高效液相色谱法测定食用植物油中5种抗氧化剂的含量
HPLC Determination of 5 Anti-oxidants in Edible Vegetable Oil
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摘要: 提出了高效液相色谱法测定食用植物油中没食子酸丙酯、特丁基对苯二酚、叔丁基羟基茴香醚、没食子酸辛酯和2,6-二叔丁基对羟基甲酚等5种抗氧化剂的方法。样品(1.000 g)用甲醇5 mL)提取2次,涡旋振荡10 min,取上清液在4 ℃冷藏,使残留脂肪析出。取其上清液浓缩后用甲醇定容为10 mL,取5 μL进样用高效液相色谱法进行测定。ZORBAX SB-C18色谱柱及不同比例混合的甲醇(A)和乙酸-水(1+99)(B)的混合液作为流动相进行梯度淋洗分离,柱温为30 ℃,流量为1.0 mL·min-1,检测波长为280 nm。结果表明:5种抗氧化剂的检出限(3S/N)依次为0.10,0.34,0.32,0.14,0.57 mg·kg-1。在3个浓度水平上(200,500,1 000 mg·kg-1)对方法做回收试验,测得回收率在90.5%~102.0%之间,相对标准偏差(n=6)在0.02%~1.35%之间。Abstract: Five synthetic anti-oxidants preferentially added to edible vegetable oil, namely propylgallate, tert-butylhydroquinone, tert-butylhydroxyanisole, octyl gallate and 2,6-di-t-butyl-p-hydroxytoluene, were determined by HPLC. The oil sample (1.000 g) was extracted twice with CH3OH (5 mL+5 mL) in a stoppered centrifuge tube by centrifuging for 10 min. The supernatant was transferred to a 25 mL glass tube which was kept at 4 ℃ to solidify and separate residual fats. The supernatant was taken, evaporated and made up to 10.0 mL with methanol. The solution (5 μL) was introduced for HPLC analysis by separation on the ZORBAX SB-C18 column (30 ℃) with gradient elution (flow rate: 1.0 mL·min-1) using mixtures of (A) methanol and (B) HOAc+H2O (1+99) in various proportions as mobile phase. UV detection with DAD at 280 nm was adopted in the determination. Detection limits (3S/N) found for the 5 antioxidants were 0.10, 0.34, 0.32, 0.14, 0.57 mg·kg-1 respectively. Recovery and precision were tested at 3 concentration levels (200, 500, 1 000 mg·kg-1), giving results of recovery ranged from 90.5% to 102.0%, and RSD′s (n=6) ranged from 0.02% to 1.35%.