1. PHYTOCHEMICAL SCREENING AND CHROMATOGRAPHIC ANALYSIS OF HENNA (LOWSONIA INERMIS) PLANT OBTAINED FROM SAMARU, ZARIA.  By Danzarami, D1; Umar, M.1; Akafyi, D.E.1; Oko, J.O.1; Yusuf S.O1. Okeh, Q.1; and Adamu, R2
Department of Science Laboratory Technology, Nigerian Institute of Leather and Science Technology, Zaria, Kaduna state, Nigeria.
Department of Microbiology, Kaduna state Polytechnic, Kaduna state, Nigeria.

2. INTRODUCTION
Henna (Lawsonia inermis) also known as (Hina tree or Mignonette tree, Lalle in Hausa) are the sole species of the Lawsonia genus. The English name “henna” comes from the Arabic word Hina means ‘dye’; the name also refers to the dye prepared from the plant (Bailey and Bailey, 1976).
The plant has been used since ages to dye skin, hair and finger nails, as well as fabrics including silk, wool and leather. The name is used in other skin and hair dyes, such as black henna and neutral henna, neither of which is derived from the henna plant (Catherine et al., 2004).

3. BACKGROUND OF THE STUDY
Medicinal plants such as Henna (Lawsonia inermis) are the richest bio-resource of drugs in traditional systems of medicine, modern medicines, pharmaceutical intermediates, nutraceuticals, food supplements, cosmetics, medicines and chemical entities for synthetic drugs.
The medicinal value of these plants lies in some chemical substances that produce definite physiological actions on the human body and these chemical substances are called phyto-chemicals (Ncube et al., 2008).

4. SIGNIFICANCE OF THE STUDY
The research study will pave a way for the determination of the phytochemical constituents of Henna leaf extracts, which are medicinally useful in traditional medicine in the treatment of different ailments, and can be used in leather industries for dyeing and preservation of tanned leather.

5. AIM
This research work targeted to extract Henna (Lawsonia inermis) plant using various solvents with view of determining the phytochemical constituents and colour contents using thin-layer chromatography analysis.

6. RESEARCH OBJECTIVES
To extract Henna (Lawsonia inermis) plant using various solvents
To determine the phytochemical constituents of the Henna plant extracts
To analyze the colour content of the extracts using thin-layer chromatography analysis

7. METHODOLOGY SAMPLE COLLECTION
Fresh plant leaves of Henna (Lawsonia inermis) were collected in June 2015, from Samaru market Zaria, and taxonomically characterized at the herbarium section of Biological Science Department, Ahmadu Bello University, Zaria with the aid of botanical keys (Arber, 1972). The leaves were thoroughly washed through running water and dried under shade for 4-6 days, and ground into powder form. The leaf powder was stored and sealed. The bioactive components were extracted using the methods of Akerele et al (2008).

8. METHODOLOGY CON’T
The method of cold maceration was used for this research work. The extraction was carried out by serial exhaustive extraction method which involves extraction with different solvents.
The extracts of the leaves were prepared by soaking 50g each in 500ml methanol, ethyl acetate and distilled water for three (3) days with frequent agitation until soluble matter is dissolved. The resulting mixture was filtered and the filtrates were concentrated by evaporation using evaporating dish to remove all solvents as adopted by Kok et al., (2004).
(Prescott, 2008).

9. METHODOLOGY Cont.
The extracts were subjected to Phytochemical analysis to detect the presence of the chemical constituents such as Tannins, Steroids, Glycosides, Saponnis, Phenol, Alkaloids, Flavonoids and Anthraquinones using standard protocol as adopted by Trease and Evans (2002); and Debela (2002).

10. METHODOLOGY Cont.
Samples were dissolved in 10mls of 70% Ethyl acetate and filtered. A standard Whitman TLC plate (TLC silica gel 60F) was used.
The spots were visualized after the TLC plate was sprayed with 10% sulphuric acid followed by heating in an oven for 2 minutes (Edeoga et al., 2005).
The number of spots developed following treating the extracts with various solvents was observed and the RF values were calculated.

11. RESULTS
Table 1: Phytochemical screening of the crude extracts and it’s fractions
CHEMICAL CONSTITUENTS
M.E
E.A.E.
W.E
Tannins + -
Flavonoid
Saponins
Alkaloids
Glycosides
Steroids
Phenols
Anthraquinones
Key: + = Present; - = Absent; ME= Methanol extract; EAE= ethyl acetate extract; WE= Water extract

12. RESULTS Cont.
Table2: TLC Analysis of the Crude Extracts and its Fractions
Sample
Number of spots
Rf values
Ethyl acetate soluble fraction 5
0.8, 0.2, 0.7, 0.1, 0.9
0.2, 0.96, 0.1, 0.3
Methanol soluble fraction 4
Water soluble fraction
Total crude extract

13. COLOURS AND THEIR RF VALUES
brown, light brown, lemon green, green, dark green, light yellow, yellow, orange and light blue

14. DISCUSSIONS
Based on the findings of this research study, the phytochemical analysis recorded various results. Table 1 shows the result of the qualitative Phytochemical screening of the crude methanol extract, ethyl acetate extract and the water extract of the leaves of Lawsonia inermis. The result of this work revealed the overall presence of tannins, glycosides, Anthraquinones, saponins, flavonoids, alkaloids, steroids, phenol in the total crude ethanol extract.

15. DISCUSSIONS Cont.
The Thin Layer Chromatography analysis revealed that henna contain nine (9) different colour components which are brown, light brown, lemon green, green, dark green, light yellow, yellow, orange and light blue. Their retention factors are as follows: (0.2, 0.96, 0.1, 0.3, 0.2, 0.8, 0.1, 0.9, and 0.7 respectively). These colours can be used in leather and cloth industries as dyes and preservatives. This concurs with the findings of Bechold and Mussak (2009), who reported that henna extract acts as an antifungal and a preservative for leather and cloth.

16. CONCLUSION
The Phytochemical screening of Henna leaves extract using different solvents demonstrated the presence of the chemical constituents which are medicinally useful in traditional medicine in the treatment of different ailments, and can be used in leather industries for dyeing and preservation of tanned leather.
TLC analysis revealed the presence of different colour components which proved that the plant extract can be used for dye and preservation purposes

17. RECOMMENDATIONS
Based on the results obtained, it is therefore recommended that henna plant could be seen as a potential source of useful drug and the continued traditional medicinal uses in the treatment of various sicknesses. These plants are encouraged to be harnessed in both orthodox and traditional medicine. Moreover, henna should be produced in large scale for use in the dye industries due to its possession of colouring agents and preservatives.

18. REFERENCES
Arber (1972): Water Plants; A Study of Aquatic Angiosperm. Welden Wisely Limited Pp. 436
Akerele, Z. O., Obasuyi, O., Ebomoyi, M. I., Oboh, I. E., and Uwumarongie. O. H. (2008). African Journal of Biotechnology, 7(2):
Bailey, L.H. and Bailey E.Z. (1976). A concise dictionary of plants cultivated in the United States and Canada Macmillan. Hortus Third A.B.C ‘Pp-54
Catherin, Cartwright-Jones (2004). Cassiaobovata Henna for hair. Journal of Ethnopharmacology
Bechold, T. and Mussak, R. (2009). Hand book of Natural Colorants. John Wiley and Sons. Pp.155
Catherin, Cartwright-Jones (2004). Cassiaobovata ‘Henna for hair .Journal of Ethnopharmacology
Debela, A. (2002). Manual for Phytochemical screening of medicinal plant. Ethiopian Health and Nutritional Research Institute Addis Ababa, Ethiopia.:
Edeoga H.O., Okwu D.G. Mbaebie B.O (2005). Phytochemical constituent of some Nigerian medicinal plants. African Journal of Biotechnology; 4;
Ncube NS, Afolayan A. And Jokoh Al (2008). Assessment techniques of antimicrobial properties of natural compounds of plant origin. African Journal of Biotechnology. 7(12):
Kok N., Ertekin M.V and Auci B. (2004) Henna induced haemolytic anaemia in siblings International Journal of Clinical Practice. 58:
Trease G.E, and Evans W.C (2002). Pharmacognosy: A physical guide to herbal medicine 15th edition Bailiere Tindal London

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