Objective: To establish a new correction method of single-point correction which was applied to the quantitative analysis of multicomponents by single-marker(QAMS) of Mori Folium. Methods: HPLC was performed on Lubex Kromasil C18 column(250 mm×46 mm,5 μm)with mobile phase consisting of acetonitrile-0.1% phosphoric acid solution(gradient elution) at the flow rate of 1.0 mL·min-1. The detection wavelength was set at 350 nm and the column temperature was 30 ℃ . The chlorogenic acid(CA) was used as internal reference. The relative correction factors(RCFs) of neochlorogenic acid,cryptochlorogenic acid,rutin,isoquercitrin, nicotiflorin,acetylisoquercitrin,astragalin,isochlorogenic acid A,isochlorogenic acid C and acetylastragalin to chlorogenic acid were calculated by slope correction(SC),multi-point correction(MC) and single-point correction (SPC),respectively. The repeatability of RCFs was investigated. The contents of 11 components in Mori Folium were determined by external standard method(ESM),standard curve method(SCM) and QAMS respectively, and the differences among the 5 methods were compared. In addition,the calibration equation(CE) was obtained by two-point correction method and relative retention time(RRT) to locate the chromatographic peaks of the components to be measured. Results: RCFs of neochlorogenic acid,cryptochlorogenic acid,rutin,isoquercitrin, nicotiflorin,acetylisoquercitrin,astragalin,isochlorogenic acid A,isochlorogenic acid C and acetylastragalin to chlorogenic acid were 0.987, 0.979,1.179,1.568,1.126,1.446,1.533,1.301,1.288,1.409,respectively (single-point correction). The reproducibility of RCFs were perfect under different conditions(RSD<3%). The contents of 11 components in 10 batches of Mori Folium were 0.377-0.889,2.522-10.567,0.576-1.483,0.542- 2.252,0.810-3.870,0.175-0.907,0.571-3.795,0.273-1.330,0.154-0.560,0.141-0.245,0.236-1.726 mg·g-1 by QAMS,respectively(single point correction). The result was consistent with that of external standard method. Two-point correction and relative retention time could not be used to locate the chromatographic peak accurately, but the combination of the two methods could achieve the accurate location of chromatographic peak,with relative error(RE) less than 2%. Conclusion: The QAMS method using chlorogenic acid as the internal reference is feasible and accurate for the determination of neochlorogenic acid,cryptochlorogenic acid,rutin,isoquercitrin, nicotiflorin,acetylisoquercitrin,astragalin,isochlorogenic acid A,isochlorogenic acid C and acetylastragalin,and it can be used for quality control of Mori Folium. Single-point correction has more advantages than slope correction and multi-point correction under certain conditions.
Key words
HPLC /
QAMS /
Mori Folium /
chlorogenic acid /
flavone /
single-point correction /
two-point correction
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Footnotes
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