aday NLRP1 drug precision and Accuracy. e QC samples in 4 concentration levels (LLOQ, low, middle, and high) and LLOQ samples have been assessed in fiveJournal of Analytical Techniques in ChemistryTable 2: Extraction recovery and PARP4 Formulation matrix impact data in the analytes in rat blood (n six).Analytes BDCQ DCQ DHCQ HCQNominal concentration (ng/mL) 2.five 2000 2.five 2000 2.5 2000 5Extraction recovery Mean SD RSD ( ) 90.27 12.97 14.37 86.64 five.45 6.29 89.59 10.98 12.26 87.14 4.06 4.66 86.42 12.24 14.16 92.00 three.67 three.99 92.14 9.86 ten.70 93.77 four.28 4.Matrix effect Imply SD RSD ( ) 66.20 three.84 5.81 83.14 two.58 three.ten 75.87 four.87 six.42 85.15 1.76 two.06 77.37 3.26 four.21 87.13 1.83 2.10 73.56 two.79 three.79 87.98 1.44 1.Table 3: Linear regression equation, variety, and correlation coefficients of 4 analytes. Analytes BDCQ DCQ DHCQ HCQ Regression equations y 0.181 x + 0.006 y 0.134 x + 0.005 y 0.110 x + 0.003 y 0.216 x + 0.009 Calibration range (ng/mL) 1.002500.00 1.002500.00 1.002500.00 2.005000.00 r2 0.987 0.994 0.990 0.Table four: Interday and intraday precision and accuracy of four analytes in rat blood (n 5). Analytes Nominal concentration (ng/mL) 1 two.5 1000 2000 1 2.5 1000 2000 1 two.5 1000 2000 two 5 2000 4000 Intraday (n five) Measured concentration Precision (mean SD, ng/mL) (RSD ) 1.03 0.005 0.56 two.65 0.05 1.99 1085.1 14.four 1.33 1797.67 91.65 five.10 1.04 0.02 two.43 2.57 0.07 2.80 1057.50 86.3 8.17 2029.26 157.29 7.75 1.04 0.03 3.37 2.61 0.09 three.77 1096.61 56.7 5.18 1815.69 23.66 1.30 2.02 0.02 1.21 5.14 0.10 two.11 2040.six 175.1 8.59 4276.65 368.71 8.62 Accuracy (RE ) 3.98 6.34 eight.51 -10.11 four.26 3.19 5.75 1.46 four.83 4.67 six.69 -9.21 1.20 2.94 two.04 six.91 Interday (n 5) Measured concentration Precision Accuracy (mean SD, ng/mL) (RSD ) (RE ) 1.02 0.02 two.59 two.45 2.54 0.15 six.24 1.82 1097.3 40.1 three.66 9.73 1758.68 59.67 3.39 -12.06 1.03 0.04 four.15 three.49 2.55 0.08 3.24 two.23 1044.80 81.4 7.80 4.48 2033.73 138.43 six.81 1.68 1.02 0.03 3.45 2.39 2.53 0.20 8.15 1.34 1097.0 58.9 five.37 9.70 1796.74 33.65 1.87 -10.16 two.06 0.06 3.13 three.02 five.05 0.20 4.14 1.11 2022.1 158.1 7.82 1.ten 4262.63 321.74 7.55 6.BDCQDCQDHCQHCQreplicates at 3 separate analytical lots to determine the intraday and interday precision and accuracy. e outcomes of interdayand intraday precision and accuracy are acceptable and are summarized in Table 4. 3.three.5. Stability. We investigated the stability of four analytes in rat blood at two concentration levels (low and higher). As a consequence, the analytes were steady in autosampler for 24 h with deviations situated inside five (RE ). Immediately after being stored at -80 for 30 days, there are no considerable deviations in QC samples. Meanwhile, no apparent deviations have been found just after three freeze-thaw cycles. Bench-top stability also presented deviations inside 5 (data not shown). Each of the benefits are shown in Table five.3.3.6. Application in HCQ Pharmacokinetic Study. An UHPLC-MS/MS technique was established and validated for determining HCQ and its three metabolites, which was then applied to a pharmacokinetic study of HCQ. HCQ (36 mg/ kg) was provided to rats by intragastric administration. e blood concentration-time curves of HCQ and its three metabolites are shown in Figure three. e pharmacokinetic parameters of HCQ and three metabolites in rats are shown in Table six. TCmax of HCQ in rats was 1440.72 298.24 g/L (mean SD) and T1/2 was 21.14 10.31 h. AUC0was 42774.94 8495.26 g/L h, and also the clearance rate was 1.52 0.38 L/h/kg. e pharmacokinetic parameters of HCQ in rats have been compared with that in mice reported in litera