木香顺气丸HPLC特征指纹图谱研究及多成分含量测定

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-C₁₈（250 mm×4.6 mm，5 μm）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 mL·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0.067~2.688 μg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×4.6 mm, 5 μm), 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, 10%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, 0.033-1.304 μg, 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^-1, 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 could be used for the quality control of MuxiangShunqi pills.

[1] 中国药典2015年版.一部[S]. 2015:610 ChP 2015. Vol Ⅰ[S]. 2015:610
[2] 赖增发,夏丽. "一测多评"测定木香顺气丸中木香内酯类成分[J].中国现代应用药学, 2014,31(4):421 LAI Z F,XIA L. Determination of costus lactonein Muxiang Shunqipills by QAMS[J]. Chin J Mod Appl Pharm,2014, 31(4):421
[3] 孙全明,朱朝德,李春雨. HPLC测定木香顺气丸中的4种有效成分[J].华西药学杂志,2008, 23(2):215 SUN QM, ZHU CD, LI CY. Determination of four effective components in Muxiang Shunqi pills by HPLC[J]. West China J Pharm Sci, 2008,23(2):215
[4] 刘彤,史浚,李婷,等. HPLC法测定木香顺气丸中甘草酸的含量[J].天津药学, 2003,15(1):12 LIU T,SHI J,LI T,et al. Determination of glycyrrhizic acid in Muxiang Shunqi pills by HPLC[J]. Tianjin Pharm,2003,15(1):12
[5] 魏清芳, 辛爱玲, 王嘉林. HPLC法测定木香顺气丸中辛弗林的含量[J].安徽医药, 2010, 14(11):1290 WEI QF, XIN AL, WANG JL. Determination of synephrinein Muxiang Shunqi pills by HPLC[J]. Anhui Med Pharm J, 2010, 14(11):1290
[6] 徐成志. HPLC法测定木香顺气丸中橙皮苷的含量[J].安徽医药, 2007, 11(1):36 XU CZ. Determination of hesperid in in Muxiang Shunqi pills by HPLC[J]. Anhui Med Pharm J, 2007, 11(1):36
[7] 李强,杜思邈,张忠亮,等.中药指纹图谱技术进展及未来发展方向展望[J].中草药, 2013, 44(22):3095 LI Q, DU SM, ZHANG ZL, et al. Progress in fingerprint technology on Chinese materiamedica and prospect of its future development[J]. Chin Tradit Herb Drugs,2013, 44(22):3095
[8] 李家春,孙兰,李红娟,等.桂枝茯苓胶囊HPLC指纹图谱研究[J].中草药,2013, 43(7):1333 LI JC, SUN L, LI HJ, et al. HPLC fingerprint of Guizhi Fuling capsules[J]. Chin Tradit Herb Drugs,2012, 43(7):1333
[9] 程红,姚志红,戴毅,等.中药复方制剂仙灵骨葆胶囊HPLC指纹图谱研究[J].中国药学杂志, 2013,48(10):772 CHENG H, YAO ZH, DAI Y, et al. HPLC fingerprint of Xianling Gubao capsules, a TCM prescription[J]. Chin Pharm J, 2013, 48(10):772
[10] 程宏英,冯启余,曹玉华,等.菊花中活性成分的高效液相色谱测定与指纹图谱研究[J].分析科学学报, 2007, 23(3):257 CHENG HY, FENG QY, CAO YH, et al. Determination of active ingredients in Flos Chrysanthemi and development of fingerprints by high performance liquid chromatography[J]. J Anal Sci, 2007, 23(3):257
[11] 潘伟东,杜义龙,赵胜男,等.韩信草的高效液相色谱指纹图谱及化学模式识别[J].药物分析杂志, 2015,35(2):250 PAN WD, DU YL, ZHAO SN, et al. HPLC fingerprint and chemical pattern recognition of Scutellariaindica[J]. Chin J Pharm Anal, 2015, 35(2):250
[12] 刘吉成,陈大建.黑草药材HPLC指纹图谱的研究[J].药物分析杂志, 2013, 33(3):424 LIU JC, CHEN DJ. HPLC fingerprint study of the herb of Buchneracruciata[J]. Chin J Pharm Anal, 2013, 33(3):424
[13] 黄琪,黄彩虹,雷鹏,等.地锦草药材HPLC指纹图谱及模式识别研究[J].中国实验方剂学杂志, 2013, 19(12):95 HUANG Q, HUANG CH, LEI P, et al. Study on HPLC fingerprint of Herba Euphorbiae Humifusae[J]. Chin J ExpTradit Med Form, 2013, 19(12):95
[14] 吴昭晖,罗佳波,游文玮.甘草药材HPLC指纹图谱研究[J].中草药, 2008, 36(12):1868 WU ZH, LUO JB, YOU WW. HPLC fingerprint spectrum of Radix Glycyrrhizae[J]. Chin Tradit Herb Drugs, 2008, 36(12):1868
[15] 朱林燕,谢建峰,孔子铭,等.藏药二十一味寒水石丸的HPLC指纹图谱研究[J].药物分析杂志, 2015, 35(8):1430 ZHU LY, XIE JF, KONG ZM, et al. Study on HPLC fingerprint of Ershiyiwei Hanshuishi pills[J]. Chin J Pharm Anal,2015,35(8):1430