Importance of measuring pharmacologically active metabolites of edoxaban: development and validation of an ultra-high-performance liquid chromatography coupled with a tandem mass spectrometry method

Siriez, Romain; Alpan, Lütfiye; Elasaad, Kossay; Devel, Philippe; Laloy, Julie; Dogné, Jean‐Michel; Douxfils, Jonathan

doi:10.1007/s11239-019-02030-5

Cited by 7 publications

(18 citation statements)

References 23 publications

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“…This increase will be even greater if there are extrinsic factors influencing the pharmacokinetics of edoxaban such as the presence of drug interactions. For example, concomitant use of ketoconazole (a P‐gp transporter and CYP3A4 inhibitor) increases total edoxaban exposure by 87%, whereas intakes of cyclosporine or erythromycin lead to increased exposures by 73% and 85%, respectively 24,28‐33 …”

Section: Discussionmentioning

confidence: 99%

“…(2.7 nM and 0.63 ng/ml) should be negligible contrary to the impact of the M4 (1.8 nM and 23.1 ng/ml) with parent edoxaban (3.0 nM and 243 ng/ml) in absence of CYP inducers. 24 Furthermore, no manufacturer has been able to offer us the M6 and M8 in sufficient quantity and with acceptable purity for UHPLC-MS/MS. In addition,…”

Section: Ponderal Concentration or Anti-xa Activity: A Point Of Debatementioning

confidence: 99%

“…The exclusion criteria and the preparation of normal pooled plasma (NPP) were the same as previously described. 24,26 The study protocol was in accordance with the Declaration of Helsinki and the recruitment of the healthy volunteers has been approved by the Ethical Committee of the CHU UCL Namur, Yvoir, Namur, Belgium (49/2009). 24,26 Briefly, blood was taken by venipuncture in the antecubital vein and collected into 0.109 M sodium citrate (9:1 v/v) tubes (Venosafe, Terumo, Belgium) using a 21-gauge needle (Terumo).…”

Section: Preparation Of Normal Pooled Plasmamentioning

confidence: 99%

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The edoxaban‐M4 metabolite and measurement of edoxaban by chromogenic assays in human plasma

Siriez

Yıldız

Bouvy³

et al. 2022

Research and Practice in Thrombosis and Haemostasis

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Introduction:Edoxaban is the only anti-Xa inhibitor metabolized in pharmacologically active moiety that could interfere with chromogenic anti-Xa assays, especially in case of drug-drug interactions or physiological disorders. Materials and methods:We evaluated the contribution of the main metabolite of edoxaban, edoxaban-M4 (M4), in 79 plasma samples from patients taking edoxaban.The total anti-Xa activity was evaluated on three different chromogenic factor Xabased assays. Results were compared with a validated ultra-high-performance liquid chromatography coupled with a tandem mass spectrometry measurement. Edoxaban and its active M4 metabolite have also been spiked separately in normal pooled plasma to assess the sensitivity of chromogenic anti-Xa assays to both molecules individually.Results: Spiked edoxaban or M4 provided different slopes of linear regression models between chromogenic and chromatographic measurement (from 0.97 for STA Liquid Anti-Xa to 1.10 for Biophen Heparin LRT Low with edoxaban and from 0.70 for Biophen DiXaI High to 0.83 for Biophen Heparin LRT High, respectively). A positive correlation is observed between the increase of the ratio M4/edoxaban with the difference between chromogenic and chromatographic measurements. Conclusion:Edoxaban and M4 do not similarly impact chromogenic assays, leading to biased chromogenic estimations of ponderal concentrations. In patient samples, this impact is even more important at low concentrations or in the case of an increase in the M4/edoxaban ratio because of hepatic or renal impairments or in case of drug interactions. This study highlights the limitations and risks of error of expressing results in ponderal concentrations instead of global activity anti-Xa.

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Section: Discussionmentioning

confidence: 99%

Section: Ponderal Concentration or Anti-xa Activity: A Point Of Debatementioning

confidence: 99%

Section: Preparation Of Normal Pooled Plasmamentioning

confidence: 99%

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The edoxaban‐M4 metabolite and measurement of edoxaban by chromogenic assays in human plasma

Siriez

Yıldız

Bouvy³

et al. 2022

Research and Practice in Thrombosis and Haemostasis

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“…This in vitro study was conducted at the hematology laboratory of the CHU UCL Namur. Tests were performed using either commercial normal pooled plasma (NPP; Cryocheck ® , Cryopep, Montpellier, France, batches A1278 & A1291; citrate levels equivalent to 109 mM (due to the margin of tolerance during manufacturing by mixing numerous bags from healthy donors)) or home‐made NPP prepared from healthy volunteers at the University of Namur, as previously described 12 . Plasma aliquots (1 mL aliquots for commercial NPP and 4mL aliquots for home‐made NPP) were stored at −70℃ and thawed for five minutes at 37℃ in a thermostatic bath before use.…”

Section: Methodsmentioning

confidence: 99%

Study of in vitro thrombin generation after neutralization of heparin

Hardy¹,

Douxfils

Morimont

et al. 2021

Int J Lab Hematology

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Introduction Thrombin generation (TG) documents hypercoagulability. TG in platelet‐poor plasma is exquisitely sensitive to heparins, which thus must be neutralized before testing. Heparinase and hexadimethrine bromide (polybrene) have been used for that purpose, but their effects per se on TG have been poorly studied so far. Methods (i) TG was studied in commercial normal pooled plasma (NPP; CryoCheck®, Cryopep) in absence or presence of neutralizing agents. (ii) NPP was spiked with increasing concentrations of unfractionated heparin (UFH; up to 1.0 IU/mL) or low‐molecular‐weight heparin (LMWH; enoxaparin up to 1.2 IU/mL) and TG studied after incubation of heparinase (Hepzyme®; 15 minutes) or polybrene (0.025 mg/mL; 10 minutes). Results (i) With ThromboScreen reagent to initiate TG, addition of heparinase was associated with increased peak, whereas polybrene caused lengthening of lag time and time to peak, compared with nonsupplemented NPP. (ii) With polybrene, TG was completely restored over the whole range of UFH and LMWH studied. By contrast, heparinase failed to fully restore TG in presence of UFH concentrations ≥0.8 IU/mL or LMWH concentrations ≥1.0 IU/mL. Those effects were matched with detectable tiny residual amounts of non‐neutralized heparin (as assessed with an anti‐Xa assay) and were less pronounced with a higher picomolar concentration of tissue factor (DrugScreen reagent). Conclusion Polybrene fully restored TG of heparinized plasma at the expense of an alteration of TG, pointing to the need to use adapted reference ranges. Heparinase failed to do so in presence of high concentrations of both heparins.

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“…Up to now, only a few methods for the qualitative and quantitative determination of DOAC exist. HPLC-MS/MS methods are most commonly used [9][10][11][12] but ultraviolet-visible spectrophotometric methods are also available [13][14][15][16][17]. However, most of these methods determine only a subset of these drugs, which is clearly a downside in an emergency toxicology screening setting.…”

Section: Introductionmentioning

confidence: 99%

Development and validation of an analytical method for the determination of direct oral anticoagulants (DOAC) and the direct thrombin-inhibitor argatroban by HPLC–MS/MS

Brückner

Beyer‐Westendorf

Tiebel

et al. 2021

J Thromb Thrombolysis

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Since direct oral anticoagulants (DOAC) are administered frequently to an elderly, co-morbid population, medical emergencies including trauma, acute bleeding or organ failure are not uncommon. In these situations, the type, dosage or the time of last intake of anticoagulants is often unknown and single substance analysis by functional tests is only possible if the substance contained in the sample is known. A reliable and validated toxicology screen of DOAC and argatroban would be helpful inform not only attending physicians in the emergency department but also law enforcement and courts of justice. After precipitation with acetone, HPLC separation was achieved on a Phenomenex Luna Pentafluorophenyl Colum using acetonitrile–water (90:10, v/v) as mobile phase system. Detection was performed using a 3200 Q Trap mass spectrometer (AB Sciex). For analysis MRM Scans (MS/MS) with positive ionization were chosen. The method was validated for blank serum as the matrix of choice. Limits of detection are between 0.5 and 1.0 ng/mL, limits of quantification are between 1.9 and 3.6 ng/mL and recoveries are above 60%. The applicability of the method was demonstrated by the determination of DOAC in body fluids from forensic cases and in therapeutic drug monitoring. The rapid simultaneous detection and quantification of apixaban, argatroban, dabigatran etexilate, dabigatran, edoxaban and rivaroxaban in body fluids by HPLC–MS/MS closes an important gap in emergency toxicology.

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Importance of measuring pharmacologically active metabolites of edoxaban: development and validation of an ultra-high-performance liquid chromatography coupled with a tandem mass spectrometry method

Cited by 7 publications

References 23 publications

The edoxaban‐M4 metabolite and measurement of edoxaban by chromogenic assays in human plasma

The edoxaban‐M4 metabolite and measurement of edoxaban by chromogenic assays in human plasma

Study of in vitro thrombin generation after neutralization of heparin

Development and validation of an analytical method for the determination of direct oral anticoagulants (DOAC) and the direct thrombin-inhibitor argatroban by HPLC–MS/MS

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