一测多评法同时测定没食子药材及制剂西帕依固龈液中5个没食子鞣质  

Simultaneous determination of five gallotannins in Turkish galls and Xipayi mouth rinse by QAMS

作　　者：霍仕霞 陈两绵 祖力皮卡尔·吾斯曼[1] 窦勤 高慧敏 王智民[4] 李治建[2,3] 邢建国 HUO Shi-xia;CHEN Liang-mian;Zulipikaer Wusiman;DOU Qin;GAO Hui-min;WANG Zhi-min;LI Zhi-jian;XING Jian-guo(School of Pharmacy,Xinjiang Medical University,Urumqi,Xinjiang 830011,China;Uygur Medical Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830049,China;Uygur Medicine Research Institute of Xinjiang Uygur Autonomous Region,Urumqi 830049,China;National Engineering Laboratory for Quality Control Technology of Chinese Materia Medica,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Medicine Research Institute of Xinjiang Uygur Autonomous Region,Urumqi 830002,China)

机构地区：[1]新疆医科大学药学院,乌鲁木齐830011 [2]新疆维吾尔自治区维吾尔医医院,乌鲁木齐830049 [3]新疆维吾尔自治区维吾尔医药研究所,乌鲁木齐830049 [4]中国中医科学院、中药研究所中药质量控制技术国家工程实验室,北京100700 [5]新疆维吾尔自治区药物研究所,乌鲁木齐830002

出　　处：《药物分析杂志》2023年第5期780-792,共13页Chinese Journal of Pharmaceutical Analysis

基　　金：新疆维吾尔自治区自然科学基金(2021D01A157);新疆维吾尔自治区天山英才项目(2022TSYCCX0021,2022TSYCLJ009);新疆维吾尔自治区卫生健康青年医学科技人才专项(WJWY-202225)。

摘　　要：目的:建立同时测定没食子药材及制剂西帕依固龈液中没食子酸(GA)、1-没食子酸酰葡萄糖(1-GG)、没食子酸甲酯(MG)、1,2,3,6-四没食子酰葡萄糖(TeGG)和1,2,3,4,6-五没食子酰葡萄糖(PGG)5个没食子鞣质的一测多评方法(QAMS)。方法:采用超高效液相色谱-四极杆-飞行时间质谱(UPLC-Q-TOF MS)法,对没食子药材的主要没食子鞣质进行定性鉴别,并确定待测成分;采用HPLC建立多成分同步测定方法,色谱柱为Phenomenex C_(18)柱(250 mm×4.6 mm,5μm),流动相为乙腈(A)-0.2%磷酸溶液(B),梯度洗脱(0~20 min,3%A→8%A;20~30 min,8%A→10%A;30~70 min,10%A→15%A;70~100 min,15%A→18%A;100~120 min,18%A→30%A),流速1.0 mL·min^(-1),柱温35℃,检测波长275 nm。以GA为内参物,采用斜率法,分别建立1-GG、MG、TeGG、PGG的相对校正因子(RCFs),并考察其耐用性。采用QAMS法和外标法(ESM)分别计算和比较9批次没食子药材和3批次西帕依固龈液中上述没食子鞣质的含量,以验证QAMS法的准确性和可行性。结果:从没食子中定性鉴别出GA、1-GG、MG、TeGG和PGG 5个含量较高且具有良好生物活性的没食子鞣质,选择作为HPLC定量分析的指标性成分。GA、1-GG、MG、TeGG和PGG的线性范围分别为8.13~520、6.25~400、8.28~530、9.00~576、9.00~576μg·mL^(-1)。1-GG、MG、TeGG和PGG的RCFs分别为1.75、0.95、1.12、1.25,对不同HPLC仪、色谱柱、流速和柱温的耐用性良好(RSD≤5.00%)。采用QAMS法测得9批次没食子药材中GA、1-GG、MG、TeGG和PGG的含量范围分别为3.69%~5.07%、1.26%~1.98%、14.0%~17.4%、19.8%~25.3%、5.93%~9.24%,3批次西帕依固龈液中GA、1-GG和MG的含量范围分别为1.89~2.40、0.0750~0.105、0.874~1.12 mg·mL^(-1)。QAMS法和ESM法测定的含量结果的相对误差均小于3.5%,无显著性差异。结论:以GA为内参物,建立的上述5个没食子鞣质的QAMS法,简便、准确、可靠,节省大量对照品,可用于没食子药材及制剂西帕依固龈液的定量分�Objective:To establish a quantitative analysis of multi-components by single marker(QAMS)method for the simultaneous determination of five gallotannins of gallic acid(GA),1-galloylglucose(1-GG),methylgallate(MG),1,2,3,6-tetragalloylglucose(TeGG)and 1,2,3,4,6-pentagalloylglucose(PGG)in Turkish galls and Xipayi mouth rinse.Methods:Ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF MS)was used to identify the main gallotannins in Turkish galls and for selecting the components for quantitative analysis.HPLC was used to establish the method for simultaneous determination of the multi-components with the chromatographic conditions of Phenomenex C_(18)(250 mm×4.6 mm,5μm)column,the mobile phase of acetonitrile(A)-0.2% phosphoric acid aqueous solution(B)for gradient elution(0-20 min,3%A→8%A;20-30 min,8%A→10%A;30-70 min,10%A→15%A;70-100 min,15%A→18%A;100-120 min,18%A→30%A),flow rate of 1.0 mL·min^(-1),column temperature of 35℃,and detection wavelength of 275 nm.Using GA as the internal reference substance,the relative correction factors(RCFs)of 1-GG、MG、TeGG and PGG were calculated by slope method,respectively,and their durability were examined.The contents of the above gallotannins in nine batches of Turkish galls and three batches of Xipayi mouth rinse were calculated and compared by using QAMS and external standard method(ESM),respectively,to verify the accuracy and feasibility of QAMS.Results:Five gallotannins of GA,1-GG,MG,TeGG and PGG,with high contents and good biological activities,were qualitatively identified from Turkish galls,and were selected as the index components for HPLC quantitative analysis.The linear ranges of GA,1-GG,MG,TeGG and PGG were 8.13-520,6.25-400,8.28-530,9.00-576 and 9.00-576μg·mL^(-1),respectively.The RCFs of 1-GG,MG,TeGG and PGG were 1.75,0.95,1.12 and 1.25,respectively.It showed good durability for different HPLC instruments,chromatographic columns,flow rates and column temperatures(RSD≤5.00%).The content ranges

关 键 词：没食子 西帕依固龈液 没食子鞣质 超高效液相色谱-四级杆-飞行时间质谱法 一测多评 定量分析 质量控制 

分 类 号：R917[医药卫生—药物分析学]