Development and validation of an LC-MS/MS method for the simultaneous quantification of milbemycin oxime and praziquantel in plasma: application to a pharmacokinetic study in cats

Shiting Xie; Yixing Lu; Jun Wang; Changcheng Lin; Peiyu Ye; Xiaolin Liu; Wenguang Xiong; Zhenling Zeng; Dongping Zeng

doi:10.3389/fvets.2023.1285932

Development and validation of an LC-MS/MS method for the simultaneous quantification of milbemycin oxime and praziquantel in plasma: application to a pharmacokinetic study in cats

Front Vet Sci. 2023 Oct 30:10:1285932. doi: 10.3389/fvets.2023.1285932. eCollection 2023.

Authors

Shiting Xie^#^{1

2}, Yixing Lu^#^{1

2}, Jun Wang^{1

2}, Changcheng Lin^{1

2}, Peiyu Ye^{1

2}, Xiaolin Liu³, Wenguang Xiong^{1

2}, Zhenling Zeng^{1

2}, Dongping Zeng^{1

2}

Affiliations

¹ Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
² National Risk Assessment Laboratory for Antimicrobial Resistance of Animal Original Bacteria, Guangzhou, China.
³ Livcare (Guangdong) Animal Health Co., Ltd, Qingyuan, China.

^# Contributed equally.

Abstract

Introduction: Milbemycin oxime (MBO) and praziquantel (PZQ) have a broad spectrum of biological activity and are commonly used to treat the parasitic infection in the veterinary clinic. In this study, a fast and efficient LC-MS/MS method was established and validated for the simultaneous determination of MBO, PZQ, cis-4-hydroxylated-PZQ (C-4-OH-PZQ) and trans-4-hydroxylated-PZQ (T-4-OH-PZQ) and in cat plasma.

Methods: Extraction of analytes and internal standards from cat plasma by acetonitrile protein precipitation, allows rapid processing of large batches of samples. MBO, PZQ, C-4-OH-PZQ, T-4-OH-PZQ, and internal standard (IS) were eluted for 13.5 min on a C₁₈ column with a 0.1% formic acid water/acetonitrile mixture as the mobile phase.

Results: Results showed that the method had good precision, accuracy, recovery, and linearity. The linearity range was 2.5-250 ng/mL for MBO, and 10-1000 ng/mL for PZQ, C-4-OH-PZQ, and T-4-OH-PZQ. The intra-day and inter-day precision CV values of the tested components were within 15%. The extraction recoveries of the four components ranged from 98.09% to 107.46%. The analytes in plasma remained stable for 6 h at room temperature, 26 h in the autosampler (4 °C), after freeze-thaw (-20°C) cycles, and 60 days in a -20°C freezer. Method sensitivity sufficed for assessing pharmacokinetic parameters of MBO, PZQ, C-4-OH-PZQ, and T-4-OH-PZQ in plasma samples with LLOQ of 2.5 ng/mL for MBO and 10 ng/mL for PZQ, C-4-OH-PZQ, and T-4-OH-PZQ.

Conclusion: In this study, a selective and sensitive LC-MS/MS method for the simultaneous quantification of MBO, PZQ, C-4-OH-PZQ, and T-4-OH-PZQ in cat plasma was developed and validated.This method had been successfully applied to evaluate the pharmacokinetics of MBO, PZQ, C-4-OH-PZQ, and T-4-OH-PZQ after a single oral administration of 8 mg MBO and 20 mg PZQ in cats.

Keywords: LC–MS/MS; cat plasma; milbemycin oxime; pharmacokinetics; praziquantel.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program (grant number 2019BT02N054).