Method For Determination Of Nicotine Research Articles

Validation of GC-MS Method for Determination of Nicotine and Cotinine in Plasma and Urine

Validation of GC-MS Method for Determination of Nicotine and Cotinine in Plasma and Urine

Enhanced Chromatographic Determination of Nicotine in Human Plasma: Applied to Human Volunteers

Development of enhanced UPLC-UV method for determination of nicotine in human plasma was achieved on a Symmetry C18 column (100 mm × 2.1 mm, 2.2 μm) applying isocratic elution based on Methanol: Acetonitrile: Phosphate Buffer (pH: 2.7) with the ratio (20:30:50, v/v/v) as a mobile phase. The ultraviolet detector was operated at 260 nm. The mobile phase was pumped through the column at a flow rate of 0.2 mL min-1. The column temperature was adjusted to 50ºC and the injection volume was 2 μL. Quinine was selected as an internal standard (IS) due to its structure similarity to nicotine having basic pyridine ring to optimize the liquid liquid extraction procedure using diethyl ether coupled with vacuum evaporation at 40°C. Validation parameters for nicotine were found to be acceptable over the concentration range of 2.5-50 ng ml-1. The application of the proposed method on four healthy human volunteers was approved by the ethical committee. The study was carried out under fasting conditions and the concerned subjects were informed about the objectives and possible risks involved in the study. The proposed method proved to be simple and fast which is a major advantage to analyze large number of samples per day using the accelerated vacuum evaporation technique. The method showed satisfactory data for all the parameters tested within the limits for bioanalytical assays. The lower limit of quantification (LLOQ) permits the application of the method for further pharmacological and clinical studies.

Determination of nicotine and three minor alkaloids in tobacco by hydrophilic interaction chromatography-tandem mass spectrometry

A reliable, sensitive and rapid method for determination of nicotine and three minor alkaloids (cotinine, anabasine and nornicotine) in tobacco by ultra-high performance liquid chromatography in hydrophilic interaction chromatography mode coupled with tandem mass spectrometry (HILIC-MS/MS) has been established. HILIC separation was performed on a BEH HILIC column using isocratic elution at 0.5 mL/min with acetonitrile:water (85:15, v/v) mobile phase containing 5 mmol/L ammonium acetate (pH 5.00). Separated analytes were determined by electrospray ionization MS/MS in the positive ion mode using multiple reaction monitoring. Alkaloids from tobacco were extracted in an ultrasonic bath for 10 min with acetonitrile:water mixture (8:2, v/v) containing 5 mmol/L ammonium acetate (pH 5.00). Limits of quantification were 10 μg/g for cotinine, 20 μg/g for anabasine and nornicotine, and 30 μg/g for nicotine. Mean recoveries from tobacco ranged between 94.8% and 104.1% for different analytes with relative standard dev...

Simple and rapid assay method for simultaneous quantification of urinary nicotine and cotinine using micro-extraction by packed sorbent and gas chromatography-mass spectrometry.

A simple and rapid method for determination of nicotine and cotinine levels in urine was developed using samples prepared by micro-extraction by packed sorbent (MEPS) and subjected to gas chromatography-mass spectrometry (GC-MS) analysis. This method provided good reproducibility, as well as good linearity of calibration curves in the range of 1-100 and 50-1000 ng/mL for quality control samples spiked with nicotine and cotinine, respectively. The detection limit of nicotine and cotinine was as low as 0.25 and 20 ng/mL, respectively. An evaporation procedure is not suitable for nicotine determination, thus an advantage of the present MEPS assay method is direct testing with GC-MS without the need for evaporation to a dry solvent. Our findings show that it may be useful for determining nicotine levels in various types of research studies.

Validation of GC–MS method for determination of nicotine and cotinine in plasma and urine

Validation of GC–MS method for determination of nicotine and cotinine in plasma and urine

An Improved Commercial Method for Determination of Nicotine in Tobacco

A high-pressure thin-layer chromatography (HPTLC) method for quantitative analysis of nicotine in Nicotiana sp. was developed using a methanol extract of leaves and stems and TLC plates (silica gel 60 GF254) with spot visualization under ultraviolet (UV) light. Scanning at 235 nm in the absorption-reflection mode produced linear calibration curves in the range of 2 to 25 μg, with a correlation coefficient of 0.991. The average recovery rate was 95% (CV % 1.35). From the present study, the lower limit of detection was 0.08 μg spot−1 for the nicotine. The validity of the method was confirmed by comparing the UV spectra of the tobacco plant samples with standards within the same Rf window.

A validated method for the determination of nicotine, cotinine, trans‐3′‐hydroxycotinine, and norcotinine in human plasma using solid‐phase extraction and liquid chromatography‐atmospheric pressure chemical ionization‐mass spectrometry

A liquid chromatographic-mass spectrometric method for the simultaneous determination of nicotine, cotinine, trans-3'-hydroxycotinine, and norcotinine in human plasma was developed and validated. Analytes and deuterated internal standards were extracted from human plasma using solid-phase extraction and analyzed by liquid chromatography/atmospheric pressure chemical ionization-mass spectrometric detection with selected ion monitoring (SIM). Limits of detection and quantification were 1.0 and 2.5 ng/ml, respectively, for all analytes. Linearity ranged from 2.5 to 500 ng/ml of human plasma using a weighting factor of 1/x; correlation coefficients for the calibration curves were > 0.99. Intra- and inter-assay precision and accuracy were < 15.0%. Recoveries were 108.2-110.8% nicotine, 95.8-108.7% cotinine, 90.5-99.5% trans-3'-hydroxycotinine, and 99.5-109.5% norcotinine. The method was also partially validated in bovine serum, owing to the difficulty of obtaining nicotine-free human plasma for the preparation of calibrators and quality control (QC) samples. This method proved to be robust and accurate for the quantification of nicotine, cotinine, trans-3'-hydroxycotinine, and norcotinine in human plasma collected in clinical studies of acute nicotine effects on brain activity and on the development of neonates of maternal smokers.

A Rapid LC-MS-MS Method for the Determination of Nicotine and Cotinine in Serum and Saliva Samples from Smokers: Validation and Comparison with a Radioimmunoassay Method

The development and validation of a rapid liquid chromatography (LC)-tandem mass spectrometry (MS-MS) method for determination of nicotine and cotinine in smokers' serum is described. The method is based on solid-phase extraction in a 96-well plate format and requires only 100 microL of serum. Using normal-phase chromatography, both analytes elute in less than 1 min, which permits high sample throughput applications. The calibrated range is 2-100 ng/mL nicotine and 20-1,000 ng/mL cotinine. For known samples, recovery is 95-116% for nicotine and 93-94% for cotinine. The method is extended to rat serum and human saliva (cotinine only) using partial validation techniques. When compared with an existing radioimmunoassay method in our laboratory, the LC-MS-MS method gives improved accuracy, precision, and sample throughput.

Improved highly sensitive method for determination of nicotine and cotinine in human plasma by high-performance liquid chromatography

A highly sensitive and reliable method for the determination of nicotine and its metabolite cotinine in human plasma by high-performance liquid chromatography was developed. Nicotine and cotinine were extracted from alkalinized plasma with dichloromethane and the volatility of nicotine was prevented by the addition of conc. HCl to the organic solvent during evaporation. The sensitivity of quantification at 260 nm absorption was improved by using a noise-base clean Uni-3 to 0.2 ng/ml nicotine and 1.0 ng/ml cotinine. The method was validated over linear ranges of 0.2–25.0 ng/ml for nicotine and 1.0–80.0 ng/ml for cotinine. The intra-day precision and accuracy were ≤15.9% relative standard variation (RSD) and 89.9–103.5% for nicotine and ≤8.0% RSD and 98.7–103.0% for cotinine. The inter-day precision and accuracy were ≤17.0% RSD and 94.2–100.9% for nicotine and ≤8.2% RSD and 98.0–105.1% for cotinine.

A solid phase extraction method for determination of nicotine in serum and urine by isotope dilution gas chromatography/mass spectrometry with selected ion monitoring.

A rapid method for measuring nicotine concentration in serum and urine is described. Deuterated nicotine is used as an internal standard. Nicotine and deuterated nicotine are extracted using a copolymeric-bonded phase silica column. The extract is analysed by gas chromatography coupled with mass spectrometry (GC/MS) operating in selected ion monitoring mode. The method has a lower limit of detection of approximately 2 micrograms/L and is linear to at least 2000 micrograms/L. Within-run percentage coefficients of variation (% CV) are < 4 in both assays over a nicotine concentration range of 10-2000 micrograms/L. Between-run % CV in the serum assay are 5.4, 5.2, 4.8 and 5.9, respectively, at nicotine concentrations of 10, 15, 25, and 50 micrograms/L. Between-run % CV in the urine assay are 5.9, 4.5, 2.7 and 5.2, respectively, at nicotine concentrations of 100, 250, 500, and 2000 micrograms/L. The absolute recovery of nicotine is 61 +/- 6% (mean +/- SD) over the range of 10-250 micrograms/L. The assay has been used to measure serum nicotine concentrations and 24-h urinary excretion of nicotine to monitor the extent of replacement in subjects receiving transdermal nicotine therapy for smoking cessation.

Simple and sensitive method for determination of nicotine in plasma by gas chromatography

Smoking and other forms of nicotine consumption are among the most important risk factors for cardiovascular disease and cancer. Many of the cessation therapies require administration of nicotine. Accordingly, precision analysis for nicotine in plasma has become increasingly important. Several of the recently published methods require elaborate sample handling and/or processing. We report a simple, rapid and reliable gas chromatography method with a high sensitivity for determination of unchanged nicotine in plasma, which can be used in the processing and quantification of large series of nicotine samples, e.g., in clinical trials of nicotine-based smoking or tobacco cessation drug delivery systems.

Selected ion monitoring method for determination of nicotine, cotinine and deuterium-labeled analogs: Absence of an isotope effect in the clearance of (S)-nicotine-3′,3′-d2 in humans

A method for simultaneous determination of nicotine, its metabolite cotinine, and the stable isotope-labeled analogs nicotine-3',3'-d2 and cotinine-4',4'-d2 in human plasma has been developed. The method utilizes capillary column gas chromatography with detection by electron impact mass spectrometry and selected ion monitoring. Sensitivity is adequate for determination of nicotine and nicotine-d2 at concentrations as low as 1 ng ml-1, and cotinine and cotinine-d2 at concentrations as low as 10 ng ml-1 with good precision and accuracy. The method has been used to compare the elimination kinetics of (S)-nicotine-3',3'-d2 with natural nicotine in human subjects. Total clearance of nicotine-3',3'-d2 was virtually identical to the total clearance of natural nicotine, which validates the use of the deuterium-labeled analog in quantitative studies of nicotine metabolic disposition.

A Rapid, Quantitative GLC Method for the Simultaneous Determination of Nicotine and Cotinine

Published gas-liquid chromatographic (GLC) methods for the determination of nicotine and cotinine have proved impractical for the analysis of a large number of clinical samples. Significant improvements over other methods have been achieved, being low sample volume (0.5 mL plasma), rapid two-step extraction from plasma, no evaporation step, and good separation. The lower limits of sensitivity for nicotine and cotinine were 1 and 5 ng/mL, respectively. The method was validated by the analysis of plasma samples from cigarette-smoking volunteers. The method described permits the quick, routine determination of nicotine and cotinine in a large number of samples.

Improved Spectrophotometric Method for Determination of Nicotine in Tobacco Smoke

Improved Spectrophotometric Method for Determination of Nicotine in Tobacco Smoke

The colorimetric bromothymol blue method for determining small quantities of nicotine

AbstractA rapid and accurate method for determination of nicotine in a large number of plant or dust samples is described. Nicotine is extracted with light petroleum from an alkaline suspension of finely powdered material. The petroleum is then shaken with dilute hydrochloric acid and the aqueous portion is made up to volume. An aliquot is treated with 0·04% bromothymol blue and a suitable buffer, and shaken with chloroform; the nicotine is determined by examination of the coloured chloroform extract in a Spekker absorptiometer, using a 601 violet filter.