Quantitative Analysis of Amoxycillin and Its Major Metabolites in Animal Tissues by Liquid Chromatography Combined with Electrospray Ionization Tandem Mass Spectrometry
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- 作者：Siegrid De Baere ; Marc Cherlet ; Kris Baert ; and Patrick De Backer
- 刊名：Analytical Chemistry
- 出版年：2002
- 出版时间：March 15, 2002
- 年：2002
- 卷：74
- 期：6
- 页码：1393 - 1401
- 全文大小：122K
- 年卷期：v.74,no.6(March 15, 2002)
- ISSN：1520-6882
文摘
A sensitive and specific method for the quantitativedetermination of amoxycillin and its major metabolites(amoxycilloic acid, amoxycillinpiperazine-2',5'-dione) inanimal tissue samples using liquid chromatography combined with positive electrospray ionization tandem massspectrometry (LC/ESI-MS/MS) is presented. A liquidextraction using an aqueous 0.01 M potassium dihydrogenphosphate solution as the extraction solvent wasperformed for a preliminary sample cleanup. The extractswere further purified by a solid-phase extraction using anoctadecyl (C18) column. Ampicillin was used as theinternal standard. Chromatographic separation of theanalytes of interest was achieved on a reversed-phaseHypersil column (100 × 3 mm i.d., dp, 5 
m), using amixture of 9.6 mM pentafluoropropionic acid in water andacetonitrile as the mobile phase. Gradient elution wasperformed. To obtain as high sensitivity and selectivity aspossible, the mass spectrometer was operated in themultiple reaction monitoring mode. The method wasvalidated for the analysis of amoxycillin and its investigated metabolites in various porcine tissues, kidney, liver,muscle, and fat, according to the requirements definedby the European Community. Calibration graphs wereprepared for all tissues, and good linearity was achievedover the concentration range tested (25-500 ng/g, r 
0.9974, and goodness of fit 
9.6). A limit of quantification of 25 ng/g was obtained for amoxycillin and itsmetabolites in all tissues, which corresponds to half themaximum residue limit for amoxycillin. Limits of detection ranged from 2.3 to 12.0 ng/g for amoxycillin andfrom 1.1 to 15.1 ng/g and 0.2 to 2.4 ng/g for amoxycilloicacid and amoxycillinpiperazine-2',5'-dione, respectively.The results for the within-day precision and the truenessfell within the ranges specified. The method has beensuccessfully used for the quantitative determination ofamoxycillin and its major metabolites in tissue samplesfrom pigs medicated via the drinking water, proving theusefulness of the developed method for application in thefield of residue analysis.
m), using amixture of 9.6 mM pentafluoropropionic acid in water andacetonitrile as the mobile phase. Gradient elution wasperformed. To obtain as high sensitivity and selectivity aspossible, the mass spectrometer was operated in themultiple reaction monitoring mode. The method wasvalidated for the analysis of amoxycillin and its investigated metabolites in various porcine tissues, kidney, liver,muscle, and fat, according to the requirements definedby the European Community. Calibration graphs wereprepared for all tissues, and good linearity was achievedover the concentration range tested (25-500 ng/g, r 0.9974, and goodness of fit 9.6). A limit of quantification of 25 ng/g was obtained for amoxycillin and itsmetabolites in all tissues, which corresponds to half themaximum residue limit for amoxycillin. Limits of detection ranged from 2.3 to 12.0 ng/g for amoxycillin andfrom 1.1 to 15.1 ng/g and 0.2 to 2.4 ng/g for amoxycilloicacid and amoxycillinpiperazine-2',5'-dione, respectively.The results for the within-day precision and the truenessfell within the ranges specified. The method has beensuccessfully used for the quantitative determination ofamoxycillin and its major metabolites in tissue samplesfrom pigs medicated via the drinking water, proving theusefulness of the developed method for application in thefield of residue analysis.