固相萃取结合HPLC-荧光法测定人血浆中维生素B_2含量  

HPLC-fluorescence determination of vitamin B_2 content in human blood plasma with solid phase extraction

作　　者：田兰[1] 孔彬[1] 杜晨松[2] 居来提.安尼瓦尔 买买提艾力.乌马尔 巴吐尔.买买提明 

机构地区：[1]新疆医科大学药学院,乌鲁木齐830011 [2]新疆医科大学基础医学院,乌鲁木齐830011 [3]新疆医科大学第一附属医院,乌鲁木齐830011

出　　处：《药物分析杂志》2012年第12期2150-2153,2121,共5页Chinese Journal of Pharmaceutical Analysis

基　　金：国家自然科学基金项目(No.8116045P)

摘　　要：目的:建立固相萃取结合HPLC-荧光法测定人血浆中维生素B2含量的方法。方法:采用SiroccoTM96孔蛋白沉淀板和正压固相萃取装置处理样品,采用SymmetryshieldTMRP-C18(250 mm×4.6 mm,5μm)色谱柱,流动相为甲醇-5 mmol.L-1乙酸铵(35∶65),流速为0.6 mL.min-1,荧光检测器检测(激发波长:450 nm,发射波长:520 nm)。结果:维生素B2线性范围为0.2～10 ng.mL-1(r=0.9937)。定量下限为0.17 ng.mL1,日内精密度(RSD)为2.0%～4.8%,日间精密度(RSD)为3.8%,方法回收率为101.5%～116.0%(n=3),提取回收率为91.8%～115.1%。结论:所建方法采用96孔蛋白沉淀装置自动处理样品,高效液相色谱-荧光法测定,可用于临床血浆样品中维生素B2含量的测定。Objectives：To establish a method for the determination of the content of vitamin B2 in human blood plasma by HPLC -fluorescence detection combined with solid phase extraction. Method：SiroccoTM96 precipitation plate and the positive pressure - 96 processor were used for sample pre - treatment. The chromatographic column was SymmetryshieldTM RP -C18 （250 mm×4.6mm,5μm） ,the mobile phase was 35% methanol and 65% 5 mmol · L^-1 ammonium acetate solution at a flow rate of 0.6 mL · min ^-1. The spectro - photofluorimeter was set at wave- length of 450 nm for excitation and 520 nm for emission. Results：The linear ranges were 0.2 -10 ng· mL^-1（ r = 0. 9937）. The lowest limit of quantification was 0. 17 ng· mL-1. The intra - day RSD were 2. 0% -4. 8% ,the in- ter - day RSD was 3.8%. The relative recoveries were between 101.5% and 116. 0% （n = 3 ）. The recovery of ex- traction were 91.8% -115.1%. Conclusions： A 96 -well plate automation- compatible precipitation plate was used to dispose the samples,which determined the content of vitamin B2 by the HPLC - fluorescence detector. This method can be used for determining of vitamin B2 in clinical plasma samples.

关 键 词：固相萃取 高效液相色谱法 荧光检测 维生素B2(核黄素) 血浆样品 生物样品前处理 

分 类 号：R917[医药卫生—药物分析学]