摘要
采用高效液相色谱法测定血浆中维生素D_3和25-OH-D_3的含量。样品经乙醇-乙腈(60+40)混合溶液提取后,提取液用氮气吹干,残渣用正己烷溶解后,过Waters silica固相萃取柱净化,净化液用氮气吹干后,将残余物溶于pH 6.5的5mmol·L-1磷酸盐溶液-乙腈-甲醇(30+15+55)缓冲溶液中。待测物在Symmetry C18色谱柱上分离,以pH 6.5的乙腈-5mmol·L^(-1)磷酸盐(20+80)混合液为流动相A,以甲醇-乙腈-四氢呋喃(65+20+15)混合液为流动相B,进行梯度洗脱;采用光电二极管阵列检测器,检测波长为265nm。维生素D_3和25-OH-D_3的线性范围均为3.0~72μg·L^(-1),检出限均为2.0μg·L^(-1)。加标回收率在88.5%~94.1%之间,测定值的相对标准偏差(n=6)小于4%。
HPLC was applied to the determination of vitamin D3 and 25 OH-D3 in human plasma. The sample was extracted with a mixture of ethanol and acetonitrile, the extract solution was evaporated to dryness under nitrogen gas stream. The residue was redissolved in hexane and the solution was purified with a Waters silica solidphase extraction column. The cleaned solution was evaporated to dryness under nitrogen gas stream. The residue was redissolved in a buffer （pH 6. 5） mixed with 5 mmol . L-1 phosphate solution, acetonitrile and methanol. Analytes were separated on a Symmetry C18 column, with a mixture （pH 6. 5） of acetonitrile and 5 mmol . L -1 phosphate solution as mobile phase A, and a mixture of methanol, acetonitrile and tetrahydrofuran as mobile phase B for gradient elution. Analytes were detected by photodiode array detector at the wavelength of 265 nmThe linearity ranges of vitamin Ds and 25-OH-D3 were both 3.0-72 μg .L =1, with detection limits of 2.0 μG. L-1. Recovery rates oblained by standard addition method were in the range of 88. 5%-94. 1% and RSDs （n=6） were less than 4%.
出处
《理化检验（化学分册）》
CAS
CSCD
北大核心
2016年第1期41-43,共3页
Physical Testing and Chemical Analysis(Part B:Chemical Analysis)
