HPLC测定混合型饲料添加剂中L-苹果酸含量
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摘要
用HPLC法测定混合型饲料添加剂中L-苹果酸(malic acid,LMA)的含量。采用C18(250 mm×4.6 mm×5μm)色谱柱,以0.02 mol/l磷酸二氢钾:乙腈=982(v:v),流速0.8 ml/min,柱温25℃,检测波长210 nm,进样量10μl。结果表明:L-苹果酸对照品和样品的保留时间均是3.9 min,其线性方程分别为Y=0.390 9X-4.578 6,R=0.999 8(n=5),在126.25~2 020μg/ml的范围内,线性关系良好,其最低检测量为12.23μg/ml。精密度的RSD为0.43%,表明此方法精密度较好;重现性的RSD为0.17%,表明重现性良好;回收率的RSD为0.61%。用此方法检测出其混合饲料添加剂中L-苹果酸含量为(99.01±0.11)%,符合产品规定的质量要求。该实验建立的HPLC方法可快速、准确测定混合饲料中L-苹果酸含量,为其质量控制提供科学依据。
The test was to determine the content of malic acid(LMA) in mixed feed additive by HPLC.The determined conditions were following: C18 column(250 mm × 4.6 mm × 5 μm); mobile phrase:0.02 mol/l potassium dihydrogen phosphateacetonitrile=98:2(v:v); flow rate: 0.8 ml/min, columntemperature: 25 ℃, detection wavelength: 210 nm, the injection volume: 10 μl. The retention times of L-malic acid reference substance and sample were 3.9 min, and the linear equations were Y=0.390 9 X-4.578 6, R=0.999 8(n=5), the linear relationship is good in the range of 126.25~2 020 μg/ml, the mini-mum detection was 12.23 μg/ml. The RSD of the precision was 0.43%, which indicated that the preci-sion of this method was good. The RSD of reproducibility was 0.17%, which indicated that the repro-ducibility was good. The RSD of the recovery was 0.61%. The content of L-malic acid in the mixedfeed additive was(99.01±0.11)%, which was in accordance with the quality requirement of the prod-uct. The HPLC method established in this experiment can be used to determine the rapid and accuratedetermination of L-malic acid in mixed feed and provide scientific basis for its quality control.
The test was to determine the content of malic acid(LMA) in mixed feed additive by HPLC.The determined conditions were following: C18 column(250 mm × 4.6 mm × 5 μm); mobile phrase:0.02 mol/l potassium dihydrogen phosphateacetonitrile=98:2(v:v); flow rate: 0.8 ml/min, columntemperature: 25 ℃, detection wavelength: 210 nm, the injection volume: 10 μl. The retention times of L-malic acid reference substance and sample were 3.9 min, and the linear equations were Y=0.390 9 X-4.578 6, R=0.999 8(n=5), the linear relationship is good in the range of 126.25~2 020 μg/ml, the mini-mum detection was 12.23 μg/ml. The RSD of the precision was 0.43%, which indicated that the preci-sion of this method was good. The RSD of reproducibility was 0.17%, which indicated that the repro-ducibility was good. The RSD of the recovery was 0.61%. The content of L-malic acid in the mixedfeed additive was(99.01±0.11)%, which was in accordance with the quality requirement of the prod-uct. The HPLC method established in this experiment can be used to determine the rapid and accuratedetermination of L-malic acid in mixed feed and provide scientific basis for its quality control.
引文
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[12]孙全,张景勍,傅亚,等.RP-HPLC法同时测定半夏药材中4种有机酸的含量[J].药物分析杂志,2015,35(6):1062-1065.
[2]吴悦,潘丽军,李兴江.生曲霉CICC40365利用木糖产L-苹果酸的发酵条件优化[J].微生物学通报,2014,41(6):1052-1062.
[3]汪多仁.L-苹果酸的开发与应用进展[J].增塑剂,2012,23(3):11-16.
[4]周丽,崔文瑕,刘中美,等.基于代谢工程策略合成L-苹果酸研究进展[J].食品工业科技,2015,36(10):383-387.
[5]汪多仁,陈体庆.L-苹果酸的开发与应用[J].饮料工业,2004,25(7):21-26.
[6]吴军林,吴清平,张菊梅.L-苹果酸的生理功能研究进展[J],食品科学,2008(29):692-695.
[7]梁静,张文举,王博,等.L-苹果酸的生理功能及其在反刍动物生产应用中的的研究进展[J].中国畜牧兽医,2016,43(7):1916-1921.
[8]卢超银,沈雨莎,张烨,等.L-苹果酸及苹果对睡眠剥夺小鼠记忆的影响[J].中国卫生产业,2016(11):16-18.
[9]李永成,钟秋平.甘蔗糖蜜预处理对产L-苹果酸的影响与培养基N源添加[J].食品工业科技,2011,32(1):178-180.
[10]刘学剑.有机酸及其复合酸在动物生产中的应用概况[J].江西饲料,2002(3):1-5.
[11]王凤荣,乔长晟,江建梅.出芽短梗霉发酵液中聚苹果酸测定方法的比较[J].食品工业科技,2010,31(6):347-350.
[12]孙全,张景勍,傅亚,等.RP-HPLC法同时测定半夏药材中4种有机酸的含量[J].药物分析杂志,2015,35(6):1062-1065.