Amlodipine In Human Plasma Research Articles

Development and Validation of a Robust LC-MS-MS Method for the Quantification of Amlodipine in Human Plasma

Development and Validation of a Robust LC-MS-MS Method for the Quantification of Amlodipine in Human Plasma

Development and Application of HPLC and UFLC Methods for the Analysis of Cardiovascular Drugs

Separation and identification of amiloride, metoprolol, hydrochlorothiazide, Carvedilol and amlodipine, in human plasma using the presented SPE and HPLC procedures was selective, efficient, robust, economical, environmentally friendly, and repeatable. Because plasma samples showed no evidence of a secondary peak, it was determined that the claimed drug-drug interaction between cardiovascular medications did not occur. Furthermore, no new peak was generated, indicating no metabolic product. This is the first report on isolating and identifying these nine cardiovascular medicines. Thus, SPE and HPLC techniques may be used to investigate these 5 cardiovascular medications. Successful monitoring of these medications in human plasma was achieved using the established SPE and HPLC technologies. The results of this inquiry were contrasted with those that had already been released. In comparison to other works in the literature, it was determined that the current study had the best complete baseline separation and lowest detection limit. The retention, separation, and resolution factors were discovered to have ranges of 0.07-9.14, 1.44-4.21, and 2.15-18.66, respectively. The SPE and UFLC techniques created and verified for this purpose were used to examine human plasma samples for the presence of cardiovascular drugs. Peak retention, separation, resolution, and symmetry all matched the reference samples' values. The results for symmetry, separation, and resolution were all in agreement with the reference samples. The methods used to separate and identify amiloride, metoprolol, hydrochlorothiazide; Carvedilol and Amlodipine in human plasma were found to be selective, effective, hardy, affordable, environmentally friendly, and reproducible, according to the authors. The selectivity of the SPE technique was validated by the absence of any secondary signal in the plasma samples. Additionally, there was no evidence of these drugs degrading in the plasma sample. This is the initial report on the simultaneous isolation and identification of these eight medications. The well-known SPE and UFLC techniques were successfully used to monitor these drugs in human plasma. Therefore, these drugs can be detected in any plasma sample using SPE or UFLC methods

Reversed phase HPLC method development and validation for the analysis of amlodipine besylate in tablets dosage form and human plasma

Introduction: Several analytical methods for quantitative determination of amlodipine either in pharmaceuticals or biological fluids have suffered different setbacks. Therefore, we aimed at developing a cost effective, simple, robust, and sensitive reversed phase high performance liquid chromatography (RP-HPLC- UV) method for quantification of amlodipine in dosage form and in human plasma. Method: The separation was performed on an analytical column 150 mm x 4.6 mm i.d., Agilent Eclipse XDB C18 Column packed with 5 μm particle size and protected by a guard column (30 mm x 4.6 mm i.d.). The mobile phase composition contained acetonitrile: phosphate buffer (25 mM KH2PO4) adjusted to pH 3.1 with orthophosphoric acid in the ratio of 45:55 v/v. The flow rate was 1.2 mL/min and the UV detector was set at a wavelength of 240 nm. The column was maintained at ambient temperature. Results: The calibration curve gave good linearity in the concentration range studied. The column efficiency was highly evident by high theoretical plates and small height equivalent to theoretical plate (HETP). The percent relative standard deviation (% RSD) was less than 3% in the inter- and intra-day o assay. Amlodipine, 50 ng/mL in plasma stored frozen at -80 C was stable There was no statistical difference in the precision of amlodipine assayed after 12 months when compared to the initial assay (P>0.05). The % recovery of amlodipine in human plasma was 97.79±0.17% to 99.52±0.63%. Conclusion: The new RP-HPLC method is sensitive, reproducible, precise, accurate and robust. We conclude that the method is suitable for estimation of amlodipine besylate in human plasma for pharmacokinetic studies and assay of pharmaceutical dosage form

DEVELOPMENT OF CAPILLARY ELECTROPHORESIS METHOD FOR DETERMINATION OF AMLODIPINE IN HUMAN PLASMA

Abstract: Background: Amlodipine, a calcium channel blocker, is commonly prescribed to treat high blood pressure. In Vietnam, there is no method of quantifying amlodipine in plasma by capillary electrophoresis. Therefore, the thesis was carried out with the objective: Developing and validating a method for quantifying amlodipine in human plasma by capillary electrophoresis. Materials and methods: Human plasma samples contain amlodipine. After optimizing the process, the method was validated according to the guidelines for validation of bioanalytical methods of US-FDA and EMA. Results: Amlodipine and nifedipine as internal standard (IS) were alkalized with sodium hydroxide and extracted from human plasma by liquid - liquid extraction technique with ethyl acetate. Electrophoresis conditions: Fused-silica capillary column (effective column length: 48.5 cm, total column length 57.5 cm, outside diameter: 75 μm, internal diameter: 75 μm), background electrolyte: 50 mM borate buffer at pH 8.6 containing 20 mM sodium dodecyl sulfate, voltage: +25 kV, current: 300 μA, column temperature: 25 ± 1 oC, sample injection: 35 mBar - 7s, detection at 237 nm. The method was validated and met US-FDA and EMA requirements. Conclusion: The method can serve as a basis in bioequivalence studies of medicinal products containing amlodipine. Key words: Amlodipin, human plasma, capillary electrophoresis

Development and Validation of Liquid Chromatography - Tandem Mass Spectrometry Method for Determination of Amlodipine in Human Plasma and Its Application

Development and Validation of Liquid Chromatography - Tandem Mass Spectrometry Method for Determination of Amlodipine in Human Plasma and Its Application

Krawtchouk image moment method for the simultaneous determination of three drugs in human plasma based on fluorescence three-dimensional spectra

The interference signals and overlapped peaks are common phenomena in fluorescence determination, which seriously influence the accuracy and reliability of analytical results. In this paper, Krawtchouk image moment method was introduced to the analysis of fluorescence three-dimensional (3D) spectra and applied to the quantitative analysis of three drugs including nicotinic acid, metoprolol and amlodipine in human plasma. Without any pretreatment to the obtained spectra, Krawtchouk moments were directly calculated on the grayscale images of 3D spectra, and the quantitative linear models for the three drugs were established by stepwise regression, respectively. The determination coefficients (R) were more than 0.9906. The correlation coefficients of leave-one-out cross-validation (Rcv) were more than 0.9256. The precisions (RSD, %) of inter-day and intra-day variations were less than 8.4% and 3.1%, separately. The recovery was from 101.84% to 107.88%. The limit of quantification and the limit of detection were 0.12μgmL−1 and 0.43μgmL−1, respectively. All the statistical parameters supported that the obtained models performed well and the proposed method was accurate and reliable. Our study indicates that Krawtchouk image moments with the powerful abilities of multi-resolution and extracting local information can be applied to the simultaneous determination of multi-target compounds in plasma based on fluorescence 3D spectra.

Determination of amlodipine in human plasma by liquid chromatography–tandem mass spectrometry and its application to pharmacokinetic & bioequivalence studies

Determination of amlodipine in human plasma by liquid chromatography–tandem mass spectrometry and its application to pharmacokinetic & bioequivalence studies

Determination of amlodipine in human plasma by liquid chromatography–tandem mass spectrometry and its application to pharmacokinetic & bioequivalence studies

Determination of amlodipine in human plasma by liquid chromatography–tandem mass spectrometry and its application to pharmacokinetic & bioequivalence studies

Simultaneous determination of rosuvastatin and amlodipine in human plasma using tandem mass spectrometry: Application to disposition kinetics

The liquid chromatography–tandem mass spectrometric assay method for the simultaneous determination of rosuvastatin and amlodipine in human plasma using deuterated analogs as internal standards has been developed and validated. The analytes were extracted from 100μL aliquots of human plasma via liquid–liquid extraction using a mixture of ethyl acetate and n-hexane (80:20, v/v) as an extraction solvent. The optimized mobile phase was composed of 0.1% formic acid in 5mM ammonium acetate, methanol, and acetonitrile (20:20:60, v/v/v) and delivered at a flow rate of 0.75mL/min. The calibration curve obtained was linear (R2⩾0.999) over the concentration range of 0.52–51.77ng/mL for rosuvastatin and 0.10–10.07ng/mL for amlodipine. A sample turnover rate of less than 2.5min makes it an attractive procedure in high-throughput bioanalysis of rosuvastatin and amlodipine. The present method was found to be applicable to clinical studies and the results were authenticated by incurred sample reanalysis.

Development and validation of a HPLC-PDA bioanalytical method for the simultaneous estimation of Aliskiren and Amlodipine in human plasma.

A simple, unique and selective HPLC-PDA method was developed and validated for the simultaneous estimation of aliskiren (ALS) and amlodipine (AML) in human plasma. Extraction of the sample was accomplished by protein precipitation. Plasma proteins were precipitated by employing acetonitrile containing hydrochlorothiazide as internal standard. The compounds were analyzed by HPLC by using PDA detector on a Hibar C18 (250 × 4.6 mm) column with a mobile phase comprising acetonitrile and phosphate buffer (pH 4.2 and 25 mm; 60:40 v/v) with a flow rate of 0.8 mL/min. Different sample pretreatment techniques were evaluated but protein precipitation was found to be satisfactory, offering good recovery values of 97.11-98.45% for ALS and 97.5-99.12% for AML. The within-day precisions for ALS were 96.66, 99.16 and 99.41% at 90, 240 and 480 ng/mL, respectively, and for AML they were 97.27, 99.54 and 99.31% at 3.3, 8.8 and 17.6 ng/mL, respectively. The between-day precisions for ALS were 96.66, 99.16 and 99.41% at 90, 240 and 480 ng/mL, respectively and the between-day precisions for AML were 98.18, 99.20 and 99.40% at 3.3, 8.8 and 17.6 ng/mL, respectively. The limit of quantitation was 30 and 1.0 ng/mL for ALS and AML respectively. Different constituents of plasma proteins did not interfere with the absolute recovery of ALS and AML.

Simultaneous determination of olmesartan and amlodipine in human plasma and urine by ultra performance liquid chromatography tandem mass spectrometry

A sensitive and rapid ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed to determine olmesartan and amlodipine levels in human plasma and urine simultaneously. Chromatographic separation was carried out on an Acquity UPLC BEH C18 column and mass spectrometric analysis was performed using a QTrap5500 mass spectrometer coupled with an electro-spray ionization (ESI) source in the positive ion mode. The MRM transitions of m/z 447→207 and 409→238 were used to quantify olmesartan and amlodipine, respectively. This assay method has been fully validated in terms of selectivity, linearity, lower limit of quantification (LLOQ), accuracy, precision, stability, matrix effect and recovery. The linearity of this method was found to be within the concentration range of 0.2-500ng/mL and 4-5000ng/mL for olmesartan in human plasma and urine and 0.1-50ng/mL and 2-1000ng/mL for amlodipine in human plasma and urine. Only 2min were needed for an analytical run. This assay was used to support a clinical study where multiple oral doses were administered to healthy Chinese subjects to investigate the pharmacokinetics of olmesartan and amlodipine.

Determination of Amlodipine in Human Plasma by LC-MS/MS and Its Bioequivalence Study in Healthy Chinese Subjects

A sensitive and reproducible liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of amlodipine in human plasma, with gliclazide as an internal standard (IS). The analyte was extracted with ethyl acetate and analyzed on a Diamond C18 (150 mm× 4.6 mm, 5 μm) column. The mobile phase was composed of methanol ?10 mM ammonium acetate with gradient flow rates and gradient conditions. Amlodipine and IS were ionized by positive ion pneumatically assisted electrospray and detected in the multi-reaction monitoring (MRM) mode using precursor→productions of m/z 409.2→238.1, 294.1 and m/z 324.2→127.3, respectively. The specificity, matrix effect, recovery, sensitivity, linearity, accuracy, precision, and stabilities were all validated over the concentration range of 0.05 - 12 ng/mL. The 90% confidence interval (CI) for AUC0-t, AUC0-∞ and Cmax ratios (test: reference) were all within 80% - 125% interval proposed by FDA. It is concluded that the validated method can successfully fulfill the requirement of clinical bioequivalence study of amlodipine in healthy Chinese subjects after administration of amlodipine/losartan combination tablets and amlodipine tablets, and the test and reference tablets were bioequivalent.

Spectrofluorimetric determination of amlodipine in human plasma without derivatization

A rapid and sensitive spectrofluorimetric method was developed for the determination of amlodipine (AD), a calcium channel blocker, in the plasma. The type of solvent, the wavelength range, and the range of AD concentration were selected to optimize the experimental conditions. The calibration curves were linear (r² >0.997) in the concentration range of 0.1-12.5 ppm of AD. The limit of quantitation and limit of detection values for the method for plasma samples were 0.1 ppm and 0.07 ppm, respectively. The precision calculated as the relative standard deviation was less than 3.5%, and the accuracy (relative error) was better than 5.5% (n=6). The method developed in this study can be directly and easily applied for the determination of AD in the plasma without derivatization in plasma.

Determination of amlodipine in human plasma using automated online solid-phase extraction HPLC–tandem mass spectrometry: Application to a bioequivalence study of Chinese volunteers

An automated method (XLC-MS/MS) that uses online solid-phase extraction coupled with HPLC-tandem mass spectrometry was reported here for the first time to quantify amlodipine in human plasma. Automated pre-purification of plasma was performed using 10 mm × 2 mm HySphere C8 EC-SE online solid-phase extraction cartridges. After being eluted from the cartridge, the analyte and the internal standard were separated by HPLC and detected by tandem mass spectrometry. Mass spectrometric detection was achieved in the multiple reaction monitoring mode using a quadrupole tandem mass spectrometer in the positive electrospray ionization mode. The XLC-MS/MS method was validated and yielded excellent specificity. The calibration curve ranged from 0.10 to 10.22 ng/mL, and both the intra- and inter-day precision and accuracy values were within 8%. This method proved to be less laborious and was faster per analysis (high-throughput) than offline sample preparation methods. This method has been successfully applied in clinical pharmacokinetic and bioequivalence analyses.

Simultaneous determination of telmisartan and amlodipine in human plasma by LC–MS/MS and its application in a human pharmacokinetic study

A rapid and sensitive liquid chromatography–tandem mass spectrometric (LC–MS/MS) assay method has been developed and fully validated for the simultaneous quantification of telmisartan and amlodipine in human plasma. Carbamazepine was used as an internal standard. Analytes and the internal standard were extracted from human plasma by solid-phase extraction technique using Waters Oasis® HLB 1cm3 (30mg) extraction cartridge. The reconstituted samples were chromatographed on a Hypurity advance C18 column (50mm×4.6mm, 5μm) using a mixture of acetonitrile–5mM ammonium acetate buffer (pH-4.0) (50:50, v/v) as the mobile phase at a flow rate of 0.8mL/min. The calibration curve obtained was linear (r≥0.99) over the concentration range of 2.01–400.06ng/mL for telmisartan and 0.05–10.01ng/mL for amlodipine. Method validation was performed as per FDA guidelines and the results met the acceptance criteria. A run time of 2.5min for each sample made it possible to analyze more than 400 human plasma samples per day. The proposed method was found to be applicable to clinical studies.

Simultaneous determination of losartan, losartan acid and amlodipine in human plasma by LC-MS/MS and its application to a human pharmacokinetic study

Introduction: A simple, rapid and sensitive liquid chromatography-tandem mass spectrometric assay method has been developed and fully validated for simultaneous quantification of losartan and its active metabolite, losartan carboxylic acid, and amlodipine in human plasma. Irbesartan was used as an internal standard. Materials and Methods: The analytes were extracted from human plasma samples by solidphase extraction technique using Oasis HLB cartridges, (Waters Corporation, Mumbai, India). The reconstituted samples were chromatographed on a C18 column by using an 85:15, v/v mixture of methanol and 0.1% v/v formic acid as the mobile phase at a flow rate of 1.0mL/min. A detailed validation of the method was performed as per the FDA guidelines. Results: The calibration curves obtained were linear (r≥0.99) over the concentration range of 0.5-1000ng/mL for losartan and for its active metabolite losartan acid and 0.05-10.1ng/mL for amlodipine. The results of the intra- and inter-day precision and accuracy studies were well within the acceptable limits. Conclusions: A run time of 2.5min for each sample made it possible to analyze more than 300 plasma samples per day. The proposed method was found to be applicable to clinical studies.

Liquid Chromatography–Mass Spectrometry Method for the Determination of Amlodipine in Human Plasma and its Application in a Bioequivalence Study

A sensitive and selective liquid chromatographic-mass spectrometric (LC-MS) method for the determination of amlodipine (CAS 88150-42-9) in human plasma has been developed. Sample preparation was based on liquid-liquid extraction using NaOH and cyclohexane. Analytical determination was carried out on a C18 column interfaced with a single quadrupole mass spectrometer. Positive electrospray ionization was employed as the ionization source. The mobile phase was 10 mmol/L ammonium acetate solution-methanol (30:70, v/v) at the flow rate of 0.2 mL/min. The method was linear in the concentration range of 0.2-20 ng/mL. The lower limit of quantification was 0.2 ng/mL. Amlodipine was sensitive to UV light. The method was validated in terms of accuracy, precision, absolute recovery, and stability. The intra- and interday relative standard deviation across three validation runs over the entire concentration range was less than 8.17%. The accuracy determined at three concentrations (0.4, 2.0 and 10 ng/mL for amlodipine maleate) was within +/- 3.17% in terms of relative error. The method herein described was successfully applied for the evaluation of pharmacokinetic profiles of amlodipine maleate tablets in 20 healthy volunteers. The results showed that AUC, Tmax, Cmax and T 1/2 between the test and reference formulation have no significant difference (P > 0.05). The relative bioavailability was 103.7 +/- 12.3%.

Simple and Validated Method for Estimation of Amlodipine by LC-MS (ESI) Using Healthy Indian Human Volunteers: and Evaluation of Pharmacokinetic Parameters

A simple and validated liquid chromatographic–mass spectrometric method (LC-MS) for amlodipine in human plasma was quanti fi ed using LC-MS (ESI). Chromatography was performed on a C 18 analytical column, the mobile phase used was Acetonitrile-10mM Ammonium acetate in the ratio of 90:10%v/v and the retention times were 0.829 and 1.281 min for azithromycin (Internal standard) and amlodipine respectively. The ionization was optimized using ESI (+) and enhanced selectivity was achieved. The method is validated as per FDA guidelines. The analyte was shown to be stable over the timescale of the whole procedure. The pharmacokinetic parameters such as peak plasma concentration (C max ), Time to peak Concentration (t max ), Area under the plasma concentration-time curve (AUC 0-t & AUC 0- ∞ ), elimination rate constant (K eli ), Elimination half-life (t ½ ) were calculated. Log transferred values were compared by Analysis of Variance (ANOVA) followed by classical 90% con fi dence interval for C max AUC 0-t .and AUC 0- ∞ and was found to be within the range. These results indicated that the Test and Reference formulation is bioequivalent.

High-Performance Liquid Chromatographic Method for Quantitative Determination of Amlodipine in Human Plasma and Pharmaceutical Dosage Form and its Application to Pharmacokinetic Studies

An accurate, sensitive, and reproducible high-performance liquid chromatographic method for the quantitation of amlodipine besylate in human plasma has been developed and validated. The drug, internal standard, and major metabolite were eluted from a C(18) hypersil HyPurity column (3 microm, 3.9 mm i.d. x 150 mm) at room temperature with a mobile phase consisting of acetonitrile-potassium dihydrogen phosphate buffer (0.05 M) and acetic acid (62:38:0.1) with the pH adjusted to 3.5 using phosphoric acid. The flow-rate was 1.8 mL/min. The limit of detection was 1.0 ng/mL, and the limit of quantification of amlodipine besylate in plasma was 10 ng/mL. The intra- and inter-day precisions showed coefficients of variation ranging from 5.98-11.4% and from 5.60-11.74%, respectively at three different levels of concentration. The averages of the absolute and relative recoveries were found to be 96.74-98.51% and 95.96-100.71%, respectively. Stability studies showed that amlodipine besylate is stable for at least 2 months in plasma after freezing at -20 degrees C. The method was successfully applied for a pharmacokinetic study and for the determination of commercial amlodipine tablet content.

Determination of nicardipine and amlodipine in human plasma using on-line solid-phase extraction with a monolithic weak cation-exchange column

An on-line solid-phase extraction (SPE)-HPLC method was developed for simultaneous screening of nicardipine and amlodipine in human plasma. A short monolithic poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) [p(GMA-EDMA)]-based weak cation-exchange (WCX) column was prepared and employed as the selective extraction sorbent, which exhibited good permeability and biocompatibility. During the on-line SPE protocol, high-abundance proteins (human serum albumin, immunoglobulin G, immunoglobulin A and transferrin) and most matrixes in plasma were fast removed while nicardipine and amlodipine were effectively trapped on this monolithic column. Furthermore, the monolithic WCX sorbent could be continuously reused more than 300 times without obvious changes in analytes extraction and proteins cleanup. The proposed method was linear over a range of 0.5–50.0 ng mL −1 for both analytes with a linear regression coefficient greater than 0.998, and the limit of detection (LOD) for each analyte was 0.2 ng mL −1. Validation assays also demonstrated acceptable precision and adequate recovery for simultaneous quantitative screening of nicardipine and amlodipine in human plasma. Real plasma samples from hypertensive patients receiving a dosing of 5 mg antagonists were examined by using the proposed method. Results indicated that the on-line SPE-HPLC method could be applied for simultaneously monitoring of nicardipine and amlodipine in clinical plasma samples.