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.
Resumen
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.
Por qué esto importa para la hirudoterapia
Este estudio desarrolló y validó un método de cromatografía líquida con espectrometría de masas para cuantificar la r-RGD-hirudina —una hirudina bifuncional recombinante— en plasma humano dentro de un rango de concentración verificado de 10–2000 ng/mL, y posteriormente lo aplicó a un estudio farmacocinético en 40 voluntarios chinos sanos. La r-RGD-hirudina es una variante recombinante de la hirudina, el potente inhibidor directo de la trombina aislado originalmente de la saliva de la sanguijuela medicinal, por lo que los métodos bioanalíticos validados para medirla en plasma humano respaldan directamente el desarrollo clínico y la monitorización terapéutica de los anticoagulantes derivados del secretoma de la sanguijuela. La advertencia honesta es que se trata de una validación de un método analítico y una caracterización farmacocinética, no de un ensayo clínico de eficacia o seguridad; establece la capacidad de medición y los parámetros de exposición al fármaco, pero no demuestra beneficio terapéutico.
Citación
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
Contexto clínico relacionado
Explore cómo esta investigación se conecta con la práctica clínica
Añadido a la biblioteca ASH: May 27, 2026 · Última actualización del sitio: 18 de junio de 2026