The use of hydrazine-based derivatization reagents for improved sensitivity and detection of carbonyl containing compounds using MALDI-MSI
The use of hydrazine-based derivatization reagents for improved sensitivity and detection of carbonyl containing compounds using MALDI-MSI
复制标题
DOI：
10.1007/s00216-014-8223-8
复制
发表时间：
2015-03-01
影响因子：
4.3
通讯作者：
Clench, Malcolm R.
中科院分区：
文献类型：
--
作者：
Flinders, Bryn;Morrell, Josie;Clench, Malcolm R.
肼基衍生试剂已被用于通过MALDI-MSI检测大鼠肺组织中含羰基糖皮质激素丙酸氟替卡松的存在。此类试剂还充当用于通过MALDI-MS进行分析的基质，并且已被称为“反应性基质”。用声学试剂点样器，用2，4-二硝基苯肼（DNPH）和4-二甲氨基-6-（4-甲氧基-1-萘基）-1，3，5-三嗪-2-肼（DMNTH）原位衍生化用一系列浓度的丙酸氟替卡松点样的大鼠肺组织（12 μ m）冷冻切片。已经证明，与DNPH相比，DMNTH给出了上级结果，并且在应用DMNTH后立即分析样品导致检测到浓度为500 ng/μ L的丙酸氟替卡松的质子化腙衍生物（[M-D + H]（+））。进一步表明，延长的反应时间（类似于48小时）将质子化腙衍生物的检测限提高到50 ng/μ L，并且当常规MALDI基质CHCA与DNPH/DMNTH反应基质结合使用时，获得了灵敏度和检测限的提高。
Hydrazine-based derivatization reagents have been used to detect the presence of the carbonyl containing glucocorticoid fluticasone proprionate in rat lung tissue by MALDI-MSI. Such reagents also act as a matrix for analysis by MALDI-MS and have been termed "reactive matrices". Cryosections of rat lung tissue (12 mu m), spotted with a range of concentrations of fluticasone proprionate, were derivatized in situ with 2,4-dinitrophenylhydrazine (DNPH) and 4-dimethylamino-6-(4-methoxy-1-naphthyl)-1,3,5-triazine-2-hydrazine (DMNTH) by the use of an acoustic reagent spotter. It has been demonstrated that DMNTH gave superior results compared to DNPH and that analysis of samples immediately after application of DMNTH resulted in the detection of the protonated hydrazone derivative ([M-D + H](+)) of fluticasone propionate at a concentration of 500 ng/mu L. It has been further shown that a prolonged reaction time (similar to 48 h) improves the detection limit of the protonated hydrazone derivative to 50 ng/mu L and that improvements in sensitivity and limits of detection are obtained when a conventional MALDI matrix CHCA is employed in conjunction with the DNPH/DMNTH reactive matrix.