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

doi:10.1016/j.jchromb.2010.03.003

Journal of Chromatography B

Volume 878, Issues 15–16, 1 May 2010, Pages 1185-1189

https://doi.org/10.1016/j.jchromb.2010.03.003 Get rights and content

Abstract

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 μL plasma sample, containing 100 μg/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 μg/mL (r² ≥ 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.

Introduction

Procaine, lidocaine, ropivacaine, tetracaine and bupivacaine are often used in combinations to achieve appropriate onset time and duration of action for local anesthesia [1], [2]. Several analytical methods have been published for simultaneous determination of some of these compounds in human plasma [3], [4], [5], [6], [7]. These five local anesthetics can be chemically divided into two groups, the amino-amides (lidocaine, ropivacaine and bupivacaine) and the amino-esters (procaine and tetracaine) [8]. Some challenges existed in simultaneous determination of these five anesthetics, such as that the amino-esters in human plasma is easy to degrade, and that the ultraviolet absorbance of the two groups peaks at 210 and 290 nm, respectively [4], [7]. As far as we know, only two published methods simultaneously determined both the amino-amides and the amino-esters [4], [7]. However, neither of the two methods covered all these five common local anesthetics, and both methods had some analytical limitations which need to be improved. The GC–MS method required a 3-h solid phase extraction procedure for sample preparation and the HPLC method demanded a 30 min gradient elution for chromatographic analysis [4], [7].

The purpose of our study is to develop a reliable, rapid, sensitive and selective analytical method for simultaneous determination of these five local anesthetics in human plasma. Compared to other methods that have been published, the advantages of our validated technique are its simplicity, rapidity, and capability to simultaneously measure these five anesthetics under the same sample preparation procedure and chromatographic conditions.

Section snippets

Regents and materials

Bupivacaine hydrochloride and ropivacaine hydrochloride were provided by Sunve Pharm (Shanghai, China) and AstraZeneca (Wuxi, China), respectively. Tetracaine hydrochloride, lidocaine and procaine hydrochloride were obtained from the China National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). Carbamazepine (internal standard, IS) was obtained from Sigma–Aldrich (St Louis, USA). Neostigmine methylsulfate was provided by Sine-Jinzhu Pharmaceutical. Co.,

Results and discussion

No visible interferences were observed at the retention times of these five analytes and IS at both 210 and 290 nm. All the commonly administrated drugs shown in Table 1 were not found to be interfering with the assay. The chromatogram of pooled blank plasma was shown in Fig. 1A and B. The analytical method presented linear response over the concentration range of 0.05–5.0 μg/mL by weighted (1/x) least-squares regression analysis. The regression coefficients (r²) were stable and all ≥0.998. The

Clinical application

This method was applied to a preliminary pharmacokinetic study in which 10 Chinese adult patients administrated with either a mixture (A) of lidocaine (2 mg/kg) and ropivacaine (2 mg/kg) or a mixture (B) of lidocaine (2 mg/kg) and tetracaine (2 mg/kg) as local anesthetics during brachial plexus block. The study protocol was approved by the Institutional Ethics Committee of Huashan Hospital and was conducted according to the recommendations described in the Declaration of Helsinki. All the patients

Conclusions

The present paper first described a simple and reliable HPLC-UV method for rapid simultaneous determination of procaine, lidocaine, ropivacaine, tetracaine and bupivacaine in human plasma. The method has been proven to be applicable in a pharmacokinetic study of procaine, lidocaine, ropivacaine, tetracaine and bupivacaine.

References (10)

R. Murtaza et al.
J. Pharmacol. Toxicol. Methods
(2001)
E. Tanaka et al.
J. Chromatogr. B: Analyt. Technol. Biomed. Life Sci.
(2006)
A. Koehler et al.
J. Chromatogr. A
(2005)
T. Ohshima et al.
J. Chromatogr. B: Biomed. Sci. Appl.
(1999)
J.S. Berry et al.
AANA J.
(1999)

There are more references available in the full text version of this article.

Cited by (73)

In situ enrichment and determination of 6 kinds of caine-type anesthetics in cosmetics and rat serum by thin layer chromatography-Raman spectroscopy
2023, Arabian Journal of Chemistry
A simple, economical and selective method has been established for simultaneous detection of 6 kinds of caine-type anesthetics in cosmetics and rat serum by in situ enrichment thin layer chromatography (TLC) and Raman spectroscopy. The target caine-type anesthetics, procaine hydrochloride (Pro), tetracaine hydrochloride (Tet), dibucaine (Dib), mepivacaine hydrochloride (Mep), lidocaine hydrochloride (Lid) and ropivacaine hydrochloride (Rop), were separated by TLC. The original spots (or position of R_f equal to that of the standard) of anesthetics on TLC plates were in situ enriched into much smaller ones driven by solvent from different directions, and followed by Raman measurement for quanlitative and quantitative analysis, resulting in the improvement of sensitivity and selectivity. Optimization and standardization of the experimental conditions, the composition and proportion of developing solvent, were conducted. Thus a robust and reliable approach which enabled effective separation, accurate identification and quantification has been developed with the lowest detectable amounts of 0.4, 0.2, 1.2, 1, 1 and 1 μg for Pro, Tet, Dib, Mep, Lid and Rop, respectively. The established method was successfully applied in anesthetics analysis in cosmetics (essence, cream and gel) and rat serum without complex sample pretreatment processes, and the results were verified by HPLC. It turned out that the in situ enrichment TLC-Raman technique was feasible and effective for separation and determination of 6 caine-type anesthetics in biological samples and cosmetics, and would have good prospects for on-site qualitative screening of prohibited goods and adulterants.
Efficacy of prolonged intravenous lidocaine infusion for postoperative movement-evoked pain following hepatectomy: a double-blinded, randomised, placebo-controlled trial
2023, British Journal of Anaesthesia
The analgesic effect of intravenous lidocaine varies with the duration of lidocaine infusion and surgery type. We tested the hypothesis that prolonged lidocaine infusion alleviates postoperative pain in patients recovering from hepatectomy over the first 3 postoperative days.
Patients undergoing elective hepatectomy were randomly assigned to receive prolonged i.v. lidocaine treatment or placebo. The primary outcome was incidence of moderate-to-severe movement-evoked pain at 24 h postoperatively. The secondary outcomes included incidence of moderate-to-severe pain during movement and at rest throughout the first 3 postoperative days, postoperative opioid consumption, and pulmonary complications. Plasma lidocaine concentration was also monitored.
We enrolled 260 subjects. Intravenous lidocaine lowered the incidence of moderate-to-severe movement-evoked pain at 24 h and 48 h postoperatively (47.7% vs 67.7%, P=0.001; 38.5% vs 58.5%, P=0.001) and reduced movement-evoked pain scores (3.7 [1.7] vs 4.2 [1.6]; mean difference 0.5 [95% confidence interval {CI}: 0.1–0.9]; P=0.018) and morphine equivalent consumption (47.2 [16.7] mg vs 52.6 [19.2] mg; mean difference 5.4 mg [95% CI: 1.0–9.8]; P=0.016) at 24 h postoperatively. Lidocaine also lowered the incidence of postoperative pulmonary complications (23.1% vs 38.5%; P=0.007). Median plasma lidocaine concentrations were 1.5, 1.9, and 1.1 μg ml⁻¹ (inter-quartile ranges: 1.1–2.1, 1.4–2.6, and 0.8–1.6, respectively) after bolus injection, at the end of the surgery, and 24 h postoperatively.
Prolonged intravenous lidocaine infusion reduced the incidence of moderate-to-severe movement-evoked pain for 48 h after hepatectomy. However, the reduction in pain scores and opioid consumption by lidocaine was below the minimal clinically important difference.
NCT04295330.
Stability-indicating HPLC-DAD and TLC-densitometry methods for the quantification of bupivacaine and meloxicam in their co-formulated mixture
2023, Microchemical Journal
The implementation of a novel drug formulation necessitates the assessment of its content stability and the designation of appropriate stability monitoring methodologies. Bupivacaine (BVC) and Meloxicam (MLX) were recently authorized under the name Zynrelef® solution. This novel combination was tested for stability under a variety of stress circumstances for the first time in this study. The basic degradation pathway was shown to affect BVC, while the acidic degradation pathway works for MLX. Identification of the acquired degradation products, including BVC impurity F (BVC IMP-F), 1-buytylpiperidine-2- 2carboxylic acid (1-BPC), MLX acid degradation product (MLX-DEG), and MLX impurity B (MLX IMP-B), was carried out using IR and MS spectroscopic techniques. HPTLC and HPLC, two chromatographic-based stability-indicating methods, have been established. The HPTLC method offered efficient separation upon the usage of silica gel 60 F₂₅₄ as a stationary phase and ethyl acetate/toluene/triethylamine/acetic acid (4:4:2:0.4, v/v/v/v) as a developing system with densitometric recording at 260 nm. The HPLC method was developed by using Zorbax® SB C₁₈ column as a stationary phase and a mobile phase consisting of acetonitrile: methanol: phosphate buffer, pH 6.5 (20:60:20 v/v/v), at a flow rate of 1.5 mL min^-1. Quantification of the analytes was performed at 265 nm using a diode array detector. The current research introduced the first stability-indicating methods for the assay of BVC and MLX which were successfully applied for the determination of the cited drugs in their combined pharmaceutical formulation. Moreover, the HPLC method was further extended for studying the degradation kinetics of the two drugs.
Electrochemically modulated SERS detection of procaine using FTO electrodes modified with silver-decorated carbon nanosphere
2021, Electrochimica Acta
Procaine belongs to a class of drugs whose excessive dosage results in cardiac arrest and several allergenic reactions. Therefore, its continuous monitoring is crucial for sustainable health management during treatments. In this study, a reliable electrochemically modulated sensing approach for the quantitative detection of procaine using in-situ surface-enhanced Raman scattering (SERS) was developed. A fluorine-doped tin oxide electrode modified using silver carbon nanosphere (AgCNS-FTO) was designed, electrochemically evaluated, and used as an efficient substrate. The investigation of the electrochemical interaction of procaine molecules towards the modified electrode revealed that upon the optimum electrochemical pre-concentration potential (-0.1 V) and time (400 s), significant enhancements in the intensity of key Raman peaks corresponding to the NH₂ bending, C=C stretching, CH₂ twisting and C-N stretching modes could be observed. The reported approach enabled a remarkable sensitivity and selectivity towards procaine, resulting in an unprecedented detection limit down to the level of 10^-13 M. The reproducibility and stability of the modified FTO electrode were ensured over a duration of one month, and a low RSD of 2.4% was achieved. First-principles calculations proposed the interactive nature between the AgCNS surface and procaine molecules, while the possible protein-ligand binding with the chemokine receptor CXCR4 predicted an anti-inflammatory tendency of procaine with a binding energy of – 5.7 kcal/mol for the pose with the highest affinity. The electrochemically modulated SERS approach developed in the present study shall facilitate the efforts towards the practical quantification of procaine in clinical samples and could be extended to other structural analogues.
Studies of mechanism, kinetic model and determination of bupivacaine and its application pharmaceutical forms
2020, Microchemical Journal
Bupivacaine, a local anesthetic that is often preferred in many wide ophthalmology applications, was investigated from different aspects such as electrochemical behavior, kinetic model, mechanism illumination, and analytical applications by cyclic voltammetry (CV) and square wave stripping voltammetry (SWSV) methods on glassy carbon electrode (GCE). Bupivacaine exhibited a well-defined anodic signal at nearly 850 mV, an irreversible and adsorption-controlled electrode process. Furthermore, a sensitive and selective SWSV method with a linear working range of 0.3–12.5 mg/L was developed for bupivacaine determination. Limits of detection (LOQ) and that of determination (LOD) were analyzed as 85 and 282 μg/L, respectively. In addition, the most probable study mechanism was carried out for the first time by this study. The selectivity of some co-existences such as heavy metals, ketotifen, moxifloxacin, ciprofloxacin, cyclopentanate and tropicamide was studied in 1 mg/L bupivacaine determination. They did not have a serious effect on bupivacaine determination at 5% tolerance limits. Otherwise, bupivacaine was successfully analyzed in matrix samples without any pretreatment by the proposed SWSV. Consequently, a fast, reliable, inexpensive, portable, eco-friendly, and sensitive new method has been developed to analyze bupivacaine in real samples.
Studying the effect of vasopressors on therapeutic drug monitoring of two local anesthetics using hybrid micelle liquid chromatography as an analysis tool
2020, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
A hybrid micelle based mobile phase was used to develop and validate a liquid chromatographic method for the separation and quantification of two local anesthetics namely; lidocaine hydrochloride (LID), and bupivacaine hydrochloride (BPV) in presence of the frequently co administered vasopressors phenyl ephrine (PHR) and ephedrine (EPH). Optimization of chromatographic separation conditions was performed applying experimental one factor at a time tool, and design of experiment, where the retention behavior of all analytes using both optimization protocols was in accordance. Chromatographic separation was carried on a C8 column operating at 40 °C at a flow rate of 1.5 mL/min. using a mobile phase consisting of 0.18 M sodium dodecyl sulphate, 10% acetonitrile, containing 0.3% triethyl amine and adjusted to pH 7 using 2 M ortho phosphoric acid, adopting UV detection at 230 nm. The proposed method was fully validated and applied to both in vitro and in vivo analysis of rat blood samples. The pharmacokinetics of both LID and BPV was followed when they were solitary injected or when co administered with either PHR or EPH. Moreover, the in vitro spiked experiment was also subjected to documented bio-analytical validation procedures.

View all citing articles on Scopus

View full text

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

Abstract

Introduction

Section snippets

Regents and materials

Results and discussion

Clinical application

Conclusions

J. Pharmacol. Toxicol. Methods

J. Chromatogr. B: Analyt. Technol. Biomed. Life Sci.

J. Chromatogr. A

J. Chromatogr. B: Biomed. Sci. Appl.

AANA J.

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