Bupivacaine In Human Plasma Research Articles

Simple HPLC-UV method for the quantification of bupivacaine in human plasma with one step protein precipitation

This research paper revolves around the process of the quantification of bupivacaine in human plasma through the selection of a sensitive high performance liquid chromatography method. The extraction of bupivacaine and ropivacaïne as an internal standard from plasma was performed through the use of hexane along with iso-propylalcohol. After liquid-liquid extraction we moved straightforward to the process of the mobile phase in which we had a combination of acetonitrile and potassium dihydrogen phosphate (65:35, v/v). The separation was carried out on a reversed phase C 18 column with UV-detector at 210 nm for bupivacaine and 230 nm for the internal standard. This particular examination exhibited excellent linearity ( r 2 = 0.999) in peak response over the concentration range of 0.1–1.5 μg/mL bupivacaine in human plasma. The mean absolute recoveries for 0.5 and 1.5 μg/mL of bupivacaine in plasma using the present extraction procedure were 97.43% and 95.03%, respectively. The intra- and inter-day coefficients of variation in the plasma were less than 12% at the lowest, and less than 10% at other concentrations, and the percent error values were less than 6%. The method displayed a high caliber of sensitivity and selectivity for therapeutic monitoring concentrations of bupivacaine in human plasma. Cet article de recherche s’articule autour du processus de quantification de la bupivacaïne dans le plasma humain à l’aide d’une méthode chromatographie liquide à haute performance. L’extraction de la bupivacaïne et de la ropivacaïne comme étalon interne à partir du plasma a été réalisée en utilisant de l’hexane avec de l’alcool isopropylique. Après l’extraction liquide-liquide, nous sommes passés directement au processus de préparation de la phase mobile qui est constituée par un mélange d’acétonitrile et de dihydrogénophosphate de potassium (65/35, v/v). La séparation a été effectuée sur une colonne C18 en phase inversée avec un détecteur UV à 210 nm pour la bupivacaïne et 230 nm pour l’étalon interne. Cette mise au point particulier a montré une excellente linéarité ( r 2 = 0,999) en réponse maximale sur la plage de concentration de 0,1–1,5 μg/mL de chlorhydrate de bupivacaïne dans le plasma humain. Les taux de récupération absolus moyens pour 0,5 et 1,5 μg/mL de chlorhydrate de bupivacaïne dans le plasma au moyen par le biais de mode d’extraction étaient respectivement de 97,43 % et 95,03 %. Les coefficients de variation dans le plasma intra et inter jours étaient inférieurs à 12 % aux valeurs les plus basses et inférieurs à 10 % aux autres concentrations, et les valeurs d’erreur en pourcentage étaient inférieures à 6 %. La méthode a montré une sensibilité et une sélectivité élevées pour le suivi thérapeutique des concentrations de bupivacaïne dans le plasma humain.

Molecularly imprinted polymer in microextraction by packed sorbent for the simultaneous determination of local anesthetics: lidocaine, ropivacaine, mepivacaine and bupivacaine in plasma and urine samples

This study presents the use of molecularly imprinted polymer (MIP) as packing material for microextraction by packed syringe (MEPS) to achieve higher extraction selectivity. Pentycaine was used as template for MIP. Development and validation of the determination of lidocaine, ropivacaine, mepivacaine and bupivacaine in human plasma and urine samples utilizing MIP-MEPS and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were carried out. The MEPS MIP-cartridge could be used for 100 extractions before it was discarded. The extraction recovery ranged from 60 to 80%. The correlation coefficients values were >0.999 for all assays using lidocaine, ropivacaine, mepivacaine and bupivacaine in the calibration range 5-2000 nmol/L. The accuracy of the studied compounds, given as a percentage variation from the nominal concentration values, ranged from -4.9 to 8.4% using plasma and urine samples. The between-batch precision, given as the relative standard deviation, at three different concentrations (quality control samples) was ranged from -4.7 to 14.0% and from 1.8 to 12.7% in plasma and urine, respectively. The lower limit of quantification and limit of detection of the studied substances were 5.0 and 1.0 nm, respectively.

Simultaneous determination of procaine, lidocaine, ropivacaine, tetracaine and bupivacaine in human plasma by high-performance liquid chromatography

A simple and sensitive high-performance liquid chromatography with ultraviolet detection (HPLC-UV) method has been developed and validated for simultaneous quantification of five local anesthetics in human plasma: procaine, lidocaine, ropivacaine, tetracaine and bupivacaine. In an ice-water bath, 500 microL plasma sample, containing 100 microg/mL neostigmine methylsulfate as anticholinesterase, was spiked with carbamazepine as internal standard and alkalized by sodium hydroxide. Liquid-liquid extraction with ethyl ether was used for plasma sample preparation. The chromatographic separation was achieved on a Kromosil ODS C18 column with a mobile phase consisting of 30 mM potassium dihydrogen phosphate buffer (0.16% triethylamine, pH adjusted to 4.9 with phosphoric acid) and acetonitrile (63/37, v/v). The detection was performed simultaneously at wavelengths of 210 and 290 nm. The chromatographic analysis time was 13 min per sample. The calibration curves of all five analytes were linear between 0.05 and 5.0 microg/mL (r(2) > or = 0.998). Precision ranged from 1.4% to 7.9% and accuracy was between 91.7% and 106.5%. The validated method is applicable for simultaneous determination of procaine, lidocaine, ropivacaine, tetracaine and bupivacaine for therapeutic drug monitoring and pharmacokinetic study.

High-Performance Liquid Chromatography Using UV Detection for the Simultaneous Quantification of Ropivacaine and Bupivacaine in Human Plasma

A specific high-performance liquid chromatography assay coupled with UV detection has been developed and validated for the simultaneous determination of ropivacaine and bupivacaine in human plasma. A liquid-liquid back extraction procedure was used to increase specificity, and very good and consistent recoveries were obtained: 93%-95% for ropivacaine and 90%-96% for bupivacaine. The lowest limit of quantification was 4 and 8 ng/mL for ropivacaine and bupivacaine, respectively. The method was sensitive, reproducible (coefficient of variation <or= 6.9%), and accurate (96%-102%) and was applied successfully to study the pharmacokinetics of ropivacaine and bupivacaine in a perioperative context.

Sensitive bioassay of bupivacaine in human plasma by liquid-chromatography-ion trap mass spectrometry

A sensitive high-performance liquid chromatography–tandem mass spectrometric (HPLC–MS–MS) method, using an ion trap spectrometer, was developed for quantitation of bupivacaine in human plasma. Bupivacaine and an internal standard (ropivacaine) were extracted in a single step from 100 μL of alkalinized plasma with diethyl-ether. The mobile phase consisted of acetonitrile with 0.1% formic acid (50:50, v/v), and was delivered at a flow rate of 0.3 mL/min. The effluent was detected by MS–MS in positive ion mode. Ionisation was performed, using an electrospray ion source, operating at 200 °C. The selected reaction monitoring transitions m/ z 289 → m/ z 140 and m/ z 275 → m/ z 126 were chosen for bupivacaine and ropivacaine, respectively. Calibration curves were linear over the concentration range of 3.90–500 μg/L with determination coefficients >0.996. The method is accurate (bias <10%) and reproducible (intra-assay and inter-assay precision <15%), with a quantitation limit of 3.90 μg/L, using only 100 μL of plasma. The high specificity and sensitivity, achieved by this fast method (total run-time <3 min), allowed the determination of bupivacaine plasma levels in pediatric patients, following epidural administration of bupivacaine.

Determination of ropivacaine and bupivacaine in human plasma by programmed temperature vaporiser-fast gas chromatography-mass spectrometry (PTV/fast GC/MS) utilising in-vial liquid-liquid extraction

In-Vial liquid-liquid extraction has been investigated as a sample-preparation method in combination with fast gas chromatography and mass spectrometric detection for analysis of ropivacaine and bupivacaine in human plasma. The method is simple, quick, and readily automated. It offers a new possibility for the GC analysis of drugs in biological fluids. Validation showed that the method is accurate and selective. The acceptance criteria for the study validation were well in line with international criteria.

Simultaneous determination of lidocaine and bupivacaine in human plasma: application to pharmacokinetics.

High-performance liquid chromatography has been applied to the simultaneous determination of lidocaine and bupivacaine levels in human plasma, using etidocaine as internal standard. The method was found to be linear in the range 10-2,000 micrograms/l for lidocaine and 20-1,000 micrograms/l for bupivacaine. Within-day and day-to-day coefficients of variation were generally lower than 12%. The method was applied to a pharmacokinetic study of lidocaine and bupivacaine during peribulbar anesthesia in ocular surgery. Both compounds displayed a slow apparent elimination rate. The method described here permitted the determination of the complete pharmacokinetic profiles of the two anesthetics, although they were administered at quite different doses.

Comparative study of the determination of bupivacaine in human plasma by gas chromatography-mass spectrometry and high-performance liquid chromatography

A comparison was made between high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) as methods for determining bupivacaine in human plasma. Both methods utilized pentycaine as an internal standard, were found to be linear in the range 5–320 ng and had acceptable precision and accuracy. The precision for the HPLC method was better than that for the GC-MS method. The limits of detection of the HPLC and GC-MS methods were ca. 1.0 and 0.1 ng, respectively. Good agreement between the HPLC and GC-MS methods was obtained for the analysis of samples taken from a patient receiving bupivacaine topically. For most purposes the HPLC method would be slightly better. However, for samples containing interfering peaks GC-MS provides a higher degree of resolution from such interferants.

Direct injection of large volumes of plasma in a column-switching system for the analysis of local anaesthetics II. Determination of bupivacaine in human plasma with an alkyldiol silica precolumn

A column-switching high-performance liquid chromatographic system was applied for the determination of bupivacaine in plasma. A 500-μl plasma sample was directly introduced onto a C 18-alkyl-diol silica (ADS) precolumn separating analytes from proteins and polar endogenous compounds. The fraction containing bupivacaine and ropivacaine (internal standard) was back-flushed and transferred to a conventional reversed-phase column (Kromasil C 18) for final separation. A single ADS precolumn could withstand more than 50 ml of plasma injections without changing analytical performance. Quantitative studies showed a broad range of linearity (0.033–3.31 μg/ml) and high recovery (95–99.9%) with coefficients of variation less than 3.1%. The advantages of the ADS material are its high capability of sample clean-up, due to rapid elution of plasma proteins and endogenous compounds to waste, and its ability to elicit a stable baseline. As a result, UV detection could be performed at 210 nm and clean chromatograms with baseline separation for desired peaks were obtained within 15 min. The detection limit of this system was 10 ng/ml defined by a signal-to-noise ratio of 3:1. The concentration of bupivacaine in patients determined by this method agreed well with the values obtained from an alternative method, making the technique applicable for pharmakokinetic studies in humans.

Determination of Bupivacaine in Human Plasma by HPLC

Abstract A rapid, sensitive, selective and reproducible reversed- phase high-performance liquid chromatographic method has been developed and validatd for the quantification of bupivacaine in human. Bupivacaine and the internal standard, lidocaine, are extracted from alkalinized plasma using n-hexane before chromatographing on a reversed-phase system. The mobile phase comprises 62 parts of 0.05 M phosphate buffer and 38 parts of methanol at pH 5.9 and it is pumped at 1.0 ml/min. The retention times of bupivacaine and lidocaine were observed to be 3.8 and 5.9 min respectively. Ultraviolet detection at 254 nm enabled a limit of detection of 25 ng/ml to be achieved. The reproducibility of the method was good at 1000 ng ml (C. V.=4.3%, n=6). The method is linear from 50 to 3200 ng/ml. The high sensitivity and the speed at which this assay can be performed makes it especially useful for estimating bupivacaine in human plasma.

The use of a packed column for the determination of bupivacaine in human plasma by gas chromatography: An application in a pharmacokinetic study of bupivacaine

The method involves a single extraction of the drug and the internal standard, etidocaine, from alkalinized plasma with n-hexane. The gas chromatograph is equipped with a glass column (2.0 m × 2 mm) packed with 3% W/W SP2250 on Chromosorb W, ( 80 100 mesh) and a nitrogen sensitive detector. The method can accurately measure plasma bupivacaine concentrations down to 0.01 μg ml −1 using a sample size of 0.5 ml. The day-to-day variation of bupivacaine at 2.0 μg mlsu−1 is 6.90% ( n = 10). The calibration graph is linear over the range 0.1–4.0 μg ml −1. The method is accurate, fast and sensitive and has been applied in a pharmacokinetic study of bupivacaine.

Sensitive determination of bupivacaine in human plasma by high-performance liquid chromatography

Sensitive determination of bupivacaine in human plasma by high-performance liquid chromatography

AN IMPROVED HPLC METHOD FOR DETERMINATION OF BUPIVACAINE IN HUMAN PLASMA

AN IMPROVED HPLC METHOD FOR DETERMINATION OF BUPIVACAINE IN HUMAN PLASMA

Determination of bupivacaine in human plasma by high-performance liquid chromatography

Determination of bupivacaine in human plasma by high-performance liquid chromatography