Abstract：
BACKGROUND: 1,2-Diacyl-sn-glycero-3-phospho-O-[N-(2-hydroxyethyl)glycines] (PHEGs) are a class of rare aminophospholipids found specifically in brown algae, including kombu seaweed. Despite their potential importance in algal physiology, a comprehensive mass spectrometry (MS) characterization, useful to understand their biological behaviour, is still lacking.
METHODS: To establish the structural regiochemical features of PHEGs, we employed hydrophilic interaction liquid chromatography (HILIC). Following separation, the isolated band of PHEGs was analyzed using MS techniques. This included multistage tandem MS experiments, performed in both positive and negative electrospray ionization modes at low and high resolution.
RESULTS: By comparing MS/MS and MS3 spectra acquired in negative ion mode, the regiochemical rules for PHEG identification were established. The most abundant PHEG species in kombu seaweed, from both Laminaria ochroleuca (European Atlantic) and Laminaria longissima (Japan), was identified as PHEG 20:4/20:4. Less abundant species included PHEG 20:4/20:5 and hydroxylated forms of both PHEG 20:4/20:4 (i.e. 40:8;O) and 20:4/20:5 (40:9;O). The presence of a lyso PHEG 20:4 was consistently detected but at very low levels.
CONCLUSIONS: This study employed MS analysis to elucidate the regiochemical patterns of PHEGs in kombu seaweed. We identified PHEG 20:4/20:4 as the dominant species, along with several less abundant variants, including hydroxylated forms. These findings provide valuable insights into the potential roles and metabolism of PHEGs in brown algae, paving the way for further investigation into their biological functions.
METHODS: To establish the structural regiochemical features of PHEGs, we employed hydrophilic interaction liquid chromatography (HILIC). Following separation, the isolated band of PHEGs was analyzed using MS techniques. This included multistage tandem MS experiments, performed in both positive and negative electrospray ionization modes at low and high resolution.
RESULTS: By comparing MS/MS and MS3 spectra acquired in negative ion mode, the regiochemical rules for PHEG identification were established. The most abundant PHEG species in kombu seaweed, from both Laminaria ochroleuca (European Atlantic) and Laminaria longissima (Japan), was identified as PHEG 20:4/20:4. Less abundant species included PHEG 20:4/20:5 and hydroxylated forms of both PHEG 20:4/20:4 (i.e. 40:8;O) and 20:4/20:5 (40:9;O). The presence of a lyso PHEG 20:4 was consistently detected but at very low levels.
CONCLUSIONS: This study employed MS analysis to elucidate the regiochemical patterns of PHEGs in kombu seaweed. We identified PHEG 20:4/20:4 as the dominant species, along with several less abundant variants, including hydroxylated forms. These findings provide valuable insights into the potential roles and metabolism of PHEGs in brown algae, paving the way for further investigation into their biological functions.
摘要：
背景：1,2-二酰基-sn-甘油基-3-磷酸-O-[N-（2-羟乙基）甘氨酸]（PHEGs）是一类罕见的氨基磷脂，特别是在褐藻中发现，包括昆布海藻.尽管它们在藻类生理学中具有潜在的重要性，全面的质谱（MS）表征，有助于了解它们的生物学行为，仍然缺乏。
方法：为了建立PHEGs的结构区域化学特征，我们采用亲水作用液相色谱(HILIC)。分离后,使用MS技术分析分离的PHEGs条带。这包括多级串联MS实验，在低和高分辨率下以正电喷雾电离模式和负电喷雾电离模式进行。
结果：通过比较在负离子模式下获得的MS/MS和MS3光谱，建立了PHEG鉴定的区域化学规则。康布海藻中最丰富的PHEG物种，来自海带（欧洲大西洋）和海带（日本），被鉴定为PHEG20:4/20:4。不太丰富的物种包括PHEG20:4/20:5和PHEG20:4/20:4(即40:8;O)和20:4/20:5(40:9;O)的羟基化形式。一致地检测到lysoPHEG20:4的存在,但处于非常低的水平。
结论：这项研究采用MS分析来阐明康布海藻中PHEGs的区域化学模式。我们确定PHEG20:4/20:4为优势种，以及几个不太丰富的变体，包括羟基化形式。这些发现为PHEGs在褐藻中的潜在作用和代谢提供了有价值的见解，为进一步研究它们的生物学功能铺平了道路。
方法：为了建立PHEGs的结构区域化学特征，我们采用亲水作用液相色谱(HILIC)。分离后,使用MS技术分析分离的PHEGs条带。这包括多级串联MS实验，在低和高分辨率下以正电喷雾电离模式和负电喷雾电离模式进行。
结果：通过比较在负离子模式下获得的MS/MS和MS3光谱，建立了PHEG鉴定的区域化学规则。康布海藻中最丰富的PHEG物种，来自海带（欧洲大西洋）和海带（日本），被鉴定为PHEG20:4/20:4。不太丰富的物种包括PHEG20:4/20:5和PHEG20:4/20:4(即40:8;O)和20:4/20:5(40:9;O)的羟基化形式。一致地检测到lysoPHEG20:4的存在,但处于非常低的水平。
结论：这项研究采用MS分析来阐明康布海藻中PHEGs的区域化学模式。我们确定PHEG20:4/20:4为优势种，以及几个不太丰富的变体，包括羟基化形式。这些发现为PHEGs在褐藻中的潜在作用和代谢提供了有价值的见解，为进一步研究它们的生物学功能铺平了道路。
