A sensitive, simple and rapid high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS） method was developed and fully validated for the simultaneous quantification of buprenorphine （BUP） and it...A sensitive, simple and rapid high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS） method was developed and fully validated for the simultaneous quantification of buprenorphine （BUP） and its N-dealkylated metabolite norbuprenorphine （NBUP） in 200 μL human plasma. Human plasma samples were prepared using liquid-liquid extraction, and then separated on a Shiseido MG C18 （5 μm, 2.0 mm × 50 mm） via 4.1 min gradient elution. Following electrospray ionization, the analytes were quantified on a triple-quadrupole mass spectrometer in multiple-reaction-monitoring （MRM） positive ion mode. Linearity was achieved from 25.0 to 10000 pg/mL for buprenorphine, from 20.0 to 8000 pg/mL for norbupre- norphine with r2〉0.99. The method was demonstrated with acceptable accuracy, precision and specificity for the detection of buprenorphine and norbuprenorphine. Recovery was 81.8-88.8 % for buprenorphine and 77.0-84.6% for norbuprenorphine, and the matrix effect was 95.6-97.4% for buprenorphine and 94.0-96.9% for norbuprenorphine; all were not concentration dependent. With validated matrix and autosampler stability data, this method was successfully applied in a bioequivalence study to support abbreviated new drug application.展开更多
A rapid and reproducible method has been developed for the simultaneous quantification of dopamine (DA), norepinephrine (NE), and serotonin (5-HT) in rat brain microdialysates. Derivatization was processed through red...A rapid and reproducible method has been developed for the simultaneous quantification of dopamine (DA), norepinephrine (NE), and serotonin (5-HT) in rat brain microdialysates. Derivatization was processed through reductive amination with butyraldehyde and sodium cyanoborohydride at 60℃ for 80 min incubation. Dibutylated monoamine neurotransmitters (MANTs) were directly analyzed with ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The chromatographic run time was shorter (2.1 min/sample) than previous studies [15] [18] [20]. Good linearity (R2 > 0.99) was obtained for DA, NE and 5-HT in the range of 25 - 5000 pg/mL, 5 - 1000 pg/mL and 2.5 - 500 pg/mL, respectively. Acceptable precision (CV, 8.5% - 13.4%) as well as accuracy (recovery, 94.1% - 106.8%) could be acquired by analysis of six batches of quality control samples (QCs) at four different concentrations, which demonstrated the reliability and reproducibility of current method. This method was successfully applied to the simultaneous determination of DA, NE and 5-HT in rat brain microdialysates, where basal levels as well as elevated levels after dosing with amphetamine (AMPH) were quantified for all three MANTs. This study provides a simple and rapid way to analyze MANTs in the biofluid in the future.展开更多
文摘A sensitive, simple and rapid high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS） method was developed and fully validated for the simultaneous quantification of buprenorphine （BUP） and its N-dealkylated metabolite norbuprenorphine （NBUP） in 200 μL human plasma. Human plasma samples were prepared using liquid-liquid extraction, and then separated on a Shiseido MG C18 （5 μm, 2.0 mm × 50 mm） via 4.1 min gradient elution. Following electrospray ionization, the analytes were quantified on a triple-quadrupole mass spectrometer in multiple-reaction-monitoring （MRM） positive ion mode. Linearity was achieved from 25.0 to 10000 pg/mL for buprenorphine, from 20.0 to 8000 pg/mL for norbupre- norphine with r2〉0.99. The method was demonstrated with acceptable accuracy, precision and specificity for the detection of buprenorphine and norbuprenorphine. Recovery was 81.8-88.8 % for buprenorphine and 77.0-84.6% for norbuprenorphine, and the matrix effect was 95.6-97.4% for buprenorphine and 94.0-96.9% for norbuprenorphine; all were not concentration dependent. With validated matrix and autosampler stability data, this method was successfully applied in a bioequivalence study to support abbreviated new drug application.
文摘A rapid and reproducible method has been developed for the simultaneous quantification of dopamine (DA), norepinephrine (NE), and serotonin (5-HT) in rat brain microdialysates. Derivatization was processed through reductive amination with butyraldehyde and sodium cyanoborohydride at 60℃ for 80 min incubation. Dibutylated monoamine neurotransmitters (MANTs) were directly analyzed with ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The chromatographic run time was shorter (2.1 min/sample) than previous studies [15] [18] [20]. Good linearity (R2 > 0.99) was obtained for DA, NE and 5-HT in the range of 25 - 5000 pg/mL, 5 - 1000 pg/mL and 2.5 - 500 pg/mL, respectively. Acceptable precision (CV, 8.5% - 13.4%) as well as accuracy (recovery, 94.1% - 106.8%) could be acquired by analysis of six batches of quality control samples (QCs) at four different concentrations, which demonstrated the reliability and reproducibility of current method. This method was successfully applied to the simultaneous determination of DA, NE and 5-HT in rat brain microdialysates, where basal levels as well as elevated levels after dosing with amphetamine (AMPH) were quantified for all three MANTs. This study provides a simple and rapid way to analyze MANTs in the biofluid in the future.
