高效液相色谱法测定小鼠脑组织中2,4-二羟基二苯甲酮

Determination of 2,4-dihydroxybenzophenone in mouse brain by high performance liquid chromatography

目的：建立和优化小鼠脑组织中2,4-二羟基二苯甲酮（2,4-dihydroxybenzophenone,BP-1）的测定方法。方法：采用正交试验设计优化样品预处理条件,Waters Symmetry C18（4.6 mm ×250 mm,5μm）色谱柱分离,等度洗脱,流动相为含3%（体积分数）乙酸的甲醇-水（体积比为3∶1）,pH为3.40,流速为1.0 mL/min,紫外检测器检测,检测波长为290 nm,高效液相色谱法测定小鼠脑组织中BP-1,保留时间定性,以内标法定量。结果：在优化的实验条件下,BP-1在0.2～10.0 mg/L浓度范围内线性关系良好,相关系数为0.9998；BP-1的加标回收率为96.8%～104.5%；日内和日间精密度分别为3.5%～5.7%和4.5%～6.4%；低、中、高3种浓度BP-1的提取回收率分别为90.5%、89.5%和97.7%；低、中、高3种浓度BP-1的基质效应分别为102.9%、102.7%和90.9%。结论：该方法简便、准确,适用于小鼠脑组织中BP-1的测定。

Objective：To optimize and establish the experimental methods for the determination of 2,4-dihydroxybenzophenone （BP-1） in mouse brain. Methods：BP-1 was determined by high performance liquid chromatography （HPLC） and separated by Waters Symmetry C18 （4. 6 mm × 250 mm, 5 μm） using isocratic elution, and the sample preparation conditions were optimized by orthogonal experiment design. The mobile phase was methanol-water （volume ratio 3∶1） containing 3% （volume fraction） ace-tic acid （pH 3. 40） at a flow rate of 1. 0 mL/min, and ultraviolet （UV） detection wavelength was set at 290 nm. Retention time was used for qualitative analysis and internal standard method for quantitative analysis. Results： Under the optimized experimental conditions, the calibration curve was linear with a correlation coefficient of 0. 999 8 over the concentration range of 0. 2-10. 0 mg/L. The recoveries of BP-1 were between 96. 8% and 104. 5%. The intra-day and inter-day precision of BP-1 were 3. 5% -5. 7% and 4. 5% -6. 4%, respectively. The extraction recoveries of BP-1 at three concentrations （0. 5, 2. 0, 8. 0 mg/L） in the mouse brain were 90. 5%, 89. 5%, and 97. 7%, and the matrix effect of BP-1 at these three concentrations were 102. 9%, 102. 7%, and 90. 9%, respectively. Conclusion：The method is simple, ac-curate, and suitable for determination of the contents of BP-1 in mouse brain.