficacy of the product. Stable lawsone will not bind to the hair because it has lost the hydrogens necessary to create a bond. Only the aglycone lawsone molecules released from henna powder by an acidic liquid have the ability to bind to keratin.

One can see that out of all of the other samples, AS (shown above) is one of the palest, nearly a yellow-green color, and almost seems to form a band below the solvent front. Virtually no dye remains near the application point. This indicates two things: first, that there was little to no oxidized lawsone in the paste sample. Second, the fact that the block of space between the paste application line and the solvent front is so “clean” suggests that there is only one type of dye at one point of chemical state. We know that this dye is lawsone in an aglycone state. The more dyes involved, the muddier the strip may appear, as it will show evidence of several observable dyes, or dyes in several molecular states. In fact, even a product that contains a nothing else but a blend of several different hennas would most likely show variation due to those dyes being just slightly different from each other.

Samples such as #2, 6, and 8 are more orange, and also show darker bands right above the sample application sites. The muddy brown band is very prominent in #6. In fact, all samples except for AS showed some level of banding that hugged the paste sample when tested with 99% isopropyl alcohol. This happens when there is a dye present which does not move well with the selected solvent and which prefers to stay low with the sample. Lack of dye movement in itself is not necessarily bad, and is simply indicative of the relationship between the dye and the solvent; however, because we can see that this did not happen with the AS sample, we can guess that those other powders contained something that AS did not.

Closeups of samples #2, #6, and #8.

Both #2 and #6 are particularly orange, which again suggests the presence of either added lawsone or another dye that comes up orange under these conditions. If we revisit the image of those powders prior to mixing, sample #2 especially is of a darker color. #6 and #8 are lighter, but still border on what would be a normal color for henna powder. It is possible that these three products have added lawsone or another orange dye.

#4 is very interesting, and will continue to be interesting as we go along. There is very clearly the presence of a red dye that is not lawsone. It is a vivid red which appears just above the application point. If you revisit the earlier sections of this article which show the powders and pastes, #4 is a vivid red color as both powder and paste.

#9 shows the presence of a green-blue dye which appears as faint streaks closer to the base of the strips. The solvent front also shows the same color near the edges. In addition, there is a deeper, brown band that stays just above the application point.

#7 also shows a faint green tint similar to #9, but there are no noticeable blue-green bands. Some green tones can be expected due to the presence of chlorophyll in the plant powders. Therefore, a green color does not guarantee the presence of an added dye. However, the blue-green streaks present in sample #9 are not consistent with chlorophyll.

1:1 Dilutions for 15 Minutes

In comparison, dye bands were more visible when pastes were tested with equal parts solvent and distilled water. This is because water causes the lawsone dye to oxidize, turning the dye a deeper orange. In fact, the dye stained the paper strip as it moved upward with the solvent.

Water alone was not an effective solvent because the dye did not travel with it quickly enough. Pure solvent, on the other hand, pulled the dye too quickly, leaving little to no banding, as was shown by the 99% isopropyl alcohol set. By combining distilled water and solvent, both dye oxidation and dye movement was achieved in a way that showed visible dye bands.

In the image above, A and B labels indicate results for 1:1 isopropyl alcohol/water and 1:1 acetone/water respectively. Both were left in the tank for 15 minutes. Results were generally similar for both solvent dilutions.

In comparison to 99% isopropyl alcohol, results from 1:1 dilutions were deeper orange with dye bands visible on most samples. While they are still faint, the banding for lawsone can be seen as stripes of deeper color just below the solvent front.

One can see that the AS sample remained relatively pale, and that the dye seemed to move more cleanly away from the application line, leaving lighter streaks where little or no dye stained the paper.

Sample #4 clearly shows the presence of a red dye along with lawsone. There is a bright red band that stays near the application line. The band moved higher in the 1:1 acetone/water sample, but both tests show separate red and orange bands.

Sample #2 shows a significantly darker dye which creates a nearly solid stain from application line to solvent front. There is a band near the top which could be lawsone. If you look closely, there is a slightly deeper orange band that stays near the application line. This suggests that there may be an additional orange dye along with lawsone. Sample #1 also shows a darker color in comparison to other samples, which may suggest the presence of additional dyes.

Another notable detail is the color of the paste samples at the application line after processing. Many are bleached out as the solvent pulls the dye from the sample. The AS application line is the palest. Samples #2 and #4, however, remained very dark. This is another reason to believe that additional dyes besides lawsone were present.

Sample #9, which show blue-green dye streaks when processed with 99% isopropyl alcohol, changed significantly when processed with 1:1 dilutions. Most likely the faint blue-green dye was overpowered by the orange stain.

Top: Sample #9 Tested with 99% isopropyl alcohol. A blue-green dye is visible. Bottom: Sample #9 tested with 1:1 isopropyl alcohol/water and 1:1 acetone and water respectively. The blue-green dye is no longer visible.

Set 2: Powder and Water Samples

It is important to note that all samples were treated as if they were 100% pure henna powder– that they contained no additional dyes or acids, and that they would require an acidic solution and dye release time. This was done to keep results consistent.

However, there was the possibility that added water-soluble dyes were present which could have been broken down by the acidic solution, or by sitting too long after mixing. Thus, the same process was repeated with paste samples made from just distilled water. These samples were tested immediately after mixing, with no wait time.

Theoretically, this set would show very little lawsone especially in the 99% isopropyl alcohol condition, as the henna would not have had much chance to release lawsone in its aglycone precursor state. The 1:1 dilutions may still provide an environment for a fast release/oxidation of lawsone, but some additional dyes may be better seen than in Set 1, if they were affected by the acidity and/or 12 hour wait.

For each sample, 1g powder was mixed with 3ml distilled water. The paste was stirred until a uniform consistency was achieved, then a small amount was immediately applied to paper chromatography strips and tested with the same solvents and times as Set 1. To repeat, the conditions were as follows:

  • 1. 99% isopropyl alcohol for 20 minutes
  • 2. 1:1 mixture of isopropyl alcohol and distilled water for 15 minutes
  • 3. 1:1 mixture of acetone and distilled water for 15 minutes

New paste was made for each solvent condition so that each sample was tested immediately after mixing with water.

Results

99% Isopropyl Alcohol for 20 Minutes

Results for powder-and-water samples immediately tested with 99% isopropyl alcohol.

The image above shows results for powders mixed with distilled water only and immediately tested in 99% isopropyl alcohol. Unlike the results of dye-released pastes tested with 99% isopropyl alcohol, samples mixed with only water showed consistent bands rising to just above the application point. Less dye traveled upward with the solvent, leaving paler tones near the solvent front. Below is the image for dye-released pastes tested with 99% isopropyl alcohol for comparison.

Notable results for this set are those of samples #2, #4, and #9.

Sample #2 appears more vividly orange than all other samples. This occurs in both the dye-released set and the water-only set. There is strong evidence for an added orange colored dye, lawsone or otherwise.

Sample #4 shows the same red dye that has been apparent in all tests. Because more red dye traveled with the solvent in the water-only set in comparison to the dye-released set, this is most likely a dye which is affected by pH and/or time.

Left: Sample #2 mixed with water and tested in 99% isopropyl alcohol. Right: Sample #4 mixed with water and tested in 99% isopropyl alcohol.

The faint green-blue dye that was noticeable in previous tests of sample #9 is particularly present in this condition. It was especially noticeable during processing. In the image below, all four strips show streaks of green-blue dye traveling upward with the solvent. The clearest is the strip second from the right.

After the strips were left in the glass tank for a full twenty minutes, the dye had collected at the solvent front. Some dyes streaks can also be seen rising just a few millimeters out of the application point. Particles of the same color can also be seen within the paste. It is clear that the green-blue dye is some form of powder or crystal mixed into the product.

1:1 Dilutions for 15 Minutes

The paper chromatography results using equal parts water and acetone, and equal parts water and isopropyl alcohol appeared very similar to the results of the dye-released pastes tested with the same dilutions. Nothing additional was revealed that was already noted in the earlier tests. This may be because the distilled water present in the diluted solvents was enough to reveal water-soluble dyes.

Discussion

The purpose of this article was first: to observe and report the visible qualities of each product, such as the product packaging, and the color, scent, and texture of the product both as a dry powder and after being mixed with liquid; and second: to determine the existence of dyes other than lawsone or lawsone precursor molecules through the use of paper chromatography.

None of the methods used in this article can definitively determine the safety or quality of a product. Thus, it is not our intention to rate, review, or suggest any product outside of the Ancient Sunrise brand. The fact that some products appeared similar to pure henna does not mean that they were void of additives or adulterants not shown by these tests. Paper chromatography cannot show the presence of non-dye chemicals such as pesticides. Given those disclaimers, let’s jump into what was observed.

The presence of red, orange, and green dyes in some samples indicate that those products included additional ingredients besides pure henna powder. While all of the samples tested showed the presence of some lawsone, many samples displayed results inconsistent with the AS sample which we know to be 100% pure henna powder. When dye-released pastes were tested with 99% isopropyl alcohol, all samples except AS showed dye that did not move with the solvent, staying low to the application line. Some samples were much darker in color, which is inconsistent with the pale yellow-green color that would appear with lawsone in a precursor state. It is unclear if some samples included oxidized lawsone, and whether that lawsone was added during manufacturing or if other conditions such as the age or storage of the product may have led to the presence of oxidized lawsone.

Sample #2 showed a very deep orange color in all solvent conditions. When tested with 99% isopropyl alcohol, the deeper color dye was visible just above the application site. In 1:1 dilution tests, the dye traveled further up the strip, staining the strip orange from application to just under the solvent front. This suggests that there is an added water-soluble orange dye present in sample #2.

Sample #4 as a powder, as a paste, and when tested with 1:1 solvent dilutions.

Blue-green dye was visible in sample #9 when tested with 99% isopropyl alcohol. In diluted solvent conditions, the blue-green dye was no longer visible. This type of dye was most likely added to give the powder a greener color, rather than to affect the color outcome on the hair.

Left: Sample #9 tested with 99% isopropyl alcohol. Blue-green dye is visible.
Right: Sample #9 tested with 1:1 dilutions. The lawsone staining overpowers blue-green dye visibility.
Comparison between sample #9 powder (left) and AS powder (right).

Because there is a marketing claim that a greener henna powder is fresher, adding green dye to henna powder is not an uncommon practice. Detailed information about henna powders “polished” with green dye can be found on page 39 of Chapter Four: Henna Science and Microscopy. Ironically enough, sample #9 was among the darker colored powders and pastes in the group, so the addition of green dye did not seem to do what it intended. Below is an image at 60x magnification of a henna powder which shows added green dye.

A sample of henna powder under a microscope. Green dye powder or dyed sand is mixed into this sample.

Limitations and Considerations

Paper chromatography is only one type of chromatography, and not as exact as methods accessible in a certified laboratory. High-performance thin-layer chromatography, for example, is capable of separating dyes into much clearer bands, and then a scientist can calculate and compare the distance of those bands between the application point and solvent front to determine more about the nature of each dye. In many cases, reactants can be applied to chromatography results to cause substances to be visible if they are not already. Access to such methods would have been beneficial. An example of high-performance thin-layer chromatography can bee seen on page 22 in Henna Science and Microscopy.

Additionally, some samples varied in age, which can affect the quality of a powder to some degree. That being said, pure henna products have a shelf life of several years as long as they are kept sealed and in a cool, dry environment. Because the products tested may have contained additional unknown ingredients, it is difficult to say whether those specific non-henna ingredients may have broken down or changed with time.

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Comparing Ancient Sunrise® Sudina Indigo and Zekhara Indigo (Video)

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How to Apply Ancient Sunrise® Henna for Hair

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Mehandi Employees’ Favorite Products

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