电感耦合等离子体质谱间接法测定蛋白质含量  

Indirect Determination of Protein Content by Inductively Coupled Plasma-Mass Spectrometry

作　　者：王萌[1] 丰伟悦[1] 陆文伟[2] 李柏[1] 汪冰[1] 朱墨桃[1] 王云[1] 赵宇亮[1] 柴之芳[1] 

机构地区：[1]中国科学院高能物理所,核分析技术重点实验室和纳米生物效应实验室 [2]上海交通大学,上海200030

出　　处：《分析化学》2008年第3期321-324,共4页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金重大项目(No.10490181);国家自然科学基金面上项目(No.20475055);中国科学院重要方向性项目(No.KJCX3.SYW.N3);中国科学院核分析技术重点实验室基金(No.K-135)资助

摘　　要：尺寸排阻色谱分离牛血清白蛋白(BSA),超氧化物歧化酶(SOD)和金属硫蛋白(MT)等3种标准蛋白的混合物后,在线使用电感耦合等离子体质谱测定这三种蛋白中的S,并根据每种蛋白质含有的S原子数,计算出3种蛋白质的相对含量。尺寸排阻色谱的流动相为0.1mol/LTris-HAc。向电感耦合等离子体质谱的六级杆加入反应气O2,使之与S反应生成SO+,间接测定32S16O+而避开直接测量32S时存在的严重干扰。测量结果与天平称量所得结果一致。方法的精密度好,每个蛋白质峰面积的RSD<3%(n=3)。BSA、SOD和MT的检出限分别为14、52和27pmol。Three standard proteins including bovine serum albumin （BSA）, superoxide dismutase （SOD） and metallothionein （MT） were separated by size exclusion chromatography and on-line determined via sulfur by inductively coupled plasma-mass spectrometry. Molar ratios of the proteins were calculated by the analytical results and the known sulfur content in proteins. The mobile phase was 0.1 mol/L Tris-HAc. In order to avoid the polyatomic interferences on the sulfur determination, oxygen was added into the collision cell and reacted with sulfur to form sulfur-oxide ion. Therefore, the intensity of ^32S^160^＋ represented sulfur content during the measurement. These results were in good agreement with the gravimetric results. The RSDs for the proteins are less than 3% and the detection limits for BSA, SOD and MT-Ⅱ are 14, 52 and 27 pmol, respectively.

关 键 词：硫 电感耦合等离子体质谱 碰撞池 尺寸排阻色谱 蛋白质定量 

分 类 号：Q51-3[生物学—生物化学]