Abstract
Objective: A study aimed at the validation of a high performance liquid chromatography (HPLC) method for detection and quantification of ivermectin (IVM) in commercial milk. Method: After a liquid-liquid extraction with acetonitrile and hexane, a derivatization with a mixture of trifluoroacetic acid and trifluoroacetic anhydride was performed. Chromatography conditions were a C8 column, a fluorescent detector (Ex. 360; Em. 470 nm), a run time of 25 minutes and an isocratic elution at 30°C. The methodology was validated in terms of selectivity, linearity, precision, recovery, limit of quantification (LoQ), limit of detection (LoD), and robustness. Results: The method showed an adequate linearity and selectivity (r2=0.999) with an elution time of 13.909 minutes. However, the repeatability and intermediate precision showed RSD values above of 20% and recovery between 37 to 79%. The calculated LoD and LoQ were 2.50 and 5.00 ng/ml respectively. Robustness showed a significant variation on the analytical method with small changes in heating time during the derivatization and flow rate of chromatography system. Conclusion: The HPLC-fluorescence method showed results partially satisfactory and could be the used for IVM detection in commercial milk samples.
Highlights
Ivermectin (IVM) is a macrocyclic lactone, a class of anti-parasitic drugs derived from the soil microorganism Streptomyces avermitilis, used against endectoparasites, especially in bovine, swine and equine production [1]
IVM separations were performed on a Waters SunFire C8 column (150 × 4.6 mm, 5 μm, Validation parameters The method for the identification of IVM in milk was validated in terms of the analytical parameters of linearity, selectivity, accuracy, recovery, limit of quantification (LoQ), limit of detection (LoD), and robustness following conventional protocols from international guidelines [16,17]
Same linearity was reported by Souza et al [15] who found r2 values between 0.98 and 0.99 in milk samples spiked with IVM
Summary
Ivermectin (IVM) is a macrocyclic lactone, , a class of anti-parasitic drugs derived from the soil microorganism Streptomyces avermitilis, used against endectoparasites, especially in bovine, swine and equine production [1] It was discovered in 1976 but the use related to control of endo-and ectoparasites in animal started only in 1981 [2,3]. The IVM concentrations found in milk are very low and aren’t related to a serious harm to consumers [8], their presence is important for public health since this product is a food for massive consumption all over the world. For these reason, different methods using chromatography have been described for the analysis of IVM. J Exp Food Chem 2: 107. doi:10.4172/2472-0542.1000107
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