LC-MS/MS method for determination of r-RGD-hirudin in human plasma and its application in pharmacokinetic study
Bioanalytical method published in Analytical Biochemistry (2022)
Abstract
OBJECTIVE: This study aimed to develop a simple, sensitive, and selective Liquid chromatography with a Mass spectroscopic method for simultaneous quantification of a recombinant bifunctional hirudin (r-RGD-Hirudin, Bifunctional Hirudin, BFH) in human plasma and verify its effectiveness. METHODS: The analytes and the internal standards from human plasma were extracted using the solid-phase extraction technique. The reconstituted samples were chromatographed on Waters C18 column (BEH 50 × 2.1 mm, 1.7 μm) using a mixture of 0.1% formic acid/acetonitrile (85%/15%, v/v) with gradient elution as the initial mobile phase at a flow rate of 0.3 mL/min. RESULTS: The effectiveness of the proposed method was verified over the concentration range of 10-2000 ng/mL for r-RGD-Hirudin. A linear calibration curve was obtained. The precision and accuracy of BFH in the intra- and inter-day runs fell within the range of ±15% at LQC, GMQC, MQC and HQC concentrations. The extraction recoveries and matrix effect at two quality control (QC) levels for BFH were confirmed to conform to the relevant requirement. CONCLUSION: The proposed method was successfully adapted to examine the pharmacokinetics of BFH in 40 Chinese healthy volunteers, respectively.
Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.
Summary
Develops and validates an LC-MS/MS assay for r-RGD-hirudin (engineered antiplatelet-anticoagulant chimera) in human plasma — applied to phase I PK study.
Why This Matters for Hirudotherapy
This study developed and validated a liquid chromatography with mass spectroscopic method for quantifying r-RGD-hirudin—a recombinant bifunctional hirudin—in human plasma across a verified concentration range of 10–2000 ng/mL, then applied it to a pharmacokinetic study in 40 healthy Chinese volunteers. r-RGD-Hirudin is a recombinant variant of hirudin, the potent direct thrombin inhibitor originally isolated from medicinal leech saliva, so validated bioanalytical methods for measuring it in human plasma directly support the clinical development and therapeutic monitoring of leech-secretome–derived anticoagulant therapeutics. The honest caveat is that this is an analytical method validation and pharmacokinetic characterization, not a clinical efficacy or safety trial; it establishes measurement capability and drug exposure parameters but does not demonstrate therapeutic benefit.
Citation
LC-MS/MS method for determination of r-RGD-hirudin in human plasma and its application in pharmacokinetic study.
Peng A et al. · Analytical biochemistry, 2022
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