A new LC-MS/MS bioanalytical method for perindopril and perindoprilat in human plasma and milk
A first of its kind, simple, rapid, and sensitive liquid chromatography mass spectrometry (LC-MS/MS) method was developed and validated for quantification of perindopril and perindoprilat in both human plasma and breast milk. The analytes and internal standards (phenazone and acetyl salicylic acid) were extracted from biological matrices by protein precipitation. A Phenomenex® C-18 column was used to provide an appropriate chromatographic separation of the analytes, followed by detection with tandem mass spectrometry. Gradient chromatographic and mass spectrometric detection conditions with mobile phases (A: 5% methanol + 0.1% formic acid in water v/v, and B: 95% methanol + 0.1% formic acid in water v/v) were developed to achieve a LOQ of 0.5 ng/mL in both human plasma and milk. The method was suitable of evaluating clinical samples. The mass transition was followed as m/z 369.10/172.00 for perindopril, m/z 339.00/168.10 for perindoprilat, m/z 188.90/55.95 for phenazone, and m/z 179.04/137.02 for acetyl salicylic acid. The developed method was optimized and validated with a linear range of 0.1–200 ng/mL (r 2 = better than 0.99 for both perindopril and perindoprilat). The precision and accuracy values were within 15% CV. The overall recovery of the analytes was 80–110%. The method has good specificity and repeatability. Stability studies were conducted in both human plasma and bovine milk for up to 3 months, at the storage conditions of 25, 4, and −80 °C.
KeywordsPerindopril Perindoprilat LC-MS/MS Human plasma Human breast milk Lactation
I would like to acknowledge the patient and clinicians at Women’s and Children’s Hospital, Adelaide, for supplying patient samples and blank human milk, Australian Red Cross for supplying blank human plasma, and Benjamin Tscharke and Ben Noll for their technical help.
Compliance with ethical standards
Conflict of interest
This authors declare that there is no conflict of interest regarding the publication of this paper. The study was approved by the Women’s and Children’s Hospital Research Ethics Committee (HREC14/WCHN/115) and University of South Australia Research Ethics Committee (0000033979). Written informed consent was obtained from all participants.
This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.
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