High-Performance Thin-Layer Chromatography Densitometric Method for Simultaneous Quantitation of Phyllanthin, Hypophyllanthin, Gallic Acid, and Ellagic Acid in Phyllanthus amarus

Abstract

Whole plant of Phyllanthus amarus Linn. is a reputed drug of the Indian systems of medicine that is used as hepatoprotective agent. A simple high-performance thin-layer chromatography (HPTLC) densitometric method has been developed for the simultaneous quantitation of phyllanthin, hypophyllanthin, gallic acid, and ellagic acid in the whole plant of P. amarus. They were found at levels of 0.37, 1.16, 0.36, and 0.17% (w/w), respectively. The method was validated for precision, repeatability, and accuracy. Instrumental precision was found to be 0.54, 0.93, 0.08, and 0.78% (coefficient of variation, CV); repeatability of the method was 1.01, 0.79, 0.98, and 1.06% (CV) for phyllanthin, hypophyllanthin, gallic acid, and ellagic acid, respectively. Accuracy of the method was determined by a recovery study conducted at 3 different levels, and the average recovery was found to be 99.09% for phyllanthin, 99.27% for hypophyllanthin, 98.69% for gallic acid, and 100.49% for ellagic acid. The proposed HPTLC method was found to be simple, precise, specific, sensitive, and accurate and can be used for routine quality control of raw material of P. amarus and formulations containing P. amarus. It also has the applicability in quantitating any of these marker compounds in other drugs.

Development of a UHPLC–MS/MS method for the quantitation of bioactive compounds in Phyllanthus species and its herbal formulations

“…Standards of quinic acid (1), gallic acid (2), protocatechuic acid (3), catechin (4), chlorogenic acid (5), epicatechin (6), chebulagic acid (7), quercetin-3,4'-di-O-glucoside (8), chebulinic acid (9), ellagic acid (10), rutin (11), naringin (12), kaempferol-3-O-rutinoside (13), eriodictyol (14), scutellarein (15), quercetin (16), luteolin (17), kaempferol (18), amentoferine (19), betulinic acid (22), and oleanolic acid (23) were purchased from Sigma-Aldrich (St. Louis, MO, USA; Supporting Information Fig. S1).…”

“…The quantitative analysis of bioactive lignans has been carried out in Phyllanthus species by traditional methods such as high-performance thin-layer chromatography (HPTLC) and HPLC [13][14][15][16][17][18][19][20][21][22][23][24][25]. Several researchers have reported fingerprinting, identification and characterization of bioactive compounds in crude extracts of Phyllanthus species using LC-MS [24][25][26][27][28][29][30][31][32][33][34].…”

Recent advances in plant hepatoprotectives: A chemical and biological profile of some important leads

“…Being important markers of the medicinal plant, several HPTLC methods have also been developed for quantitative estimation of these molecules. 129…”

HPTLC method Development & Validation for quantification of Markers of Dhatrinisha churna

“…[1,2] From such polyherbal formulations separation, identification and estimation of chemical components is very difficult. [3] Literature survey revealed that the above mentioned marker compound have various pharmacological properties. The rhizome and roots of Curcuma species are frequently used Curcumin and Ellagic acid of different concentration (100-600 ng/band) were applied to the HPTLC plate to prepare a calibration plot and to check for reproducibility.…”