Analysis of Gallic Acid and Kaempferol in Chloroform and Ethanol Extract of Cassia hirsuta Seeds by High-Performance Liquid Chromatography

Background: Cassia hirsuta plant belongs to the family Caesalpiniaceae and is commonly called stinking cassia and hairy senna. It is an alternative to coffee and is also used for stomach troubles, dysentery, abscesses, rheumatism, fever, and other diseases. High-performance liquid chromatography (HPLC) with spectrophotometric detection has been widely used for the identification and quantification of phenolics and flavonoids in plant extracts.

Aim: The present study's aim was to analyze the gallic acid and kaempferol in chloroform and ethanol extract of Cassia hirsuta seeds by high-performance liquid chromatography (HPLC) method.

Materials and Methods: The plant was collected from Bagalkot, Karnataka, India. In this study, the HPLC analysis of gallic acid and kaempferol was done by selected HPLC methods and these were validated for linearity, precision, limit of detection, limit of quantitation, repeatability, and reproducibility. The corresponding concentration of gallic acid and kaempferol against respective peak area values was determined using the gallic acid and kaempferol calibration curves, respectively.

Results: A calibration curve was established for gallic acid and kaempferol by injecting 4, 8, 12, 16, 20, and 24
g/mL of standard solution. In the results, it was found that the linearity was good with regression equations y=36934x-10162 and y=61050x-9395.5 for gallic acid and kaempferol, respectively, and the correlation coefficient (R2) was 0.9982 and 0.9992, respectively. Percentage relative standard deviation of 1.6% and 0.71% for gallic acid and kaempferol, respectively, indicates the high precision. Ethanol extract of C. hirsuta shows a high content of both gallic acid and kaempferol as compared to chloroform extract of C. hirsuta.

Conclusion: In the present study, gallic acid and kaempferol content were determined in ethanol and chloroform extract of C. hirsuta. The developed HPLC methods have enabled rapid, linear, accurate, and reproducible analysis in these two extracts of C. hirsuta. The developed method can be used for quantitative analysis and quality control of extracts and commercial samples of other species containing gallic acid and kaempferol.