高效液相色谱荧光测定及质谱鉴定土壤和苔藓中的脂肪酸

以吖啶酮-9-乙基对甲苯磺酸酯（AETS）作荧光衍生化试剂，建立了灵敏、简单的游离脂肪酸反相高效液相色谱测定方法。在Eclipse XDB—C8色谱柱上，实现了19种游离脂肪酸（FFA）衍生物的完全基线分离。选取AETS摩尔数为脂肪酸的6倍，以DMF作溶剂，在85℃条件下以K2CO3作催化剂可获得稳定的荧光产物，条件温和、衍生产率高。利用柱后在线串联质谱以APCI大气压化学电离源正离子模式实现了各组分的质谱定性。对土壤和3种苔藓植物中（树藓、狭叶绢藓、曲尾藓）FFA组分定量结果表明，苔藓植物从土壤中富集了大量的游离脂肪酸。荧光检测的激发和发射波长分别为404nm和440nm。绝大多数脂肪酸的线性相关系数大于0．9996，检出限为12．3～43．7fmol。本方法具有良好的重现性，用于实际样品测定，结果满意。

HPLC-APCI-MS determination of free fatty acids in Tibet folk medicine Lomatogonium rotatum with fluorescence detection and mass spectrometric identification

A simple and sensitive method for the determination of free fatty acids (FFAs) using acridone-9-ethyl-p-toluenesulfonate (AETS) as a fluorescence derivatization reagent by high performance liquid chromatography (HPLC) has been developed. Free fatty acid derivatives were separated on an Eclipse XDB-C-8 column with a good baseline resolution and detected with the fluorescence of which excitation and emission wavelengths of derivatives were set at lambda(ex) 404 and lambda(em) 440 nm, respectively. Identification of 19 fatty acid derivatives was carried out by online post-column mass spectrometry with an atmospheric pressure chemical ionization (APCI) source under positive-ion detection mode. Nineteen FFAs from the extract of Lomatogonium rotatum are sensitively determined. The results indicate that the plant Lomatogonium rotatum is enriched with an abundance of FFAs and FFAs of higher contents, which mainly focus on even carbon atoms, C-14, C-16, and C-18. The validation of the method including linearity, repeatability, and detection limits was examined. Most linear correlation coefficients for fatty acid derivatives are > 0.9989, and detection limits (at signal-to-noise of 3: 1) are 12.3-43.7 fmol. The relative standard deviations (RSDs) of the peak areas and retention times for 19 FFAs standards are < 2.24% and 0.45%, respectively. The established method is rapid and reproducible for the separation determination of FFAs from the extract of Lomatogonium rotatum with satisfactory results.