Moreau R A, Powell M J, Osman S F, Whitaker B D, Fett W F, Roth L, O'Brien D J
Eastern Regional Research Center, United States Department of Agriculture, Philadelphia, Pennsylvania 19118.
Anal Biochem. 1995 Jan 1;224(1):293-301. doi: 10.1006/abio.1995.1043.
Hopanoids and other lipids were extracted from Zymomonas mobilis and quantitatively analyzed by high-performance liquid chromatography. Previous methods for hopanoid analysis required derivatization of the hopanoids via periodate oxidation or acetylation. The current method employs a normal-phase silica gel column, a ternary gradient of hexane-isopropanol-water-triethylamine, and detection with a flame ionization detector. Three major hopanoid classes were separated and quantified by this new method, and together they comprised 30 to 40% of the total lipid in these cells. Mass spectrometry confirmed that chemical structures of these hopanoids were consistent with those previously proposed. In addition, several other common lipid classes (free fatty acids and six classes of common phospholipids), comprising the remaining 60 to 70% of the total lipids, were also separated and quantified. This new chromatographic method represents the first technique for the analysis and purification of intact hopanoids. This method should be useful for the analysis and purification of hopanoids from other bacterial species.
从运动发酵单胞菌中提取类胡芦巴烷型化合物和其他脂质，并通过高效液相色谱进行定量分析。以前用于类胡芦巴烷型化合物分析的方法需要通过高碘酸盐氧化或乙酰化对类胡芦巴烷型化合物进行衍生化。目前的方法采用正相硅胶柱、己烷 - 异丙醇 - 水 - 三乙胺的三元梯度洗脱，并使用火焰离子化检测器进行检测。通过这种新方法分离并定量了三类主要的类胡芦巴烷型化合物，它们总共占这些细胞中总脂质的30%至40%。质谱分析证实这些类胡芦巴烷型化合物的化学结构与先前提出的结构一致。此外，还分离并定量了其他几种常见的脂质类别（游离脂肪酸和六类常见的磷脂），它们占总脂质的其余60%至70%。这种新的色谱方法是第一种用于完整类胡芦巴烷型化合物分析和纯化的技术。该方法对于从其他细菌物种中分析和纯化类胡芦巴烷型化合物应该是有用的。
