Vol 7(1) January-February 2008 45

IntroductionFrom ancient days various

materials from plants like Henna,Chamomile, Indigo, etc. are used to dyethe grey hair to get natural black colour.But instead of getting black colour peopleget red to copper colour. Loss of colourin hair is due to varied reasons likegenetic influence, effect of environmentalfactors, use of alcoholic preparations,etc.1. Though permanent synthetic hair dyesare available in varied colour and rangesretain natural lustre, they have the chiefdisadvantage of producing hypersensitivereactions in some individuals. Some hairdyes marketed as natural dye, contain 1-3% of phenylenediamine which is asynthetic hair dye and stain the skin andclothes during use2.

A need was felt to formulate aproduct containing only plant products,which is safe for use and does not havethe problems of staining skin during useand hypersensitive reactions. In the present

investigation the formulations aredeveloped to get natural black colourusing combination of different plantmaterials. This investigation was plannedbased on experiences of several people whowere using different plant products forcolouring their hair without having anyproblems of irritancy, allergy or sensitivity.

The main aim of the presentinvestigation is to formulate natural andsafe hair colorants.

Materials and MethodsThe plants used in the study are:

Henna (Lawsonia inermis Linn.)3,Indigo (Indigofera tinctoria Linn.)4,5,Bhringraj [Eclipta alba (Linn.)Hassk.]6, Amla (Emblica officinalisGaertn.)7, Shoe flower (Hibiscus rosa-sinensis Linn.)8,9, Black catechu(Acacia catechu Willd.)3, Cinnamon(Cinnamomum zeylanicumBreyn.), Tea [Camellia sinensis(Linn.) O. Kuntze]3, and Brahmi

[Centella asiatica (Linn.) Urban]10.

Preparation of natural hair colorantsThe leaves of henna, indigo,

brahmi, tea and bhringraj, fruits ofamla, shoe flowers, and Cinnamon barkwere collected from different places anddried in shade. They were made intopowders and used for the preparation ofhair colorants.

Collection of unpigmented hairThe human hair was collected

from barber shops and temples fromwhich white hairs were separated andused for the studies.

Preliminary preparation of naturalhair colorants

One gram each of henna, indigo,amla, bhringraj, hibiscus, brahmi,catechu, cinnamon and tea powder wastaken separately and added 200mg (20%)of urea in each sample. A smooth pastewas made with water separately. The pasteswere kept aside for 1 h for imbibition.The hair was kept in the above pastes for1h and then washed with water andobserved for its colouring.

Selection of suitable combinationwith henna

One gram of henna was mixedwith same proportion of indigo, tea,amla, bhringraj and 20% urea and

Formulation and evaluation of commonlyused natural hair colorants

Y Madhusudan Rao*, Shayeda and P SujathaUniversity College of Pharmaceutical Sciences, Kakatiya University

Warangal-506 009, Andhra Pradesh, India*Correspondent author, E-mail: ymrao123@yahoo.com

Received 8 December 2006; Accepted 9 October 2007

AbstractLoss of colour in hair is due to varied reasons like genetic influence, effect of environmental

factors, use of alcoholic preparations, etc. Though the permanent synthetic hair dyes are availablein different colour ranges and retain natural lustre, they have the chief disadvantage of producinghypersensitive reactions in some individuals. In the present investigation various combinations ofpowdered leaves, fruits and flowers of dye yielding and hair care plants like Henna, Indigo,Bhringraj, Amla, Catechu and Centella were evaluated for their colouring and sensitivity reactions.Henna and Indigo formulation was found to be suitable natural colorant.

Keywords: Natural hair colorants, Dye yielding plants, Hair care plants, Henna, Indigo.

IPC code; Int. cl.8 A61K 8/97, A61Q 5/10, C09B 61/00

Research PaperNatural Product Radiance, Vol. 7(1), 2008, pp.45-48

Natural Product Radiance46

water was added to that in order to makea smooth paste. The paste was kept asidefor 1 h for imbibition. The hair was keptin above paste for 1 hour. After that it waswashed with water and observed for itscolouring.

Selection of suitable combination ofhenna and indigo

Henna was mixed with indigo indifferent ratios from 1:1 to 1:5, mixedwith water to make smooth paste andprocessed similarly as above to observecolouring.

Effect of pH on the selectedcombination on henna and indigo

1:3 Ratios of henna and indigowere selected because it produced alteredcolour and effect of pH was studied. Tothe paste of above ratio, dilute ammoniasolution was added in order to obtain thepH of 6, 7, 8, 9. Further process wassame for observing its effect.

Effect of successive application ofselected combination of henna andindigo

1 : 3 Ratios of henna and indigowere selected and to that successiveapplication of paste was studied. Thehenna and indigo paste was applied2 to 4 times and washed repeatedly withshampoo and observed for its colouring.

Effect of modifying agents over haircolorants

One gram of henna and threegrams of indigo powders were weighedand taken in a dish; 20% of urea and 1%of ferric chloride were added and madeinto paste. The paste was kept for 1 hour.Similarly, preparations containing

3 and 6% of ferric chloride were alsoprepared. The above pastes were appliedto the white hair, waited for 1 hour. Thehair colour was observed undermicroscope after washing with water.Similarly, same concentrations of coppersulphate were also prepared and observed.

Comparative evaluation of theformulation with marketed brands

For comparison with theprepared paste with 1:1 henna and indigowith 20% of urea prepared paste twomarketed herbal hair colorants[containing para phenylenediamine (PPD)in their formulation]. The above threepastes were applied to the hair, waitedfor 1 h and observed after washing.

Sensitivity testPrimary irritation testing:

This test11 was conducted on two albinorabbits to detect the potential primaryirritants present in the developedformulation. The animal ethicalcommittee approval was also taken. It wasascertained that the animals did not haveany skin sensitization or irritation history.The right side of back of each rabbit wasclipped with sharp blade, which wassterilized previously. The henna and indigowas applied on the lightly scarified skinof the right side and the left side of therabbit's back and the areas were coveredwith closed patches. The covered patcheswere kept in position for 24 hours. Theapplied areas were examined at intervalsof four hours for assessment of changeson the skin associated with irritation.

Predictive sensitivitytesting: The ascertainment of the irritantand sensitizing potential of formulationis prophetic or predictive sensitivity

testing. Under this test the ingredients ofthe formulation and the formulation as awhole were tested for their irritation andsensitivity12. This testing was conductedon 8 female volunteers hence, humanethical committee approval was obtained(No. is UCPSC/s/2006-01). Theformulation was applied to 1 cm2 area ofcotton gauze. The gauze was backed withadhesive tape in order to hold it inposition. Then 8 female volunteers wereasked to apply this patch to the innersurface of forearm. The purpose ofselection of this site for application wasto enable the volunteer to readily removethe patch if any irritation is encountered.The volunteers were asked to remove patchimmediately, if they feel itching orburning. Further they asked to keep thepatch site dry, avoid rubbing or scratchingtest site. The patch was kept in positionfor 24 hours. After 24 h, the patch wasremoved. The test site was observed upto 40 min from the time of removal. Thistime interval is necessary so that the skinmay recover from the effects of pressureof certain patch test substances.

Results and DiscussionIn the preliminary selection

individual powders (Henna, Indigo,Bhringraj, Amla, Shoeflower,Cinnamon, Black Catechu, Tea, andBrahmi) were tested for their haircolouring property. The colour producedby these plant products can be seen inFig. 1. From the figure it can be seen thatamong these dyes henna and indigoproduced good coloration because hennagives an orange-red colour and indigo givesan intense violet colour. By using thesetwo powders a suitable brown colour canbe developed. The other plants did not

Research Paper

Vol 7(1) January-February 2008 47

produce the desired coloration to the hair.Microscopic pictures of black and whitehairs and white hairs dyed with henna andindigo are shown in Fig. 2.

Henna and indigo were madeinto paste in different ratios as 1:1. 1:2,1:3, 1:4, and 1:5. In these preparations,1:1, 1:2 gives yellowish orange shades 1:4,1:5 gives bluish violet shades whereas 1:3ratio gives a brown shade. Hence 1:3 ratioof henna and indigo was selected forfurther study.

The effect of pH was studied on

Fig. 2 : Microscopic observation ofdifferent hairs

Fig. 3: Effect of pH on colour produced withhenna and indigo

Henna:Indigo(A*)-1:1 Henna:Indigo(A*)-1:2 Henna:Indigo(A*)-1:3

A* - Aged

Research Paper

Fig. 1 : The different colours produced by greyhair (white) after treatment with different herbal

powders in the form of paste

Henna Indigo Bhringraj Amla Shoe flower

Cinnamon Tea Powder Brahmi

Black hair White (grey hair) Henna dyed Indigo dyed white hair white hair

pH-6 pH-7 pH-8 pH-9

Fig. 4 : Microscopical observation ofcolour produced after repeated application of

henna and indigo 3 times

Fig. 6 : Microscopical observation of the effect ofmodifier (1% ferric chloride) on colour

modification with henna and indigo

Henna:Indigo(A*) -1:1 Henna:Indigo(A*) -1:2 Henna:Indigo(A*) -1:3

A*- Aged

Fig. 7 : Effect of ageing of the indigo leaves oncolour produced with 1% ferric chloride in

1hour after each application withrepeated 3 times of application

I Time II Time III Time

Henna:Indigo(not aged) (1:3) with 1% ferric chloride

I Time II Time III Time

Henna : Indigo(Aged) (1:1) without 1% ferric chloride

I Time II Time III Time I Time II Time III Time I Time II Time III Time Henna:Indigo (1:1) Henna:Indigo (1:2) Henna:Indigo (1:3)

Henna:Indigo (1:1) Henna:Indigo (1:2) Henna:Indigo (1:3)

After 6 Shampooings after 3 repeated applications

Fig. 5 : Effect of repeated application of hennaand indigo on colour development

Henna:Indigo (A*) 1:1 Marketed Product

A*-Aged

Fig. 8 : Comparison of the colour produced bythe herbal dye with marketed product

Henna:Indigo (A*) 1:1 Marketed Product

Microscopical observation

this ratio of henna and indigo. The colourwhich was obtained at pH 7 was betterthan that of pH 6, 8 and 9 as can be seenfrom the Fig. 3. Hence, all the pasteswere adjusted to pH 7 for further studies.

The henna and indigo pasteswhen applied repeatedly i.e., 3 times or4 times, there was an increase in intensityof brown coloration. The microscopicalobservation of this can be seen in Fig. 4.When hair was washed with shampoo 6times after repeated application of hennaand indigo pastes 3 times and every time

the hair was soaked in the paste for 1hand then washed with water, their browncolour was retained as such which can beseen Fig. 5.

The effect of modifier on haircoloration was studied by using 1% ferricchloride. The effect of 1% ferric chloridewas studied for the preparation containingwith and without ferric chloride whichwas subjected to the shampoo washings.After adding 1% ferric chloride, it wasobserved that there was an increase inthe intensity of brown colour and colour

Natural Product Radiance48

retaining property was also increased.Penetration of colour intensity of hennaand indigo with different ratios 1:1, 1:2and 1:3 with 1% ferric chloride wasstudied. There was an increase in intensityof hair colour observed microscopically,which can be seen in Fig. 6.

There was an increase in violetcolouring property, when the indigo leaveswere subjected to ageing.The henna andindigo aged and tender leaves powderswere taken in the ratios 1:1, 1:2, 1:3 andmade into paste, kept aside for 1h forimbibition. Then these pastes wereapplied on to grey hair and after 1hthe hairs were washed with water andcolour of the hair was observed. Thebrown colour obtained with hennaand indigo(not aged) (1:3) with 1% ferricchloride was same as that obtainedwith henna and indigo (aged) 1:1 ratiowithout 1% ferric chloride itself whichcan be seen in Fig. 7. The browncoloration was more in the later one.Hence a dark brown colour was obtainedwith pure plant products itself in theratio of 1:1.

The preparation thus made wascompared for the intensity of colour withthat of marketed products, which containsPPD in their formulation. The dark browncolour which was obtained with henna,indigo 1:1 after 3 applications wascomparable with the marketed productas shown in Fig. 8. The penetration efficacyof hair colour into the cortex was studiedmicroscopically. There was not muchdifference between henna and indigo andthe marketed product which can be seenin Fig. 8. The intensity of colourpenetration was also studied aftersubjecting to 6 shampooing. There is nodifference between marketed product and

henna, indigo combination as seen inFig. 8.

Henna and indigo paste wassubjected to predictive test andpreliminary sensitivity test. These testswere carried out initially on the albinorabbits. No irritation or hypersensitivity,reactions were observed. The preliminarysensitivity test, performed on humanvolunteers also shown no sign of irritationor hypersensitivity.

ConclusionIt can be concluded from the

investigation that by changing theproportion of henna and indigo a suitablebrown colour could be obtained for hair.A pH of 7 was best for penetration of haircolorant. Repeated application of thehenna and indigo product gives an increasein the colour intensity. Addition of ferricchloride enhance the colour retainingproperty. It was observed that when indigoleaves are subjected to ageing give morecolour intensity. No influence of ageingwas observed in colour intensity withhenna powder.

Advantage of this natural haircolorant is that it does not cause anyirritation. Staining of nails or fingers isnot seen while preparing the hair colorantformulation or paste which is the mainproblem with marketed products. At thesame time the colour does not stick tothe clothes which come into contact withthe product. Hence it is a suitable naturalhair colorant and can be recommendedfor further studies and use.

References1. Marlen McHugh- Pratt, Milady's Standard

textbook of Cosmetology, Delmer Publishers,USA, 2000, pp. 265-304.

2. Brown Keith, Hair colourants, J SocCosmetic Chem, 1982, 33, 375-383.

3. Wallis TE, Text Book of Pharmacognosy, 5thEdn, CBS Publishers & Distributors, Delhi,1997.

4. www.life.uiuc.edu : Sieglev David S, Plants andtheir uses, Department of Biology, Universityof Illinois, USA.

5. The Ayurvedic Pharmacopoeia of India,Ministry of Health and Family Welfare, Govt.of India, Dept of Indian System of Medicineand Homeopathy, Part I, Vol. II, 1st Edn,1999, pp. 120-121.

6. Chatterjee Asima and Pakrashi SatyeshChandra, The Treatise on Indian MedicinalPlants, Publications and InformationDirectorate, New Delhi, vol. 3, 1994,pp. 33-35.

7. Kirtikar KR and Basu BD, Indian MedicianlPlants, 3rd Edn, Vol. 2, International BookDistributors, Dehra Dun, 1988, p. 1361.

8. Chatterjee Asima and Pakrashi SatyeshChandra, The Treatise on Indian MedicinalPlants, Publications and InformationDirectorate, New Delhi, Vol. 2, 1992,p.180-181.

9. Sivarajan VV and Balachandran Indira,Ayurvedic drugs and other plant sources,Oxford and IBH Publishing Co. Pvt. Ltd.,New Delhi, 1994, pp. 28-29.

10. Kokate CK, Purohit AP and Gokhale SB,Pharmacognosy, 19th Edn, Nirali Prakashan,Pune, 2002, p. 220.

11. Philips L, Steinberg M, Maibach HI and AkersWA, Comparasion of rabbit and human skinresponses to certain irritants, Toxicol ApplPharamacol, 1972, 21, 639.

12. Koehler Paul B, Clinical aspects of safetytesting of cosmetic products, J SocCosmetic Chem, 1980, 31, 213-218.

Research Paper