摘要
目的建立测定血清中25-羟基D2和25-羟基D3含量的高效液相色谱一质谱法。方法采用固相萃取柱对血清样品进行处理和净化，以O．1％甲酸水溶液-0．1％甲酸的甲醇溶液（15：85）为流动相，检测波长为240nm，柱温为50℃，流速为0．5mL／min为色谱条件，以ES4-APCI组合离子源，正离子模式；离子源喷射电压5000V，离子源温度250℃，离子源喷雾流速5L／min为质谱条件。结果建立的分析方法25一羟基D2和25-羟基D3质量浓度线性范围在5～100ng／mL范围与峰面积有良好线性关系（R2=0．9992，0．9995），日内精密度均低于5％，日间精密度均低于10％，回收率在96．19％～108．23％之间，血清中检测限和定量限分别是2ng／mL和3ng／mL。结论所建立的方法准确、灵敏、可重复，可用于检测血清中25-羟基D2和25-羟基D3的含量。
Objective To establish a HPLC-MC method for determining the content of serum 25-hydroxyl D2 and 25-hydroxyl D3. Methods The solid phase extraction column was adopted to treat and purify the serum samples with 0. 1% formic acid aqueous solution -0.1% methanol formic acid solution （15 ： 85） as the mobile phase,the detecting wavelength of 240 rim,the column temperature of 50℃ , the flow velocity of 0.5 mL/min as the chromatographic condition, with ESI ＋ APCI combination ion source, the positive ions mode and the ion source jet voltage of 5 000 V,the ion source temperature of 250℃ and the ion source spray velocity of 5 L/min as the mass spectrometry conditions. Results By the established analysis method, the linear range of 25- hydroxyl D2 and 25- hydroxyl D3 was 5- 100 ng/mL （R2 =0. 999 2,0. 999 5）,the intra-day precision 〈 5% ,the inter-day precision 〈 10%, and the recovery rate was 96. 19% - 108.23% ;the detection limit and the limit of quantitation（LOQ） in serum were 2 ng/mL and 3 ng/mL respectively. Conclusion The established method is accurate,sensitive and repeative,and can be used to determine the content of 25-hy- droxyl D2 and 25- hydroxyl D3 in serum.
出处
《中国药业》
CAS
2013年第20期17-18,共2页
China Pharmaceuticals
