Products Cited in Publications Worldwide Development and Validation of a Quantification Method for Direct Oral Anticoagulants from Capillary Blood Using Volumetric Absorptive Microsampling and Online Spe-Lc-Ms

Development and Validation of a Quantification Method for Direct Oral Anticoagulants from Capillary Blood Using Volumetric Absorptive Microsampling and Online Spe-Lc-Ms

Patrick Opitz; Isabel Waltering; Georg Hempel

DOI:

Abstract

The number of prescriptions for new direct oral anticoagulants (DOACs) apixaban, edoxaban, rivaroxaban and dabigatran has increased exponentially in recent years, increasingly replacing the old gold standard, vitamin-K-antagonists. Due to their wide therapeutic range, therapeutic drug monitoring (TDM) is not required, although it has been proven that this could significantly reduce side effects. In order to develop a cost-efficient and simple method for the simultaneous detection of the DOACs and phenprocoumon, a new technology for sample preparation from capillary blood in the ambulant sector named VAMS® was integrated and an LC-MS detector with on-line solid phase extraction (SPE) applying a Turboflow HTLC CycloneTM 1.0x50 mm column was used. The mobile phase consisted of methanol with water (3/97 v/v) and 0.1% ammonia solution with a flow rate of 2.5 mL/min. For the chromatographic separation, a Phenomenex LTD Kinetex 2.6 µm C18 100 Å, 100x3.0 mm column with a flow rate of 0.3 mL/min in gradient mode was utilized. The mobile phase consisted of acetonitrile, water and formic acid (A: 10:90:0.1 v/v and B: 95:05:0.1 v/v). The method was fully validated in the therapeutic range of the substances according to current guidelines. The LLOQ ranged from 3.5 µg/L for rivaroxaban to 88 µg/L for phenprocoumon and the intra-day and inter-day precision was less than 13% and 12%, while the accuracy was within a range of 85.7-113% and 88.7-106%, respectively. Samples could be stored in the mitra® devices for at least seven days at room temperature except of dabigatran. Because the mitras® were used, no significant haematocrit effect could be observed. A reliable, simple and cost-effective extraction and analysis LC-MS method could be developed and validated. This method is therefore applicable in ambulatory care.

Keywords

therapeutic drug monitoring (TDM) ; anticoagulant ; volumetric absorptive microsampling (VAMS) ; LC-MS ; Validation ; On-line solid phase extraction (SPE)

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Rivaroxaban

A454849366789-02-898%

Rivaroxaban

18.0/50mg

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