柱前衍生化-HPLC法同时测定注射用沙棘籽油中脂肪酸的含量

Determination of the content of fatty acids in Hippophae rhamnoides L. seed oil by HPLC with precolumn derivatization

目的建立沙棘籽油中脂肪酸的HPLC测定法。方法以2，4＇-二溴苯乙酮为衍生化试剂，18-冠醚-6为相转移催化剂，采用Kromasil C8反相色谱柱（250mm×4．6mm，5μm），检测波长254nm，以甲醇-乙腈-（体积比60：30：10）为流动相等度洗脱，柱温为室温，流速为1．0mL·min%^-1.七烷酸为内标，一次基线分离5种脂肪酸，建立注射用沙棘籽油的脂肪酸含量测定的方法。结果α-亚麻酸的线性范围为2．02～20．22ng，相关系数为0．9991；亚油酸的线性范围为2．90～20．90ng，相关系数为0．9991；棕榈酸的线性范围为1．27-12．67ng，相关系数为0．9990；油酸的线性范围为1．92—19．16ng，相关系数为0．9990；硬脂酸的线性范围为0．30—3．04ng，相关系数为0．9991。α-亚麻酸、亚油酸、棕榈酸、油酸、硬脂酸的平均回收率分别为106．07％、104．13％、100．81％、99．43％、95．62％，RSD分别为2．64％、2．42％、3．14％、1．18％、2．24％（n=9）。结论本方法重现性好，定量准确，可作为注射用沙棘籽油中脂肪酸测定的定量方法，用于沙棘籽油的质量控制。

Objective To develop a method for the determination of fatty acid in Hippophae rhamnoides L. seed oil for injection by precolumn derivation HPLC. Methods The fatty acids were derivatized with 2,4＇-dibromoacetophenone as derivative and 18-crown-6 as catalyst. Kromasil C8 column （250 mm ×4. 6 mm, 5 μm）was used, methanol-acetonitrile-water（ V： V： V = 60： 30：10 ）was used as eluent and the internal standard was heptadecanoic acid. The detection wavelength was set at 254 nm. Column temperature was fixed at room temperature, and the flow rate was 1.0 mL. min-1. Results The standard curves of α-linolenic, linoleic,palmitic, oleic, and steraric acid were linear within the range of 2. 02- 20. 22 ng, 2.90- 20. 90 ng, 1.27- 12. 67 ng,1. 92-19. 16 ng and 0. 30-3.04 ng,respectively. The five fatty acid recoveries were 106.07%, 104. 13% ,100. 81% ,99.43% ,95.62% and the RSD were 2. 64% ,2. 42% ,3. 14%, 1.18% ,2. 24% ,respectively. Conclusions Five fatty acids are separated within 40 min during a single run. The present method is reliable and relatively simple for the determination of fatty acid in Hippophae rhamnoides L. seed oil.