Concurrent NP-HPTLC Determination of Shikonin and β,β-Dimethylacryl Shikonin in Arnebia benthamii.
Naphthoquinones are important class of molecules found as a natural red color pigments in roots of Arnebia benthamii (Wall. ex G. Don) L M. Johnston. The aim of present investigation is to develop and validates a simple, cost-effective and reliable method for quantification of these compounds. Therefore, a normal phase-high performance thin-layer chromatography (NP-HPTLC) method for concurrent determination of shikonin and β,β-dimethylacryl shikonin in A. benthamii was established. Method development of naphthoquinones in the methanol extract was done using hexane-ethyl acetate-methanol (40:7.5:2.5, v/v/v) solvent system at 520 nm. The developed method showed good band separation for shikonin (Rf, 0.37) and β,β-dimethylacryl shikonin (Rf, 0.58). The linearity ranged between 100 and 8,000 ng spot(-1) with an average recovery of >97% in both cases. The results showed reproducible intraday and interday precision (<2.0% RSD) in quantification of naphthoquinones. The limits of detection are 12.96 and 14.65 ng spot(-1) while the limits of quantification are 39.27 and 44.39 ng spot(-1) for shikonin and β,β-dimethylacryl shikonin, respectively. The developed method is reliable, fast, easy to follow and economic in concurrent assessment of shikonin and β,β-dimethylacryl shikonin in A. benthamii root samples. In addition, it seems to be first report for identification and quantification of β,β-dimethylacryl shikonin from the A. benthamii.
Division of Biotechnology, CSIR-Institute of Himalayan Bioresource Technology, Palampur, HP 176 061, India.
Preeti
P
Katoch
Division of Biotechnology, CSIR-Institute of Himalayan Bioresource Technology, Palampur, HP 176 061, India Academy of Scientific and Innovative Research, CSIR-Institute of Himalayan Bioresource Technology, Palampur, HP 176 061, India.
Shalika
S
Rana
Academy of Scientific and Innovative Research, CSIR-Institute of Himalayan Bioresource Technology, Palampur, HP 176 061, India Natural Product Chemistry and Process Development Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur, HP 176 061, India sharmadinesh82@gmail.com dineshkumar@ihbt.res.in.
Dinesh
D
Kumar
Natural Product Chemistry and Process Development Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur, HP 176 061, India.
Shiv
S
Kumar
Division of Biotechnology, CSIR-Institute of Himalayan Bioresource Technology, Palampur, HP 176 061, India Academy of Scientific and Innovative Research, CSIR-Institute of Himalayan Bioresource Technology, Palampur, HP 176 061, India.
Shashi
S
Bhushan
D028082:Boraginaceae / Q000737:chemistry*; D002623:Chemistry Techniques, Analytical / Q000379:methods* / Q000592:standards; D002855:Chromatography, Thin Layer; D057230:Limit of Detection; D009285:Naphthoquinones / Q000032:analysis*; D015203:Reproducibility of Results; D012997:Solvents / Q000737:chemistry
D009285:Naphthoquinones; D012997:Solvents; C016101:shikonin