目的 建立小鼠血浆中苯并三唑类紫外线(ultraviolet, UV)吸收剂UV-327和UV-328的高效液相色谱测定方法。方法 在小鼠血浆中加入正己烷-丙酮溶液(体积比1 ∶1)涡旋混匀提取,上清液于50 ℃下氮气吹干,残渣用甲醇溶解后离心过滤,采用高效液相色谱-紫外检测器测定小鼠血浆中UV-327和UV-328的浓度。分析柱为Waters Symmetry^?C₁₈柱 (250 mm×4.6 mm,5 μm),流动相为100%甲醇,等度洗脱,流速1.0 mL/min,紫外检测波长为340 nm,以UV-320为内标,保留时间定性,内标法定量。 结果 UV-327、UV-328在0.05~10.0 mg/L范围内呈现良好的线性关系(r=0.999 7),检出限为0.01 mg/L,定量限为0.03 mg/L。小鼠血浆中,低、中、高3种浓度(0.50、1.00、2.00 mg/L)UV-327和UV-328的平均回收率分别为91.7%~101.0%和97.5%~103.9%,日内精密度(n=6)分别为2.9%~6.6%和2.7%~7.4%,日间精密度(n=3)分别为6.0%~9.3%和6.6%~8.6%,提取回收率分别为98.8%~103.8%和99.8%~100.9%。血浆样品室温下放置6 h和-40 ℃放置15 d,UV-327测定值的相对偏差分别为0.9%~3.5%和7.4%~15.0%,UV-328测定值的相对偏差分别为2.0%~4.3%和2.1%~13.8%。3种加标浓度的小鼠血浆样品室温下至少可以放置6 h,-40 ℃冰箱中可以保存15 d。结论 高效液相色谱法简便、快速,准确度、精密度和灵敏度较高,可应用于小鼠血浆中UV-327和UV-328的分析测定。

Objective: To establish a high performance liquid chromatography (HPLC) method for the determination of ultraviolet (UV) absorbers UV-327 and UV-328 in mouse plasma.Methods: N-hexane-acetone (volume ratio 1 ∶1) was added to a mouse plasma sample as the extraction solvent for vortex extraction, and the supernatant was dried at 50 ℃ with nitrogen. Thereafter the residue was redissolved with methanol, centrifuged and filtered. The separation was performed on a Waters Symmetry^?C₁₈ column (250 mm×4.6 mm, 5 μm), and the concentrations of UV-327 and UV-328 in the mouse plasma were determined by HPLC with an UV detector. The elution was isocratic at a flow rate of 1.0 mL/min with a mobile phase composed of 100% methanol, and the UV detection wavelength was 340 nm. The retention time was used for qualitative analysis, and the internal standard method was used for quantitative analysis using UV-320 as the internal standard. Results: The calibration curves of UV-327 and UV-328 were linear with correlation coefficients of 0.999 7 over the concentration range of 0.05 to 10.0 mg/L. The limit of detection was 0.01 mg/L, and the limit of quantitation was 0.03 mg/L. The average recoveries at low, medium and high three concentrations (0.50, 1.00, 2.00 mg/L) in the mouse plasma were 91.7%-101.0% for UV-327, and 97.5%-103.9% for UV-328. The intra-day precisions (n=6) of UV-327 were 2.9%-6.6%, and 2.7%-7.4% for UV-328. The inter-day precisions (n=3) of UV-327 were 6.0%-9.3%, and 6.6%-8.6% for UV-328. The extraction recoveries of UV-327 were 98.8%-103.8%, and 99.8%-100.9% for UV-328. The measured relative deviations of UV-327 in the mouse plasma samples placed at room temperature for 6 hours and -40 ℃ for 15 days were 0.9%-3.5% and 7.4%-15.0%, and the measured relative deviations of UV-328 were 2.0%-4.3% and 2.1%-13.8%, respectively. The mouse plasma samples could be stored at room temperature for 6 hours at least and -40 ℃ for 15 days at three spiked concentration levels.Conclusion: The method was simple and fast with high accuracy, precision and sensitivity, and could be applied to the determination of UV-327 and UV-328 in mouse plasma.