Mochamad Lazuardi, Hermanto Bambang
Department of Basic Science, Veterinary Pharmacy Subdivision, Faculty of Veterinary Medicine, Airlangga University, Surabaya, Indonesia.
Department of Pharmacology, Medical Faculty Airlangga University, Prof. Dr. Moestopo 47, Pacar Kembang, Surabaya, Indonesia.
Vet World. 2017 Aug;10(8):932-938. doi: 10.14202/vetworld.2017.932-938. Epub 2017 Aug 17.
The objective of the current study is to determine the concentration of aflatoxin B1 using high-performance liquid chromatography (HPLC) with a photodiode array (PDA) detector.
Aflatoxin B certified reference grade from Trilogy Analytical Laboratory dissolved acetonitrile (ACN) at 10 µg/mL was using standard assessment. HPLC instruments such as ultraviolet-PDA detector used a Shimadzu LC-6AD pump with DGU-20A5 degasser, communication module-20A, and PDA detector SPD-M20A with FRC-10A fraction collector. The HPLC was set isocratic method at 354 nm with a reverse-phase ODS C18 column (LiChrospher 100 RP-18; diameter, 5 µm) under a 20°C controlled column chamber. Rheodyne sample loops were performed in 20 µL capacities. The mobile phase was performed at fraction 63:26:11 HO: methanol:ACN at pH 6.8. A total of 1 kg of feed contained 10% bread crumbs and 30% concentrated, 40% forage, and 20% soybean dregs were using commercials samples. Samples were extracted by ACN and separated with solid phase extraction ODS 1 mL than elution with mobile phase to collect at drying samples performed. The samples were ready to use after added 1 mL mobile phase than injected into the system of HPLC.
We found that the retention time of aflatoxin B was approximately 10.858 min. Linearity of 0.01-0.08 µg/mL aflatoxin B dissolved in mobile phase was obtained at R=0.9. These results demonstrate that these methods can be used to analyze aflatoxin B and gain 89-99% recovery. The limit of detection of this assay was obtained at 3.5 × 10 µg/mL.
This method was easy to apply and suitable to analyzing at small concentrations of aflatoxin B in formulated product of feed cattle.
本研究的目的是使用配备光电二极管阵列（PDA）检测器的高效液相色谱法（HPLC）测定黄曲霉毒素B1的浓度。
使用Trilogy Analytical Laboratory认证的黄曲霉毒素B标准参考级，将其溶解于乙腈（ACN）中，浓度为10 μg/mL，进行标准评估。HPLC仪器，如紫外-PDA检测器，使用岛津LC-6AD泵、DGU-20A5脱气机、通讯模块-20A以及带有FRC-10A馏分收集器的PDA检测器SPD-M20A。HPLC采用等度洗脱法，在354 nm波长下，使用反相ODS C18柱（LiChrospher 100 RP-18；直径5 µm），柱温控制在20°C。使用20 μL定量环的Rheodyne进样阀。流动相为体积比63:26:11的水:甲醇:乙腈，pH值为6.8。总共1 kg饲料包含10%面包屑、30%浓缩料、40%草料和20%豆渣，使用商业样品。样品用乙腈提取，通过1 mL ODS固相萃取分离，然后用流动相洗脱，收集干燥后的样品。加入1 mL流动相后注入HPLC系统即可进行分析。
我们发现黄曲霉毒素B的保留时间约为10.858分钟。溶解于流动相中的黄曲霉毒素B在0.01 - 0.08 μg/mL范围内呈线性，R = 0.9。这些结果表明这些方法可用于分析黄曲霉毒素B，回收率为89 - 99%。该测定方法的检测限为3.5×10 μg/mL。
该方法易于应用，适用于分析牛饲料配方产品中低浓度的黄曲霉毒素B。
