Kallem Raja Reddy, Trivedi Ravi Kumar, Mullangi Ramesh, Srinivas Nuggehally R
Department of Drug Metabolism and Pharmacokinetics, Discovery Research, Dr Reddy's Laboratories Ltd, Miyapur, Hyderabad-500 049, India.
Biomed Chromatogr. 2006 Dec;20(12):1289-94. doi: 10.1002/bmc.674.
For pharmacokinetic and toxicokinetic purpose a simple HPLC-UV method has been developed and validated for the estimation of DRF-4848, a novel COX-2 inhibitor in rat plasma. A liquid-liquid extraction was used to extract DRF-4848 and internal standard (IS, DRF-4367) from rat plasma. The analysis was performed on a C(18) column with UV detection at 285 nm. The isocratic mobile phase, 0.01 M potassium dihydrogen ortho phosphate (pH 3.2) and acetonitrile (50:50, v/v) was run at a flow rate of 1 mL/min. The retention times of DRF-4848 and IS were 6.8 and 11.2 min, respectively. Absolute recovery for analyte and IS was >80% from rat plasma. A linear response was observed over a concentration range 0.1-20 microg/mL. The lower limit of quantification (LLOQ) of DRF-4848 was 0.1 microg/mL. The inter- and intra-day precisions in the measurement of quality control (QC) samples, 0.1, 0.3, 8.0 and 15.0 microg/mL, were in the range 1.74-8.70% relative standard deviation (RSD) and 0.75-8.43% RSD, respectively. Accuracy in the measurement of QC samples was in the range 93.29-116.51% of the nominal values. Analyte and IS were stable in the battery of stability studies viz., benchtop, autosampler, long-term and freeze/thaw cycles.
为了进行药代动力学和毒代动力学研究，已开发并验证了一种简单的高效液相色谱 - 紫外检测法，用于测定大鼠血浆中新型环氧化酶 - 2抑制剂DRF - 4848。采用液 - 液萃取法从大鼠血浆中提取DRF - 4848和内标（IS，DRF - 4367）。分析在C(18)柱上进行，于285 nm处进行紫外检测。等度流动相为0.01 M磷酸二氢钾（pH 3.2）和乙腈（50:50，v/v），流速为1 mL/min。DRF - 4848和IS的保留时间分别为6.8分钟和11.2分钟。大鼠血浆中分析物和内标的绝对回收率均大于80%。在0.1 - 20 μg/mL的浓度范围内观察到线性响应。DRF - 4848的定量下限（LLOQ）为0.1 μg/mL。质量控制（QC）样品0.1、0.3、8.0和15.0 μg/mL的日间和日内精密度，相对标准偏差（RSD）分别在1.74 - 8.70%和0.75 - 8.43%范围内。QC样品测量的准确度在标称值的93.29 - 116.51%范围内。在一系列稳定性研究中，即台式、自动进样器、长期和冻融循环中，分析物和内标均稳定。
