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Separation and quantification of nine bioactive compounds in traditional Unani formulations by High Performance Liquid Chromatography–Photodiode Array Detector

Kamal Y. T., Ahmad Sayeed, Mahadevan Nanjaian, Alam Prawez, Salam Shahana, Asiri Yahya, Muhsinah Abdullatif Bin, Alsayari Abdulrhman

Acta Chromatographica

2021

Vol. 33, no. 1

3--10

A new High Performance Liquid Chromatography–Photodiode Array Detector (HPLC–PDA) method has been developed for the chromatographic separation and simultaneous quantitative determination of nine bioactive compounds, i.e. four phenolic (gallic acid, ellagic acid, chebulinic acid, and tannic acid), two flavanoids (rutin and quercetin), two anthraquinones (sennoside A and B) and one oxygenated hydrocarbon (vitamin C) in a well-known Unani polyherbal formulation namely Itrifal’s. Separation was accomplished on a C18 LiChrospher 100 column (5 mm, 250 3 4.6 mm) with a gradient elution and recorded at 254 nm. The results demonstrated that the proposed method is reproducible, accurate, economic, and suitable for the quality control of traditional polyherbal Unani formulations containing complex compounds with different structures such as Itrifals.

Validated HPLC method development for simultaneous analysis of withaferin-A and 6-gingerol

Jain V., Thakur A, Soman G, Laddha K. S.

Acta Chromatographica

2010

Vol. 22, no. 1

153--159

A simple, rapid, and specific reversed-phase HPLC method has been developed for simultaneous analysis of withaferin-A and 6-gingerol in a polyherbal formulation containing Withania somnifera and Zingiber officinalis extracts. HPLC analysis was performed on a C 18 column using a 40:60 ( v / v ) mixture of acetonitrile and water as isocratic mobile phase at a flow rate of 1.5 mL min −1 . UV detection was at 227 nm for withaferin-A and 278 nm for 6-gingerol. The method was validated for accuracy, precision, linearity, specificity, and sensitivity in accordance with International Conference on Harmonization guidelines. Validation revealed the method is specific, accurate, precise, reliable and reproducible. Good linear correlation coefficients ( r 2 > 0.9996) were obtained for calibration plots in the ranges tested. Limits of detection were 0.2 and 0.4 μg and limits of quantification were 0.5 and 1.0 μg for withaferin-A and 6-gingerol, respectively. Intra and inter-day RSD of retention times and peak areas were less than 2.1%. Recovery was between 94.5 and 98.8% for withaferin-A and 94.2 and 102.4% for 6-gingerol. The established HPLC method is appropriate and the two markers are well resolved, enabling efficient quantitative analysis of withaferin-A and 6-gingerol. The method was successfully used for quantitative analysis of these two marker constituents in a marketed polyherbal formulation.