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首页> 外文期刊>Journal of mass spectrometry: JMS >Sodiation as a tool for enhancing the diagnostic value of MALDI-TOF/TOF-MS spectra of complex astaxanthin ester mixtures from Haematococcus pluvialis
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Sodiation as a tool for enhancing the diagnostic value of MALDI-TOF/TOF-MS spectra of complex astaxanthin ester mixtures from Haematococcus pluvialis

机译：盐渍化作为增强血红球菌复杂虾青素酯混合物MALDI-TOF / TOF-MS光谱诊断价值的工具

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摘要

The microalga Haematococcus pluvialis produces the pigment astaxanthin mainly in esterified form with a multitude of fatty acids, which results in a complex mixture of carotenol mono- and diesters. For rapid fingerprinting of these esters, matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF/TOF-MS) might be an alternative to traditional chromatographic separation combined with MS. Investigation of ionization and fragmentation of astaxanthin mono- and diester palmitate standards in MALDI-TOF/TOF-MS showed that sodium adduct parent masses [M + Na]~+ gave much simpler MS~2 spectra than radical / protonated [M]~(+●) / [M+H]~+ parents. [M + Na]~+ fragments yielded diagnostic polyene-specific eliminations and fatty acid neutral losses, whereas [M]~(+●) / [M+H]~+ fragmentation resulted in a multitude of non-diagnostic daughters. For diesters, a benzonium fragment, formed by polyene elimination, was required for identification of the second fatty acid attached to the astaxanthin backbone. Parents were forced into [M + Na]~+ ionization by addition of sodium acetate, and best signal-to-noise ratios were obtained in the 0.1 to 1.0mM range. This method was applied to fingerprinting astaxanthin esters in a crude H. pluvialis extract. Prior to MALDI-TOF/TOF-MS, the extract was fractionated by normal phase Flash chromatography to obtain fractions enriched in mono- and diesters and to remove pheophytin a, which compromised monoester signals. All 12 types of all-trans esterified esters found in LC were identified with MALDI-TOF/TOF-MS, with the exception of two minor monoesters.
机译：微藻类雨生红球菌主要以酯化形式与多种脂肪酸生成虾青素色素，这导致了类胡萝卜素单酯和二酯的复杂混合物。为了对这些酯进行快速指纹分析，基质辅助激光解吸电离飞行时间质谱仪（MALDI-TOF / TOF-MS）可能是与MS结合的传统色谱分离方法的替代方法。在MALDI-TOF / TOF-MS中虾青素单棕榈酸酯和二酯棕榈酸酯标准品的电离和裂解研究表明，钠加合物母体[M + Na]〜+给出的MS〜2谱比自由基/质子化[M]〜（ +●）/ [M + H]〜+父母。 [M + Na]〜+片段产生诊断性的多烯特异性消除和脂肪酸中性损失，而[M]〜（+●）/ [M + H]〜+片段导致大量无法诊断的子代。对于二酯，需要通过多烯消除形成的苯甲片段来鉴定连接到虾青素骨架上的第二种脂肪酸。通过添加乙酸钠迫使亲本进入[M + Na]〜+电离，并且在0.1至1.0mM的范围内获得最佳的信噪比。该方法用于指纹图谱中粗虾嗜血杆菌提取物中虾青素酯的指纹图谱。在MALDI-TOF / TOF-MS之前，将提取物通过正相快速色谱法进行分馏，以获得富含单酯和二酯的馏分，并去除破坏单酯信号的脱镁叶绿素a。 LC中发现的所有12种全反酯化酯均用MALDI-TOF / TOF-MS鉴定，除了两种次要的单酯。

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