Center for Systems Biology, University of Iceland, Sturlugata 8, IS 101 Reykjavik, Iceland.
Anal Bioanal Chem. 2012 Dec;404(10):3145-54. doi: 10.1007/s00216-012-6434-4. Epub 2012 Oct 2.
A fast method for quantification and identification of carotenoid and chlorophyll species utilizing liquid chromatography coupled with UV detection and mass spectrometry has been demonstrated and validated for the analysis of algae samples. This method allows quantification of targeted pigments and identification of unexpected compounds, providing isomers separation, UV detection, accurate mass measurements, and study of fragment ions for structural elucidation in a single run. This is possible using parallel alternating low- and high-energy collision spectral acquisition modes, which provide accurate mass full scan chromatograms and accurate mass high-energy chromatograms. Here, it is shown how this approach can be used to confirm carotenoid and chlorophyll species by identification of key diagnostic fragmentations during high-energy mode. The developed method was successfully applied for the analysis of Dunaliella salina samples during defined red LED lighting growth conditions, identifying 37 pigments including 19 carotenoid species and 18 chlorophyll species, and providing quantification of 7 targeted compounds. Limit of detections for targeted pigments ranged from 0.01 ng/mL for lutein to 0.24 ng/mL for chlorophyll a. Inter-run precision ranged for of 3 to 24 (RSD%) while inter-run inaccuracy ranged from -17 to 11.
一种利用液相色谱-紫外检测-质谱联用技术快速定量和鉴定类胡萝卜素和叶绿素种类的方法已经过验证，可用于藻类样品的分析。该方法可以定量目标色素，并鉴定意外化合物，提供异构体分离、紫外检测、精确质量测量以及片段离子研究，以在单次运行中进行结构解析。这可以通过平行交替的低能和高能碰撞光谱采集模式来实现，该模式提供精确质量全扫描色谱图和精确质量高能色谱图。本文展示了如何通过在高能模式下鉴定关键的特征片段来确认类胡萝卜素和叶绿素的种类。该方法成功应用于在定义的红色 LED 光照生长条件下分析杜氏盐藻样品，鉴定了 37 种色素，包括 19 种类胡萝卜素和 18 种叶绿素，并对 7 种目标化合物进行了定量。目标色素的检测限范围为 0.01ng/mL（叶黄素）至 0.24ng/mL（叶绿素 a）。运行间精密度范围为 3%至 24%（RSD%），而运行间准确度范围为-17%至 11%。
