木香顺气丸HPLC特征指纹图谱研究及多成分含量测定

Fingerprint and multi-components determination of Muxiang Shunqi pills by HPLC

目的:建立木香顺气丸的指纹图谱,并进行多成分定量分析,为评价其质量提供依据。方法:采用SB-C_(18)(250 mm×4.6 mm,5μm)色谱柱,以甲醇(A)-乙腈(B)-0.2%磷酸水溶液(C)为流动相,梯度洗脱(0~5 min,10%A,10%B;5~20 min,10%A→20%A,10%B→20%B;20~40 min,20%A→50%A,20%B→10%B;40~60 min,50%A,10%B),流速1.0 m L·min^(-1),检测波长225 nm,柱温35℃。通过相似度评价结合聚类分析对15批次木香顺气丸指纹图谱进行质量评价,并对指认的5个指标成分进行定量测定研究。结果:在特征图谱研究中,共确定木香顺气丸HPLC指纹图谱8个共有峰,通过与对照品比较指认其中5个共有峰分别为木香烃内酯、去氢木香内酯、和厚朴酚、厚朴酚和甘草酸,利用相似度软件对15批样品指纹图谱进行分析,各批样品相似度均在0.90以上。木香烃内酯、去氢木香内酯、和厚朴酚、厚朴酚和甘草酸进样量分别在0.067~2.688、0.057~2.264、0.033~1.304、0.032~1.264和0.067~2.688μg范围内线性关系良好,r均大于0.998 0;平均加样回收率为98.6%~99.6%,RSD在1.0%~1.9%。15批样品中木香烃内酯、去氢木香内酯、和厚朴酚、厚朴酚和甘草酸含量范围分别为3.93~8.15、0.52~9.73、8.00~13.67、2.21~5.96、1.99~5.01 mg·g^(-1)。通过聚类分析,15批样品聚成三类。结论:所建立的木香顺气丸HPLC指纹图谱和含量测定分析方法可用于木香顺气丸的质量控制。

Objective： To establish fingerprint and multi-components determination method of Muxiang Shunqi pills, and to provide reference for quality evaluation. Methods： The analysis was performed on a C18 column （ 250 mm ×4.6mm,5μm ）, and the mobile phase consisted of methanol （ A ）, acetonitrile （ B ） and 0.2% phosphoric acid （ C ） with gradient elution （ 0-5 min, 10%A and 10%B ; 5-20 min, lO%A → 20%A and 10%B →20%B ; 20-40 min, 20%A → 50%A and 20%B →10%B ; 40-60 min, 50%A and 10%B ） at a flow rate of 1.0 mL min-1. The detection wavelength was 225 nm and the column temperature was 35 ℃. Similarity evaluation combined with cluster analysis （ CA ） was used to evaluate the fingerprint of 15 batches of Muxiang Shunqi pills, and the five markers were quantified. Results： The common mode of the fingerprint showed 8 common peaks, 5 of which were identified by comparison with the reference substances. They were costunolide, dehydrocostuslactone, honokiol, magnolol and glycyrrhizic acid. The similar degrees of 15 batches of samples were above 0.9. The linear ranges were 0.067-2.688 μg, 0.057-2.264 μg, 0.033-1.304μg, 0.032-1.264 μg and 0.067-2.688 ng for costunolide, dehydrocostuslactone, honokiol, magnolol and glycyrrhizic acid, respectively, with r〉0.998 0. The average recoveries varied from 98.6%-99.6% with RSDs of 1.0%-1.9%. The content ranges of costunolide, dehydrocostuslactone, honokiol, magnolol and glycyrrhizic acid in fifteen batches of samples were 3.93-8.15 mg g-i, 0.52-9.73 mg g-1, 8.00-13.67 mg g-1 , 2.21-5.96 mg g-1 and 1.99-5.01 mg g-1 , respectively. All 15 batches of samples could be classified into three clusters. Conclusion： The established method eould be used for the quality control of MuxiangShunqi pills.