LC-MS/MS法测定升麻葛根汤颗粒在犬体内的药代动力学研究
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摘要
[目的]建立液相色谱-质谱联用(LC-MS/MS)的方法测定犬灌胃给药升麻葛根汤颗粒后葛根素、芍药苷、异阿魏酸的药代动力学。[方法]采用Waters ACQUITY UPLCBEH C18柱(2.1 mm×50 mm,1.7μm),流动相:0.05%甲酸水(A)-乙腈(B)梯度洗脱;流速:0.2 mL/min;柱温:25℃;进样量:5μL。以甲苯磺丁脲为内标,采用电喷雾离子源(ESI离子源),在负离子检测方式下进行多反应离子检测(MRM),检测离子对分别为m/z 415.2→294.9(葛根素)、m/z 525.0→449.0(芍药苷)、m/z 193.05→133.9(异阿魏酸)、m/z 269.09→169.9(甲苯磺丁脲,内标)。[结果]葛根素、芍药苷、异阿魏酸在10～4 000 ng/mL范围内线性关系良好。血浆中内源性物质不干扰测定,方法回收率、基质效应、精密度、准确度和稳定性均符合生物样品的测定要求。[结论]该方法准确度强,专属性好,灵敏度高,可用于检测葛根素、芍药苷、异阿魏酸的药代动力学。
[Objective] LC-MS/MS was developed for the determination of the pharmacokinetics of puerarin,paeoniflorin and isoferulic acid in dogs after oral Shengma Gegen decoction granule. [Methods] The column ofWaters ACQUITY UPLCBEH C18(1.7 μm,2.1 mm×50 mm)and a mixture of 0.05% formic acid water solution(A)-acetonitrile(B) as the mobile phase at the flow-rate of 0.2 mL/min,the column temperature was 25 ℃ and the injection volume was 5 μL.Tolbutamide was used as the internal standard,detection was by negative ion electrospray ionization(ESI) mass spectrometry with multiple-reaction monitoring(MRM). Detection of ion pairs respectively are m/z 415.2 →294.9 for puerarin,m/z 525.0 →449.0 for paeoniflorin,m/z 193.05 →133.9 for isoferulic acid,m/z269.09 →169.9 for tolbutamide. [Results] For the puerarin,paeoniflorin and isoferulic acid,the linear calibration curves were obtained over the concentration range 10 ～4 000 ng/mL.Endogenous substances in the plasma do not interfere with the determination,the recovery rate,matrix effect,precision,accuracy and stability of the method are in accordance with the requirements of biological samples. [Conclusion] The method has the advantages of strong accuracy,good specificity and high sensitivity,and can be used to detect the pharmacokinetics of puerarin,paeoniflorin and isoferulic acid.
[Objective] LC-MS/MS was developed for the determination of the pharmacokinetics of puerarin,paeoniflorin and isoferulic acid in dogs after oral Shengma Gegen decoction granule. [Methods] The column ofWaters ACQUITY UPLCBEH C18(1.7 μm,2.1 mm×50 mm)and a mixture of 0.05% formic acid water solution(A)-acetonitrile(B) as the mobile phase at the flow-rate of 0.2 mL/min,the column temperature was 25 ℃ and the injection volume was 5 μL.Tolbutamide was used as the internal standard,detection was by negative ion electrospray ionization(ESI) mass spectrometry with multiple-reaction monitoring(MRM). Detection of ion pairs respectively are m/z 415.2 →294.9 for puerarin,m/z 525.0 →449.0 for paeoniflorin,m/z 193.05 →133.9 for isoferulic acid,m/z269.09 →169.9 for tolbutamide. [Results] For the puerarin,paeoniflorin and isoferulic acid,the linear calibration curves were obtained over the concentration range 10 ～4 000 ng/mL.Endogenous substances in the plasma do not interfere with the determination,the recovery rate,matrix effect,precision,accuracy and stability of the method are in accordance with the requirements of biological samples. [Conclusion] The method has the advantages of strong accuracy,good specificity and high sensitivity,and can be used to detect the pharmacokinetics of puerarin,paeoniflorin and isoferulic acid.
引文
[1]李冀.方剂学[M].北京:中国中医药出版社,2012:37.
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[5]李秀荣,李世海,马琪.升麻葛根汤合四物汤配合针灸治疗面瘫30例[J].甘肃中医,2011,24(4):51.
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[7]王本祥,刘媛.升麻葛根汤在耳鼻喉科的运用[J].四川中医,2002,20(1):70.
[8]李德珍,安德明,王抗战,等.升麻葛根汤治疗免疫清除期慢性乙型肝炎临床研究[J].中西医结合肝病杂志,2015,25(2):90-91.
[9]李光,宋德新,蒋凯.升麻葛根汤加复方丹参注射液治疗丙肝78例[J].河南中医药学刊,2001,16(2):36-37.
[10] Shiyi Ou,Kin-Chor Kwork. Ferulic acid:Pharmaceutical functions,preparation and applications in foods[J].Science of Food and Agriculture,2004,84(11):1261-1269.
[11]韩斐,王婷.阿魏酸钠联合贝那普利治疗早期糖尿病肾病的疗效研究[J].新疆医科大学学报,2012,35(7):942-945.
[12]柯雪红,陈为,陈锦富,等. HPLC法测定不同厂家补中益气丸中阿魏酸和异阿魏酸的含量[J].安徽医药,2010,14(7):768-770.
[13]刘颖,张小茜.升麻质量标准的研究[J].中草药,2005,36(9):126-128.
[14]陈斌,刘训红,蔡宝昌.葛根高效液相色谱指纹图谱的研究[J].时珍国医国药,2005,16(5):384-385.
[15] Huang TK. Handbook of the Composition and Pharmacology of Common Chinese Drugs[Z].Beijing:China Medico-Pharmaceutical Sciences and Technology Publishing House,1994,25(4):745-749.
[16]韩珍,贺弋.白芍总苷的药理作用及其毒性研究进展[J].宁夏医学院学报,2008,30(4):538-554.
[17] Kobayashi M,Ueda C,Aoki S,et al. Anticholinergic action Paeony root and its active constituents[J].Yakugaku-Zasshi,1990,110(12):964.
[18]姚少姿,常梦春,卢鹏,等. HPLC法同时测定升麻葛根汤中3个有效成分的含量[J].天津中医药,2017,34(5):349-352.
[19]彭晓一,可成友,王立立,等. HPLC法同时测定升麻葛根汤中四种有效成分的含量[J].中国药师,2010,13(6):835-837.
[20]赵莹莹,蔡皓,刘晓,等.白芍硫磺熏蒸后导致升麻葛根汤HPLC指纹图谱变化的研究[J].南京中医药大学学报,2014,30(4):365-369.
[21]郭珉,李珠华,张志勤,等. RP-HPLC法同时测定复方升麻颗粒中异阿魏酸、葛根素和芍药苷[J].中成药,2012,34(6):1081-1084.
[22]李煦颖,赵妍,王琳,等.葛根素及葛根提取物在大鼠体内药代动力学研究[J].中国生化药物杂志,2009,30(6):383-386.
[23]姜清华,邓晶晶,吴琼,等.芍药苷的药代动力学研究进展[J].实用药物与临床,2010,13(6):451-453+478.
[24]王晓明,张帆,刘莹,等. LC-MS/MS法测定升麻中咖啡酸、阿魏酸、异阿魏酸含量的研究[J].天津中医药,2014,31(11):686-689.
[2]国家药典委员会.中华人民共和国药典(一部)[M].北京:中国医药科技出版社,2015,73.
[3]董德翠,王璐瑜.升麻葛根汤加味治疗带状疱疹后遗神经痛26例疗效观察[J].中国民族民间医药,2009,18(5):103-104.
[4]温红伟,李岳军.“加味升麻葛根汤”治疗后循环缺血37例临床观察[J].江苏中医药,2010,42(6):36-37.
[5]李秀荣,李世海,马琪.升麻葛根汤合四物汤配合针灸治疗面瘫30例[J].甘肃中医,2011,24(4):51.
[6]张团昌.升麻葛根汤临床应用举隅[J].山西中医学院学报,2003,1(4):36.
[7]王本祥,刘媛.升麻葛根汤在耳鼻喉科的运用[J].四川中医,2002,20(1):70.
[8]李德珍,安德明,王抗战,等.升麻葛根汤治疗免疫清除期慢性乙型肝炎临床研究[J].中西医结合肝病杂志,2015,25(2):90-91.
[9]李光,宋德新,蒋凯.升麻葛根汤加复方丹参注射液治疗丙肝78例[J].河南中医药学刊,2001,16(2):36-37.
[10] Shiyi Ou,Kin-Chor Kwork. Ferulic acid:Pharmaceutical functions,preparation and applications in foods[J].Science of Food and Agriculture,2004,84(11):1261-1269.
[11]韩斐,王婷.阿魏酸钠联合贝那普利治疗早期糖尿病肾病的疗效研究[J].新疆医科大学学报,2012,35(7):942-945.
[12]柯雪红,陈为,陈锦富,等. HPLC法测定不同厂家补中益气丸中阿魏酸和异阿魏酸的含量[J].安徽医药,2010,14(7):768-770.
[13]刘颖,张小茜.升麻质量标准的研究[J].中草药,2005,36(9):126-128.
[14]陈斌,刘训红,蔡宝昌.葛根高效液相色谱指纹图谱的研究[J].时珍国医国药,2005,16(5):384-385.
[15] Huang TK. Handbook of the Composition and Pharmacology of Common Chinese Drugs[Z].Beijing:China Medico-Pharmaceutical Sciences and Technology Publishing House,1994,25(4):745-749.
[16]韩珍,贺弋.白芍总苷的药理作用及其毒性研究进展[J].宁夏医学院学报,2008,30(4):538-554.
[17] Kobayashi M,Ueda C,Aoki S,et al. Anticholinergic action Paeony root and its active constituents[J].Yakugaku-Zasshi,1990,110(12):964.
[18]姚少姿,常梦春,卢鹏,等. HPLC法同时测定升麻葛根汤中3个有效成分的含量[J].天津中医药,2017,34(5):349-352.
[19]彭晓一,可成友,王立立,等. HPLC法同时测定升麻葛根汤中四种有效成分的含量[J].中国药师,2010,13(6):835-837.
[20]赵莹莹,蔡皓,刘晓,等.白芍硫磺熏蒸后导致升麻葛根汤HPLC指纹图谱变化的研究[J].南京中医药大学学报,2014,30(4):365-369.
[21]郭珉,李珠华,张志勤,等. RP-HPLC法同时测定复方升麻颗粒中异阿魏酸、葛根素和芍药苷[J].中成药,2012,34(6):1081-1084.
[22]李煦颖,赵妍,王琳,等.葛根素及葛根提取物在大鼠体内药代动力学研究[J].中国生化药物杂志,2009,30(6):383-386.
[23]姜清华,邓晶晶,吴琼,等.芍药苷的药代动力学研究进展[J].实用药物与临床,2010,13(6):451-453+478.
[24]王晓明,张帆,刘莹,等. LC-MS/MS法测定升麻中咖啡酸、阿魏酸、异阿魏酸含量的研究[J].天津中医药,2014,31(11):686-689.