Recent advances in the analysis of electronic cigarette liquids and aerosols: Sample preparation and chromatographic characterization.

Electronic cigarette (e-cigarette) usage has risen dramatically worldwide in recent years. It has been publicized as a safer alternative to the conventional combustible cigarette. This, however, has not yet been supported by robust toxicological research evidence. Analysis of the chemical compositions of e-liquids and generated aerosols is an important step in evaluating the toxicity effects of e-cigarettes. Currently, a broad spectrum of analytical methods have been employed for qualitative and quantitative analysis of chemical compositions of e-cigarette liquids and aerosols. The aim of this article is to review the advances in the chromatographic characterization of chemical composition of the latter in the recent five years. In addition, sample preparation methods for e-liquids and aerosols are surveyed and discussed. A study of the relevant literature indicates that, expectedly, gas chromatography and liquid chromatography with a variety of detection systems, particularly mass spectrometry, have been the main analytical techniques used in this field. Sample preparation procedures primarily include headspace sampling, dilute-and-shoot approach, liquid-liquid extraction and sorbent-based extraction for e-liquids and for aerosols (the latter usually with laboratory-built collection devices). Some challenges of current e-cigarette analytical research, and an overview on prospective work are also presented.

Comparison of supercritical fluid chromatography-tandem mass spectrometry and liquid chromatography-tandem mass spectrometry for the stereoselective analysis of chlorfenvinphos and dimethylvinphos in tobacco.

This study used reversed-phase liquid chromatography-tandem mass spectrometry and supercritical fluid chromatography-tandem mass spectrometry for determination of the stereoisomers of chlorfenvinphos and dimethylvinphos in tobacco. Tobacco samples were extracted and purified with a modified quick, easy, cheap, effective, rugged, and safe technique using spherical carbon. The performance of both methodologies was comprehensively compared in terms of methods validation parameters (separation efficiency, linearity, selectivity, recovery, repeatability, sensitivity, matrix effect, etc.). Under optimized conditions, the calibration curves of the stereoisomers of chlorfenvinphos and dimethylvinphos in the range of 10-500 ng/mL showed excellent linearity with R2 ≥ 0.997 in both methods. The adequate recoveries of analytes from three different spiked tobaccos were obtained using reversed-phase liquid chromatography-tandem mass spectrometry (86.1-95.7%) as well as supercritical fluid chromatography-tandem mass spectrometry (86.5-94.0%). The relative standard deviations for spiked samples were all below 7.0%. Compared with supercritical fluid chromatography-tandem mass spectrometry, lower matrix effects and LODs can be obtained in reversed-phase liquid chromatography-tandem mass spectrometry.

Development of a colloidal gold immunochromatographic test strip for the rapid detection of iprodione.

Iprodione is a dicarboximide fungicide that is widely used in agriculture around the world. A reliable and rapid detection method is needed for the on-site monitoring of iprodione residues in a variety of agricultural products. Herein, a colloidal gold immunochromatographic test strip was developed based on a selected coating antigen and a specific monoclonal antibody against iprodione. The particle size of colloidal gold, the preparation technique of the conjugate pad, the composition of the loading buffer, and the extraction solvent were comprehensively optimized for the test strip. A cut-off value of 0.9 mg kg-1 (50 ng mL-1) and a visual limit of detection of 0.09 mg kg-1 (5 ng mL-1) were achieved in a complex matrix of tobacco. No cross-reactivity was observed for iprodione metabolite and four other widely used pesticides during tobacco growth. Furthermore, the developed colloidal gold immunochromatographic test strip was applied to determine iprodione residues in tobacco samples, and the obtained results were in good agreement with those obtained by liquid chromatography tandem mass spectrometry. Additionally, the test strip was found to be stable afterlong-term storage at 37 °C for two months. The developed colloidal gold immunochromatographic test strip showed excellent accuracy, sensitivity, specificity, and stability, therefore, it is suitable for the rapid detection of iprodione residues in complex matrices.

Highly sensitive time-resolved fluoroimmunoassay for the quantitative onsite detection of Alternaria longipes in tobacco.

AIMS: Alternaria longipes is a causal agent of brown spot of tobacco, which remains a serious threat to tobacco production. Herein, we established a detection method for A. longipes in tobacco samples based on the principle of time-resolved fluoroimmunoassay, in order to fulfil the requirement of rapid, sensitive and accurate detection in situ. METHODS AND RESULTS: A monoclonal antibody against A. longipes was generated, and its purity and titration were assessed using western blot and ELISA. The size of europium (III) nanospheres was measured to confirm successful antibody conjugation. The method described here can detect A. longipes protein lysates as low as 0.78 ng ml-1 , with recovery rates ranging from 85.96% to 99.67% in spiked tobacco. The specificity was also confirmed using a panel of microorganisms. CONCLUSIONS: The fluorescent strips allow rapid and sensitive onsite detection of A. longipes in tobacco samples, with high accuracy, specificity, and repeatability. SIGNIFICANCE AND IMPACT OF THE STUDY: This novel detection method provides convenience of using crude samples without complex procedures, and therefore allows rapid onsite detection by end users and quick responses towards A. longipes, which is critical for disease control and elimination of phytopathogens.

Separation of N'-nitrosonornicotine isomers and enantiomers by supercritical fluid chromatography tandem mass spectrometry.

N'-nitrosonornicotine (NNN) is one of the most prevalent and toxic tobacco-specific nitrosoamines. A chiral center at its 2'-position results in R and S enantiomers, the partial double bond character of the NN = O group also results in E and Z isomers, therefore, NNN can form a total of four absolute configurations (E-(R)-NNN, E-(S)-NNN, Z-(R)-NNN, and Z-(S)-NNN). This study investigated the resolution of R/S enantiomers and E/Z isomers of NNN by supercritical fluid chromatography tandem mass spectrometry (SFC-MS/MS). The baseline separation of E/Z-(R,S)-NNN isomers/enantiomers was accomplished through the optimization of chiral columns and co-solvents. Due to the lack of single standard of E/Z isomers, only R-NNN (sum of E-(R)-NNN and Z-(R)-NNN) and S-NNN (sum of E-(S)-NNN and Z-(S)-NNN) were further examined. Through the comprehensive optimization of SFC-MS/MS conditions, R-NNN and S-NNN were separated with a run time of 5 min, the developed method was validated, and its applicability to the determination of NNN enantiomers in burley tobacco samples was demonstrated. This study could be applied to preparative separation of single enantiomer and/or isomer of NNN, and could provide potential benefits to biologic activity studies on these enantiomers and isomers.

A green and effective method for the determination of metalaxyl enantiomers in tobacco and soil by supercritical fluid chromatography-tandem mass spectrometry.

We reported a new methodology for the stereoselective determination of metalaxyl enantiomers in tobacco and soil. The QuEChERS (quick, easy, cheap, effective, rugged, and safe) method was used for the extraction and clean-up of the tobacco and soil samples. Separation of the metalaxyl enantiomers was performed on an ACQUITY UPC2 Trefoil CEL1 chiral column coupled with supercritical fluid chromatography with tandem mass spectrometry (SFC-MS/MS), and the run time was only 5 minutes. Under the optimized conditions, the recoveries for the enantiomers were between 78.2% and 93.3% with intraday relative standard deviations (RSDs) ranging from 1.1% to 5.4%. The limit of detection (LOD) for the enantiomers in tobacco and soil varied from 0.005 to 0.007 mg/kg, and the limit of quantitation (LOQ) ranged from 0.017 to 0.020 mg/kg. In this method, only a small amount of methanol was consumed to obtain a rapid stereoselective separation. This proposed method showed good accuracy and precision and might be suitable for fast enantioselective determination of metalaxyl in food and environmental samples. The developed method was further validated by application to the analysis of authentic samples.

Use of autoclave extraction and liquid chromatography with tandem mass spectrometry for determination of maleic hydrazide residues in tobacco.

Maleic hydrazide has been extensively used as an effective growth regulator in tobacco sucker control. After application, maleic hydrazide distributes itself throughout the tobacco plant where it can exist as free, or forms glucoside conjugates with glucose, or becomes bound with lignin. Among them, free maleic hydrazide and its glucoside conjugates are extractable under conventional solvent extraction, while lignin bound maleic hydrazide is claimed to be non-extractable. Herein, an autoclave extraction method has been developed to extract maleic hydrazide effectively, in which tobacco samples are extracted in an autoclave at 130°C for 1 h using 4 M hydrochloric acid. Under such pressurized hot acidic water conditions, lignin bound maleic hydrazide can be released. Meanwhile, glucoside conjugates are hydrolyzed. Total maleic hydrazide is detected by liquid chromatography coupled with tandem mass spectrometry, and the quantitative results coincide well with that obtained from the international standard method. The proposed autoclave extraction with liquid chromatography and tandem mass spectrometry method exhibits excellent linearity in the range of 5-200 mg/kg (R2  = 0.9998), the matrix matched limit of detection and limit of quantification is 0.68 and 2.27 mg/kg, respectively. This method is simple and improves sample capacity, providing an effective approach to monitoring maleic hydrazide residues in tobacco.

Use of autoclave extraction-supercritical fluid chromatography/tandem mass spectrometry to analyze 4-(methylintrosamino)-1-(3-pyridyl)-1-butanone and N'-nitrosonornicotine in tobacco.

4-(methylintrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N'-nitrosonornicotine (NNN) are the most prevalent and toxic tobacco specific nitrosamines (TSNAs). Due to their carcinogenicity, knowledge of the composition of NNK and NNN in tobacco is necessary. Herein, a sensitive and rapid method, which employs autoclave extraction-supercritical fluid chromatography/tandem mass spectrometry (SFC-MS/MS), has been developed for the analysis of NNK and NNN in tobacco. Both water-soluble and matrix-bound NNK and NNN were extracted with 100 mM ammonium acetate in an autoclave (130 °C, 4 h), and the aqueous extract was subjected to solvent replacement prior to SFC-MS/MS analysis. NNK and NNN were effectively separated within 5 min by using supercritical CO2 as the main mobile phase coupled with a co-solvent of methanol. Excellent linearity was obtained with coefficients of determination (R2) greater than 0.9997 in the range of 1-160 ng/mL and 5-800 ng/mL for NNK and NNN, respectively. The recoveries were in the range of 92.5-110.0% at different spiked levels of real samples. 12 tobacco samples which include 3 typical tobacco varieties of burley, flue-cured, and oriental tobaccos had been analyzed, and the fraction of matrix-bound NNK was determined as well. In addition, a comparison between the proposed SFC-MS/MS method and a validated liquid chromatography tandem mass spectrometry (LC-MS/MS) internal standard method was conducted. Both techniques exhibit comparable analysis results, but peak splitting of NNN was observed by LC-MSMS due to the existence of E/Z isomers, while SFC-MS/MS offers great improvement through elution condition optimization, demonstrating the applicability of SFC-MS/MS as an alternative tool for NNK and NNN analysis.

Ultra-performance convergence chromatography with tandem mass spectrometry-assisted method for rapid enantioseparation and determination of fluazifop-butyl in tobacco and soil.

A quick, green, and sensitive method for chiral separation and determination of fluazifop-butyl enantiomers in tobacco and soil was established by ultra-performance convergence chromatography with tandem mass spectrometry (UPC2 -MS/MS). The baseline separation was obtained on an ACQUITY UPC2 Trefoil CEL2 column in 4 minutes with CO2 and methanol as mobile phase. Column temperature, auto back pressure regulator pressure (ABPR), and modifier solvent were optimized to obtain the best separation efficiency. Under the optimal conditions, the recoveries of both enantiomers were 82.8% to 99.5% with relative standard deviations (RSDs) less than 5.5% at three different concentration levels in two matrices. Good coefficients of determination (R2  ≥ 0.9976) were achieved over the concentration range of 10 to 500 ng/mL. The limits of detection (LODs) for all enantiomers in the two matrices varied from 1.6 to 2.1 µg/kg, and the limits of quantification (LOQs) did not exceed 7.0 µg/kg. The proposed method was then successfully applied to analyze authentic samples, confirming that it was a green, convenient, and reliable strategy for the analysis of fluazifop-butyl enantiomers in tobacco and soil.

An environmentally friendly method for the enantioseparation and determination of benalaxyl in tobacco and soil by ultra-performance convergence chromatography with tandem mass spectrometry.

For the purpose of chiral separation and determination of benalaxyl enantiomers in tobacco and soil, we developed a rapid, green, and sensitive method using ultra-performance convergence chromatography with tandem mass spectrometry. The samples were extracted and purified by the quick, easy, cheap, effective, rugged, and safe method before injection. The baseline separation was obtained on a chiral column in 5 min with carbon dioxide and ethanol as mobile phase. Separation parameters were optimized for the best separation efficiency. Under optimal conditions, the recoveries of both enantiomers were 77.1-98.4% with relative standard deviations <5.0% at spiked level of 0.1, 2.0, and 5.0 mg/kg in two matrices. Good coefficients of determination were achieved over the concentration range of 10-250 ng/mL. The limit of detection and the limit of quantification for all enantiomers ranged from 0.43 to 0.72 µg/kg and from 1.25 to 2.15 µg/kg, respectively. The results show that ultra-performance convergence chromatography with tandem mass spectrometry provides a reliable, green, and rapid method for the separation and determination of benalaxyl enantiomers in tobacco and soil. This method has important theoretical significance for studying the enantioselectivity and bioactivity of benalaxyl in the environment and in organisms.

Rapid fluorescence immunoassay of benzo[a]pyrene in mainstream cigarette smoke based on a dual-functional antibody-DNA conjugate.

Benzo[a]pyrene (BaP) is considered as one of the most carcinogenic pollutants in cigarette smoke. The development of simple and sensitive BaP screening methods can help assess the risk of cigarette exposure to the human body rapidly. In this report, a rapid fluorescence immunoassay (RFIA) method for the detection of BaP is proposed, the core of which is the synthesis of bifunctional covalent antibody-DNA conjugates for target recognition and signal amplification. Based on the optimization of the SYBR Green I and PAH-BSA concentrations, as well as DNA-antibody immune complex's dilution in the RFIA system, a serial dilution of BaP was tested with this method. The results showed that the linear working range of the RFIA for BaP is 0.46 to 333 ng mL-1, which is much wider than traditional ELISA. The detection limit was 0.32 ng mL-1, which was more sensitive than other methods such as the redox-labeled electrochemical immunoassay method and the competitive piezoelectric biosensor. Then the cross-reactions (CR) of other PAHs in cigarette smoke were evaluated using this RFIA and found that the cross-reactions of naphthalene, anthracene, and pyrene were very low (<1%). The cross-reaction in this RFIA system can be reduced by improving the specificity of the antibody. To the best of our knowledge, this is the first time that the BaP in mainstream cigarette smoke was tested; the RFIA demonstrates fast and simple experimental manipulations and better working curves and sensitivity.

Enantioseparation of nornicotine in tobacco by ultraperformance convergence chromatography with tandem mass spectrometry.

Nornicotine, an alkaloid constituent of tobacco, is a precursor to the carcinogen N-nitrosonornicotine that is produced during the curing and processing of tobacco. Accumulating evidence reveals that nornicotine enantiomers have different neurochemical and behavioral effects. In the present study, an accurate and rapid method was developed for the enantioseparation of (R)-(+)-nornicotine and (S)-(-)-nornicotine enantiomers in tobacco by ultra-performance convergence chromatography with tandem mass spectrometry. Chromatographic conditions were investigated to achieve the optimal resolution of two enantiomers. Results indicated that (R)-(+)-nornicotine and (S)-(-)-nornicotine could be separated within 5 min when ammonium hydroxide was added into the cosolvent, and the best resolution (Rs  = 4.76) was achieved on a immobilized cellulose tris-(3,5-dichlorophenylcarbamate) chiral stationary phase. The proposed method was validated and was finally applied to analyze the compositions of (R)-(+)-nornicotine and (S)-(-)-nornicotine in three typical types of tobaccos (flue-cured, burley, and oriental). It was found that, enantiomer fraction of nornicotine (the proportion of (S)-(-)-nornicotine in the nornicotine pool) in burley tobacco samples was relatively high and constant compared with flue-cured and oriental tobaccos. The effective and rapid enantioseparation of nornicotine may help the understanding of alkaloid metabolites in different tobacco varieties and may also benefit pharmacological studies of alkaloid enantiomers.

Rapid determination of 9 aromatic amines in mainstream cigarette smoke by modified dispersive liquid liquid microextraction and ultraperformance convergence chromatography tandem mass spectrometry.

Aromatic amines in mainstream cigarette smoke have long been monitored due to their carcinogenic toxicity. In this work, a reliable and rapid method was developed for the simultaneous determination of 9 aromatic amines in mainstream cigarette smoke by modified dispersive liquid liquid microextraction (DLLME) and ultraperformance convergence chromatography tandem mass spectrometry (UPC2-MS/MS). Briefly, the particulate phase of the cigarette smoke was captured by a Cambridge filter pad, and diluted hydrogen chloride aqueous solution is employed to extract the aromatic amines under mechanical shaking. After alkalization with sodium hydroxide solution, small amount of toluene was introduced to further extract and enrich aromatic amines by modified DLLME under vortexing. After centrifugation, toluene phase was purified by a universal QuEChERS cleanup kit and was finally analyzed by UPC2-MS/MS. Attributing to the superior performance of UPC2-MS/MS, this novel approach allowed the separation and determination of 9 aromatic amines within 5.0min with satisfactory resolution and sensitivity. The proposed method was finally validated using Kentucky reference cigarette 3R4F, and emission levels of targeted aromatic amines determined were comparable to previously reported methods At three different spiked levels, the recoveries of most analytes were ranged from 74.01% to 120.50% with relative standard deviation (RSD) less than 12%, except that the recovery of p-toluidine at low spiked level and 3-aminobiphenyl at medium spiked level was 62.77% and 69.37% respectively. Thus, this work provides a novel alternative method for the simultaneous analysis of 9 aromatic amines in mainstream cigarette smoke.

Influence of measurement uncertainty on the world health organization recommended regulation for mainstream cigarette smoke constituents.

The World Health Organization Study Group on Tobacco Product Regulation (WHO TobReg) proposed mandated ceilings on 9 prioritized mainstream cigarette smoke constituents determined from the market-specific median of nicotine-normalized yield distributions. Considering the requirements for assessing and reporting of compliance with ceilings, it is of great importance to estimate the measurement uncertainty. To have a better understanding of influence of measurement uncertainty on the WHO recommended regulation for cigarette smoke constituents, in the present study, the measurement uncertainties were evaluated systematically based on series of collaborative studies reported by three different authorities over the years from 2012 to 2016, according to the approaches guided in ISO/TS 21748. Furthermore, the compliance assessment of 20 representative cigarette samples with proposed ceilings was conducted by taking measurement uncertainty into account. This work demonstrated that measurement uncertainty had great influence on the implementation of the regulated mandated lowering of toxic smoke constituents, both on the setting of ceilings and the compliance assessment as well.

Determination of Aromatic Amines Released from Azo Dyes in Paper Packaging by Liquid Chromatography-Tandem Mass Spectrometry.

An LC-tandem MS (LC-MS/MS) method for the determination of 21 kinds of carcinogenic aromatic amines released from azo dyes in food wrappers was used in this research. Sodium dithionite was added to a citric acid buffer medium to reduce and decompose possible azo dyes. The extract was analyzed after liquid-liquid extraction (LLE) and dispersive SPE (d-SPE). The conditions for chromatographic separation, mass spectrum, LLE, and d-SPE were optimized. Under optimal conditions, the LOD was in the range of 0.13-0.35 mg/kg and LOQ in the range of 0.38-1.05 mg/kg, with the addition of standard recoveries of most aromatic amines being ≥80% and RSDs ≤10%. The recoveries for 2,4-diaminotoluene and 2,4-diaminoanisole were significantly lower, being ≤40%. The method was successfully used to analyze 30 practical samples, and the results showed that it is user-friendly, with high sensitivity, rapid control, and low matrix interference.

Analysis of pesticide residues in tobacco with online size exclusion chromatography with gas chromatography and tandem mass spectrometry.

An ultrasensitive method for the simultaneous analysis of pesticides residues in tobacco was developed with online size exclusion chromatography with gas chromatography and tandem mass spectrometry. Tobacco samples were extracted with the solvent mixture of cyclohexane and acetone (7:3, v/v) and centrifuged. Then, the supernatant liquors were injected directly into the online size exclusion chromatography with gas chromatography and tandem mass spectrometry without any other purification procedures after being filtered with a 0.22 µm organic phase filter. The matrix interferences were effectively removed and recoveries of most pesticides were in the range of 72-121%. Especially, for chlorothalonil, the analysis efficiency of this method was much more favorable than that of the general method, in which dispersive solid-phase extraction was used as an additional purified procedure. In addition, the limits of quantitation of this method were from 1 to 50 µg/kg. Therefore, a rapid, cost-effective, labor-saving method was proposed in the present work, which was suitable for the analysis of 41 pesticide residues in tobacco.

An environmentally friendly method for the determination of pentachlorophenol in paper packaging materials using ultrasonic-assisted micellar extraction.

Ultrasonic-assisted micellar extraction coupled with high-pressure liquid chromatography has been developed for the determination of pentachlorophenol (PCP) in paper packaging materials. A non-ionic surfactant, polyethylene glycol sorbitan monolaurate (Tween 20), was used for ultrasonic extraction and the method was applied to different paper packaging materials under optimized conditions. The recoveries for spiked samples were 83-90.1% and relative standard deviations were <5.0%. The limit of detection and the limit of quantification for PCP were 0.9 and 2.0 µg/g, respectively. Results confirmed that this relatively simple procedure proved satisfactory for the determination of PCP in paper packaging materials.

Determination of multiresidues of three acid herbicides in tobacco by liquid chromatography/tandem mass spectrometry.

A method to determine residues of the three acid herbicides, 2,4,5-trichlorophenoxyacetic acid, 2,4-dichlorophenoxyacetic acid, and 3,6-dichloro-2-methoxybenzoic acid (dicamba), in tobacco using LC/MS/MS is presented. Because these herbicide residues in tobacco might exist in different forms (free acid, salt, and ester), tobacco samples were first pretreated by alkaline hydrolysis and then the pH was adjusted in order to convert the residues completely to their free acid forms. Dichloromethane extraction and dispersive SPE using C18 sorbent were carried out before LC/MS/MS analysis, and quantification was performed using the internal standard method. Linearity was good for all analytes (R(2) ≥ 0.999) in the concentration range of 0.02 to 0.5 mg/kg. LODs were below 0.05 mg/kg. Recoveries ranged from 80.4 to 93.5%, and RSD was below 10%. This simple, efficient, and sensitive method can be applied to the determination of residues of the three acid herbicides in tobacco.

Simultaneous determination of herbicide residues in tobacco using ultraperformance convergence chromatography coupled with solid-phase extraction.

A time-saving and organic solvent efficient method to simultaneously determine six kinds of herbicide residues in tobacco using solid-phase extraction for sample clean-up and preconcentration and the highly sensitive ultraperformance convergence chromatography method was developed. Parameters for ultraperformance convergence chromatography, including the choice of stationary phase and modifiers, autobackpressure regulator pressure, column temperature, and the flow rate of mobile solvents, were optimized. The herbicide residues of napropamide, alachlor, quizalofop-ethyl, diphenamid, metolachlor, and clomazone in tobacco samples were successfully separated and detected at levels as low as 0.0043-0.0086 mg/kg within 5 min using a nonpolar high strength silica C18 selectivity for bases column and methanol as the cosolvent of the mobile phase of carbon dioxide (75-99.9%, v/v). Analysis of tobacco samples had recoveries of 69.8-95.0%, limit of quantitation of 0.0127-0.0245 mg/kg, limit of detection of 0.0043-0.0086 mg/kg, and correlation coefficient of >0.9990. Results support this method as an efficient alternative to current methodologies for the determination of herbicide residues in tobacco.

Environmentally friendly method for the determination of acrylamide and trimethylolpropane in paper packaging materials by liquid chromatography with tandem mass spectrometry.

A simple, rapid, and environmentally friendly method was developed for the determination of acrylamide and trimethylolpropane in paper packaging materials. No organic solvent was used and the matrix effect was investigated. The extract was directly analyzed by liquid chromatography with tandem mass chromatography for quantification and confirmation. The chromatographic separations were performed on a ZORBAX HILIC Plus (2.1 mm × 150 mm, 3µm; Agilent, USA) column with only one mobile phase (100% water). Calibration curves for acrylamide and trimethylopropane were achieved with concentrations ranging from 0.4 to 20 mg/kg and the corresponding r(2) values were 0.998 and 0.999, respectively. The recoveries were >85% with relative standard deviations <10%. The validated method was applied to the analysis of 50 real samples, and positive results were obtained for 30 samples. The result indicated that trimethylolpropane is associated with inks and printing activity and acrylamide is widely used as a papermaking additive in many paper packages. The concentrations of acrylamide and trimethylolpropane ranged from 0.41 to 7.5 and 0.50 to 8.8 mg/kg, respectively. The results of this study revealed that this method could be used accurately and precisely.