一种测定氟尼辛葡甲胺含量新方法的建立
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摘要
为了建立高效液相色谱法测定复方庆大霉素注射液中氟尼辛葡甲胺的含量,选用色谱柱Thermo Accucore C18 100×2.1 mm,2.6 m;流动相A为0.1%三氟乙酸水溶液,B为乙腈;流动相A与B之间的比例为9∶1;用DAD检测器检测波长为254 nm;流速0.4 mL/min;柱温30℃,进样体积2.00 L。化合物的色谱图采集和积分由软件Chemstation进行处理。用内标法计算待测样品中氟尼辛葡甲胺的含量。结果,氟尼辛葡甲胺在该色谱条件下具有良好的系统适应性。回归方程的结果如下:y=2.545 35x+0.020 695 2。线性相关系数为r=0.999 91,氟尼辛葡甲胺的浓度在31.25～1000 g/mL范围内线性相关性良好。平均回收率在102.71%～108.33%之间,相对标准偏差为2.35%,小于3%。精密度试验中所有相对标准偏差都低于5%。复方庆大霉素注射液中氟尼辛葡甲胺的平均含量为212.8 g/mL,为标示量的106.4%,符合《中华人民共和国兽药典一部》的规定。上述结果表明,该方法可用于复方庆大霉素注射液中氟尼辛葡甲胺含量的测定。
In order to establish a new method of high performance liquid chromatography for the content determination of flunixin meglumine in compound gentamicin injection,thermo accucore C18(100×2.1 mm,2.6 m) is used as one of the chromatographic conditions.Mobile phase A is 0.1% aqueous solution of trifluoroacetic acid and Mobile phase B is acetonitrile.The ratio between mobile phase A and B is nine to one.The wavelength tested by DAD detector is 254 nm.The velocity is 0.4 mL/min,The temperature of column is 30 ℃,and the injection volume is 2.00 L.Chromatogram acquisition and integration of compounds were processed by the software Chemstation.The content of flunixin meglumine in the sample was calculated by internal standard method.In result,flunixin meglumine has good system adaptability under this chromatographic condition.The result of regrsssion equation is as follow: y=2.545 35x+0.020 695 2.Linear correlation coefficient is equal to 0.999 91,and the concentration of flunixin meglumine has strong linear correlation in the range from 31.25 g/mL to 1 000 g/mL.The average recovery rate is between 102.71% and 108.33%.The relative standard deviation which is 2.35% is less than3%.All of the relative standard deviation in the precision test are lower than 5%.The average content of flunixin meglumine in the compound gentamycin injection is 212.8 g/mL.And the average content of flunixin meglumine is 106.4% of the labeled amount.The results meet the Chinese animal pharmacopoeia.The above-mentioned results showed that this method can be used to carry on the content determination of flunixin meglumine in compound gentamicin injection.
In order to establish a new method of high performance liquid chromatography for the content determination of flunixin meglumine in compound gentamicin injection,thermo accucore C18(100×2.1 mm,2.6 m) is used as one of the chromatographic conditions.Mobile phase A is 0.1% aqueous solution of trifluoroacetic acid and Mobile phase B is acetonitrile.The ratio between mobile phase A and B is nine to one.The wavelength tested by DAD detector is 254 nm.The velocity is 0.4 mL/min,The temperature of column is 30 ℃,and the injection volume is 2.00 L.Chromatogram acquisition and integration of compounds were processed by the software Chemstation.The content of flunixin meglumine in the sample was calculated by internal standard method.In result,flunixin meglumine has good system adaptability under this chromatographic condition.The result of regrsssion equation is as follow: y=2.545 35x+0.020 695 2.Linear correlation coefficient is equal to 0.999 91,and the concentration of flunixin meglumine has strong linear correlation in the range from 31.25 g/mL to 1 000 g/mL.The average recovery rate is between 102.71% and 108.33%.The relative standard deviation which is 2.35% is less than3%.All of the relative standard deviation in the precision test are lower than 5%.The average content of flunixin meglumine in the compound gentamycin injection is 212.8 g/mL.And the average content of flunixin meglumine is 106.4% of the labeled amount.The results meet the Chinese animal pharmacopoeia.The above-mentioned results showed that this method can be used to carry on the content determination of flunixin meglumine in compound gentamicin injection.
引文
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[13]中国兽药典委员会.中华人民共和国兽药典一部[M].北京:化学工业出版社,2015.Chinese Veterinary Pharmacopoeia Commission.The Veterinary Pharmacopoeia of the People's Republic of China.Part 1[M].Beijing:Chemical Industry Press,2015.(in Chinese)
[14]黄诚,卢丽明,刘国平.内标法和外标法测定血液中乙醇含量的比较分析[J].生命科学仪器,2008,6(3):41-43.HUANG Cheng,LU Li-ming,LIU Guo-ping.Comparative analysis of determination of ethanol content in blood by internal standard method and external standard method[J].Life Science Instruments,2008,6(3):41-43.(in Chinese)
[15]余祖功,蒋春茂,郭永刚,等.猪静注、肌注氟尼辛葡甲胺的药物动力学研究[J].中国农业科学,2007,40(9):2068-2072.YU Zu-gong,JIANG Chun-mao,GUO Yong-gang,et al.Pharmacokinetics of intravenous injection and intramuscular injection of flunixin meglumine[J].China Agricultural Science,2007,40(9):2068-2072.
[16] LUO Y,RUDY J A,UBOHA C E,et al.Quantification and confirmation of flunixin in equine plasma by liquid chromatography-quadrupole time-of-flight tandem mass spectrometry[J].Journal of Chromatography B,2004,801(2):173-184.
[17] KISSELL L W,BRINSON P D,GEHRING R,et al.Pharmacokinetics and tissue elimination of flunixin in veal calves[J].American Journal of Veterinary Research,2016,77(6):634-640.
[18] KNYCH H K,ARTHUR R M,MCKEMIE,D S,et al.Pharmacokinetics and effects on thromboxane B2 production following intravenous administration of flunixin meglumine to exercised thoroughbred horses[J].Journal of Veterinary Pharmacology&Therapeutics,2015,38(4):313-320.
[19] KISSELL L W,SMITH G W,LEAVENS T L,et al.Plasma pharmacokinetics and milk residues of flunixin and5-hydroxy flunixin following different routes of admistration in dairy cattle[J].American Dairy Science,2012,12(95):7151-7157.
[20]孙嘉一,蒋红,钟文诗,等.用响应面法优化四黄止痢复方中药提取工艺[J].中国兽医科学,2019,49(01):119-126.SUN Jia-yi,JIANG Hong,ZHONG Wen-shi,et al.Optimization of Chinese traditional medicine extraction process of Sihuang Zhizhi compound by response surface methodology[J].Chinese Veterinary Science,2019,49(01):119-126.(in Chinese)
[21]汪正范.色谱定性与定量[M].化学工业出版社,2007.WANG Zheng-fan.Chromatographic characterization and quantification[M].Chemical Industry Press,2007.(in Chinese)
[2] VANE J R,BOTTING R M.Mechanism of action of antiinflammatory drugs[J].Advances in Experimental Medicine&Biology,1997,25(sup102):9-21.
[3]吴文学,王中杰,李金祥.新兽药氟尼辛葡甲胺的研究[J].中国兽医杂志,2006,42(1):51-53.WU Wen-xue,WANG Zhong-jie,LI Jin-xiang.Study on the new veterinary drug flunixin meglumine[J].Chinese Journal of Veterinary Medicine,2006,42(1):51-53.(in Chinese)
[4] HIGGINS A J,LEES P,TAYLOR J B,et al.Flunixin meglumine:quantitative determination in and effects on composition of equine inflammatory exudate[J].British Veterinary Journal,1986,142(2):163-169.
[5]蒋美琳,艾晓杰,李银生,等.兽药对猪肝细胞CYP450酶系影响的研究进展[J].畜牧与兽医,2014,46(9):116-120.JIANG Mei-lin,AI Xiao-jie,LI Yin-sheng,et al.Advances in research on effects of veterinary drugs on CYP450 enzymes in porcine hepatocytes[J].Animal Husbandry and Veterinary Medicine,2014,46(9):116-120.(in Chinese)
[6] KEEGAN K G,MESSER N T,REED S K.Effectiveness of administration of phenylbutazone alone or concurrent administration of phenylbutazone and flunixin meglumine to alleviate lameness in horses[J].American Journal of Veterinary Research,2008,69(2):167-173.
[7] THIRY J,GONZALEZ-MARTAN J V,ELVIRA L,et al.Treatment of naturally occurring bovine respiratory disease in juvenile calves with a single administration of a florfenicol plus flunixin meglumine formulation[J].Veterinary Record,2014.174(17):430-438.
[8] PAIRIS-GARCIA M D,JOHNSON A K,STALDER K J,et al.Measuring the efficacy of flunixin meglumine and meloxicam for lame sows using nociceptive threshold tests[J].Animal Welfare,2014,23:219-229.86.lib.dr.iastate.edu.
[9] LUO Y,RUDY J A,UBOHA C E,et al.Quantification and confirmation of flunixin in equine plasma by liquid chromatography-quadrupole time-of-flight tandem mass spectrometry[J].Journal of Chromatography B,2004,801(2):173-184.
[10] LUGOBONI B,BARBAROSSA A,GAZZOTTI T E,et al.A Quick LC-MS-MS method for the determination of flunixin in bovine muscle[J].Journal of Analytical Toxicology,2014,38(2):173-184.
[11] KISSELL L W,SMITH G W,LEAVENS T L,et al.Plasma pharmacokinetics and milk residues of flunixin and5-hydroxy flunixin following different routes of adminstratrion in dairy cattle[J].Americn Dairy Science,2012,12(95):7151-7157.
[12] ELMAS M,YAZAR E,UNEY K,et al.Pharmacokinetics of enrofloxacin and flunixin meglumine and interactions between both drugs after intravenous co-adminstratrion in healthy and endotoxacmic[J].The Veterinary Journal,2008,177(3):418-424.
[13]中国兽药典委员会.中华人民共和国兽药典一部[M].北京:化学工业出版社,2015.Chinese Veterinary Pharmacopoeia Commission.The Veterinary Pharmacopoeia of the People's Republic of China.Part 1[M].Beijing:Chemical Industry Press,2015.(in Chinese)
[14]黄诚,卢丽明,刘国平.内标法和外标法测定血液中乙醇含量的比较分析[J].生命科学仪器,2008,6(3):41-43.HUANG Cheng,LU Li-ming,LIU Guo-ping.Comparative analysis of determination of ethanol content in blood by internal standard method and external standard method[J].Life Science Instruments,2008,6(3):41-43.(in Chinese)
[15]余祖功,蒋春茂,郭永刚,等.猪静注、肌注氟尼辛葡甲胺的药物动力学研究[J].中国农业科学,2007,40(9):2068-2072.YU Zu-gong,JIANG Chun-mao,GUO Yong-gang,et al.Pharmacokinetics of intravenous injection and intramuscular injection of flunixin meglumine[J].China Agricultural Science,2007,40(9):2068-2072.
[16] LUO Y,RUDY J A,UBOHA C E,et al.Quantification and confirmation of flunixin in equine plasma by liquid chromatography-quadrupole time-of-flight tandem mass spectrometry[J].Journal of Chromatography B,2004,801(2):173-184.
[17] KISSELL L W,BRINSON P D,GEHRING R,et al.Pharmacokinetics and tissue elimination of flunixin in veal calves[J].American Journal of Veterinary Research,2016,77(6):634-640.
[18] KNYCH H K,ARTHUR R M,MCKEMIE,D S,et al.Pharmacokinetics and effects on thromboxane B2 production following intravenous administration of flunixin meglumine to exercised thoroughbred horses[J].Journal of Veterinary Pharmacology&Therapeutics,2015,38(4):313-320.
[19] KISSELL L W,SMITH G W,LEAVENS T L,et al.Plasma pharmacokinetics and milk residues of flunixin and5-hydroxy flunixin following different routes of admistration in dairy cattle[J].American Dairy Science,2012,12(95):7151-7157.
[20]孙嘉一,蒋红,钟文诗,等.用响应面法优化四黄止痢复方中药提取工艺[J].中国兽医科学,2019,49(01):119-126.SUN Jia-yi,JIANG Hong,ZHONG Wen-shi,et al.Optimization of Chinese traditional medicine extraction process of Sihuang Zhizhi compound by response surface methodology[J].Chinese Veterinary Science,2019,49(01):119-126.(in Chinese)
[21]汪正范.色谱定性与定量[M].化学工业出版社,2007.WANG Zheng-fan.Chromatographic characterization and quantification[M].Chemical Industry Press,2007.(in Chinese)