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目的:建立一种同时测定小鼠股骨中钙和磷含量的方法。方法:以反相高效液相色谱-蒸发光散射检测器(HPLCELSD)法,采用Prevail C18色谱柱(250 mm×4.6 mm i.d.,5?m),以乙腈与0.7%三氟醋酸溶液(含5.0 mmol/L)为流动相进行洗脱,洗脱时间为10 min。结果:氯化钙、磷酸峰面积的自然对数与相应浓度的自然对数呈良好的线性关系,其方程为氯化钙:Y=1.206 0X+8.445 8,r=0.999 8,线性范围0.10~0.30 mg/ml,磷酸:Y=1.428 9X+9.074 0,r=0.999 7,线性范围0.027~0.081 mg/ml。氯化钙、磷酸平均回收率分别为100.5%,97.9%,RSD分别为1.8%,2.3%。结论:该法快速、简便、准确,对判断和研究骨矿物质的含量多有重要的意义。
Objective: To establish a method for simultaneous determination of calcium and phosphorus in the femur of mice. Methods: A reversed phase high performance liquid chromatography coupled with evaporative light scattering detector (HPLCELSD) was used on a Prevail C18 column (250 mm × 4.6 mm id, 5 μm) with acetonitrile and 0.7% trifluoroacetic acid mmol / L) as mobile phase, the elution time was 10 min. Results: The natural logarithm of the peak areas of calcium chloride and phosphate showed a good linear relationship with the natural logarithm of the corresponding concentration. The equation was calcium chloride: Y = 1.206 0X + 8.445 8, r = 0.999 8, linear range 0.10 ~ 0.30 mg / ml, phosphoric acid: Y = 1.428 9X + 9.074 0, r = 0.999 7, linear range 0.027 ~ 0.081 mg / ml. The average recoveries of calcium chloride and phosphoric acid were 100.5% and 97.9%, respectively, with RSDs of 1.8% and 2.3%, respectively. Conclusion: The method is rapid, simple and accurate. It is of great significance to judge and study the content of bone mineral.