Quantification of treprostinil concentration in rat and human using a novel validated and rapid liquid chromatography-tandem mass spectrometry method: Experimental and clinical applications in ischemia–reperfusion injury

Overview of biological sample preparation and quantification of treprostinil by LC-MS/MS. [Display omitted] •Renal ischemia–reperfusion injury (IRI) is inevitable during kidney transplantation with no approved therapy.•Treprostinil, a prostacyclin analog, is a potential therapy to treat renal IRI.•Analytical methods are required to quantify treprostinil in biological samples.•This novel LC-MS/MS method was developed and validated to quantify treprostinil in animal and human serum and plasma. Treprostinil (Remodulin®) is a Food and Drug Administration (FDA) approved prostacyclin analog to treat pulmonary arterial hypertension. Recently, treprostinil has been investigated to reduce ischemia–reperfusion injury (IRI) during transplantation, which currently has no approved treatment. A validated analytical method is necessary to measure treprostinil concentrations in biological specimens. Here, a novel, sensitive, and specific method to measure treprostinil concentrations in rat serum, human serum, and human plasma has been developed using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Biological samples were processed by protein precipitation before chromatography and 6-keto Prostaglandin F1α-d4 was used as an internal standard. A gradient method was established with a total run time of 4 min. The assay was linear over the range of 0.25–75.0 ng/ml with accuracy (92.97–107.87 %), intra-assay precision (1.16–3.34 %), and inter-assay precision (1.11–4.58 %) in all biological matrices, which are within FDA acceptance criteria. No significant variation in treprostinil or 6-keto Prostaglandin F1α-d4 concentrations were observed under the investigated storage conditions. This novel, sensitive, and specific LC/MS-MS method is cost-effective and suitable for measuring treprostinil concentrations in animal studies and human biological samples for clinical applications.