Establishment of characteristic chromatograms of TLC and HPLC and determination of six components in water-extract of Inula japonica
Objective To establish the characteristic chromatograms of thin-layer chromatography(TLC)and high-performance liquid chromatography(HPLC)of the aqueous-extract of Inula japonica Thunb.,and to determine the contents of six components(chlorogenic acid,caffeic acid,isoquercitrin,1,5-dicaffeoylquinic acid,4,5-dicaffeoylquinic acid and inulicin)in the aqueous-extract.Methods(1)The HF254 TLC silica gel plate,developing solvents of n-butanol-acetone-pyridine-water-glacial acetic acid(1∶5∶3∶1∶10µL)and 10%sulphuric acid-ethanol were used before the chromatographic spots were examined at 365 nm.(2)The Sino Chrom ODS-BP column(ϕ4.6 mm×250 mm,5µm)and a mobile phase of 0.05%phosphoric acid solution and acetonitrile were used.The flow rate was 0.8 mL/min,the column temperature 35℃and detection wavelength 205 nm.(3)The analytic conditions of the six components were similar to those described in method(2),but the wavelengths were respectively 203 nm(inulicin),256 nm(isoquercitrin)and 327 nm(caffeic acid,chlorogenic acid,1,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid).Results(1)The TLC chromatograms showed fine properties before and after the colour developer was sprayed.(2)Twenty-eight common peaks were marked in the HPLC characteristic chromatogram,so were ten components(chlorogenic acid,caffeic acid,1,5-dicaffeoylquinic acid,4,5-dicaffeoylquinic acid,taxifolin,rutin,quercitrin,isoquercitrin,luteolin and inulicin).The aqueous-extracts of eleven batches of materials of Inula japonica shared much similarity based on similarity and cluster analysis.(3)There were good linear relationships,and the linear ranges of chlorogenic acid,caffeic acid,isoquercitrin,1,5-dicaffeoylquinic acid,4,5-dicaffeoylquinic acid and inulicin were 3.25-65.00µg/mL,4.00-80.00µg/mL,1.60-32.00µg/mL,17.75-355.00µg/mL,2.50-50.00µg/mL and 3.75-75.00µg/mL,respectively.The contents of chlorogenic acid,caffeic acid,isoquercitrin,1,5-dicaffeoylquinic acid,4,5-dicaffeoylquinic acid and inulicin in elven batches of water-extract from Inula japonica were 3.38-6.66µg/mg,2.13-7.35µg/mg,0.51-1.57µg/mg,15.81-24.87µg/mg,0.50-1.08µg/mg and 0.77-4.73µg/mg,respectively.Conclusion The specific TLC and HPLC characteristic chromatograms can help ensure accurate quantitative determination of six components of the water-extract of Inula japonica,which can be used for quality control of the water-extract of Inula japonica.
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