摘要
采用一种简便、灵敏度高的高效液相色谱-荧光检测法（HPLC—FLD）测定蜂王幼虫中游离和总色氨酸的含量。以水（用0．1mol／L盐酸调节pH至6．3）和乙腈作为提取溶液，超声提取蜂王幼虫中的游离色氨酸；用4mol／LNaOH为水解溶液，在110℃、无氧条件下水解蜂王幼虫样品20h后用于总色氨酸含量分析。色谱条件：SymmetryC18柱（4．6×250，5μm）；水（含0．1％三氟乙酸）：甲醇（75：25，V／V）为流动相；流速0．9mL／min；柱温30℃；荧光检测器激发波长为280nm，发射波长为340nm。其线性范围为0．0625—1．0μg／mL，相关系数F2〉0．9998，精密度结果表明重现性和重复性的相对标准偏差RSD（％）均在可接受的范围内。蜂王幼虫中游离色氨酸和总色氨酸含量分别为（0．367±0．008）mg／g（0．037％）和（1．390±0．025）mg／g（0．139％）。
A convenient and sensitive method was developed for determination of free and total tryptophan in bee queen larvae by liquid chromatography with fluoreseence detection. The free tryptophan of bee queen larvae sample was extracted with water （ pH value was adjusted to 6.3 with 0.1 mol/L hydrochloric acid solution） and acetonitrile, and followed by sonicating treatment ,while the total tryptophan sample was prepared by alkaline hydrolyzing the bee queen larvae with 4 mol/L NaOH at 110 ℃ for 20 h in the anaerobic condition. The operating conditions were Symmetry Cls column （4.6×250,5 μm） ,water （0.1% trifluoroacetic acid） ：methanol（75：25 v/v）as the mobile phase at a flow rate of 0.9 mL/ min under 30 ℃. The fluorescence detector was operated at an excitation wavelength of 280 nm and an emission wavelength of 340 nm. A linear response （ r^2 〉 0.9998 ） was obtained in the range of 0.0625 - 1.0 μg,/mL. The precision results showed that the relative standard deviation of the repeatability and reproducibility were reasonable. The free and total tryptophan contents in bee queen larvae were （ 0. 367 ± 0. 008 ） mg/g （ 0. 037% ） and （ 1. 390± 0. 025 ） mg/g（0. 139% ） respectively.
出处
《天然产物研究与开发》
CAS
CSCD
2009年第B05期117-120,共4页
Natural Product Research and Development
基金
基金项目：质检公益专项项目基金（200810897）
