柱前衍生HPLC法测定鸡脂酶解物中游离脂肪酸含量

Determination of Free Fatty Acids in Enzymatic Hydrolysate of Chicken Fat by Pre-column Derivation HPLC

建立以2，4-二溴苯乙酮为衍生化试剂，18-冠-6醚为相转移催化剂，同时测定鸡脂酶解物中四种游离脂肪酸的柱前衍生高效液相色谱法。利用Zorbax SB—C18柱（250min×4．6min i．d．，5μm），检测波长254nm，乙腈和水为流动相进行梯度洗脱，柱温25℃，体积流量1．6mL／min，外标法定量分析。结果表明：以2，4-二溴苯乙酮为衍生化试剂测定的肉豆蔻酸（15．6～78．1mg/L）、亚油酸（27．2～136．0mg／L）、油酸（73．4-367．2mg／L）和硬脂酸（31．7～158．4mg／L）在测定范围内呈现良好相关性，相关系数在0．9993～0．9998之间。柱前衍生HPLC法简便准确，可用于水解鸡脂中游离脂肪酸含量定量分析。

A method was developed to determine the free fatty acid simultaneously in the enzymatic hydrolysate of chicken fat using pre - column derivation HPLC. Fatty acids were derived with p - bromophenacylbromide as derivative and 18 - crown - 6 as catalyst. Zorbax SB - C18 reverse column was（250min×4.6min i.d.,5μm）and an acetonitrile - wa- ter gradient eluent was used to separate the free fatty acids. The detection wavelength was 254 nm, while the flow rate was constant at 1.6 mL/min and the column temperature was maintained at 25℃. Results show the correlation coefficients of calibrations of myristic acid（15.6- 78.1 rag/L）, linoleic acid（27.2- 136.0 mg/L）, oleic acid（73.4- 367.2 mg/L）, and stearic acid（31.7 - 158.4 mg/L） range from 0.999 3 to 0.999 8, which indicates the linearity of the four fatty acids is good. It is demonstrated that the method is simple, accurate and suitable for determination of free fatty acids simultaneously in the hydrolysate of chicken fat.