A simultaneous method was successfully established and validated for the separation and determination of bu- prenorphine （BP）, its primary metabolite, nor-buprenorphine （NBP） and a proposed co-formulate, naloxone ...A simultaneous method was successfully established and validated for the separation and determination of bu- prenorphine （BP）, its primary metabolite, nor-buprenorphine （NBP） and a proposed co-formulate, naloxone （NLX） in human plasma. The method used buprenorphine-d4 （BP-D4）, nor-buprenorphine-d3 （NBP-D3）, naltrexone （NTX） as internal standards （ISs）. 100 μL of plasma sample fortified with the ISs was cleaned up by solid-phase extraction （SPE）, and was then separated on a Waters AcquityTM BEH C18 column with gradient elution using methanol and water （containing 0.2% formic） at a flow rate of 0.25 mL·min^-1. The mass spectrometer was used for detection and was operated in the positive electrospray ionization with multiple reaction monitoring （MRM） mode. The three compounds were effectively separated in 5 min. The linear ranges of the compounds were 0.1--25, 0.25--25 and 0.05--25 ng·mL^-1 for BP, NBP and NLX, respectively, with r≥0.9935. The method had high sensitivity （the lim- its of detection were 0.02, 0.1 and 0.01 ng.mL-1 for BP, NBP and NLX, respectively） and high recoveries （≥97.6%）. The result was shown to be linear and satisfactorily met current acceptance criteria for validation of bio- analytical method： intra and inter assay precisions within the required limits of ≤25% RSD. The LOQs fulfilled the LOQ requirements： precision≤25% RSD, and was fully validated according to the State Food and Drug Administration （SFDA） regulations. The results demonstrated that ultra-high performance liquid chromatography- tandem mass spectrometer （UPLC-MS/MS） with SPE was a powerful detection tool and contributed to pharmaceutical analysis in biological matrices.展开更多
Ultra performance liquid chromatography(UPLC) coupled with orthogonal acceleration time-of-flight(oaTOF) mass spectrometry is becoming an effective and high-throughput analytical technique in metabonomics research.In ...Ultra performance liquid chromatography(UPLC) coupled with orthogonal acceleration time-of-flight(oaTOF) mass spectrometry is becoming an effective and high-throughput analytical technique in metabonomics research.In this paper,an UPLC-oaTOF-MS system was employed to investigate the serum profiles of 40 diabete mellitus(DM)/diabetic nephropathy(DN) patients and 25 healthy volunteers in an effort to find potential biomarkers.The UPLC system produced information-rich chromatograms with typical measured peak widths of 4 s,and generated peak capacities of 225 in 15 min.After the UPLC-oaTOF-MS analysis,principal component analysis(PCA) was used for group differentiation and marker selection.In the projection of positive mode,DM/DN patients and the controls were obviously divided into three groups.Furthermore,two potential markers were identified as phytosphingosine and sphinganine,respectively.Phytosphingosine and sphinganine with decreased concentrations in the patients,were closely related to the sphingolipid metabolism.展开更多
文摘A simultaneous method was successfully established and validated for the separation and determination of bu- prenorphine （BP）, its primary metabolite, nor-buprenorphine （NBP） and a proposed co-formulate, naloxone （NLX） in human plasma. The method used buprenorphine-d4 （BP-D4）, nor-buprenorphine-d3 （NBP-D3）, naltrexone （NTX） as internal standards （ISs）. 100 μL of plasma sample fortified with the ISs was cleaned up by solid-phase extraction （SPE）, and was then separated on a Waters AcquityTM BEH C18 column with gradient elution using methanol and water （containing 0.2% formic） at a flow rate of 0.25 mL·min^-1. The mass spectrometer was used for detection and was operated in the positive electrospray ionization with multiple reaction monitoring （MRM） mode. The three compounds were effectively separated in 5 min. The linear ranges of the compounds were 0.1--25, 0.25--25 and 0.05--25 ng·mL^-1 for BP, NBP and NLX, respectively, with r≥0.9935. The method had high sensitivity （the lim- its of detection were 0.02, 0.1 and 0.01 ng.mL-1 for BP, NBP and NLX, respectively） and high recoveries （≥97.6%）. The result was shown to be linear and satisfactorily met current acceptance criteria for validation of bio- analytical method： intra and inter assay precisions within the required limits of ≤25% RSD. The LOQs fulfilled the LOQ requirements： precision≤25% RSD, and was fully validated according to the State Food and Drug Administration （SFDA） regulations. The results demonstrated that ultra-high performance liquid chromatography- tandem mass spectrometer （UPLC-MS/MS） with SPE was a powerful detection tool and contributed to pharmaceutical analysis in biological matrices.
文摘Ultra performance liquid chromatography(UPLC) coupled with orthogonal acceleration time-of-flight(oaTOF) mass spectrometry is becoming an effective and high-throughput analytical technique in metabonomics research.In this paper,an UPLC-oaTOF-MS system was employed to investigate the serum profiles of 40 diabete mellitus(DM)/diabetic nephropathy(DN) patients and 25 healthy volunteers in an effort to find potential biomarkers.The UPLC system produced information-rich chromatograms with typical measured peak widths of 4 s,and generated peak capacities of 225 in 15 min.After the UPLC-oaTOF-MS analysis,principal component analysis(PCA) was used for group differentiation and marker selection.In the projection of positive mode,DM/DN patients and the controls were obviously divided into three groups.Furthermore,two potential markers were identified as phytosphingosine and sphinganine,respectively.Phytosphingosine and sphinganine with decreased concentrations in the patients,were closely related to the sphingolipid metabolism.
