Determination of ng/mL Levetiracetam using Ultra-High-Performance Liquid Chromatography-Photodiode Absorbance

Oláh, Erzsébet; Bacsói, Gy.; Fekete, Judit; Sharma, Virender K.

doi:10.1093/chromsci/bmr053

Cited by 31 publications

(26 citation statements)

References 16 publications

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“…Three sample preparation procedures (liquid–liquid extraction, solid‐phase extraction and protein precipitation with organic solvents) were described by Pucci et al . () and Oläh et al (). In this study methanol was used for protein precipitation and resulted in a clean reliable sample for injection.…”

Section: Introductionmentioning

confidence: 94%

“…Guo et al () and Matar () reported method development for the determination of LEV on liquid chromatography tandem mass spectrometry (LCMS). Ultra‐high performance chromatography (UHPLC) (Oläh et al, ; Kuhn and Knabbe, ) and capillary electrophoresis (Shihabi et al, ) have also been described for measuring LEV in human plasma. Several spectrophotometric methods (Srinivasu et al, ; El‐Yazbi et al, ) have also been reported.…”

Section: Introductionmentioning

confidence: 99%

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A simple and cost‐effective HPLC‐UV method for the detection of levetiracetam in plasma/serum of patients with epilepsy

Engelbrecht

Grobler

Rheeders

2017

Biomedical Chromatography

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A simple, fast and cost-effective method was developed and validated for the determination of levetiracetam (LEV) in plasma/serum of patients using high performance liquid chromatography (HPLC) with ultraviolet detection. The stability of LEV plasma/serum samples over time and in different blood collection tubes was evaluated. Serum/plasma samples were deproteinized by methanol spiked with the internal standard, gabapentin. HPLC was carried out on a Venusil XBP C , 250 × 4.6 mm, 5 μm column, at a flow rate of 1.0 mL/min and with mobile phase consisting of 50 mm potassium dihydrogen phosphate-acetonitrile at a pH of 5.5. The UV detector was set at 205 nm and 10 μL was injected. Total runtime was 15 min. Calibration curves were linear (correlation coefficient = 0.999) over a concentration range of 1-60 μg/mL. Relative standard deviation values for both the inter-day and intra-day precision and accuracy were <5% for the concentration range. The influence of different collection tubes and the effect of time on the stability of LEV was investigated. These factors may cause inaccuracies owing to drug-protein binding and interference in the matrix. This method is simple, fast, cost-effective, reliable and accurate with minimal sample preparation for daily routine use in therapeutic drug monitoring.

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Section: Introductionmentioning

confidence: 94%

Section: Introductionmentioning

confidence: 99%

A simple and cost‐effective HPLC‐UV method for the detection of levetiracetam in plasma/serum of patients with epilepsy

Engelbrecht

Grobler

Rheeders

2017

Biomedical Chromatography

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“…Since only the S-enantiomer has anticonvulsant activity, levetiracetam was developed and is administered as a single pure enantiomer instead of the racemic mixture. Some achiral analytical techniques, including HPLC and GC, have been employed for the routine therapeutic monitoring of levetiracetam in human biological samples comprising plasma, serum, saliva and urine (Pucci et al, 2004;Guo et al, 2007;Matar, 2008;Oláh et al, 2012;Kuhn and Knabbe, 2013). However, to the best of our knowledge, there are only two papers published reporting the development and validation of levetiracetam and its R-enantiomer: one employed a GC technique for quantification of both enantiomers in dog plasma and urine (Isoherranen et al, 2000), while the second one used HPLC for separation and quantification of the unwanted enantiomer (R-enantiomer) in bulk materials and in the final dosage form of levetiracetam (Keppra®, tablets; Rao et al, 2004).…”

Section: Levetiracetammentioning

confidence: 99%

“…In contrast those with lipophilic solvents hamper the levetiracetam extraction owing to the low molecular mass and the chemical structure of the analyte while CN cartridges do not retain levetiracetam and more than 80% of the compound is lost upon loading the sample through the cartridge. Furthermore, it is important to highlight that, for GC-MS analysis, tje solid-phase extraction procedure is preferred in relation to plasma protein precipitation, even though this is the most accurate and simplest procedure for levetiracetam quantification by HPLC-UV techniques (Pucci et al, 2004;Oláh et al, 2012). In contrast, plasma protein precipitation procedures have the disadvantages of diluting the biological sample and decreasing the LLOQ of capillary GC techniques, which have restricted sensitivity owing to the small injection volume.…”

Section: Levetiracetammentioning

confidence: 99%

Chiral chromatographic resolution of antiepileptic drugs and their metabolites: a challenge from the optimization to the application

Fortuna

Alves

2013

Biomedical Chromatography

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A large number of the antiepileptic drugs (AEDs) presently available for clinical practice are chiral compounds while others, although achiral, may originate pharmacologically active chiral metabolites in vivo. The well-known implications of chirality in pharmacokinetics and pharmacodynamics demand the investigation of pharmacological properties for a racemic mixture and each enantiomer. To achieve these objectives, appropriate chiral analytical methods must be available. This article provides the first review of the current state of the art in chiral chromatographic methods available for quantifying enantiomers of AEDs in distinct matrices. Particular attention is paid to the methodological aspects and optimization strategies that successfully allow enantiomeric chromatographic separation of chiral AEDs and/or metabolites. Furthermore, the relevance of these methods in supporting the discovery and development of chiral AEDs is emphasized. In parallel and whenever available, the principal validation parameters are herein considered and related to the stage of drug discovery and development. In an attempt to optimize anticonvulsant activity and simultaneously diminish toxic effects, many pharmaceutical companies have started to manufacture single enantiomers. Therefore, chiral chromatographic techniques will be essential and the information herein compiled can be used as a framework for developing them.

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“…Some of these assay methods however, may require large sample volumes [14], [15], [17], [39], tedious extraction procedures using solid-phase extraction [13], [14], [16], [33] or liquid-liquid extraction [9], [14], [19], [22], [39] or a lengthy chromatographic run time of 10 minutes or longer, for an analysis of a single analyte [9], [16], [17], [19], [22]. Moreover, these assay methods mainly focus on the quantification of LEV, either alone or together with other antiepileptic drugs.…”

Section: Introductionmentioning

confidence: 99%

Rapid and Simultaneous Quantification of Levetiracetam and Its Carboxylic Metabolite in Human Plasma by Liquid Chromatography Tandem Mass Spectrometry

Yeap

2014

PLoS ONE

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A simple liquid chromatography tandem mass spectrometry method was developed and validated according to the guidelines of the US Food and Drug Administration and the European Medicines Agency for a simultaneous quantification of levetiracetam (LEV) and its metabolite, UCB L057 in the plasma of patients. A 0.050 mL plasma sample was prepared by a simple and direct protein precipitation with 0.450 mL acetonitrile (ACN) containing 1 µg/mL of internal standard (IS, diphenhydramine), then vortex mixed and centrifuged. A 0.100 mL of the clear supernatant was diluted with 0.400 mL water and well mixed. A 0.010 mL of the resultant solution was injected into an Agilent Zorbax SB-C18 (2.1 mm×100 mm, 3.5 µm) column with an isocratic elution at 0.5 mL/min using a mixture of 0.1% formic acid in water and ACN (40∶60 v/v). Detection was performed using an AB Sciex API 3000 triple quadrupole mass spectrometer, equipped with a Turbo Ion Spray source, operating in a positive mode: LEV at transition 171.1>154.1, UCB L057 at 172.5>126.1, and IS at 256.3>167.3; with an assay run time of 2 minutes. The lower limit of quantification (LLOQ) for both LEV and UCB L057 was validated at 0.5 µg/mL, while their lower limit of detection (LOD) was 0.25 µg/mL. The calibration curves were linear between 0.5 and 100 µg/mL for both analytes. The inaccuracy and imprecision of both intra-assay and inter-assay were less than 10%. Matrix effects were consistent between sources of plasma and the recoveries of all compounds were between 100% and 110%. Stability was established under various storage and processing conditions. The carryovers from both LEV and UCB L057 were less than 6% of the LLOQ and 0.13% of the IS. This assay method has been successfully applied to a population pharmacokinetic study of LEV in patients with epilepsy.

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Determination of ng/mL Levetiracetam using Ultra-High-Performance Liquid Chromatography-Photodiode Absorbance

Cited by 31 publications

References 16 publications

A simple and cost‐effective HPLC‐UV method for the detection of levetiracetam in plasma/serum of patients with epilepsy

A simple and cost‐effective HPLC‐UV method for the detection of levetiracetam in plasma/serum of patients with epilepsy

Chiral chromatographic resolution of antiepileptic drugs and their metabolites: a challenge from the optimization to the application

Rapid and Simultaneous Quantification of Levetiracetam and Its Carboxylic Metabolite in Human Plasma by Liquid Chromatography Tandem Mass Spectrometry

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