Romeu-Nadal M, Morera-Pons S, Castellote A I, López-Sabater M C
Department Nutrició i Bromatologia, Centre de Referència en Tecnologia dels Aliments (CeRTA), Facultat de Farmàcia, Universitat de Barcelona. Avda. Joan XXIII s/n, E-08028 Barcelona, Spain.
J Chromatogr A. 2006 May 5;1114(1):132-7. doi: 10.1016/j.chroma.2006.02.049. Epub 2006 Mar 30.
A rapid direct method (Method I) for measuring gamma- and alpha-tocopherols in human milk was developed and validated using reversed-phase high-performance liquid chromatography with ultraviolet/visible (UV-vis) detection. Human milk, with an internal standard (alpha-tocopherol acetate) added, was diluted in hexane. The chromatographic system consisted of a short column (50 mm x 2.1mm I.D., 3 microm particle size) that allowed the separation of the gamma- and alpha-tocopherols in less than 6 min. The new direct method (Method I) was compared with other methods. Method II (saponification with ultraviolet/visible detection) determined 24% and 22% less gamma- and alpha-tocopherols, respectively. Method III (saponification with evaporative light scattering detection) gave the same values for alpha-tocopherol content as Method II. However, the amount of sample used in the application of Method III was higher than that used in Method II. Furthermore, Method I uses smaller amounts of solvents, and it is simpler and faster than Methods II or III. Only a small volume of sample is needed, which is an additional advantage for biological assays.
开发了一种快速直接的方法（方法I），用于测定人乳中的γ-生育酚和α-生育酚，并采用反相高效液相色谱-紫外/可见光（UV-vis）检测进行了验证。向人乳中加入内标（α-生育酚醋酸酯）后，用己烷稀释。色谱系统由一根短柱（50 mm×2.1mm内径，3微米粒径）组成，该短柱可在不到6分钟的时间内分离出γ-生育酚和α-生育酚。将新的直接方法（方法I）与其他方法进行了比较。方法II（皂化-紫外/可见光检测）测定的γ-生育酚和α-生育酚分别比方法I少24%和22%。方法III（皂化-蒸发光散射检测）测定的α-生育酚含量与方法II相同。然而，方法III应用时所用的样品量高于方法II。此外，方法I使用的溶剂量较少，并且比方法II或III更简单、更快。只需要少量样品，这对生物测定来说是一个额外的优点。
