High-performance thin-layer chromatography densitometric determination of beta-sitosterol in Phyllanthus species.
An HPTLC densitometric method was established for the determination of beta-sitosterol in in vitro cultures of some species of the genus Phyllanthus. Two derivatization reagents commonly used in TLC for the visualization and detection of sterols, namely, anisaldehyde reagent and 5% phosphomolybdic acid, were compared with vanillin reagent. The densitometric quantification of beta-sitosterol was carried out on HPTLC Si60 F254 plates with the mobile phase chloroform-hexane-methanol (65 + 30 + 5, vlv/v) at 525 nm. The method was validated for each derivatization reagent in terms of linearity, precision, repeatability, intra- and interday precision, LOD, and LOQ. The presence of beta-sitosterol was revealed in all analyzed plant material. The concentrations of beta-sitosterol determined ranged from 0.48 to 2.75 mg/g (dry weight). In addition, traces of beta-amyrin were detected in some plant samples.
Sparzak
Barbara
B
Medical University of Gdańsk, Department of Pharmacognosy with the Medicinal Plants Garden, Hallera 107, 80-416 Gdańsk, Poland.
Krauze-Baranowska
Mirosława
M
Pobiocka-Olech
Loretta
L
D001547:Benzaldehydes / Q000032:analysis; D002623:Chemistry Techniques, Analytical; D002626:Chemistry, Pharmaceutical / Q000379:methods; D002725:Chloroform / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002855:Chromatography, Thin Layer / Q000379:methods*; D003720:Densitometry / Q000379:methods*; D006586:Hexanes / Q000737:chemistry; D000432:Methanol / Q000737:chemistry; D008956:Models, Chemical; D009828:Oleanolic Acid / Q000031:analogs & derivatives / Q000737:chemistry; D029803:Phyllanthus / Q000378:metabolism*; D010944:Plants / Q000378:metabolism; D015203:Reproducibility of Results; D012855:Sitosterols / Q000737:chemistry*
D001547:Benzaldehydes; D006586:Hexanes; D012855:Sitosterols; D009828:Oleanolic Acid; D002725:Chloroform; C100058:vanillin; C036380:beta-amyrin; D000432:Methanol