利用 HPLC-DAD 技术对青贮饲料和饲料中甲醛进行定量的内部验证方法。

In-house validation method for quantification of formaldehyde in silage and feedingstuffs with the use HPLC-DAD technique.

机构信息

Department of Hygiene of Animal Feedingstuffs, National Veterianry Research Institute, Partyzantów 57 Avenue, 24-100 Puławy, Poland.

出版信息

Pol J Vet Sci. 2020 Dec;23(4):553-561. doi: 10.24425/pjvs.2020.134705.

DOI:10.24425/pjvs.2020.134705

PMID:33480494

Abstract

An HPLC-DAD method was developed for the determination of formaldehyde in animal feed and silage. The method is based on the determination of the product of chemical reaction between formaldehyde and 2,4-dinitrophenylhydrazine. A 3 g of feed or silage were extracted with Milli-Q water with phosphoric acid and next formaldehyde was derivative with the use 2,4-dinitrophenyl- hydrazine in acetronitrile solution. The extract was purified with 0.45 µm syringe filters and separeted on Zorbax Eclipse XDB C18 column and detection was carried out at 360 nm. Formal- dehyde was eluted with a mobile phase consisting of acetonitrile/water in isocratic elution. This method provided average recoveries of 90.6% to 102.2%, with CVs of 2.6% to 6.4% for feed and from 91.3% to 108.7% with CVs of 1.1% to 4.1% for silage in the ranged of 50 to 1000 mg/kg feeds and silage. The LOD and LOQ for formaldehyde in feed and silage ranged from 1.6 to 2.6 and 2.7 to 5.7 mg/kg, respectively. The methodology was applied for the analysis of feed and silage samples collected from poultry, pigs and cows farms.

摘要

建立了高效液相色谱-二极管阵列检测法（HPLC-DAD）用于测定动物饲料和青贮料中的甲醛。该方法基于甲醛与 2,4-二硝基苯肼（DNPH）反应产物的测定。取 3g 饲料或青贮料，用 Milli-Q 水和磷酸提取，然后用乙腈溶液中的 2,4-二硝基苯肼将甲醛衍生化。提取液用 0.45µm 注射器过滤器纯化，在 Zorbax Eclipse XDB C18 柱上分离，在 360nm 处检测。甲醛用乙腈/水组成的流动相在等度洗脱条件下洗脱。该方法在 50 至 1000mg/kg 饲料和青贮料范围内，对饲料的平均回收率为 90.6%至 102.2%，变异系数（CV）为 2.6%至 6.4%，对青贮料的平均回收率为 91.3%至 108.7%，CV 为 1.1%至 4.1%。饲料和青贮料中甲醛的检出限（LOD）和定量限（LOQ）分别为 1.6 至 2.6mg/kg 和 2.7 至 5.7mg/kg。该方法应用于禽畜养殖场采集的饲料和青贮料样品的分析。