A MARKER-BASED STABILITY INDICATING HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHIC METHOD FOR VITEX TRIFOLIA

Abstract

Casticin, chrysoplenol-D, p-hydroxy benzoic acid, and p-methoxy benzoic acid are representative flavonoids and benzoic acid derivatives in Vitex trifolia, a well-known traditional Chinese, as well as ayurvedic medicine, with a wide range of biological activities. In the present paper, the aforementioned four markers were isolated and simultaneously quantified by TLC densitometric method. In the present study, a novel stability-indicating high-performance thin-layer chromatography (HPTLC) method for quantitative determination of the aforementioned four markers has been developed and validated on silica using solvent dichloromethane–methanol–formic acid (9.6:0.4:0.06 v/v/v) in the absorbance mode at 254 nm. The method showed a good linear relationship (r² = 0.999) in the concentration range 100–2400 ng per band. It was found to be linear, accurate, precise, specific, robust, and stability-indicating and can be applied for quality control and standardization of other multi-component formulations based on Vitex trifolia.

Keywords:

• densitometric
• flavonoids
• method validation
• stability
• thin-layer chromatography
• Vitex trifolia

ACKNOWLEDGMENTS

The authors thank Director, CIMAP, for keen interest during the course of work and the Council of Scientific and Industrial Research (CSIR), New Delhi for providing financial supports. One of the authors, Neerja Tiwari, is thankful to CSIR (New Delhi) for the award of the Junior Research Fellowship.

Notes

^a Cold percolation method at 40°C for 48 hr.

^b Plant dry weight basis.

Number of concentration levels (n) = 5.

For all equations the significance levels p < 0.0001.

^a n = 5.

^a Correlation of spectrum at start of peak with spectrum at the center of peak c.

^b Correlation of spectrum at center of peak with spectrum at the end of peak.