Zhu M Q, Cui R
Department of Occupational and Environmental Health Science, Peking University School of Public Health, Beijing 100191, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2020 Jun 18;52(3):591-596. doi: 10.19723/j.issn.1671-167X.2020.03.030.
To establish a high performance liquid chromatography (HPLC) method for the determination of ultraviolet (UV) absorbers UV-327 and UV-328 in mouse plasma.
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 SymmetryC 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.
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 (=6) of UV-327 were 2.9%-6.6%, and 2.7%-7.4% for UV-328. The inter-day precisions (=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.
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.
建立高效液相色谱（HPLC）法测定小鼠血浆中紫外线（UV）吸收剂UV - 327和UV - 328的含量。
向小鼠血浆样品中加入正己烷 - 丙酮（体积比1∶1）作为萃取溶剂进行涡旋萃取，上清液于50℃用氮气吹干。然后将残渣用甲醇重新溶解，离心并过滤。采用Waters SymmetryC柱（250 mm×4.6 mm，5μm）进行分离，用紫外检测器通过HPLC测定小鼠血浆中UV - 327和UV - 328的浓度。以100%甲醇为流动相，等度洗脱，流速为1.0 mL/min，紫外检测波长为340 nm。采用保留时间进行定性分析，以内标法进行定量分析，选用UV - 320作为内标。
UV - 327和UV - 328的校准曲线在0.05至10.0 mg/L浓度范围内呈线性，相关系数为0.999 7。检测限为0.01 mg/L，定量限为0.03 mg/L。小鼠血浆中低、中、高三个浓度（0.50、1.00、2.00 mg/L）的UV - 327平均回收率为91.7% - 101.0%，UV - 328为97.5% - 103.9%。UV - 327的日内精密度（n = 6）为2.9% - 6.6%，UV - 328为2.7% - 7.4%。UV - 327的日间精密度（n = 3）为6.0% - 9.3%，UV - 328为6.6% - 8.6%。UV - 327的萃取回收率为98.8% - 103.8%，UV - 328为99.8% - 100.9%。置于室温6小时和 - 40℃15天的小鼠血浆样品中UV - 327的测定相对偏差分别为0.9% - 3.5%和7.4% - 15.0%，UV - 328的测定相对偏差分别为2.0% - 4.3%和2.1% - 13.8%。在三个加标浓度水平下，小鼠血浆样品至少可在室温保存6小时，在 - 40℃保存15天。
该方法简便快速，具有较高的准确度、精密度和灵敏度，可用于测定小鼠血浆中UV - 327和UV - 328的含量。
