Feb 21, 2022
Pharmacokinetics and bioequivalence of two imidocarb formulations in cattle after subcutaneous injection
- Anonymous1,
- Chen Chen1,
- Maolin Liu1,
- Xiaojie Chen1,
- Chunshuang Liu1,
- Yanyan Feng2,
- Xinbo Yan3,
- Yiming Liu1,
- Xiubo Li1,
- Honglei Wang1
- 1National Feed Drug Reference Laboratories, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China;
- 2Qilu Animal Health Products Corp. LTD Shangdong province, China;
- 3College of Veterinary Medicine, China Agricultural University, Beijing, China
Protocol Citation: Anonymous, Chen Chen, Maolin Liu, Xiaojie Chen, Chunshuang Liu, Yanyan Feng, Xinbo Yan, Yiming Liu, Xiubo Li, Honglei Wang 2022. Pharmacokinetics and bioequivalence of two imidocarb formulations in cattle after subcutaneous injection. protocols.io https://dx.doi.org/10.17504/protocols.io.b5c6q2ze
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it’s working
Created: February 21, 2022
Last Modified: February 21, 2022
Protocol Integer ID: 58494
Keywords: IMD, Cattle, Pharmacokinetics, Bioequivalence
Funders Acknowledgements:
2016YFD0501305
Grant ID: the National key R & D Program “Drug Resistance Database Construction and Ecological Risk Assessment in Livestock and Poultry Breeding Environment”
Innovation Project of Chinese Academy of Agricultural Sciences
Grant ID: （CAAS-FRI-06)
Disclaimer
The authors thank farmers in Beijing suburban district for supply with 48 cattle.
Abstract
Imidocarb (IMD) is commonly used for treatment of eperythrozoon, babesia, piroplasma and trypanosoma in animals, but there are few studies on its pharmacokinetics in cattle. The purpose of this study was to obtain pharmacokinetic parameters and assess the bioequivalence of subcutaneous injections of two IMD formulations in cattle. Forty-eight healthy cattle, 24 males and 24 females, were randomly divided into two groups (test group and reference group) with 12 males and 12 females per group. The generic IMD was injected subcutaneously with a single dose of 3.0 mg/kg in the test group. Reference group animals were given one injection of the marketedIMDat the same dosage. The limit of detection (LOD) and limit of quantification (LOQ) for IMD in cattle plasma were 0.05 μg/Land 0.1 μg/L, respectively. The recoveries ranged from 88.50% to 92.42%, and the equation of this calibration curve was Y=13672.1X+187.43. The pharmacokinetics parameters of the test group showed that the maximum concentration of 2257.5±273.62 µg/L was obtained at 2.14±0.67 h, AUC0-t 14553.95±1946.85 µg·h/L, AUC∞15077.88±1952.19 µg·h/L, T1/2 31.77±25.75 h, CL/F 0.14±0.02 L/h/kg, and Vz/F 6.53±5.34 L/kg. There was no significant difference in AUC0-t, AUC∞ and Cmax between the test group and the reference group (P＞0.05). The 90% confidence interval of AUC0-t,AUC0-∞ andCmax in the test group was included in 80%–125% AUC0-t, AUC0-∞and 70%–143% Cmax in the reference group, respectively. Based on these results, the two preparations were found to be bioequivalent.
Materials
Animals
Twenty-four healthy male cattle and 24 healthy female cattle, aged almost 6 months and weighing 180±15 kg, were selected. The cattle received no treatment for several months prior to the study and were housed in open-air pens. The cattle had free access to water and were fed with a conventional feed without antibiotics during the study. These 48 cattle were divided into 2 groups (test group and reference group) with 24 cattle (12 males and 12 females) in each group. The animals in the test groupwere treated subcutaneously by a single dose injection with a generic IMD formulation, at the recommended dosage 3.0 mg/kg. Animals in the referencegroup were injected subcutaneously with a marketed IMD formulation in the same dose as the test group. All cattle experiment procedures were approved and performed in accordance with the Animal Use and Care Committee of Feed Research Institute, Chinese Academy of Agricultural Sciences （number:FRI-CAAS20150811）. There was no anesthesia and euthanasia in our study. All cattle were alive and healthy after the whole experiment.
Drug formulation
The test drug (the generic IMD, 100 mL:85 mg) was manufactured and provided by Qilu Animal Health ProductsCorp. LTD (Shangdong province, China).
The reference drug (the marketed IMD, 100 mL:85 mg) was manufactured and provided by AKZO-NOBEL Corp. (Boxmeer, Netherlands).
Sample collection
Blood samples (10–15 mL) were collected from the jugular vein at 0 h before administration and 10 min, 30 min, 1, 2, 4, 6, 8, 10, 12, 24, 48, 72 and 96 h after subcutaneous administration, and were drawn in vacutainers containing disodium EDTA as anticoagulant. The samples were immediately centrifuged at 1500 g for 10 min. All the plasma samples in plastic vials were stored at −20°C until they were analyzed.
Sample preparation
We used a weak cation-exchange solid phase extraction procedure described by Tarbin [10], and made some modifications to determine the amount of IMD in the plasma. Briefly, 1.0 mL plasma samples were added to plastic centrifuge tubes, adding 3 mL methanol/acetonitrile (90/10, v/v), vertexing for 1 min, ultrasonicating for 15 min, and centrifuging for 10 min at 7000 g. To improve IMD recoveries, the residues were extracted twice. The second extracted solution was combined with the first. The combined extracted solution was passed through SPE columns (Waters Oasis WCX, 3cc 60 mg, Waters Company, USA) conditioned with 3 mL methanol and 3 mL water. The loaded cartridge was washed with 3 mL methanol/water (50/50, v/v), and was eluted with 3 mL methanol/formic acid (96/4, v/v). The analyte was evaporated under a nitrogen stream at 40°C, reconstituted in 1 mL methanol/water (15/85, v/v), and filtered through a 0.22 µm nylon syringe filter before analysis by UPLC-MS/MS.