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Comparative tissue stainability and localization of active staining principle in fractions of Lawsonia inermis (Henna) leaves
Author(s) -
Alawa Judith Nkiruka,
Kauche Eunice,
Ahmed Bilkisu,
Adetiba Bamidele
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.532.4
Subject(s) - staining , ethyl acetate , distilled water , haematoxylin , chemistry , butanol , chromatography , lawsone , eosin , uranyl acetate , stain , biochemistry , biology , ethanol , organic chemistry , genetics
The crude aqueous and ethanolic extracts of the leaves of Lawsonia inermis (Henna) have demonstrated cytoplasmic staining properties in animal tissues. However, to improve the staining efficacy on tissues and to localize the active dyestuff, the extracts were fractionated sequentially in n‐hexane, ethyl acetate, n‐butanol and distilled water using column chromatography. The soluble fractions were subjected to thin layer chromatography to identify the active compounds. Tissue stainability of the fractions was evaluated in formalin‐fixed and paraffin embedded tissues (liver, kidney, cardiac muscle, testis, lungs, brain and intestine) obtained from Wistar rats. Ethyl acetate was the most effective and showed better tissue stainability compared to n‐Butanol, characterized by the deep brown colour intensity on the tissues and remarkable contrast with haematoxylin comparable to haematoxylin and eosin stain. This corroborated the TLC analysis which demonstrated that the ethyl acetate and n‐butanol fractions showed brownish spots which were absent in the hexane and distilled water fractions. Ethyl acetate spot showed deeper colouration than the n‐Butanol spot suggesting that the active staining principle known as lawsone is localized in these fractions. The n‐hexane and distilled water fractions showed weak to no staining activity on tissues. This indicates that the active staining principle of Henna is localized in the ethyl acetate and n‐butanol fractions and these will serve as good solvents for the further purification of the active staining compound. Grant Funding Source : A.B.U Zaria