Characterization of polysorbate 80 with liquid chromatography mass spectrometry and nuclear magnetic resonance spectroscopy: Specific determination of oxidation products of thermally oxidized polysorbate 80
Characterization of polysorbate 80 with liquid chromatography mass spectrometry and nuclear magnetic resonance spectroscopy: Specific determination of oxidation products of thermally oxidized polysorbate 80
复制标题
DOI：
10.1016/j.jpba.2011.12.009
复制
发表时间：
2012-03-25
影响因子：
3.4
通讯作者：
Dyrstad, Knut
中科院分区：
文献类型：
--
作者：
Hvattum, Erlend;Yip, Wai Lam;Dyrstad, Knut
已采用H-1 NMR、LC-UV/MS和MS/MS对聚山梨酯80（PS 80）中的聚山梨酯物质进行了表征。MS采用负离子和正离子电喷雾电离（ESI）进行操作。发现PS 80含有聚氧乙烯（POE）脱水山梨醇、POE异山梨醇和POE的单脂肪酸酯和二脂肪酸酯。此外，相同的物种也存在不含脂肪酸酯。用C12：0、C14：0、C16：1、C16：0、C18：2、C18：1和C18：0脂肪酸酯化聚山梨酯物质。主要物质为C18：1的聚山梨酯。C18：2聚山梨酯物质通过H-1 NMR显示具有较大的脂肪酸组分，该脂肪酸组分具有ZE和/或EZ构型的共轭双键。聚山梨酯物质的正ESI质谱显示每种脂肪酸酯的特定源内脂肪酸片段。使用源内脂肪酸片段的质量色谱图测定PS 80中特定聚山梨酯物质的降解。在40 ℃空气气氛下储存8周后，C18：2聚山梨酯物质完全降解，而主要C18：1聚山梨酯物质减少至约100%。80-86%伴随着短链POE C18：1物种的增加。发现用C18：1-OH、C18：1酮和C18：0环氧酸酯化的聚山梨酯物质是在40 ℃空气中储存8周的PS 80的降解产物。C18：1-OH、C18：1酮基和C18：0环氧酸被认为是C18：1的氧化产物。在当前条件下，C18：1-OH和C18：0环氧聚山梨酯物质的正ESI质谱显示出与C18：2聚山梨酯物质相同的离子，这是由于质子化分子中H2O容易源内损失。使用负离子ESI MS确定了C18：1-OH和C18：0环氧酸的聚山梨酯酯的存在。通过皂化和提取，随后对游离脂肪酸进行负离子ESI LC-MS分析，也证实了降解PS 80中存在氧化脂肪酸。(C)2011爱思唯尔有限公司版权所有。
The polysorbate species in polysorbate 80 (PS 80) have been characterized with H-1 NMR, LC-UV/MS and MS/MS. The MS was operated with both negative and positive electrospray ionization (ESI). PS 80 was found to contain mono- and di-fatty acid esters of polyoxyethylene (POE) sorbitan, POE isosorbide and of POE. In addition, the same species were also present without fatty acid esters. The polysorbate species were esterified with C12:0, C14:0, C16:1, C16:0, C18:2, C18:1 and C18:0 fatty acids. The main species were polysorbate esters of C18:1. The C18:2 polysorbate species were shown, by H-1 NMR, to have a large component of fatty acids with a conjugated double bond of ZE and/or EZ configuration. The positive ESI mass spectra of the polysorbate species displayed a specific in-source fatty acid fragment for each fatty acid ester. The mass chromatograms of the in-source fatty acid fragments were used to determine the degradation of specific polysorbate species in PS 80. The C18:2 polysorbate species were completely degraded after 8 weeks at 40 degrees C under an atmosphere of air, while the main C18:1 polysorbate species were reduced to ca. 80-86% accompanied by an increase of short-chain POE C18:1 species. Polysorbate species esterified with C18:1-OH, C18:1 keto and C18:0 epoxy acids were found as degradation products of PS 80 stored at 40 degrees C for 8 weeks under air. C18:1-OH, C18:1 keto and C18:0 epoxy acids were believed to be oxidation products of C18:1. With the present conditions, the positive ESI mass spectra of C18:1-OH and C18:0 epoxy polysorbate species displayed identical ions to the C18:2 polysorbate species due to a facile in-source loss of H2O from the protonated molecules. The presence of polysorbate esters of C18:1-OH and C18:0 epoxy acids were established using negative ESI MS. The presence of oxidized fatty acids in degraded PS 80 was also confirmed by saponification and extraction followed by negative ESI LC-MS analysis of the free fatty acids. (C) 2011 Elsevier B.V. All rights reserved.