超高效液相色谱-飞行时间质谱法鉴定蛇胆汁特征成分及其应用研究  

Identification of snake bile characteristic components by ultra-high performance liquid chromatography-time of flight mass spectrometry

作　　者：黄博 黄清泉 白桂昌 罗轶 朱雪妍 HUANG Bo;HUANG Qing-quan;BAI Gui-chang;LUO Yi;ZHU Xue-yan(Guangxi Institute for Food and Drug Control,Key Laboratory for Quality Monitoring and Evaluation of Traditional Chinese Medicine,Materia Medica,NMPA,Nanning 530021)

机构地区：[1]广西壮族自治区药品检验所国家药品监督管理局中药材质量监测与评价重点实验室,南宁530021

出　　处：《中南药学》2022年第6期1301-1308,共8页Central South Pharmacy

基　　金：广西重点研发计划(桂科AB17292069);广西自然科学基金项目(No.2017GXNSFAA198367)。

摘　　要：目的建立以特征离子为目标的鉴别蛇胆汁的方法,并用于检查蛇胆川贝液。方法采用超高效液相色谱-四极杆-飞行时间质谱(UPLC-Q-TOF-MS)测定蛇胆汁和其他动物胆汁的质谱数据,并进行主成分分析(PCA),找出蛇胆汁的特征成分。再用高效液相色谱-串联质谱(HPLC-MS-MS)的多反应检测(MRM)定性和验证特征成分。针对蛇胆川贝液处方中的药材,制订供试品制备方法,并将蛇胆汁的MRM条件应用于蛇胆川贝液的检测中。结果UPLC-Q-TOF-MS的液相方法条件为:甲醇(A)-10 mmol·L-1乙酸铵溶液(B),梯度洗脱(0~5 min,30%→50%A;5~20 min,50%→90%A;20~25 min,90%A;25~26 min,90%→30%A;26~30 min,30%A);质谱条件为:电喷雾电离源(ESI-),干燥气温度:220℃,干燥气流速8 L·min-1,雾化气压力2 bar,毛细管电压3500 V,喷嘴电压500 V,扫描范围50~1000,扫描频率1 Hz。共找出两个蛇胆汁的专属性成分,分别为牛磺胆酸硫酸酯(SOTC)和牛磺去氧胆酸硫酸酯(SOTDC)。建立了SOTC和SOTDC的HPLCMS-MS MRM方法,条件为:甲醇(A)-10 mmol·L^(-1)乙酸铵溶液(B),梯度洗脱(0~10 min,40%→50%A;10~25 min,50%→80%A;25~25.5 min,80%→40%A;25.5~30 min,40%A),质谱条件为:电喷雾电离源(ESI-),SOTC的离子对为594.2→514.3,594.2→124.0;SOTDC为578.2→498.3,578.2→124.0。将建立的方法应用于41批蛇胆川贝液制剂中,发现40批样品均含有蛇胆汁。结论该方法可有效鉴别蛇胆汁和检查蛇胆川贝液中是否含蛇胆汁。Objective To establish a method to identify snake bile with characteristic ions,and to examine Shedan Chuangbei decotion.Methods Ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS)was used to determine the mass spectrometry data of snake bile and other animal bile.The principal component analysis was used for detecting the characteristic components in snake bile.The conponents were established and verified by multiple reaction monitoring(MRM)of high performance liquid chromatography-tandem mass spectrometry.In this paper,the preparation method was established for the medicinal materials in the prescription of Shedan Chuangbei decoction and MRM condition was applied in the detection of snake bile.Results The liquid phase conditions of UPLC-Q-TOF-MS were as follows:methanol(A)-10 mmol·L^(-1) ammonium acetate(B),gradient elution(0-5 min,30%→50%A;5-20 min,50%→90%A;20-25 min,90%A;25-26 min,90%→30%A;26-30 min,30%A).The conditions of MS were as follows:electrospray ionization source(ESI-),drying temperature at 220℃,drying gas flow rate at 8 L·min^(-1),atomizing gas pressure of 2 bar,capillary voltage of 3500 V,nozzle voltage of 500 V,scanning range of 50-1000,and scanning frequency of 1 Hz.Two characteristic components in the snake bile were found,namely sulfate of taurocholate(abbreviated as SOTC,molecular formula C_(26)H_(45)NO_(10)S_(2))and sulfate of taurodeoxycholate(abbreviated as SOTDC,formula C_(26)H_(45)NO_(9)S_(2)).The MRM methods of SOTC and SOTDC were established,the liquid chromatographic conditions were as follows:methanol(A)-10 mmol·L^(-1) ammonium acetate(B),gradient elution(0-10 min,40%→50%A;10-25 min,50%→80%A;25-25.5 min,80%→40%A;25.5-30 min,40%A),mass spectrometry conditions were:negative mode electrospray ionization source(ESI-),the ion pair of SOTC was 594.2→514.3,594.2→124.0;SOTDC was 578.2→498.3,578.2→124.0.The method was applied for 41 batches of Shedan Chuangbei decoction,and snake bile was found in 40 batches.Conclusion The

关 键 词：超高效液相色谱-飞行时间质谱 蛇胆汁 鉴别 主成分分析 蛇胆川贝液 

分 类 号：R284[医药卫生—中药学]