Volume : 2, Issue : 6, JUN 2016

SEPERATION AND QUANTIFICATION OF PHARMACOLOGICALLY ACTIVE MARKERS OLEANOLIC ACID, 20-HYDROXYECDYSONE AND ?- SITOSTEROL FROM ACHYRANTHES ASPERA AND FROM MARKETED FORMULATION BY HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHY

Mansee M. Pathak, Vikas V. Vaidya, Saurabh H. Patil

Abstract

Objective: To develop simple, accurate, precise and reproducible High Performance Thin Layer Chromatographic (HPTLC) method for simultaneous quantification of Oleanolic acid, 20-hydroxyecdysone and β-sitosterol in the whole plant extract of Achyranthes aspera.
Methodology: In the present work a precise, accurate and reproducible HPTLC method is developed and validated for simultaneous quantification of three pharmacologically active markers Oleanolic acid, 20-hydroxyecdysone and β-sitosterol from the whole plant of Achyranthes aspera and from marketed Cystone® tablets of Himalaya herbal healthcare. These markers were extracted from the whole plant and from Cystone® tablets using Soxhlet extraction followed by chromatographic separation on HPTLC silica gel 60 F254 pre-coated plates using double development method. The analysis was carried out using a double development method. The first development up to 50 mm was carried out using a mixture of dichloromethane- ethanol (8:2) (v/v) as mobile phase and the second development up to 80 mm using a mixture of petroleum ether- ethyl acetate- methanol (8.5:1:1) (v/v/v), as mobile phase with chamber saturation of 20 minutes. Quantification was carried out at 540 nm for all the three markers. Results: Linear responses for Oleanolic acid, 20-hydroxyecdysone and β-sitosterol were obtained over the concentration ranges of 20-400 μg/mL , 40-400 μg/mL and 40-400 μg/mL respectively. Conclusion: This HPTLC method can be used as a quality control tool for quantification of these markers simultaneously from raw material as well as marketed formulation.

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References

1. ICH Harmonised Tripartite Guideline, Validation of Analytical Procedures: Text and
Methodology Nov. 2005; Q2 (R1).
2. Agharkar SP. Medicinal plants of Bombay presidency, PBI, Scientific Publishers, Jodhpur
(India), 1991, 230.
3. Pagliarussi R., Freitas L., Bastos J., A quantitative method for the analysis of xanthine
alkaloids in Paullinia cupana (guarana) by capillary column gas chromatography. J.
Sep Sci 2002;25:371-378.
4. Girach RD, Aminuddin A, Khan SA. Ethnomedicinal uses of A aspera in Orissa (India).
Int J Pharmacog1992; 30: 113-115.
5. Jeong HG. Inhibition of cytochrome P450 2E1 expression by oleanolic acid:
hepatoprotective effects against carbon tetrachlorideinduced hepatic injury. Toxicol
Lett 1999; 105: 215-222.