乙醇萃取-HPLC法快速分析亚麻籽油中四种环肽研究

Determination of four cyclolinopeptides content in flaxseed oil through ethanol extraction-HPLC method

本文旨在利用乙醇快速富集亚麻籽油环肽,以期替代柱层析富集环肽法。在常温下,95%乙醇与亚麻籽油以料液比(g/mL)为5∶8震荡混合5 min,随后,将其在6 000 r/min下离心5 min,重复富集3次,收集上层清液并在40℃旋蒸,除去溶剂,用甲醇定容,根据HPLC外标法建立标准曲线,对环肽A、环肽C、环肽D和环肽E进行定量分析。通过加样回收试验可得,环肽C、环肽E、环肽D和环肽A的标准样品加样回收率分别为:99.5%、104.6%、109.7%、97.6%,相对标准偏差RSD分别为:1.4%,2.0%,2.5%和1.4%,证明了乙醇富集亚麻籽油环肽的方法准确可靠。此外,乙醇萃取法与传统的正相SPE硅胶柱分离法相比,操作简单,溶剂用量节省,富集效果更好。

Ethanol extraction- HPLC method was established to extract cycolinopeptide A （CLA）, eycolinopeptide C （CLC）, cycolinopeptide D （CLD） and cycolinopeptide E （CLE） from flaxseed oils. The flaxseed oils were extracted with 95% ethanol with oil to ethanol mass ratios of 5 ： 8 （g/mL） at room temperature. After shaking, thoroughly for 5 min, the mixed solution was cen- trifuged at 6 000 rpm for 5 min, and the superna- rants were collected. The process was repeated 3 times. After that, the collected solution was evap- orated at 40 ℃ to remove the solvent and dissolved in methanol. The concentration of CLA, CLC,CLD and CLE were measured by external standard HPLC method. The average recovery of standard addition of CLA, CLC, CLD and CLE are as follows： 99. 5%, 104. 6%, 109. 7% and 97.6%, and the relative standard deviation are respectively 1.4%, 2.0%, 2. 5% and 1.4%, which shows that the ethanol extraction- HPLC method is accurate and effective. Additionally, when compared with the solid phase extraction （SPE） method, ethanol extraction- HPLC meth- od is much easier and can reduce the solvent greatly.