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Simultaneous densitometric determination of shikonin, acetylshikonin, and β‐acetoxyisovaleryl‐shikonin in ultrasonic‐assisted extracts of four Arnebia species using reversed‐phase thin layer chromatography
Author(s) -
Sharma Nandini,
Sharma Upendra K.,
Gupta Ajai P.,
Sinha Arun K.,
Lal Brij,
Ahuja Paramvir S.
Publication year - 2009
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200900129
Subject(s) - chromatography , chemistry , calibration curve , formic acid , high performance thin layer chromatography , methanol , chloroform , thin layer chromatography , absorbance , detection limit , organic chemistry
A simple, precise, and rapid high‐performance thin‐layer chromatographic (HPTLC) method for the simultaneous quantification of pharmacologically important naphthoquinone shikonin ( 1 ) together with its derivatives acetylshikonin ( 2 ), and β‐acetoxyisovalerylshikonin ( 3 ) in four species of genus Arnebia ( A. euchroma , A. guttata , A. benthamii , and A. hispidissima ) from the Indian subcontinent has been developed. In addition, the effect of solvents with varying polarity (hexane, chloroform, ethyl acetate, and methanol) for the extraction of these compounds was studied. HPTLC was performed on precoated RP‐18 F 254S TLC plates. For achieving good separation, mobile phase consisting of ACN/methanol/5% formic acid in water (40:02:08 v/v/v) was used. The densitometric determination of shikonin derivatives was carried out at 520 nm in reflection/absorption mode. The method was validated in terms of linearity, accuracy, precision, robustness, and specificity. The calibration curves were linear in the range of 100–600 ng for shikonin and acetylshikonin, and 100–1800 ng for β‐acetoxyisovalerylshikonin. Lower LOD obtained for compounds 1 – 3 were 18, 15, and 12 ng, respectively, while the LOQ obtained were 60, 45, and 40 ng, respectively.