PHARMACOGNOSTIC AND CHROMATOGRAPHIC PROFILING OF KHADIRA ( ACACIA CATECHU

* Dr. Vandana Venugopalan 1 , Dr. M. A Shajahan 2 and Dr. V. C Indulekha 3 . 1. PG Scholar, Dept of Dravyagunavijnanam, Govt Ayurveda College, Thiruvananthapuram. 2. Professor, Dept of Dravyagunavijnanam, Govt Ayurveda College, Thiruvananthapuram. 3. Assistant Professor, Dept of Dravyagunavijnanam, Govt Ayurveda College, Thiruvananthapuram. ...................................................................................................................... Manuscript Info Abstract ......................... ........................................................................ Manuscript History

Khadira (Acacia catechu (Linn.f.) Willd) is a drug widely used in the pharmaceutical industry. It is an ingredient of several ayurvedic formulations like Khadirarishtam, Kanakabindvarishtam, Katakakhadiradi kashayam, Musalikhadiradi Kashayam etc 3 . Ascertaining the identity, genuineness and purity of herbal drugs has an important role in the maintenance of the quality of Ayurvdic formulations. This indicates the need to assess the genuineness of Khadira through pharmacognostical and phytochemical evaluation.
The objective of the present study is to evaluate the preliminary pharmacognostical phyto-chemical characters of the drug.

Materials And Methods:-
Sample collection-The sample drug Khadira (Acacia catechu (Linn.f.) Willd) was collected from herbal garden at Pharmacognosy Unit, Govt. Ayurveda College, Thiruvananthapuram and the sample was authenticated by Botanist, Drug Standardisation Unit, Govt. Ayurveda College Thiruvananthapuram, Kerala.

Study settings
Drug Standardisation Unit, Govt. Ayurveda College Thiruvananthapuram, Kerala.

Macroscopic evaluation-
Fresh log of wood of Acacia catechu (Linn.f.) Willd was collected. The bark and the sapwood were removed and the heartwood was collected, shade dried and stored. The heartwood of Acacia catechu (Linn.f.) Willd was subjected to organoleptic evaluation and the observed macroscopic characters were recorded.

Microscopic evaluation-
Microscopic evaluation is used for studying the histological features of transverse and longitudinal section of heartwood of Acacia catechu (Linn.f.) Willd. A portion of the heartwood was selected, soaked in water overnight and enough number of sections were taken. Transverse and longitudinal sections were prepared and stained as per standard procedure 4 . The sections were carefully transferred to a petri dish containing water and few thin sections that floated in water were selected. A stained section was carefully transferred on a clean glass slide using thin brush. With the help of forceps and needle a clean cover slip was gently placed over the section. Excess water was wiped with a blotting paper and the slide was placed on the stage of a digital microscope (Olympus digital-CS41, Japan, with CCD camera) for histological examination and direct images were taken.

Preliminary physical and phytochemical evaluation
Preliminary physicochemical and phytochemical analysis of genuine sample of the study were conducted. Physicochemical analysis included the evaluation of parameters like foreign matter, moisture content, ash values, fibre content, volatile oil content, sugar content and different extractive values, were done. It also included the qualitative tests for the detection of phenols, steroids, alkaloids, flavonoids, tannins and glycosides. The procedures were done as per procedures mentioned in ICMR 5 and Ayurvedic Pharmacopoeia of India 6 .
Heavy metal analysis by Atomic Absorption Spectroscopy and chromatographic techniques like TLC, HPTLC were also conducted.

Results And Discussion:-
The organoleptic evaluation and the microscopy of heartwood of Acacia catechu were conducted and the observations were noted. The data was analysed and results were interpreted from it.

Macroscopic evaluation
The observed macroscopical characters of heart wood of Acacia catechu (Khadira) are given in Table 1.

II. Microscopic evaluation
The microscopic evaluation of Acacia catechu was carried out. Transverse section of heart wood showed vessels occurring isolated or in small groups. Vessels were often blocked with tyloses. Biseriate to multi serriate medullary rays were seen. Medullary rays were seen as vertically running linear bands. Multiseriate medullary rays and pitted border vessels were seen.

Results of preliminary physicochemical evaluation
The screening of physical and phytochemical parameters of Acacia catechu were done as per the standard procedures. Physical characters like moisture content, total ash, acid insoluble ash, water extractive and alcohol soluble extractives were analysed.  Saponin +

V. Results of TLC and HPTLC
The spots obtained in TLC gives an idea about the plant constituents. The best separation was achieved using Toluene: Ethyl acetate: Formic acid in the ratio 6:3:1. The plates were first viewed through UV-fluorescence viewing cabinet (365 nm) and the Rf values of the spots were noted (Table No: 4). HPTLC was done and the HPTLC profile was shown in figure 4 and its 3D scan profile was shown in figure 5. Total ash (%) 1.9 +/-0.217 5 Acid insoluble ash (%) 1.1 +/-0.012 6 Water soluble extractive (%) 6.1 +/-0.047 7 Alcohol soluble extractive (%) 1.5 +/-0.

VI. Results of Atomic Absorption Spectroscopy
Atomic absorption spectroscopy is used in the determination of heavy metal elements and some non-metal elements in atomic state and the results are shown in Table No: 5. Four heavy metals-copper, cadmium, iron and lead contents were analysed and found within permissible limits. Hence the drug is not contaminated by heavy metals and can be used safely for internal administration.
Track I -Alcoholic extract of heartwood of Acacia catechu 422

Conclusion:-
Macroscopy and microscopy along with the preliminary phytochemical evaluation of heartwood of A catechu confirmed the identity, quality and purity of the plant. On physico chemical analysis the heartwood was found with water soluble extractive value higher than alcohol soluble extractive value (6 % and 1.5 % respectively). The total ash value was 2.9 %. It showed high fibre content (45.24%) The preliminary phytochemical screening showed the presence of different phytoconstituent groups such as flavonoids, saponins, alkaloids and tannins. AAS evidenced that the drug is not contaminated by heavy metals and can be used safely for internal administration. Here the observations and results obtained are useful for further pharmacological and therapeutical evaluation.