Vecka M, Tvrzická E, Stanková B, Zák A
4th Department of Medicine, 1st Faculty of Medicine, Charles University, Prague, Czech Republic.
J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Apr 25;770(1-2):91-9. doi: 10.1016/s1570-0232(01)00630-4.
Four capillary columns (A: CP-WAX 52 CB 25 m x 0.25 mm; B: CP WAX 52 CB 30 m x 0.25 mm; C: CP-WAX 58 CB 25 m x 0.25 mm, Chrompack; D: OMEGAWAX 320 30 m x 0.32 mm, Supelco) and two integration software (Mosaic v.5.10, Chrompack and CSW v.1.7, Data Apex5) were compared for analysis of fatty acids. Column A was mounted stepwise in two different instruments. Fatty acids of blood plasma phosphatidylcholine and standard mixture of saturated fatty acids were analysed as methyl esters under identical chromatographic conditions. Both integrating software did not differ significantly in most results; differences were observed only for minor components: 16:1n9 (0.10+/-0.020 vs. 0.17 +/- 0.005 M%, P < 0.0001, column Al; 0.09 +/- 0.011 vs. 0.16 +/- 0.007 M%, P< 0.0001, column A2; 0.09 +/- 0.010 vs. 0.17 +/- 0.003 M%, P < 0.0001, column C; 0.09 +/ -0.008 vs. 0.19 +/- 0.003 M%, P < 0.0001, column D), 20:0 (0.10+0.001 vs. 0.06 +/- 0.005 M%, P < 0.05, column C) and 20:2n6 (0.43 +/- 0.030 vs. 0.91 +/- 0.016 M%, P < 0.0001, column A2). Increased values for 16:1n9 and 20:2n6 integrated by MOSAIC are caused by cointegration of two poorly resolved peaks: fatty acid and impurity from sample matrix. Lower values for 20:0 are caused by incomplete integration of minor peak. Differences between columns were observed mostly for minor fatty acids. The results indicate that CSW is more suitable software for integration of complicated chromatograms. Linear calibration dependences measured with standard mixture of saturated fatty acids (carbon number 10-24) were observed in wide range of concentrations (three orders). Slope close to unity and minimal value of intercept confirmed theoretical relations when analyses are run under optimal conditions. Use of one column is advisable in small intervention or experimental metabolic studies.
比较了四根毛细管柱（A：CP-WAX 52 CB 25 m×0.25 mm；B：CP WAX 52 CB 30 m×0.25 mm；C：CP-WAX 58 CB 25 m×0.25 mm，Chrompack；D：OMEGAWAX 320 30 m×0.32 mm，Supelco）和两种积分软件（Mosaic v.5.10，Chrompack和CSW v.1.7，Data Apex5）用于脂肪酸分析。将柱A逐步安装在两台不同的仪器中。在相同的色谱条件下，将血浆磷脂酰胆碱的脂肪酸和饱和脂肪酸标准混合物作为甲酯进行分析。两种积分软件在大多数结果上没有显著差异；仅在次要成分上观察到差异：16:1n9（0.10±0.020对0.17±0.005 M%，P<0.0001，柱A1；0.09±0.011对0.16±0.007 M%，P<0.0001，柱A2；0.09±0.010对0.17±0.003 M%，P<0.0001，柱C；0.09±0.008对0.19±0.003 M%，P<0.0001，柱D），20:0（0.10+0.001对0.06±0.005 M%，P<0.05，柱C）和20:2n6（0.43±0.030对0.91±0.016 M%，P<0.0001，柱A2）。MOSAIC积分得到的16:1n9和20:2n6值增加是由于两个分辨率较差的峰共积分：脂肪酸和样品基质中的杂质。20:0值较低是由于小峰积分不完全。柱之间的差异主要在次要脂肪酸上观察到。结果表明，CSW是更适合复杂色谱图积分的软件。在宽浓度范围（三个数量级）内观察到用饱和脂肪酸标准混合物（碳数10 - 24）测得的线性校准依赖性。当在最佳条件下进行分析时，接近1的斜率和最小的截距值证实了理论关系。在小型干预或实验性代谢研究中建议使用一根柱。
