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高效液相色谱法测定复方非泼罗尼滴剂中抗氧剂的含量
姚路路,程婵婵,张聪,魏轲,杨会鲜,周德刚
0
(洛阳惠中兽药有限公司)
摘要:
建立高效液相色谱法(HPLC)测定复方非泼罗尼滴剂中抗氧剂叔丁基对羟基茴香醚(BHA)与二丁基羟基甲苯(BHT)含量。采用高效液相色谱法,色谱柱为Agilent Zorbax SB-C18(250×4.6 mm,5 μm),流动相A为乙腈-甲醇-水(47∶ 21∶32),混合前在水中加入1%冰醋酸,流动相B为乙腈,线性梯度洗脱;检测波长为285 nm;流速为0.9 mL/min;柱温为30 ℃;进样量为20 μL;采用外标法计算BHA、BHT含量。在建立的色谱条件下,空白溶液、阴性样品对BHA、BHT的测定均无干扰;BHA、BHT的检测限分别为0.08 μg/mL和0.19 μg/mL,定量限分别为0.31 μg/mL和0.77 μg/mL,分别在5.13 ~ 15.39 μg/mL与2.56 ~ 7.66 μg/mL范围内线性关系良好;平均回收率分别为98.5%和99.9%(n=9)。所建立的方法简单易行、专属性强、准确度高、灵敏度好,可用于复方非泼罗尼滴剂中抗氧剂的含量测定。
关键词:  复方非泼罗尼滴剂  叔丁基对羟基茴香醚  二丁基羟基甲苯  抗氧剂  含量
DOI:
投稿时间:2022-02-22修订日期:2022-07-22
基金项目:
Determination of Antioxidant in Compound Fipronil Spot-On Solutions by HPLC
(Luoyang Huizhong Animal Medicine Co.,Ltd)
Abstract:
To establish an HPLC method for the content determination of 3-ter-butyl-4-hydroxyanisole (BHA) and 2,6-di-tert-butyl-4-methylphenol (BHT) in compound fipronil spot-on solutions, the analysis was performed on an Agilent Zorbax SB-C18 column (250 × 4.6 mm, 5 μm). The gradient mobile phase was using acetonitrile-methanol-water(47∶ 21∶ 32), and 1% acetic acid was added to water before mixing as mobile phase A, acetonitrile as mobile phase B. The detection wavelength was 285 nm, the flow rate was 0.9 mL/min, the column temperature was 30 ℃, and the injection volume was 20 μL. Under the proposed chromatographic conditions, the diluent and the negative samples didn’t interfere with the determination of BHA and BHT. The limit of dection of BHA and BHT were 0.08 μg/mL and 0.19 μg/mL respectively, and the limit of quantification of BHA and BHT were 0.31 μg/ml and 0.77 μg/mL respectively. The linear relationship was good in the ranges of 5.13~15.39 μg/mL for BHA and 2.56 ~7.66 μg/mL for BHT. The average recoveries of BHA and BHT were 98.5% and 99.9% respectively. The method is simple and accurate with high specificity, which can be used to determine the content of BHA and BHT in compound fipronil spot-on solutions.
Key words:  Compound Fipronil Spot-On Solutions  BHA  BHT  antioxidant  content determination

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