幼泻宁颗粒HPLC指纹图谱研究

目的：建立幼泻宁颗粒的HPLC指纹图谱，对市售幼泻宁颗粒的质量进行控制。方法：采用Agilent Zorbax-C₁₈色谱柱（4.6 mm×250 mm，5 μm），以乙腈（A）-0.3%磷酸水溶液（B）为流动相进行梯度洗脱，流速1.0 mL·min^-1，检测波长350 nm（0~40 min，检测木犀草苷及木犀草素）、220 nm（40~60 min，检测白术内酯Ⅰ、白术内酯Ⅱ和白术内酯Ⅲ），柱温30℃，进样量10 μL。采用国家药典委员会《中药色谱指纹图谱相似度评价系统2004A版》对11批市售幼泻宁颗粒进行相似度评价，通过比对化学分离对照品的保留时间对主要特征峰进行明确化学指认并初步确定组方中药来源。结果：共确定幼泻宁颗粒HPLC指纹图谱21个共有峰，通过与对照品比较指认其中5个指标成分分别是木犀草苷（5号峰）、木犀草素（6号峰）、白术内酯Ⅰ（17号峰）、白术内酯Ⅱ（18号峰）和白术内酯Ⅲ（19峰），利用相似度软件对11批制剂指纹图谱进行分析，各批样品相似度均在0.95以上。建立的幼泻宁颗粒的HPLC指纹图谱的精密度、专属性、稳定性和重复性均良好。结论：该研究建立的幼泻宁颗粒HPLC指纹图谱基本能全面反映该中药复方制剂中各味组方的整体特征，可用于幼泻宁颗粒的质量控制和评价。

Objective: To establish the HLC fingerprint of Youxiening granules,and to control the quality of commercial Youxiening granules. Methods: The separation was developed on an Agilent Zorbax-C₁₈ column (250 mm×4.6 mm,5 μm)by gradient elution of acetonitrile (A)-0.3% phosphoric acid (B)at a flow rate of 1.0 mL·min^-1. The detection wavelength was 350 nm (0-40 min)for luteolin-7-glucoside and luteolin,and 220 nm (40-60 min)for atractylenolide Ⅰ,atractylenolide Ⅱand atractylenolide Ⅲ. The column temperature was 30℃ and the sample volume was 10 μL. The similarities of 11 batches of commercially available Youxiening granules were evaluated by the National Pharmacopoeia Committee's "Chinese medicine fingerprint similarity evaluation system 2004A". The main characteristic peaks were identified by comparing with the retention time of the chemically separated reference substances and the origin of the prescription was preliminarily determined. Results: 21 common peaks were selected in the fingerprint. By comparing with the mixed reference substances,5 indexes were identified as luteolin-7-glucoside (peak 5),luteolin (peak 6),atractylenolide Ⅰ (peak 17),atractylenolide Ⅱ (peak 18)and atractylenolide Ⅲ (peak 19). The similarities of all samples were above 0.95. The precision,specificity,stability and reproducibility of the method were satisfactory. Conclusion: The HPLC fingerprint of Youxiening granules established in this study can basically reflect the overall characteristics of each prescription in the compound preparation of Chinese medicine,and can be used for the quality control and evaluation of Youxie Ning granules.

[1] 卫生部药品标准.中药成方制剂.第5册[S].1993:49 Drug Specifications Promulgated by the Ministry of Public Health,P R China.Chinese Medicine Prescription Preparation.Vol 5[S].1993:49
[2] 戚雁飞,谢升谷.幼泻宁颗粒质量标准研究[J].浙江中医药大学学报,2010,34(5):773 QI YF,XIE SG.Quality standard study on Youxiening granules[J].J Zhejiang Univ Tradit Chin Med,2010,34(5):773
[3] 杨柳,李婷,范彦博,等.妇科洗剂的HPLC指纹图谱研究[J].中国药师,2017,20(3):463 YANG L,LI T,FAN YB,et al.Study on the HPLC fingerprint of gynecological lotion[J].China Pharm,2017,20(3):463
[4] 刘东方,赵丽娜,李银峰,等.中药指纹图谱技术的研究进展及应用[J].中草药,2016,47(22):4085 LIU DF,ZHAO LN,LI YF,et al.Research progress and application in fingerprint technology on Chinese materia medica[J].Chin Tradit Herb Drugs,2016,47(22):4085
[5] 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
[6] 阎克里,朱秀卿,赵丽.HPLC法同时测定白术挥发油中5种抗肿瘤成分含量[J].药物分析杂志,2011,31(6):1106 YAN KL,ZHU XQ,ZHAO L.HPLC simultaneous determination of five anti-tumor components in Atractylodes macrocephala Koidz.volatile oil[J].Chin J Pharm Anal,2011,31(6):1106
[7] 封传华,任琦,汤小林,等.车前草中总苯乙醇苷及毛蕊花糖苷的含量测定[J].中国药师,2017,20(8):1477 FENG CH,REN Q,TANG XL,et al.Determination of total phenylethanold glycosides and acteoside in Plantago Herba[J].China Pharm,2017,20(8):1477
[8] 黄桃芬,卢丹选,喻芳君,等.HPLC法同时测定车前草中4种成分的含量[J].中药材,2017,40(3):645 HUANG TF,LU DX,YU FJ,et al.Simultaneous determination of 4 components in Plantago asiatica by HPLC[J].J Chin Med Mater,2017,40(3):645
[9] 夏玲红,金冠钦,孙黎,等.车前草的化学成分与药理作用研究[J].中国药师,2013,16(2):294 XIA LH,JIN GQ,SUN L,et al.Studies on chemical constituents and pharmacological effects of Plantago asiatica[J].China Pharm,2013,16(2):294
[10] 杨亚军,周秋贵,曾红,等.车前草化学成分及新生物活性研究进展[J].中成药,2011,33(10):1771 YANG YJ,ZHOU QG,ZENG H,et al.Advances in studies on chemical constituents and bioactivities of Plantago asiatica[J].Chin Tradit Pat Med,2011,33(10):1771
[11] 冯文林,伍海涛.白术治疗肠道疾病的作用机制探讨[J].辽宁中医杂志,2016,43(1):125 FENG WL,WU HT.Mechanism of atractylenolide macrocephala on intestinal diseases[J].Liaoning J Tradit Chin Med,2016,43(1):125
[12] 陈晓萍,张长林.白术不同化学成分的药理作用研究概况[J].中医药信息,2011,28(2):124 CHEN XP,ZHANG CL.Research progress on pharmacological effects of different chemical constituents of Atractylodes macrocephala[J].Inf Tradit Chin Med,2011,28(2):124
[13] 董海燕,董亚琳,贺浪冲,等.白术抗炎活性成分的研究[J].中国药学杂志,2007,42(14):1055 DONG HY,DONG YL,HE LC,et al.Studies on constituents and anti-inflammatory activity of Rhizoma Atractylodis Macrocephalae[J].Chin Pharm J,2007,42(14):1055