MICRO-ELECTROSPRAY MASS-SPECTROMETRY - ULTRA-HIGH-SENSITIVITY ANALYSIS OF PEPTIDES AND PROTEINS
MICRO-ELECTROSPRAY MASS-SPECTROMETRY - ULTRA-HIGH-SENSITIVITY ANALYSIS OF PEPTIDES AND PROTEINS
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DOI：
10.1016/1044-0305(94)85001-1
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发表时间：
1994-07-01
影响因子：
3.2
通讯作者：
CAPRIOLI, RM
中科院分区：
文献类型：
--
作者：
EMMETT, MR;CAPRIOLI, RM
开发了一种“微电喷雾”电离源，显著提高了传统电喷雾源的灵敏度。这是通过优化源来实现的，以适应从300到800 nl /min的纳升流速，直接从毛细管针喷射，用于分析肽，含有C18液相色谱填料作为集成的浓度脱盐装置。因此，从毛细管柱喷雾针中解吸出总共1 fmol的蛋氨酸脑啡肽，装入10 μ l的100 μ l/ μ l溶液。质谱显示在M /z 574.2处存在[M + H]+离子，信噪比大于5:1，色谱峰宽度约为12 s。500 amol注射液中蛋氨酸脑啡肽获得了窄范围（15-u）串联质谱，50 fmol注射液获得了全扫描串联质谱。对于蛋白质，注射2.5 fmol的无肌红蛋白时，平均质量测量精度约为100-200 ppm，注射200 fmol时，平均质量测量精度约为20-40 ppm。碳酸酐酶B和牛血清白蛋白具有相似的质量测量精度。
A ''micro-electrospray'' ionization source has been developed that markedly increases the sensitivity of the conventional electrospray source. This was achieved by optimization of the source to accommodate nanoliter flow rates from 300 to 800-nL/min spraying directly from a capillary needle that, for the analysis of peptides, contained C18 liquid chromatography packing as an integrated concentration-desalting device. Thus, a total of 1 fmol of methionine enkephalin was desorbed from the capillary column spray needle, loaded as a 10-muL injection of 100-amol/muL solution. The mass spectrum showed the [M + H]+ ion at m/z 574.2 with a signal-to-noise ratio of better than 5:1 from a chromatographic peak with a width of about 12 s. A narrow range (15-u) tandem mass spectrum was obtained for methionine enkephalin from the injection of 500 amol, and a full-scan tandem-mass spectrum was obtained from 50 fmol. For proteins, the average mass measurement accuracy was approximately 100-200 ppm for the injection of 2.5 fmol of apomyoglobin and 20-40 ppm for 200 fmol. Carbonic anhydrase B and bovine serum albumin showed similar mass measurement accuracies.