Abstract
Porcine relaxin is a 6 kDa peptide hormone of pregnancy with important physiological and pharmacological effects. It contains a number of analogs of which porcine relaxin B29 is one of the most important. To support the development of porcine relaxin B29 as a new drug, we established an UPLC-MS/MS method for its quantitation in dog plasma. Sample preparation by protein precipitation and ion exchange solid phase extraction was followed by UPLC on an XBridge™ BEH300 C18 column at 40 °C in a run time of only 5.5 min. Detection was performed on a Qtrap 6500 mass spectrometer using ESI in the positive ion mode with MRM of the transitions at m/z 831.7 [M+7H]7+ → 505.4 and m/z 1162.4 [M+5H]5+ → 226 for pRLX B29 and internal standard (recombinant human insulin), respectively. The method was linear over the concentration range 30–2000 ng/mL with no matrix effects. Intra- and inter-day precisions were < 15% with accuracies in the range 98.8–100.6%. The method was successfully applied to a pharmacokinetic study in beagle dogs after administration of a 0.15 mg/kg intravenous dose.
Sample preparation and detection procedure.
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The authors gratefully acknowledge financial support from the National Natural Science Foundation of China (Grant Nos. 81430087, 81473142, 81673396, 81673502, and 81603182).
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All procedures involving animals in this study were approved by the ethics committee of Jilin University and were performed in accordance with the Guidelines for the Care and Use of Laboratory Animals of Jilin University.
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Su, C., Sun, H., Yang, H. et al. A validated UPLC-MS/MS method coupled with protein precipitation and ion exchange solid phase extraction for the quantitation of porcine relaxin B29 in dog plasma and its application to a pharmacokinetic study. Anal Bioanal Chem 409, 6559–6565 (2017). https://doi.org/10.1007/s00216-017-0604-3
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DOI: https://doi.org/10.1007/s00216-017-0604-3