| 摘要: | Monitoring pyrazinamide (PZA) and its metabolites during anti-TB treatment is important to prevent PZA-associated liver injury. This study explores the potential of using liquid chromatography-tandem mass spectrometry (LC-MS/MS) combined with dried plasma spots (DPS) as an assay for simultaneous measurement of PZA and its three metabolites, i.e., pyrazinoic acid (POA), hydroxypyrazinamide (OH-PZA), and hydroxypyrazinoic acid (OH-POA). The DPS method employed a micro-sampling strategy, requiring a 15 μL of plasma volume for each sample. Sample extraction from DPS was achieved using a 40 second of 400-watt microwave-assisted process with 60% methanol solution. All analytes were separated with a C18 column (100 mm × 2.1 mm, 2.6 μm, 100 ?). A multiple reaction monitoring detection was used with positive electrospray ionization mode. The correlation coefficients of the calibration curves were > 0.999 for all analytes. The quantification ranges were 0.31 to 120 μg/mL for PZA, and 0.06 to 24 μg/mL for POA, OH-PZA, and OH-POA. Intra-assay precision (n=5) exhibited a maximum relative standard deviation (RSD) of 3.00% and a minimum of 0.7%, with accuracy ranging from 89.2% to 113.1%. Inter-assay precision ranged from 1.5% to 12.1% of RSD, with accuracy between 95.1% and 111.8% (n=3). Following validation, we applied the developed method to 35 paired DPS and plasma samples from five TB patients for therapeutic drug monitoring and pharmacokinetic study. Plasma and DPS showed a high correlation with correlation of coefficients (rho) were 0.988, 0.994, 0.993, and 0.995 for PZA, POA, OH-PZA, and OH-POA, respectively. This method has the potential to significantly benefit TB patients management for personalized PZA treatment.
Monitoring pyrazinamide (PZA) and its metabolites during anti-TB treatment is important to prevent PZA-associated liver injury. This study explores the potential of using liquid chromatography-tandem mass spectrometry (LC-MS/MS) combined with dried plasma spots (DPS) as an assay for simultaneous measurement of PZA and its three metabolites, i.e., pyrazinoic acid (POA), hydroxypyrazinamide (OH-PZA), and hydroxypyrazinoic acid (OH-POA). The DPS method employed a micro-sampling strategy, requiring a 15 μL of plasma volume for each sample. Sample extraction from DPS was achieved using a 40 second of 400-watt microwave-assisted process with 60% methanol solution. All analytes were separated with a C18 column (100 mm × 2.1 mm, 2.6 μm, 100 ?). A multiple reaction monitoring detection was used with positive electrospray ionization mode. The correlation coefficients of the calibration curves were > 0.999 for all analytes. The quantification ranges were 0.31 to 120 μg/mL for PZA, and 0.06 to 24 μg/mL for POA, OH-PZA, and OH-POA. Intra-assay precision (n=5) exhibited a maximum relative standard deviation (RSD) of 3.00% and a minimum of 0.7%, with accuracy ranging from 89.2% to 113.1%. Inter-assay precision ranged from 1.5% to 12.1% of RSD, with accuracy between 95.1% and 111.8% (n=3). Following validation, we applied the developed method to 35 paired DPS and plasma samples from five TB patients for therapeutic drug monitoring and pharmacokinetic study. Plasma and DPS showed a high correlation with correlation of coefficients (rho) were 0.988, 0.994, 0.993, and 0.995 for PZA, POA, OH-PZA, and OH-POA, respectively. This method has the potential to significantly benefit TB patients management for personalized PZA treatment. |
