Objective: To establish two standards for determination of multiple components(TSDMC) in quantitative analysis of hesperidin, tangeretin, nobiletin, 3,3’, 4, 4’, 5, 6, 7-heptamethoxyflavone and narirutin in Citrus Reticulate Pericarpium. Methods: The peak area and retention time of the five flavonoids in Citrus Reticulate Pericarpium were measured for quantitative and qualitative analysis with three different brands of HPLC instruments and 13 C18 columns of different brands and types. For the quantitative analysis, hesperidin was used as the reference for determination of relative correction factors of tangeretin, nobiletin, 3, 3’, 4, 4’,5,6,7-heptamethoxyflavone and narirutin. For the qualitative analysis, the average retention time of the five components on each column was regarded as the standard retention time. Both hesperidin and tangeretin were used as linear calibration standards to calculate the retention time of the other three components. Meanwhile, the contents of hesperidin, tangeretin, nobiletin, 3, 3’, 4, 4’, 5, 6, 7-heptamethoxyflavone and narirutin were determined by external standard method,and the deviation of two methods was used to verify the accuracy and feasibility. Results: For quantitative results, the relative correction factors were reproducible with different HPLC instruments, different brands and types of columns, as well as a certain variation range of proportion of the wavelength, mobile phase, column temperature and flow rate. The relative correction factors of tangeretin, nobiletin, 3,3’, 4, 4’, 5,6,7-heptamethoxyflavone and narirutin were 0.86, 0.50, 1.11, 0.59, and their RSDs were less than 2.0%. For qualitative results, the absolute deviations of the predicted values of the linear calibration using two reference substances wereless than 0.5 min while those of the relative retention time method were more than 1.5 min, which indicated that the linear calibration using two reference substances was more accurate than the relative retention time method for chromatographic peak identification on 13 columns.The relative average deviations of the contents of the 5 components determined by TSDMC and the external standard method were less than 4%, which showed no significant difference. Conclusion: TSDMC for simultaneous determination of 5 components in Citrus Reticulate Pericarpium is feasible and durable.
LI Dan, WAN Lin-chun, ZHAO-Wen, YU Jian-dong, XU Chun-liang
. Determination of five flavonoids in Citrus Reticulate Pericarpium by dual-standard multi-measurement method*[J]. Chinese Journal of Pharmaceutical Analysis, 2022
, 42(9)
: 1643
-1651
.
DOI: 10.16155/j.0254-1793.2022.09.18
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