目的:建立HPLC法同时测定复方苦参注射液中7个成分(甲基氧化偶氮甲醇樱草糖苷、苦参碱、槐果碱、槐定碱、氧化槐果碱、氧化苦参碱、番石榴酸)的含量及指纹图谱研究,为其质量控制提供依据。方法:采用Waters Xselect CSHTM C18色谱柱(250 mm×4.6 mm,5 μm),以甲醇-0.2%磷酸二氢钾为流动相,梯度洗脱,流速0.6 mL·min-1,柱温30 ℃,检测波长211 nm。结果:在建立的液相色谱条件下,测定的甲基氧化偶氮甲醇樱草糖苷、苦参碱、槐果碱、槐定碱、氧化槐果碱、氧化苦参碱、番石榴酸7个成分的平均加样回收率分别为95.7%、101.3%、100.8%、101.7%、102.6%、102.5%、99.5%,RSD分别为2.0%、0.72%、0.90%、0.74%、1.4%、0.96%、1.8%;10批次样品中7个成分的含量分别为0.53~0.73、2.66~4.22、0.75~1.11、0.70~0.89、2.20~2.84、7.62~9.95、1.63~2.20 mg·mL-1。在相同液相色谱条件下建立的指纹图谱测定方法确立了9个共有峰,生成对照图谱,10批样品的相似度测定结果在0.99以上。结论:本研究同时对复方苦参注射液的主要成分进行定量分析和指纹图谱进行定性分析,含量测定准确,指纹峰特征性强,该方法稳定可靠,可为复方苦参注射液的质量控制与评价提供参考。
Objective: To establish an HPLC method for simultaneous determination of seven components (macrozamin, matrine, sophocarpine, sophoridine, oxysophocarpine, oxymatrine and piscidic acid) in compound Kushen injection and the fingerprint, providing the evidence for the quality control and evaluation of compound Kushen injection. Methods: Waters Xselect CSHTM C18 column(250 mm×4.6 mm, 5 μm) was adopted, the mobile phase consisted of methanol-0.2% potassium dihydrogen phosphate with gradient elution. The flow rate was 0.6 mL·min-1, the detection wavelength was set at 211 nm and the column temperature was set at 30 ℃. Results: Under the above chromatographic conditions, the average recovery rates of macrozamin, matrine, sophocarpine, sophoridine, oxysophocarpine, oxymatrine and piscidic acid were 95.7%, 101.3%, 100.8%, 101.7%, 102.6%, 102.5% and 99.5% with RSDs of 2.0%, 0.72%, 0.90%, 0.74%, 1.4%, 0.96% and 1.8%, respectively. The contents of 7 components in 10 batches of samples were 0.53-0.73, 2.66-4.22, 0.75-1.11, 0.70-0.89, 2.20-2.84, 7.62-9.95 and 1.63-2.20 mg·mL-1, respectively. The HPLC fingerprints of compound Kushen injection were established and 9 peaks were distinguished by developed HPLC method. And the similarities of HPLC fingerprints of 10 different batches were all above 0.99. Conclusion: The established method can perform the quantitative analysis of the main components and the qualitative analysis of the fingerprint simultaneously. The assay method is accurate and the fingerprint peaks are characteristic. The method is stable and reliable, which can provide supporting evidence for quality control and evaluation of CKI.
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