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首页> 外文期刊>Rapid Communications in Mass Spectrometry: RCM >Comparison of high-performance liquid chromatography/tandem mass spectrometry and high-performance liquid chromatography/photo-diode array detection for the quantitation of carotenoids, retinyl esters, α-tocopherol and phylloquinone in chylomicron-rich fractions of human plasma
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Comparison of high-performance liquid chromatography/tandem mass spectrometry and high-performance liquid chromatography/photo-diode array detection for the quantitation of carotenoids, retinyl esters, α-tocopherol and phylloquinone in chylomicron-rich fractions of human plasma

机译：高效液相色谱/串联质谱法与高效液相色谱/光电二极管阵列检测法定量分析人血浆中富含乳糜微粒的类胡萝卜素，视黄酯，α-生育酚和叶醌的比较

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RATIONALE Bioavailability of essential lipophilic micronutrients and carotenoids is of utmost interest for human health, as the consumption of these compounds may help alleviate major nutritional deficiencies, cardiovascular disease, and cancer. High-performance liquid chromatography/photo-diode array detection (HPLC-PDA) and high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) were compared for the quantitative analysis of α- and β-carotene, β-cryptoxanthin, lutein, lycopene, α-tocopherol, phylloquinone, and several retinyl esters from chylomicron-containing triglyceride rich lipoprotein (TRL) fractions of human plasma obtained from two clinical trials. METHODS After selecting an efficient extraction method for the analytes, both the HPLC/PDA and the HPLC/MS/MS methods were developed and several parameters validated using an HP 1200 series HPLC system interfaced with a HP 1200 series diode-array detector (Agilent Technologies, Santa Clara, CA, USA) and a QTRAP 5500 (AB Sciex, Foster City, CA, USA) via an atmospheric pressure chemical ionization (APCI) probe operated in positive ion mode. RESULTS For lycopene, α- and β-carotene, HPLC/MS/MS was up to 37 times more sensitive than HPLC-PDA. PDA detection was shown to be up to 8 times more sensitive for lutein. MS/MS signals were enhanced by matrix components for lutein and β-cryptoxanthin, as determined by referencing to the matrix-independent PDA signal. In contrast, matrix suppression was observed for retinyl palmitate, α-carotene, and β-carotene. Both detectors showed similar suitability for α-tocopherol, lycopene and retinyl palmitate (representing ～73% of total retinyl esters). MS/MS exclusively allowed the quantitation of minor retinyl esters, phylloquinone, and (Z)-lycopene isomers. CONCLUSIONS HPLC/MS/MS was more sensitive than HPLC-PDA for six of the eight analytes and represents a powerful tool for the analysis of chylomicron samples and potentially other biological samples of limited sample size. When internal standards are available for the target carotenoid, employing MS/MS detection may reduce the necessary blood sample volume, which is particularly advantageous for minimizing risk and discomfort to human subjects during clinical studies.
机译：基本的亲脂性微量营养素和类胡萝卜素的生物利用度对人类健康至关重要，因为食用这些化合物可能有助于缓解主要的营养缺乏症，心血管疾病和癌症。比较了高效液相色谱/光电二极管阵列检测（HPLC-PDA）和高效液相色谱/串联质谱（HPLC / MS / MS）进行α-和β-胡萝卜素，β-隐黄质的定量分析，叶黄素，番茄红素，α-生育酚，叶醌和几种含乳糜微粒的富含甘油三酸酯的脂蛋白（TRL）馏分的几种视黄酯，它们来自两项临床试验。方法在为分析物选择一种有效的提取方法后，开发了HPLC / PDA和HPLC / MS / MS方法，并使用与1200系列二极管阵列检测器连接的HP 1200系列HPLC系统验证了一些参数（Agilent Technologies （美国加利福尼亚州圣塔克拉拉市）和QTRAP 5500（美国加利福尼亚州福斯特城的AB Sciex公司）通过以正离子模式运行的大气压化学电离（APCI）探针。结果对于番茄红素，α-和β-胡萝卜素，HPLC / MS / MS的灵敏度是HPLC-PDA的37倍。 PDA检测对叶黄素的敏感度提高了8倍。叶黄素和β-隐黄质的基质成分增强了MS / MS信号，这是通过参考与基质无关的PDA信号确定的。相反，观察到棕榈酸视黄酯，α-胡萝卜素和β-胡萝卜素的基质抑制作用。两种检测器均显示出对α-生育酚，番茄红素和棕榈酸视黄酯的相似适用性（占视黄酯总量的约73％）。 MS / MS仅允许定量次要的视黄酯，叶醌和（Z）-番茄红素异构体。结论HPLC / MS / MS对八种分析物中的六种比HPLC-PDA更为灵敏，是分析乳糜微粒样品和有限样品量的其他生物样品的有力工具。当内标可用于目标类胡萝卜素时，采用MS / MS检测可减少所需的血液样本量，这对于在临床研究期间最大程度地降低人类受试者的风险和不适感特别有利。

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