Development of a Rapid and Simultaneous Detection Method for Buprenorphine, Norbuprenorphine and Naloxone in Human Plasma Using Ultra-high Performance Liquid Chromatography- tandem Mass Spectrometer with Solid-phase Extraction.pdfVIP

FULLPAPER． DevelopmentofaRapidandSimultaneousDetectionMethod forBuprenOrphine，NOrbuprenOrphineandNaloxoneinHuman PlasmaUsingUltra-highPerformanceLiquidChromatography— tandem MassSpectrometerwithSolid-phaseExtraction Guo，Qiaozhen 郭巧珍 Du，Zhenxia 杜振霞 FacultyofScience，BeijingUniversityofChemicalTechnology，Beijing100029，China A simultaneousmethodwassuccessfullyestablishedandvalidatedfortheseparationanddeterminationofbu— prenorphine BP ，itsprimraymetabolite，nor—buprenorphine NBP andaproposedco—formulate，naloxone NLX inhumanplasma．Themethodusedbuprenorphine—d4 BP—D4 ，nor—buprenorphine—d3 NBP—D3 ，naltrexone NTX asinternalstandards ISs ．100uLofplasmasamplefortifiedwiththeISswascleanedupbysolid—phaseextraction fSPE ．andwasthenseparatedonaWatersAcquity BEHC18columnwithgradientelutionusingmethanoland waterfcontaining0．2％ ofrmic1ataflowrateof0．25mL~rain ．Thcmassspectrometerwasusedfordetectionand wasoperatedinthepositiveelectrosprayionizationwiht multiplereaction monitoring fMRM mode．Thethree compoundswereeffectivelyseparatedin5rain．The1inera rangesofthecompoundswere0．1— 25．0．25— 25and 0．05— 25ng-mL一。ofrBP，NBPandNLX．respectively．withr≥0．9935．Themethodhadhighsensitivity thelim— itsofdetectionwere0．O2，0．1and0．01ng．mL ofrBP，NBP andNLX，respectively andhighrecoveries f≥97．6％ ．Theresultwasshowntobe1inera andsarisfactorilymetcurrentacceptancecriteriaofrvalidationofbio— analytica1method：intraandinterassayprecisionswithinhterequired1imitsof≤25％RSD．TheLOQsfulfilled hteLOQ requirements：precision≤25％RSD．andwasfullyvalidatedaccordingtotheStateFoodandDrugAd— ministration SFDA regulations．Theresultsdemonstratedthatultra—highperfomr anceliquidchromatography— tandem massspectrometerfUPLC—MS／MS1wim SPEwasapowerfuldetectiont