用高效液相色谱法和气相色谱法同时测定鱼油中顺式和反式棕榈油酸。
Comparing the simultaneous determination of cis- and trans-palmitoleic acid in fish oil using HPLC and GC.
机构信息
College of Pharmacy, Nanjing University of Chinese Medicines, Nanjing, 210023, People's Republic of China.
Third Institute of Oceanography, Ministry of Natural Resources, Xiamen, 361005, People's Republic of China.
出版信息
Lipids Health Dis. 2019 Apr 4;18(1):86. doi: 10.1186/s12944-019-1033-4.
BACKGROUND
Cis- and trans-palmitoleic acids (Cis-POA and trans-POA) are isomers of palmitoleic acid, a monounsaturated fatty acid which affects glucose and lipid metabolism, and reduces insulin resistance. Trans-POA is used as a biomarker for indicating the risk of type II diabetes and coronary heart disease, but no methods of analysis or distinguishing between cis-POA and trans-POA have yet been reported.
METHOD
An accurate and precise HPLC method was developed to determine cis- and trans-POA simultaneously, and compared with results from a GC method. Cis- and trans-POA were analyzed by HPLC on a reverse-phase BDS-C18 column, equilibrated and eluted with acetonitrile (A) and water (B). In the established and validated GC method used for comparison, potassium hydroxide ester exchange was chosen to derivatize the cis- and trans-POA, before being determined.
RESULTS
The calibration curves for cis- and trans-POA were linear over the range 0.05 to 500 μg/mL. The HPLC method exhibited good sensitivity, precision and accuracy. The limits of detection (LOD) for cis- and trans-POA were 0.2 and 0.05 μg/mL, respectively. The method successfully determined cis- and trans-POA in fish oil. For the GC method, the contents of cis-POA quantified were similar to those from the HPLC method, but the contents of trans-POA revealed significant variation between the two methods.
CONCLUSIONS
After a comprehensive consideration of the characteristics of the saponification and methyl esterification methods which have been tested and verified, the HPLC method was found to be suitable for determining cis- and trans-POA contents in fish oil. It was also suggested that in natural fish oil, cis-POA may be in the glyceride state, and trans-POA almost completely in the free acid form. In comparison with the GC method, the HPLC method provided a simpler process and faster analyses for identifying and determining cis- and trans-POA. The study has also provided technical support for studying the pharmacological differences and relationship between structure and activity of cis- and trans-POA. This could help physicians to analyze patients' samples more quickly in 10 min and therefore provide a more rapid diagnosis of problems relating to the risk of type II diabetes and coronary heart disease.
背景
顺式和反式棕榈油酸（Cis-POA 和 trans-POA）是棕榈油酸的异构体，棕榈油酸是一种单不饱和脂肪酸，可影响葡萄糖和脂质代谢，并降低胰岛素抵抗。反式棕榈油酸可用作表明 II 型糖尿病和冠心病风险的生物标志物，但尚未报道用于分析或区分顺式和反式棕榈油酸的方法。
方法
开发了一种准确、精密的 HPLC 方法，可同时测定顺式和反式棕榈油酸，并与 GC 方法的结果进行比较。通过反相 BDS-C18 柱在 HPLC 上分析 Cis-POA 和 trans-POA，用乙腈（A）和水（B）平衡和洗脱。在用于比较的已建立和验证的 GC 方法中，选择氢氧化钾酯交换来衍生 Cis-POA 和 trans-POA，然后进行测定。
结果
顺式和反式棕榈油酸的校准曲线在 0.05 至 500μg/mL 的范围内呈线性。HPLC 方法表现出良好的灵敏度、精密度和准确性。顺式和反式棕榈油酸的检测限（LOD）分别为 0.2 和 0.05μg/mL。该方法成功测定了鱼油中的顺式和反式棕榈油酸。对于 GC 方法，定量的 Cis-POA 含量与 HPLC 方法相似，但两种方法之间的 trans-POA 含量存在显著差异。
结论
在全面考虑已测试和验证的皂化和甲酯化方法的特点后，发现 HPLC 方法适用于测定鱼油中顺式和反式棕榈油酸的含量。还表明在天然鱼油中，Cis-POA 可能处于甘油酯状态，而反式棕榈油酸几乎完全处于游离酸形式。与 GC 方法相比，HPLC 方法为识别和测定 Cis-POA 和 trans-POA 提供了更简单的过程和更快的分析。该研究还为研究 Cis-POA 和 trans-POA 的药理学差异和结构与活性关系提供了技术支持。这可以帮助医生在 10 分钟内更快地分析患者样本，从而更快地诊断与 II 型糖尿病和冠心病风险相关的问题。
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