Development and validation of LC–MSMS assay for the determination of the prodrug dabigatran etexilate and its active metabolites in human plasma

Nouman, Eman G.; Al‐Ghobashy, Medhat A.; Lotfy, Hayam M.

doi:10.1016/j.jchromb.2015.02.042

Cited by 33 publications

(23 citation statements)

References 23 publications

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“…In contrast to the present study, Tan et al used nine individual samples for frozen long-term stability, 15 individual samples for short-term stability, and three pooled samples for short-term stability, autosampler stability, and freeze-thaw stability [13]. Nouman et al used one pooled sample that was prepared by pooling samples from a subject at multiple sampling time points [14]. Because an acceptable number of samples for ISS tests can vary for practical reasons, it is advisable to determine the number on a case-by-case basis.…”

Section: Issmentioning

confidence: 89%

“…To my knowledge, few studies [13,14] have considered ISS in the presence of AG. Studies of ISS under general situation (i.e.…”

Section: Discussionmentioning

confidence: 99%

“…Because the test items in the present study included stability during sample collection and extraction, the reference value was acquired from samples for which the times required for sample collection and extraction were minimized as much as possible. This experimental procedure was not employed in studies by either Tan et al [13] or Nouman et al [14], possibly because human incurred samples were used, and because it is generally difficult in a clinical setting to start an analysis immediately after blood collection. Sample H (positive control) helped qualify the use of Sample A as a reference value, because the ASP3258 concentration of Sample H was higher than that of Sample A, suggesting that ASP3258-AG was not completely converted to ASP3258 in Sample A.…”

Section: Issmentioning

confidence: 99%

“…Because an acceptable number of samples for ISS tests can vary for practical reasons, it is advisable to determine the number on a case-by-case basis. Tan et al and Nouman et al chose samples to cover multiple sampling time points [13,14], likely because they planned to use their methods to evaluate full pharmacokinetic profiles. When both Cmax and elimination phase are important in a pharmacokinetic study, it is reasonable to select samples accordingly, because the abundance of AG relative to the analyte can vary depending on the sampling times and the ISS results therefore can change as well.…”

Section: Issmentioning

confidence: 99%

“…Given that GBC [5], the 7th WRIB [10], and 7th GCC [6] recommended evaluating ISS when metabolites are likely to affect assay results, it is important to discuss the test items and study design of ISS experiments in the presence of AG. Unfortunately, to my knowledge, ISS in the presence of AG has been addressed by only two case studies [13,14]. Further, both test items and the design of the two studies were inconclusive.…”

Section: Introductionmentioning

confidence: 96%

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Incurred sample stability of ASP3258 in the presence of its acyl glucuronide

Ohtsu¹

2017

J Appl Bioanal

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The acyl glucuronide (AG) of ASP3258 was abundant in incurred monkey samples, and because an AG generally back-converts to its aglycone, accurate quantification of ASP3258 in monkey plasma was a challenge. To prevent the back-conversion of ASP3258-AG, cooling and acidification were incorporated during sample collection, storage, and extraction. To demonstrate that the AG did not affect the determination of ASP3258, the present study used incurred samples to examine whole blood stability, short-term stability, freeze-thaw stability, frozen stability, and stability during extraction. The concentration changes were within −11.4% to 15.0% compared with a reference value and were therefore judged acceptable. The present study presents a detailed account of test items, a reference value, sample numbers, sample selection, and an equation for assessment in the incurred sample stability tests. This bioanalytical method was applied successfully to a study of the toxicokinetics of ASP3258 in monkeys.Keywords: incurred sample stability, acyl glucuronide, ASP3258, animal plasma, toxicokinetics. IntroductionThe potent phosphodiesterase 4 inhibitor ASP3258 (Figure 1) is a novel therapeutic agent for asthma and chronic obstructive pulmonary disease [1]. We previously developed and validated a bioanalytical method to quantify ASP3258 in rat plasma [2]. Because ASP3258 metabolite was not detected in rat plasma [3], this bioanalytical method was developed without encountering issues associated with the metabolite. Subsequently, monkey toxicity studies were planned that required an appropriate bioanalytical method. When ASP3258 was orally administered to monkeys in a preliminary study, the acyl glucuronide (AG) metabolite of ASP3258 was detected in abundance in monkey plasma (data on file), in contrast to our findings using rat plasma [3]. In general, AGs readily back-convert to aglycones, causing overestimation of

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Section: Issmentioning

confidence: 89%

“…To my knowledge, few studies [13,14] have considered ISS in the presence of AG. Studies of ISS under general situation (i.e.…”

Section: Discussionmentioning

confidence: 99%

Section: Issmentioning

confidence: 99%

Section: Issmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 96%

See 3 more Smart Citations

Incurred sample stability of ASP3258 in the presence of its acyl glucuronide

Ohtsu¹

2017

J Appl Bioanal

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A synergy of liquid chromatography with high‐resolution mass spectrometry and coagulation test for determination of direct oral anticoagulants for clinical and toxicological purposes

Maier

Slavík

Ondra

2021

Biomedical Chromatography

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Direct oral anticoagulants are an alternative to anticoagulants based on vitamin K antagonists. Monitoring of direct oral anticoagulant concentration levels is necessary in specific cases (e.g. in emergency conditions, for determination of the cause of bleeding, adverse effects, risk of drug-direct oral anticoagulants interaction); therefore, a sensitive and specific method is needed. A methanol protein precipitation method followed by liquid chromatography with high-resolution mass spectrometry was developed for simultaneous separation and determination of apixaban, betrixaban, edoxaban, dabigatran, rivaroxaban and ximelagatran. The proposed method was fully validated in terms of linearity, the limits of detection and quantification, intra-and inter-day trueness and precision, recovery, matrix effect, process efficiency and stability. The method shows a strong correlation (Pearson's correlation coefficients > 0.92) with coagulation assays of apixaban, dabigatran and rivaroxaban (dilute thrombin time for gatrans and anti Xa factor (anti-Xa) activity for xabans). In addition, the developed method was applied for the identification and determination of apixaban and dabigatran in post-mortem serum samples. The developed method is a good alternative to coagulation tests which may show various interferences.

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Development and validation of a ultraperformance liquid chromatography–tandem mass spectrometry method for quantifying free and total dabigatran in human plasma: An application for a bioequivalence study

Abd-Allah

Almrasy

Abdelhmaid³

et al. 2022

Biomedical Chromatography

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Dabigatran etexilate mesylate (DABE), a prodrug, quickly changes into dabigatran (DAB) after its oral administration. Accordingly, detecting DABE in plasma is practically unmanageable. An ultraperformance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) technique was developed and validated to compute free DAB in participants. For the first time, the central composite design, a type of response surface methodology, was applied for optimizing variables affecting the cleavage of glucuronide bond. In addition, the pharmacokinetic parameters of generic medication (okanadab) were determined, and the obtained outcomes were compared with those of the branded drug (pradaxa). The sample preparation was done using methanol as a protein precipitant and the separation was achieved using an ACQUITY UPLC BEH C18 column (2.1 × 50 mm, 1.7 μm). The elution was isocratically conducted using 10 mM ammonium formate:methanol (72:28, v/v) as a mobile phase and the flow rate was 0.25 mL/min. Multiple reaction monitoring and positive electrospray ionization were used. The determination was performed within 1 min, and the calibration growth curve was established over a range of 1.19–475 ng/mL using DAB‐d3 as a tagged internal standard. Bioequivalence research was validated following the US Food and Drug Administration (US FDA) guidelines for bioanalytical procedures and acceptable outcomes were achieved. The outcomes for okanadab and pradaxa did not differ significantly.

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Development and validation of LC–MSMS assay for the determination of the prodrug dabigatran etexilate and its active metabolites in human plasma

Cited by 33 publications

References 23 publications

Incurred sample stability of ASP3258 in the presence of its acyl glucuronide

Incurred sample stability of ASP3258 in the presence of its acyl glucuronide

A synergy of liquid chromatography with high‐resolution mass spectrometry and coagulation test for determination of direct oral anticoagulants for clinical and toxicological purposes

Development and validation of a ultraperformance liquid chromatography–tandem mass spectrometry method for quantifying free and total dabigatran in human plasma: An application for a bioequivalence study

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