Quantification Of Carvedilol Research Articles

Simultaneous Determination of Carvedilol, Enalaprilat, and Perindoprilat in Human Plasma Using LC\u2013MS/MS and Its Application to a Pharmacokinetic Pilot Study

A method for the extraction and quantification of carvedilol, enalaprilat, and perindoprilat in 50 µL human plasma, using high-performance liquid chromatography with tandem mass spectrometry (LC–MS/MS) detection was developed and validated. Samples were prepared via protein precipitation with chromatographic separation on a Restek Ultra II Biphenyl column using gradient elution at a corresponding flowrate of 300 µL/min. Electrospray ionisation with mass detection at unit resolution in the multiple reaction monitoring (MRM) mode on an AB Sciex API 5500 mass spectrometer was used. Accuracy, precision, selectivity, sensitivity, matrix effects, recovery, process efficiency, and stability were assessed over the validation period. The assay was validated over the calibration range 0.2–200 ng/mL for all three analytes. The inter- and intra-day precision expressed as the coefficient of variation (CV) and accuracy (%Nom) all fell within acceptable limits. The overall recovery was calculated as 72.9%, 77.1%, and 77.0% for carvedilol, enalaprilat, and perindoprilat respectively, with the recovery being shown to be reproducible at the low, medium and high end of the calibration range for all three analytes. The method proved to be specific for all three analytes with no significant matrix effects observed. The validated method facilitated the analysis of carvedilol, enalaprilat, and perindoprilat in human plasma collected from adults as part of a pilot pharmacokinetic study. This validated analytical method lays the foundation for determining adherence in heart failure patients prescribed with carvedilol, enalapril and perindopril.

Development and Validation of a Rapid Analytical Method for the Simultaneous Quantification of Metabolic Syndrome Drugs by HPLC-DAD Chromatography

Worldwide, 25% of the population suffers from metabolic syndrome (MetS). The treatment of patients with MetS regularly includes drugs prescribed simultaneously to treat several disorders that manifest at the same time, such as hypercholesterolemia, arterial hypertension, and diabetes. To the authors’ best knowledge, there is no previous published analytical method for the simultaneous quantification of drugs used in the treatment of these diseases. In the present study, a rapid high-performance liquid chromatography with a diode-array detector HPLC-DAD methodology was developed for simultaneous quantification of carvedilol (CVD), telmisartan (TEL), bezafibrate (BZT), gliclazide (GZD), and glimepiride (GMP) in bulk and pharmaceutical form. The chromatographic separation of the five pharmaceuticals was achieved on a Hypersil GOLD C18 Selectivity (5 µm, 150 × 4.60 mm2) using a mobile phase of acetonitrile (50%) and 0.02 M KH2PO4, pH 3 (50%) at a flow rate of 1 mL/min and at 25 °C. The total separation time was 9 min. The analytical method was validated following the International Conference on Harmonization guidelines. A reproducible method was obtained with acceptable limits of detection (LOD) and quantification (LOQ) for CVD (0.012 and 0.035 μg mL−1), TEL (0.103 and 0.313 μg mL−1), BZT (0.025 and 0.076 μg mL−1), GZD (0.039 and 0.117 μg mL−1), and GMP (0.064 and 0.127 μg mL−1). The validated method allowed the determination of these drugs in commercial pharmaceutical products both individually and simultaneously. The present method was found to be suitable for simultaneous quantification of the five drugs that are most commonly used in the simultaneous treatment of the metabolic syndrome.

Development and validation of UPLC method for simultaneous quantification of carvedilol and ivabradine in the presence of degradation products using DoE concept

ABSTRACTA methodical design-of-experiments were performed by applying quality-by-design concepts to establish a design-space for simultaneous and rapid quantification of Carvedilol and Ivabradine by UPLC in the presence of degradation products. Response-surface, central-composite design, and quadratic model were employed for statistical assessment of experimental data using the Design-Expert software. Response variables such as resolution and retention time were analyzed statistically for chromatographic screening. During DoE study, various plots such as perturbation, contour, 3D and design-space plots were considered for method optimization. The method was developed using C8 [100 × 2.1 mm, 1.8 µ] UPLC column, mobile phase comprising 0.5% triethylamine buffer [pH 6.4] and acetonitrile in the ratio of 50:50 v/v, the flow rate of 0.4 mL minute−1 and UV detection at 285 nm for both Carvedilol and Ivabradine. The method was developed with a short run time of two minutes. The method was found to be linear in the range of 25.0–199.9 µg mL−1 and 8.9–21.3 µg mL−1 for Carvedilol and Ivabradine, respectively with a correlation coefficient of 0.9998 in each case. The recovery values were found in the range of 99.7–100.8% and 98.9–100.9% for Carvedilol and Ivabradine, respectively. The method was validated according to ICH Q2 (R1) guidelines.

HPLC/Fluorometric Detection of Carvedilol in Real Human Plasma Samples Using Liquid-Liquid Extraction.

A simple, rapid and sensitive high-performance liquid chromatography (HPLC) method has been developed to quantify carvedilol in human plasma using an isocratic system with fluorescence detection. The method included a single-step liquid-liquid extraction with diethylether and ethylacetate mixture (3 : 1, v/v). HPLC separation was carried out by reversed-phase chromatography with a mobile phase composed of 20 mM phosphate buffer (pH 7)-acetonitrile (65 : 35, v/v), pumped at a flow rate of 1.0 mL/min. Fluorescence detection was performed at 240 nm (excitation) and 330 nm (emission). The calibration curve for carvedilol was linear from 10 to 250 ng/mL. Intra- and interday precision values for carvedilol in human plasma were <4.93%, and accuracy (relative error) was better than 4.71%. The analytical recovery of carvedilol from human plasma averaged out to 91.8%. The limits of detection and quantification of carvedilol were 3.0 and 10 ng/mL, respectively. Also, the method was successfully applied to three patients with hypertension who had been given an oral tablet of 25 mg carvedilol.

Application of UPLC-MS/MS for separation and quantification of carvedilol and 4-hydroxyphenyl carvedilol and comparative bioavailability of two carvedilol formulations in healthy volunteers

Application of UPLC-MS/MS for separation and quantification of carvedilol and 4-hydroxyphenyl carvedilol and comparative bioavailability of two carvedilol formulations in healthy volunteers

Determination of Carvedilol in Human Plasma by Gas Chromatography-Mass Spectrometry Method

An analytical procedure was developed and validated for the quantitation of carvedilol in human plasma. Carvedilol and atenolol (internal standard) were extracted from human plasma with a mixture of diethylether and ethylacetate at basic pH with liquid‐liquid extraction. The extracts were derivatized with nMethyl-n-(trimethylsilyl)trifluoroacetamide and analyzed by gas chromatography-mass spectrometry (GC‐MS). Calibration curves were linear over the concentration range 15‐500 ng/mL. Intra- and inter-day precision, expressed as the relative standard deviation were less than 8.0%, and accuracy (relative error) was better than 11.0%. The limits of detection and quantification of carvedilol were 5.0 and 15 ng/mL, respectively. Also, the method was successfully applied to three patients with hypertension who had been given an oral tablet of 25 mg carvedilol.

Quantification of carvedilol in human plasma by liquid chromatography using fluorescence detection: Application in pharmacokinetic studies

A simple, rapid and sensitive isocratic reversed-phase HPLC method with fluorescence detection using a monolithic column has been developed and validated for the determination of carvedilol in human plasma. The separation was performed on a Chromolith Performance (RP-18e, 100 mm × 4.6 mm) column with an isocratic mobile phase consisting of 0.01 M disodium hydrogen phosphate buffer–acetonitrile (40:60, v/v) adjusted to pH 3.5. The sample preparation involves protein precipitation procedure and analytical recovery was complete. Letrozole was used as internal standard. The assay enables the measurement of carvedilol for therapeutic drug monitoring with a minimum quantification limit (LOQ) of 1 ng ml −1. The excitation and emission wavelengths were set at 240 and 340 nm, respectively. The calibration curve was linear over the concentration range 1–80 ng ml −1. The coefficients of variation for inter-day and intra-day assay were found to be less than 8.0%.

Quantification of carvedilol in human plasma by high-performance liquid chromatography coupled to electrospray tandem mass spectrometry: Application to bioequivalence study

A rapid, sensitive and specific method to quantify carvedilol in human plasma using metoprolol as the internal standard (IS) is described. The analyte and the IS were extracted from plasma by liquid–liquid extraction using a diethyl-ether solvent. After removed and dried the organic phase, the extracts were reconstituted with a fixed volume of acetonitrile–water (50/50; v/v). The extracts were analyzed by a high performance liquid chromatography coupled to electrospray tandem mass spectrometry (HPLC–MS/MS). Chromatography was performed isocratically on Alltech Prevail C 18 5 μm analytical column, (150 mm × 4.6 mm i.d.). The method had a chromatographic run time of 3.5 min and a linear calibration curve over the range 0.1–200 ng ml −1 ( r 2 > 0.997992). The limit of quantification was 0.1 ng ml −1. This HPLC–MS/MS procedure was used to assess the bioequivalence of two carvedilol 25 mg tablet formulations (carvedilol test formulation from Laboratórios Biosintética Ltda and Coreg ® from Roche Químicos e Farmacêuticos S.A standard reference formulation). A single 25 mg dose of each formulation was administered to healthy volunteers. The study was conducted using an open, randomized, two-period crossover design with a 2-week wash-out interval. Since the 90% CI for C max and AUCs ratios were all inside the 80–125% interval proposed by the US Food and Drug Administration Agency, it was concluded that carvedilol formulation elaborated by Laboratórios Biosintética Ltda is bioequivalent to Coreg ® formulation for both the rate and the extent of absorption.

HPLC quantification of carvedilol in small plasma volumes from children

A sensitive high-performance liquid chromatographic method has been developed for the determination of the β-receptor blocker carvedilol in small volumes of plasma from pediatric patients. Carvedilol and the internal standard naftopidil were extracted from only 300 μL plasma with solid-phase extraction columns and eluted with a mixture of 80% (v/v) acetonitrile and 20% (v/v) potassium acetate buffer (0.1 M, pH 5). Analysis of the extracts was performed on a Spherisorb C6 column with a mobile phase of 65% acetonitrile, 35% potassium acetate buffer (0.25 M, pH 4) and fluorescence detection. Carvedilol and the internal standard showed recoveries of 87.0±13.9% and 97.7±18.0% respectively (n=75, each; values are means±standard deviations). The analysis of carvedilol was linear over a range of 0.40–242 ng mL−1, the limit of quantification was 0.40 ng mL−1. As an application of the assay, a profile of plasma concentrations versus time of a six-month-old patient is reported.