目的:建立高效液相色谱-质谱联用法测定止咳平喘类中成药中非法添加的磺胺嘧啶、吡哌酸、甲氧苄啶、氧氟沙星、诺氟沙星、磺胺二甲嘧啶、磺胺甲噁唑7个化学抗菌类物质。方法:以Venusil MP C18（4.6 mm×250 mm, 5 μm）为色谱柱, 乙腈-甲醇（1∶1）-甲酸铵溶液为流动相, 梯度洗脱, 柱温为30 ℃, 流速为1 mL•min-1; 质谱配置电喷雾电离源（ESI）, 干燥气温度325 ℃, 干燥气流速5 L•min-1, 雾化压力103.4 kPa, 扫描方式为正离子模式进行一级质谱全扫描、二级质谱全扫描, 扫描范围为m/z 50~650。结果:7个化学抗菌药检测下限均小于0.04 μg, 回收率为94.4%~98.9%; 在1.4~17.6 μg•mL-1 范围内, 线性关系良好（r>0.999 6）; 3 批阴性样品呈阴性, 3批模拟阳性样品呈阳性, 方法专属性强; 用本文建立的方法对假药A~E 进行了检验, 检出假药A、C、D 含甲氧苄啶、磺胺甲噁唑。结论:本方法专属性强, 灵敏度高, 可用于7个化学抗菌类非法添加物的验证试验。
Objective：To establish an HPLC-MS/MS method for simultaneous determination of seven antimicrobial agents (sulfadiazine, pipemidic acid, trimethoprim, sulfamethazine, ofloxacin, norfloxacin and sulfamethoxazole) in adulterated Chinese patent medicines for relieving cough and asthma. Methods：The analysis was performed on a Venusil MP C18 column (4.6 mm×250 mm, 5 μm) with a gradient elution of acetonitrile-methanol (1∶1) -0.3% ammonium formate solution at a flow rate of 1 mL·min-1. The column temperature was 30 ℃. The target compounds were analyzed with LC-MS in two scan modes including full scan MS and full scan MS2. The scan range was m/z 50~650. The drying gas temperature was 325 ℃, the drying gas flow rate was 5 L·min-1, and the drying gas pressure was 103.4 kPa. Results：Sulfadiazine, pipemidic acid, trimethoprim, sulfamethazine, ofloxacin, ofloxacin and sulfamethoxazole were all less than low 0.04 μg and had good linearities in the range of 1.4-17.6 μg•mL-1 (r>0.999 6). The recoveries of high, median, and low concentrations control samples were 94.4%-98.9%. The test result of three batches of negative samples was negative, and the test result of three batches of analog positive samples was positive. Trimethoprim and sulfamethoxazole were found in three batches of counterfeit drugs. Conclusion：The method can be used for examining seven antimicrobial agents in adulterated Chinese patent medicines.
[1] 徐硕,徐文峰,金鹏飞,等. 止咳平喘类中成药和保健品中非法添加化学药物检测技术的研究进展[J].中南药学,2017,15(6): 798
XU S,XU WF,JIN PF,et al.Analytical technologies for illegal adulterated chemical substances in antitussive and antiasthmatic Chinese patent medicines and health foods[J].Cent South Pharm, 2017,15(6):798
[2] 李皓,朱青霞,柴逸峰,等.TLC-SERS 法快速检测止咳平喘类中成药中4 种添加化学成分[J].药物分析杂志,2014,34(4):693
LI H,ZHU QX,CHAI YF,et al.Rapid detection of four kinds of chemicals added in Chinese patent medicine for relieving cough and asthma by TLC-SERS[J].Chin J Pharm Anal,2014,34(4): 693
[3] 中华人民共和国药典2015 年版. 二部[S] .2015:205,472, 1137,1196,1555
ChP2015. VolⅡ[S].2015:205,472,1137,1196,1555
[4] 沈于兰,申兰慧,苏嘉焱.HPLC 法同时测定止咳平喘类中成药中非法添加的9 种化学成分[J].中国药品标准,2016,17(1):37
SHEN YL,SHEN LH,SU JY.HPLC determination of 9 chemical compositions illegally added in traditional Chinese medicine for relieving cough and asthma[J].Drug Stand China,2016,17(1): 37
[5] 孔卫东,周世玉.高效液相色谱法检测止咳平喘类中成药中非法添加的醋酸泼尼松、茶碱[J].中国药业,2013,22(14):55
KONG WD,ZHOU SY.Determination of metacortandracin acetate and theophylline illegally added in Chinese patent medicines of relieving cough and asthma by HPLC[J].China Pharm,2013,22(14):55
[6] 孙晓,李卓,荀英,等.HPLC-MS/MS 法同时测定止咳平喘类中成药中非法添加化学药物[J]. 中成药,2019,41(10):2405
SUN X,LI Z,XUN Y,et al.Simultaneous determination of illegally added chemical drugs in cough-relieving Chinese traditional patent medicines by HPLC-MS/MS[J].Chin Tradit Pat Med,2019,41(10):2405
[7] 高辉,李冰,王杰.HPLC-DAD-MS/MS 法用于止咳平喘类中成药中添加14 种肾上腺素皮质激素的筛查和定量研究[J].药物分析杂志,2015,35(8):1458
GAO H,LI B,WANG J.Application of HPLC-DAD-MS/MS to the screening and quantification of 14 adrenocortical hormones adulterated in Chinese patent drugs[J].Chin J Pharm Anal,2015,35(8):1458
[8] 李奕婧,李晓琳,韩家忠,等.HPLC-DAD-MS/MS 检测止咳平喘类中药制剂中非法添加的12 种化学药品[J].药学学报,2017, 52(6):959
LI YJ,LI XL,HAN JZ,et al.Screening for 12 mixed illegally cough-relieving chemical composition in anti-cough and antiasthmatic traditional Chinese medicines by HPLC-DAD-MS/MS[J].Acta Pharm Sin,2017,52(6):959
[9] 郭宇鹏,潘军. 中成药中非法添加10 种喹诺酮类药物的LC-MS/MS 检测方法研究[J]. 中国药师,2015,18(12):2162
GUO YP,PAN J.Study on LC-MS/MS method for the detection of 10 quinolone drugs illegally added in health foods and Chinese traditional patent medicines[J].China Pharm,2015,18(12): 2162
[10] 彭耀文,申兰慧,王丽,等.UPLC/Q-TOF-MS 法检测2 种中成药中土霉素、马来酸氯苯那敏和醋酸泼尼松[J]. 中成药,2015,37(9):1959
PENG YW,SHEN LH,WANG L,e t a l .Detection of oxytetracycline,chlorpheniramine maleate and prednisone acetate in two Chinese traditional patent medicines by UPLC/Q-TOF-MS[J]. Chin Tradit Pat Med,2015,37(9):1959
[11] 林伟杰,梁祈,詹若挺. 止咳平喘类中成药中 9 种非法添加化学成分的定性定量检验方法研究[J]. 广州中医药大学学报, 2017,34(3):418
LIN WJ,LIANG Q,ZHAN RT.Qualitative and quantitative method for detection of 9 chemicals illegally added into cough-checking and asthma-relieving Chinese patent medicines[J]. J Guangzhou Univ Tradit Chin Med,2017,34(3):418
[12] 陈晓虎,秦剑,苏晶,等.UPLC-Q-TOF 检测止咳平喘类中成药中非法添加的 8 种化学药品[J].中国实验方剂学杂志,2015,21(4):64
CHEN XH,QIN J,SU J,et al.Determination of 8 kinds of chemical medicines illegally added in traditional Chinese medicines for relieving cough and asthma by UPLC-Q-TOF[J].Chin J Exp Tradit Med Form,2015,21(4):64
[13] 陈繁华,曾玉梅,刘佳,等. 胶体金免疫层析法快速检测非法添加磺胺甲噁唑的研究[J].解放军药学学报,2014,30(2):145
CHEN FH,ZENG YM,LIU J,et al.Study of colloidal gold-immunochromatography assay for rapid detection of illegally-added sulfamethoxazole[J] .Pharm J Chin PLA,2014,30(2): 145
[14] 郭亚格,李珺沬,刘圆圆,等. 微波提取–电喷雾电离–离子迁移谱法快速筛查止咳平喘化痰类中成药中4种化学成分[J]. 分析科学学报,2020,36(6):863
GUO YG,LI JM, LI YY,et al.Rapid screening of 4 chemical compositions in traditional Chinese medicines for antitussive, antiasthmatic and phlegm-resolving by microwave-assisted extraction-electrospray ionization-ion mobility spectrometry[J].J Anal Sci,2020,36(6):863
[15] 药品检验补充检验方法和检验项目批件[S]. 2009
China Food and Drug Administration.Instructions of drug testing supplementary inspection method and program[S].2009
