目的:建立高效液相色谱法同时测定通窍耳聋丸中龙胆苦苷、栀子苷、黄芩苷、汉黄芩苷、黄芩素、木香烃内酯、大黄酚的含量。方法:供试品以70% 甲醇水回流提取1 h, 采用Waters Atlantis T3-C18 色谱柱（250 mm×4.6 mm, 5 μm）, 流动相为甲醇-0.15% 磷酸水溶液, 梯度洗脱, 流速1.0 mL·min-1, 柱温30 ℃, 检测波长240 nm（检测龙胆苦苷和栀子苷）、278 nm（检测黄芩苷、汉黄芩苷和黄芩素）、224 nm（检测木香烃内酯和大黄酚）, 进样量10 μL。对16 批样品进行聚类分析。结果:龙胆苦苷、栀子苷、黄芩苷、汉黄芩苷、黄芩素、木香烃内酯、大黄酚的线性范围分别为3.256~32.56 μg·mL-1（r=0.999 9）、5.303~53.03 μg·mL-1（r=0.999 8）、23.74~273.4 μg·mL-1（r=0.999 7）、5.721~57.21 μg·mL-1（r=0.999 9）、2.174~21.74 μg·mL-1（r=0.999 9）、2.283~22.83 μg·mL-1（r=0.999 9）、1.053~10.53 μg·mL-1（r=0.999 8）; 平均加样回收率（n=6）分别为100.9%、101.6%、101.1%、98.5%、100.7%、98.4%、98.8%, RSD 分别为1.3%、1.3%、1.9%、2.0%、2.6%、1.4%、2.5%。16 批样品中龙胆苦苷、栀子苷、黄芩苷、汉黄芩苷、黄芩素、木香烃内酯、大黄酚的含量分别为0.390 0~2.038、1.912~3.560、10.40~14.34、2.118~3.745、0.189 8~1.324、0.291 4~0.784 3、0.230 0~0.545 9 mg·g-1, 16批样品可分为2类。结论:该方法准确, 重复性和耐用性好, 可用于通窍耳聋丸的质量控制和评价。
Objective：To develop an HPLC method for simultaneous determination of gentiopicroside, geniposide, baicalin, wogonoside, baicalein, costunolide and chrysophanol in Tongqiao Erlong pills. Methods：The sample was extracted with 70% methanol by refluxing for 1h. The analysis was performed on a Waters Atlantis T3-C18 column (250 mm×4.6 mm, 5 μm) by gradient elution of acetonitrile-0.15% phosphoric acid solution at a flow rate of 1.0 mL·min-1. The column temperature was set at 30 ℃, the injection volume was 10 μL, and the detection wavelengths were set at 240 nm (detecting gentiopicroside and geniposide), 278 nm (detecting baicalin, wogonoside and baicalein) and 224 nm (detecting costunolide and chrysophanol) . Cluster analysis was conducted in 16 batches of samples. Results：The linear ranges of gentiopicroside, geniposide, baicalin, wogonoside, baicalein, costunolide and chrysophanol were 3.256-32.56 μg·mL-1 (r=0.999 9), 5.303-53.03 μg·mL-1 (r=0.999 8), 23.74-273.4 μg·mL-1 (r=0.999 7), 5.721-57.21 μg·mL-1 (r=0.999 9), 2.174-21.74 μg·mL-1 (r=0.999 9), 2.283-22.83 μg·mL-1 (r=0.999 9) and 1.053-10.53 μg·mL-1 (r=0.999 8), respectively. The average recoveries were 100.9%, 101.6%, 101.1%, 98.5%, 100.7%, 98.4% and 98.8%, and the RSDs were 1.3%, 1.3%, 1.9%, 2.0%, 2.5%, 1.4% and 2.5%, respectively. The contents of the seven components in 16 batches of samples were 0.390 0-2.038, 1.912-3.560, 10.40-14.34, 2.118-3.745, 0.189 8-1.324, 0.291 4-0.784 3 and 0.230 0-0.545 9 mg·g-1, respectively. 16 batches of samples were clustered into two categories. Conclusion：This accurate, reproducible and durable method can be used for quality control and quality evaluation of Tongqiao Erlong pills.
[1] 潘旭,朱鹤云,史健艺,等.高效液相色谱法同时测定龙胆中5个环烯醚萜苷成分的含量研究[J].中药新药与临床药理,2020, 31(10):1223
PAN X,ZHU HY,SHI JY,et al.Simultaneous determination of five iridoid glycosides in Gentiance Radix et Rhizoma by HPLC[J]. Tradit Chin Drug Res Clin Pharm,2020,31(10):1223
[2] 李振雨,魏梅,孙冬梅,等.基于HPLC 指纹图谱结合化学计量学的龙胆和坚龙胆药材鉴别及质量评价[J].中国中医药信息杂志,2020,27(9):75
LI ZY,WEI M,SUN DM,et al.Identification and quality evaluation of Gentiana scabra Bge.and Gentiana rigescens Franch.based on HPLC fingerprints combined with chemometrics[J].Chin J Inf Tradit Chin Med,2020,27(9):75
[3] 徐鑫,戚华文,高德嵩,等.基于化学模式识别的栀子UPLC 定量指纹图谱研究[J].中国中药杂志,2020,45(18):4416
XU X,QI HW,GAO DS,et al.UPLC quantitative fingerprint research on Gardeniae Fructus based on chemical pattern recognition technology[J].China J Chin Mater Med,2020,45(18):4416
[4] 雷磊,王玉,霍志鹏,等.LCMS-IT-TOF 分析栀子炒焦前后化学成分的变化[J].中国实验方剂学,2019,25(17):89
LEI L,WANG Y,HUO ZP,et al.Variations of chemical constituents in Gardeniae Fructus before and after stir-frying by LCMS-IT-TOF[J]. Chin J Exp Tradit Med Form,2019,25(17):89
[5] 何艳,胡小祥.HPLC 法同时测定小儿解表颗粒中6 种成分[J]. 中成药,2020,42(2):305
HE Y,HU XX.Simultaneous determination of six constituents in Xiao’er Jiebiao Granules by HPLC[J].Chin Tradit Pat Med,2020, 42(2):305
[6] 陈韶彬,曹玲,张莉,等.功劳去火胶囊中多种指标性成分的同时测定[J].药物分析杂志,2020,40(11):1994
CHEN SB,CAO L,ZHANG L,et al.Simultaneous determination of index components in Gonglao Quhuo capsules[J].Chin J Pharm Anal,2020,40(11):1994
[7] 胡小祥,何艳.HPLC 法同时测定骨折挫伤胶囊中7 种成分的含量[J].中药新药与临床药理,2020,31(2):224
HU XX,HE Y.Simultaneous determination of seven components in Guzhe Cuoshang capsules by HPLC[J].Tradit Chin Drug Res Clin Pharm,2020,31(2):224
[8] 王云,张雪,麻印莲,等.熟大黄的炮制、药效及临床应用研究进展[J].中国实验方剂学,2018,24(24):219
WANG Y,ZHANG X,MA YL,et al.Advances in processing, pharmacodynamics and clinical application of prepared Rhei Radix et Rhizoma[J].Chin J Exp Tradit Med Form,2018,24(24):219
[9] 陈会朋,王银,陈香玲,等.液质联用法同时测定加味左金丸中13 种有效成分[J].中草药,2020,51(14):3693
CHEN HP,WANG Y,CHEN XL,et al.Simultaneous determination of 13 bioactive components in Jiawei Zuojin Pill by HPLC-MS/MS[J].Chin Tradit Herb Drugs,2020,51(14):3693
[10] 徐僮,杜欢,李琪,等.HPLC 法同时测定六味木香丸中4 种成分[J].中成药,2019,41(10):2315
XU T,DU H,LI Q,et al.Simultaneous determination of four constituents in Liuwei Muxiang pills by HPLC[J].Chin Tradit Pat Med,2019,41(10):2315
[11] 高长清,张素云,张洪波.反相高效液相色谱法测定通窍耳聋丸中黄芩苷的含量[J].中国药业,2003,12(12):40
GAO CQ,ZHANG SY,ZHANG HB.HPLC determination of baicalin in Tongqiao Erlong pills[J].Chin Pharm,2003,12(12): 40
[12] 夏文. 通窍耳聋丸的质量标准[J].中国药师,2010,13(11): 1670
XIA W.Study on quality standard of Tongqiao Erlong pills[J]. Chin Pharm,2010,13(11):1670
[13] 郑婕,伍名兰,谭丽盈.HPLC 法测定通窍耳聋丸中芦荟苷含量[J].亚太传统医药,2014,10(2):15
ZHENG J,WU ML,TAN LY.HPLC Determination of aloin in Tongqiaoerlong pills[J].Asia-Pacific Tradit Med,2014,10(2): 15
[14] 陈叶青,陈凡凡,金佩芬,等.HPLC 法测定通窍耳聋丸中橙皮苷和黄芩苷的含量[J].解放军药学学报,2015,31(4):324
CHEN YQ,CHEN FF,JIN PF,et al.HPLC determination of hesperidin and baicalin in Tongqiaoerlong pills[J].Pharm J Chin PLA,2015,31(4):324
[15] 王艺纯,唐秋竹.通窍耳聋丸的质量标准研究[J].中国当代医药,2017,24(3):116
WANG YC,TANG QZ.Study on quality standard of Tongqiao Erlong pills[J].China Mod Med,2017,24(3):116
