Optimization of immobilized enzyme hydrolysis combined with high-performance liquid chromatography/thermospray mass spectrometry for the determination of neuropeptides.
Optimization of immobilized enzyme hydrolysis combined with high-performance liquid chromatography/thermospray mass spectrometry for the determination of neuropeptides.
复制标题
优化固定化酶水解联合高效液相色谱/热喷雾质谱测定神经肽的方法。
DOI：
10.1016/0003-2697(90)90529-i
复制
发表时间：
1990
影响因子：
2.9
通讯作者：
Swaisgood,HE
中科院分区：
文献类型：
--
作者：
Voyksner,RD;Chen,DC;Swaisgood,HE
Peptidases, including chymotrypsin, thermolysin, trypsin, V8protease, and carboxypeptidases A, B, and Y, were immobilized for use in conjunction with HPLC/thermospray MS for the analysis of neuropeptides. The optimal operating conditions for each immobilized enzyme bioreactor were determined. Optimal hydrolysis usually occurred at the highest percentage of aqueous solution in the mobile phase at pH 7–8 and 40–50°C. Often post-HPLC column addition of aqueous solutions before the bioreactor could improve activity and thermospray sensitivity without changing the HPLC separation. Enzymatic hydrolysis requirements were compatible under conditions for HPLC separation and thermospray MS detection of the selected neuropeptides. Synthetic α-, β-, and γ-endorphins were the primary neuropeptides used to evaluate on-line immobilized enzyme bioreactor/MS. HPLC followed by peptidase hydrolysis produced characteristic hydrolysis products for confirming the peptides' identity using thermospray MS detection. Furthermore, the peptide formed from enzymatic hydrolysis resulted in a MS ion current 10–40 times higher than that of the [M + 2H]2+ion for unhydrolyzed β-endorphin. The increased sensitivity achieved for detecting the hydrolysis products permits detection and quantitation of synthetic peptides down to 800 fmol.
登录
查看更多内容
通过酶热喷雾液相色谱/质谱法进行快速蛋白质测序
DOI：
--
发表时间：
1988
期刊：
影响因子：
--
作者：
K. Stachowiak;C. Wilder;M. Vestal;D. Dyckes
通讯作者：
D. Dyckes
高效液相色谱/热喷雾质谱法半定量高分子量神经肽。
DOI：
10.1002/bms.1200181009
发表时间：
1989
期刊：
Biomedical & environmental mass spectrometry
影响因子：
--
作者：
Voyksner,RD;Pack,TW
通讯作者：
Pack,TW
通过热喷雾 LC/MS 分析中等质量肽。
DOI：
--
发表时间：
1988
期刊：
Biomedical & environmental mass spectrometry
影响因子：
--
作者：
P. Rudewicz
通讯作者：
P. Rudewicz
DOI：
--
发表时间：
1980
期刊：
影响因子：
--
作者：
James B. Taylor;H. Swaisgood
通讯作者：
H. Swaisgood
热喷雾高效液相色谱/质谱联用技术的优化及应用
DOI：
--
发表时间：
1985
期刊：
影响因子：
--
作者：
R. Voyksner;C. A. Haney
通讯作者：
C. A. Haney