scholarly journals Determination of Clobazam and Its Major Metabolite N-desmethylclobazam in Human Plasma with High-Performance Liquid Chromatography

Analytica ◽  
2021 ◽  
Vol 2 (3) ◽  
pp. 57-65
Author(s):  
Fadumo Ahmed Isse ◽  
Sherif Hanafy Mahmoud
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Human Plasma ◽  
High Performance ◽  
Practical Method ◽  
Therapeutic Drug ◽  
Reverse Phase ◽  
Simple Method ◽  
Metabolite Concentrations

Clobazam (CLB) is a benzodiazepine that is used in many types of epilepsy. Although therapeutic drug monitoring (TDM) of CLB is not routine, there is evidence that TDM may be of value in conditions where pharmacokinetic alterations are suspected. Therefore, determination of both CLB and its active metabolite concentrations is essential for TDM. Herein, we present a simple and practical method for determination of CLB and N-desmethylclobazam (NDMCLB) in human plasma by high-performance liquid chromatography (HPLC). The drugs were extracted by hexane:dichloromethane (1:1, v/v) from 0.3 mL plasma. The separation was carried out with a C18 reverse phase column using a mobile phase of water:acetonitrile (57:43, v/v) pumped at 0.8 mL/min. The analytes were detected at 228 nm. The method was linear over the concentration range 20–500 ng/mL for CLB and 200–3000 ng/mL for NDMCLB. The intra-day coefficient of variation (CV) was <10% for CLB and <6% for NDMCLB, while the inter-day CV for CLB was <16%. The metabolite inter-day CV was <6%. The accuracy of intra- and inter-day assessments determined for CLB and NDMCLB was within ±10%. This paper describes a rapid, reliable, and simple method for measuring CLB and its metabolite NDMCLB in human plasma. This UV-HPLC procedure offers acceptable precision and accuracy to quantify CLB and its metabolite in human plasma.

scholarly journals Simultaneous Determination of Eight β-Lactam Antibiotics, Amoxicillin, Cefazolin, Cefepime, Cefotaxime, Ceftazidime, Cloxacillin, Oxacillin, and Piperacillin, in Human Plasma by Using Ultra-High-Performance Liquid Chromatography with Ultraviolet Detection

2016 ◽  
Vol 60 (8) ◽  
pp. 4734-4742 ◽  
Author(s):  
Tiphaine Legrand ◽  
Dominique Vodovar ◽  
Nicolas Tournier ◽  
Nihel Khoudour ◽  
Anne Hulin
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Simultaneous Determination ◽  
Human Plasma ◽  
High Performance ◽  
Internal Standard ◽  
Therapeutic Drug ◽  
Simple Method ◽  
Validation Parameters

ABSTRACTA simple and rapid ultra-high-performance liquid chromatography (UHPLC) method using UV detection was developed for the simultaneous determination of eight β-lactam antibiotics in human plasma, including four penicillins, amoxicillin (AMX), cloxacillin (CLX), oxacillin (OXA), and piperacillin (PIP), and four cephalosporins, cefazolin (CFZ), cefepime (FEP), cefotaxime (CTX), and ceftazidime (CAZ). One hundred-microliter samples were spiked with thiopental as an internal standard, and proteins were precipitated by acetonitrile containing 0.1% formic acid. Separation was achieved on a pentafluorophenyl (PFP) column with a mobile phase composed of phosphoric acid (10 mM) and acetonitrile in gradient elution mode at a flow rate of 500 μl/min. Detection was performed at 230 nm for AMX, CLX, OXA, and PIP and 260 nm for CFZ, FEP, CTX, and CAZ. The total analysis time did not exceed 13 min. The method was found to be linear at concentrations ranging from 2 to 100 mg/liter for each compound, and all validation parameters fulfilled international requirements. Between- and within-run accuracy errors ranged from −5.2% to 11.4%, and precision was lower than 14.2%. This simple method requires small-volume samples and can easily be implemented in most clinical laboratories to promote the therapeutic drug monitoring of β-lactam antibiotics. The simultaneous determination of several antibiotics considerably reduces the time to results for clinicians, which may improve treatment efficiency, especially in critically ill patients.

scholarly journals Ultra High Performance Liquid Chromatography Method for the Determination of Two Recently FDA Approved TKIs in Human Plasma Using Diode Array Detection

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Marwa Fouad ◽  
Maxime Helvenstein ◽  
Bertrand Blankert
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Tyrosine Kinase ◽  
Human Plasma ◽  
Tyrosine Kinase Inhibitors ◽  
High Performance ◽  
Kinase Inhibitors ◽  
Therapeutic Window ◽  
Therapeutic Drug

Generally, tyrosine kinase inhibitors have narrow therapeutic window and large interpatient variability compared to intrapatient variability. In order to support its therapeutic drug monitoring, two fast and accurate methods were developed for the determination of recently FDA approved anticancer tyrosine kinase inhibitors, afatinib and ibrutinib, in human plasma using ultra high performance liquid chromatography coupled to PDA detection. Diclofenac sodium was used as internal standard. The chromatographic separation was achieved on an Acquity UPLC BEH C18 analytical column using a mobile phase combining ammonium formate buffer and acetonitrile at a constant flow rate of 0.4 mL/min using gradient elution mode. AµSPE (solid phase extraction) procedure, using Oasis MCXµElution plates, was processed and it gave satisfying and reproducible results in terms of extraction yields. Additionally, the methods were successfully validated using the accuracy profiles approach (β= 95% and acceptance limits = ±15%) over the ranges 5–250 ng/mL for afatinib and from 5 to 400 ng/mL for ibrutinib in human plasma.

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