HPLC-QAMS法同时测定复方罗布麻片Ⅰ中8个活性成分的含量

目的：建立同时测定复方罗布麻片Ⅰ中8个活性成分含量的一测多评法（QAMS）。方法：采用HPLC法，以氢氯噻嗪为内参物，分别计算其与金丝桃苷、粉防己碱、防己诺林碱、绿原酸、蒙花苷、盐酸异丙嗪、硫酸双肼屈嗪的相对校正因子，通过相对校正因子计算复方罗布麻片Ⅰ中7个活性成分的含量（计算值），同时采用外标法测定8个活性成分的含量（实测值），比较计算值与实测值的差异。色谱柱为Kromasil C₁₈（250 mm×4.6 mm，5μm），流动相为乙腈-0.5%磷酸溶液（梯度洗脱），流速0.7 mL·min^-1，检测波长260 nm（氢氯噻嗪、盐酸异丙嗪、硫酸双肼屈嗪、金丝桃苷）、280 nm（粉防己碱、防己诺林碱）、325 nm（绿原酸、蒙花苷），柱温30℃，进样量10μL。结果：金丝桃苷、粉防己碱、防己诺林碱、绿原酸、蒙花苷、氢氯噻嗪、盐酸异丙嗪、硫酸双肼屈嗪检测质量浓度的线性范围分别为1.082~19.48、3.016~54.29、1.350~24.30、1.102~19.84、1.623~29.21、19.49~350.8、11.04~198.7、13.03~234.5μg·mL^-1（r为0.9995~0.9998）；平均加样回收率分别为98.3%、99.1%、98.3%、97.5%、97.9%、96.5%、96.3%、97.8%（RSD<2.0%，n=9）；检测下限分别为3.05、2.96、4.25、3.21、2.62、2.95、3.26、4.08 ng，定量下限分别为9.18、8.94、12.95、9.71、7.95、8.81、9.86、12.24 ng；精密度、重复性、稳定性（48 h）试验的RSD<2.0%（n=6）。金丝桃苷、粉防己碱、防己诺林碱、绿原酸、蒙花苷、盐酸异丙嗪、硫酸双肼屈嗪的相对校正因子分别为0.662、1.253、1.548、1.190、0.979、1.236、1.654，其计算值和实测值之间无显著性差异。结论：该方法简单、有效，结果准确，节约成本，可用于复方罗布麻片Ⅰ中上述8个活性成分的同时测定。  

Objective: To establish the quantitative analysis of multi-components by single marker (QAMS)for simultaneous determination of 8 active constituents in Kendir leaves tabletsⅠ. Methods: Using hydrochlorothiazide as internal standard, HPLC method was used to calculate relative correction factor (RCF)of hyperoside, tetrandrine, fangchinoline, chlorogenicacid, buddleodide, promethazine, dihydralazine sulfate. The contents of above-mentioned 7 active components in Kendir leaves tablets Ⅰ were calculated by using RCF (calculation value). The external standard method (ESM)was used to determine the contents of 8 constituents (measured value). The differences between calculation value and measured value were compared. The determination was performed on Kromasil C₁₈ (250 mm×4.6 mm, 5 μm)column with mobile phase consisted of acetonitrile-0.5% phosphoric acid (gradient elution)at the flow rate of 0.7 mL·min^-1. The detection wavelength was set at 260 nm for hyperoside, hydrochlorothiazide;promethazine;dihydralazine sulfate, 280 nm for tetrandrine, fangchinoline, 325 nm for chlorogenic acid, buddleodide.The column temperature was set at 30℃ and the injection volume was 10 μL. Results: The linear range was 1.082-19.48 μg·mL^-1 for hyperoside, 3.016-54.29 μg·mL^-1 for tetrandrine, 1.350-24.30 μg·mL^-1 for fangchinoline, 1.102-19.84 μg·mL^-1 chlorogenicacid, 1.623-29.21 μg·mL^-1 forbuddleodide, 19.49-350.8 μg·mL^-1 for hydrochlorothiazide, 11.04-198.7 μg·mL^-1 forpromethazine, 13.03-234.5 μg·mL^-1 for dihydralazine sulfate (r=0.999 5-0.999 8), respectively. Average recoveries were 98.3%, 99.1%, 98.3%, 97.5%, 97.9%, 96.5%, 96.3%, 97.8% (RSD<2.0%, n=9). Limits of detection were 3.05, 2.96, 4.25, 3.21, 2.62, 2.95, 3.26, 4.08 ng;limits of quantitation were 9.18, 8.94, 12.95, 9.71, 7.95, 8.81, 9.86, 12.24 ng. RSDs of precision, reproducibility and stability tests (48 h)were lower than 2.0% (n=6). RCFs of hyperoside, tetrandrine, fangchinoline, chlorogenicacid, buddleodide, promethazine, dihydralazinesulfate were 0.662, 1.253, 1.548, 1.190, 0.979, 1.236, 1.654. There was no statistical significance between calculated values and measured ones. Conclusions: The method is simple, effective, accurate and cost saving. It can be used for simultaneous determination of 8 active constituents in Kendir leaves tablets Ⅰ. 

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