Ethanol In Alcoholic Beverages Research Articles

Analysis of Methanol and Ethanol Content in Illegal Alcoholic Beverages using Headspace Gas Chromatography: Case Studies at Rwanda Forensic Institute

Illegal alcoholic production in Rwanda has become a critical social concern due to its widespread prevalence and associated health risks, stemming from the inexpensive distillation process employed by illicit producers. This often results in the unintentional production of toxic methanol instead of safe ethyl alcohol, presenting a substantial hazard in the context of illegal alcohol production. Methanol, when consumed, transforms into toxic formic acid in the body, causing severe health complications by disrupting mitochondrial respiration. Between 2021 and 2022, 183 forensic cases related to methanol and ethanol poisoning were collected nationwide and examined at the Rwanda Forensic Institute (RFI). Utilizing Headspace Gas Chromatography, the study aimed to quantitatively assess the extent of the problem and compare results to Rwanda’s allowable limits for methanol and ethanol in alcoholic beverages. The analysis demonstrated a significant variation in ethanol content (3.8% to 98.9% v v-1) and methanol levels (32% to 58.3% v v-1), with 6.6% of samples exceeding the methanol limit (0.5% v v-1) and 16.9% surpassing the ethanol limit (45% v v-1). The City of Kigali emerged as the primary contributor to non-compliance, notably associated with specific brands like K’bamba, African Buffalo, Merry Cane, Royal Castle, and unbranded alcoholic beverages. Importantly, none of the samples tested positive for both methanol and ethanol simultaneously, emphasizing the urgency of monitoring and regulating Rwanda’s alcoholic beverages market to ensure compliance with acceptable methanol and ethanol levels and safeguard public health.

Development of a Fluorescent Sensor Based on Resazurin and Hydrotalcite for the Determination of Ethanol in Alcoholic Beverages

Development of a Fluorescent Sensor Based on Resazurin and Hydrotalcite for the Determination of Ethanol in Alcoholic Beverages

Reduced production of Ethyl Carbamate in wine by regulating the accumulation of arginine in Saccharomyces cerevisiae

Ethyl carbamate (EC), a multisite carcinogenic compound, is naturally produced from urea and ethanol in alcoholic beverages. In order to reduce the content of EC in wine, the accumulation of arginine in Saccharomyces cerevisiae was regulated by genetic modifying genes involved in arginine transport and synthesis pathways to reduce the production of urea. Knockout of genes encoding arginine permease (Can1p) and amino acid permease (Gap1p) on the cell membrane as well as argininosuccinate synthase (Arg1) respectively resulted in a maximum reduction of 66.88% (9.40 µg/L) in EC, while overexpressing the gene encoding amino acid transporter (Vba2) reduced EC by 52.94% (24.13 µg/L). Simultaneously overexpressing Vba2 and deleting Arg1 showed the lowest EC production with a decrease of 68% (7.72 µg/L). The yield of total higher alcohols of the mutants all decreased compared with that of the original strain. Comprehensive consideration of flavor compound contents and sensory evaluation results indicated that mutant YG21 obtained by deleting two allele coding Gap1p performed best in must fermentation of Cabernet Sauvignon with the EC content low to 9.40 μg/L and the contents of total higher alcohols and esters of 245.61 mg/L and 41.71 mg/L respectively. This study has provided an effective strategy for reducing the EC in wine.

A New Highly Sensitive Electrochemical Biosensor for Ethanol Detection Based on Gold Nanoparticles/Reduced Graphene Oxide/Polyallylamine Hydrochloride Nanocomposite.

A highly sensitive electrochemical biosensor for ethanol based on a screen-printed electrode modified with gold nanoparticles-electrochemically reduced graphene oxide-poly (allylamine hydrochloride) nanocomposite (AuNPs-ERGO-PAH) is reported in this work. Ethanol was oxidized in the presence of the oxidized form of the nicotinamide adenine dinucleotide (NAD+) in a reaction catalyzed by alcohol dehydrogenase (ADH) immobilized in sol-gel. The AuNPs-ERGO-PAH nanocomposite was used as a transducer for the electrocatalytic oxidation of the reduced form the nicotinamide adenine dinucleotide (NADH) produced in the enzyme reaction. Under the optimal conditions, the ethanol biosensor exhibits a wide dynamic range from 0.05 to 5 mM with a low detection limit of 10 µM (S/N = 3) and a high sensitivity of 44.6 ± 0.07 µA/mM·cm2 for the linear range between 0.05 and 0.2 mM. The biosensor response was stable for up to 6 weeks. Furthermore, the developed biosensor has been used to detect ethanol in alcoholic beverages with good results, suggesting its potential application in various fields, including fermentation processes and food quality control.

Direct determination of ethanol in alcoholic beverages based on its anti-aggregation of melamine-silver nanoparticle assembly

We report a new method for determination of ethanol based on anti-aggregation of silver nanoparticles (AgNPs) in the presence of melamine. In the system, ethanol and melamine act as protecting and aggregating agents, respectively. Melamine can induce citrate-stabilized AgNPs to aggregate, leading to a color change from yellow to green. However, if the AgNPs are pre-incubated in ethanol, ethanol readily surrounds the particles by forming hydrogen bonds with the citrate stabilizer. An external nanoshell of ethanol hinders particle aggregation caused by melamine. Minor aggregation of AgNPs was observed, the solution color maintained its yellow-orange color. Higher ethanol concentrations result in a lower degree of particle aggregation. The colorimetric response of AgNPs was monitored using a UV–vis spectrophotometer at 390 nm. The current method could determine ethanol concentrations over a wide dynamic range of 5–80% (v/v), with a detection limit of 3.1% (v/v) (3SD of blank/slope). This method was applied for direct quantification of ethanol in alcoholic drinks without sample pretreatment and the results are well correlated with those of gas chromatography. Our method is convenient and cost effective, making it auspicious for ethanol monitoring in alcoholic drink manufacture and control.

Home-Made Membraneless Vaporization Gas-Liquid Separator for Colorimetric Determination of Ethanol in Alcoholic Beverages.

This work utilized the simplicity of a so-called membraneless vaporization (MBL-VP) unit as a gas separator for the colorimetric determination of ethanol in alcoholic beverages. A beverage sample with a volume of 1 mL was directly injected into a small container which was hung from a lid inside a closed 40 mL reused glass bottle without pretreatment such as distillation. An acidified potassium dichromate (Cr2O72−) acceptor solution, preadded to the glass bottle, was reduced to Chromium (III) ion by the diffusion of vaporized ethanol from the sample. After 5 min, the absorbing solution was collected for colorimetric detection at 590 nm. The unit manually quantifies ethanol in the range 1.0–90% (v/v) with satisfactory interday precision but without matrix effect (recovery 89−109%). The method was validated with the conventional distillation/pycnometer method which showed no significant difference of ethanol contents between those two methods and the declared values of 12 alcoholic beverages, indicating sufficient accuracy. Analyses of alcoholic beverages using this method were successful with benefits of simplicity, cheapness, and less energy consumption.

Reusable alcohol oxidase-nPtCu/alginate beads for highly sensitive ethanol assay in beverages.

Nanozymes (NZs) are nanoparticles that mimic the catalytic properties of natural enzymes. The present work aimed to obtain effective peroxidase mimetics (PO-like NZs), to characterize their morphological properties, estimate the kinetic parameters of NZs and evaluate the prospects of their application in analysis of ethanol. Herein, we have proposed a convenient spectrophotometric method for ethanol assay using reusable alginate beads enriched with alcohol oxidase (AO) and nanoparticles of PtCu (nPtCu) as PO-like NZs, and 3,3′,5,5′-tetramethylbenzidine (TMB) as a chromogen. The linear range for the proposed nPtCu–AO/alginate beads/TMB-based method is from 0.01 mM to 0.15 mM with a limit of detection of 3.3 μM ethanol. The method is used for the quantitative determination of ethanol in alcoholic beverages. The obtained results proved to be in a good correlation with the enzymatic reference method. These results highlight the potential of the nPtCu with PO-like activity in bioanalytical applications. The proposed method, being sensitive, economical and suitable for routine and micro-volume formats, can be used in clinical diagnostics for the detection of ethanol.

A rapid method for determination of ethanol in alcoholic beverages using capillary gas chromatography

A simple and rapid method was developed to determine ethanol content in alcoholic beverages using megapore polar column (CP-Wax 58 CB, 30 m × 0.53 mm) with direct injection gas chromatography. Contrary to packed GLC method, distillation and/or stepwise dilution of samples were not necessary by the method developed in this study. Ethanol in sample was injected directly into GC for analysis, after adding suitable amount of internal standard, acetonitrile solution. Using this method, less than 8 min was required to obtain result since sample preparation, and the limit of quantitation (LOQ) was about 0.5 μg/mL. Recovery studies were performed using 0.5 mL of red wine and whisky. Each was spiked with ethanol at 50 and 100 mg, respectively. The recoveries were found in the range of 99-104% and 99-101%, respectively. The coefficients of variation were less than 3.4%. Comparisons of the AOAC method (AOAC 969.12 and 920.57) with current method showed no significant difference. These results suggested that precision of direct injection GC method was higher than that of AOAC methods. Several commercial alcohol beverages, including distilled and non-distilled spirits, were analyzed by the current method. The ethanol content of distilled and non-distilled spirit were found as: 165.2 ± 4.9-415.7 ± 17.6 and 28.2 ± 0.8-141.2 ± 4.9 mg/mL, respectively. Using this GC method, we could change the concentration in gravimetric percentage (%, w/v) to volumetric percentage (%, v/v) by the equation (%, w/v) = 0.814 (%, v/v) with linear coefficient R2, higher than 0.999.

Visual chemo-chronometric assay for quantifying ethanol in alcoholic drinks by the colorimetric Belousov-Zhabotinsky oscillator

A simple chemo-chronometric method utilizing the Belousov-Zhabotinsky (BZ) oscillating reaction for the efficient detection of ethanol in alcoholic beverages has been developed. This visual detecting strategy only required a smartphone, without implemented instrumental detection systems, to video a unique colorimetric alternating (red-to-blue) feature of the BZ solution system containing 0.50 M bromate, 0.12 M malonic acid, 0.03 M bromide, 0.10 M sulfuric acid, and 0.2 mM ferroin under the perturbation of ethanol. The RGB intensity signals, termed the colorimetric oscillating signals (COS), could be drawn from recorded files through the image processing. We found from the COS characteristics that the oscillation period was inversely proportional to the ethanol levels as a consequence of its reaction with bromate in the studied system. Based on the established calibration, the ethanol content was effectively evaluated with good precision (<2.8% RSD). Also, the assay was successful with 5% error for the quantification of ethanol in commercial drink samples, signifying its accuracy and potential applicability.

Gravity-assisted distillation on a chip: Fabrication, characterization, and applications

Distillation is widely used in industrial processes and laboratories for sample pre-treatment. The conventional apparatus of flash distillation is composed of heating source, distilling flask, condenser, and receiving flask. As disadvantages, this method shows manual and laborious analyses with high consumption of chemicals. In this paper, all these limitations were addressed by developing a fully integrated microscale distiller in agreement with the apparatus of conventional flash distillation. The main challenge facing the distillation miniaturization is the phase separation since surface forces take over from the gravity in microscale channels. Otherwise, our chip had ability to perform gravity-assisted distillations because of the somewhat large dimensions of the distillation chamber (roughly 900 μL) that was obtained by 3D-printing. The functional distillation units were integrated into a single device composed of polydimethylsiloxane (PDMS). Its fabrication was cost-effective and simple by avoiding the use of cleanroom and bonding step. In addition to user-friendly analysis and low consumption of chemicals, the method requires cost-effective instrumentation, namely, voltage supply and analytical balance. Furthermore, the so called distillation-on-a-chip (DOC) eliminates the use of membranes and electrodes (usually employed in microfluidic desalinations reported in the literature), thus avoiding drawbacks such as liquid leakage, membrane fouling, and electrode passivation. The DOC promoted desalinations at harsh salinity (NaCl 600.0 mmol L−1) with high throughput and salt removal efficiency (roughly 99%). Besides, the method was used for determination of ethanol in alcoholic beverages to show the potential of the approach toward quantitative purposes.

Hydrogel-based piezoresistive sensor for the detection of ethanol

Abstract. This article describes a low-cost sensor for the detection of ethanol in alcoholic beverages, which combines alcohol-sensitive hydrogels based on acrylamide and bisacrylamide and piezoresistive sensors. For reproducible measurements, the reversible swelling and deswelling of the hydrogel were shown via microscopy. The response time of the sensor depends on the swelling kinetics of the hydrogel. The selectivity of the hydrogel was tested in different alcohols. In order to understand the influence of monomer and crosslinker content on the swelling degree and on the sensitivity of the hydrogels, gels with variable concentrations of acrylamide and bisacrylamide were synthesized and characterized in different aqueous solutions with alcohol contents. The first measurements of such hydrogel-based piezoresistive ethanol sensors demonstrated a high sensitivity and a short response time over several measuring cycles.

Total and beverage-specific alcohol intake and the risk of aggressive prostate cancer: a case-control study.

Ethanol in alcoholic beverages is a known carcinogen, but its association with aggressive prostate cancer (APC) is uncertain. Recent studies have shown a modest increase in risk of APC associated with heavy alcohol intake while association for beverage types remain inconsistent. Using a case-control design and self-administered questionnaire, we examined the association between APC (high grade and/or advanced stage) and frequency and quantity of alcohol intake 2 years prior to enrolment. Furthermore, we delineated the relationships for beverage-specific intakes of beer, red wine, white wine and spirits. The study included 1282 APC cases and 951 controls. Beer intake frequency of ⩾5 days per week was associated with increased risk compared with no beer intake (odds ratio=1.66, 95% confidence interval: 1.12-2.48) whereas wine was protective at all frequencies of consumption compared with those with no wine intake. For every 10 g per week ethanol intake from beer increase, the odds of advanced PC rose by 3% (OR=1.03, 95% CI: 1.02-1.05). No such increased risk was observed for red or white wine while a marginal dose-response relationship was found for spirits (OR=1.03, 95% CI: 0.99-1.07). Heavy beer and possibly spirits consumption is associated with increased risk while no dose-response relationship was found for red or white wine. Wine drinkers at all frequencies have a decreased risk of APC compared with those who did not drink wine.

Lifetime alcohol intake is associated with an increased risk of KRAS+ and BRAF-/KRAS- but not BRAF+ colorectal cancer.

Ethanol in alcoholic beverages is a causative agent for colorectal cancer. Colorectal cancer is a biologically heterogeneous disease, and molecular subtypes defined by the presence of somatic mutations in BRAF and KRAS are known to exist. We examined associations between lifetime alcohol intake and molecular and anatomic subtypes of colorectal cancer. We calculated usual alcohol intake for 10-year periods from age 20 using recalled frequency and quantity of beverage-specific consumption for 38,149 participants aged 40-69 years from the Melbourne Collaborative Cohort Study. Cox regression was performed to derive hazard ratios (HRs) and 95% confidence intervals (CIs) for the association between lifetime alcohol intake and colorectal cancer risk. Heterogeneity in the HRs across subtypes of colorectal cancer was assessed. A positive dose-dependent association between lifetime alcohol intake and overall colorectal cancer risk (mean follow-up = 14.6 years; n = 596 colon and n = 326 rectal cancer) was observed (HR = 1.08, 95% CI: 1.04-1.12 per 10 g/day increment). The risk was greater for rectal than colon cancer (phomogeneity = 0.02). Alcohol intake was associated with increased risks of KRAS+ (HR = 1.07, 95% CI: 1.00-1.15) and BRAF-/KRAS- (HR = 1.05, 95% CI: 1.00-1.11) but not BRAF+ tumors (HR = 0.89, 95% CI: 0.78-1.01; phomogeneity = 0.01). Alcohol intake is associated with an increased risk of KRAS+ and BRAF-/KRAS- tumors originating via specific molecular pathways including the traditional adenoma-carcinoma pathway but not with BRAF+ tumors originating via the serrated pathway. Therefore, limiting alcohol intake from a young age might reduce colorectal cancer originating via the traditional adenoma-carcinoma pathway.

Alcoholic Beverage Consumption and Chronic Diseases.

Epidemiological and experimental studies have consistently linked alcoholic beverage consumption with the development of several chronic disorders, such as cancer, cardiovascular diseases, diabetes mellitus and obesity. The impact of drinking is usually dose-dependent, and light to moderate drinking tends to lower risks of certain diseases, while heavy drinking tends to increase the risks. Besides, other factors such as drinking frequency, genetic susceptibility, smoking, diet, and hormone status can modify the association. The amount of ethanol in alcoholic beverages is the determining factor in most cases, and beverage types could also make an influence. This review summarizes recent studies on alcoholic beverage consumption and several chronic diseases, trying to assess the effects of different drinking patterns, beverage types, interaction with other risk factors, and provide mechanistic explanations.

New CNT/poly(brilliant green) and CNT/poly(3,4-ethylenedioxythiophene) based electrochemical enzyme biosensors

A combination of the electroactive polymer poly(brilliant green) (PBG) or conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) with carbon nanotubes to obtain CNT/PBG and CNT/PEDOT modified carbon film electrodes (CFE) has been investigated as a new biosensor platform, incorporating the enzymes glucose oxidase (GOx) as test enzyme, alcohol oxidase (AlcOx) or alcohol dehydrogenase (AlcDH). The sensing parameters were optimized for all biosensors based on CNT/PBG/CFE, CNT/PEDOT/CFE platforms. Under optimized conditions, both GOx biosensors exhibited very similar sensitivities, while in the case of AlcOx and AlcDH biosensors, AlcOx/CNT/PBG/CFE was found to give a higher sensitivity and lower detection limit. The influence of dissolved O2 on oxidase-biosensor performance was investigated and was shown to be different for each enzyme. Comparisons were made with similar reported biosensors, showing the advantages of the new biosensors, and excellent selectivity against potential interferents was successfully demonstrated. Finally, alcohol biosensors were successfully used for the determination of ethanol in alcoholic beverages.

An Ethanol Biosensor Based on Simple Immobilization of Alcohol Dehydrogenase on Fe3O4@Au Nanoparticles

AbstractAn ethanol biosensor based on alcohol dehydrogenase (ADH) attached to Au seeds decorated on magnetic nanoparticles (Fe3O4@Au NPs) is presented. ADH was immobilized on Fe3O4@Au NPs, which were subsequently fixed by a magnet on a carbon paste electrode modified with 5 % (m : m) MnO2. Optimum conditions for the amperometric determination of ethanol with the biosensor were as follows: working potential +0.1 V (vs. Ag/AgCl); supporting electrolyte: 0.1 M phosphate buffer solution at pH 6.8 containing 0.25 mM of the coenzyme (NAD+); working electrode: carbon paste with magnetically attached Fe3O4@Au NPs (0.012 mg ⋅ cm−2 electrode area) with immobilized alcohol dehydrogenase (120 units per cm2 of electrode area). Linearity between signal and concentration was found for the range from 0.1 to 2.0 M ethanol (r2=0.995) with a detection limit of 0.07 M, a sensitivity of 0.02 µA ⋅ mM−1 ⋅ cm−2, a reproducibility of 4.0 % RSD, and a repeatability of 2.7 % RSD. The results for the determination of ethanol in alcoholic beverages showed good agreement with gas chromatography (GC) with recovery of 96.0 – 108.8 %.

Detection of ethanol in alcoholic beverages or vapor phase using fluorescent molecules embedded in a nanofibrous polymer.

An alcohol sensor was developed using the solid-state fluorescence emission of terphenyl-ol (TPhOH) derivatives. Admixtures of TPhOH and sodium carbonate exhibited bright sky-blue fluorescence in the solid state upon addition of small quantities of ethanol. A series of terphenol derivatives was synthesized, and the effects of solvent polarities and the structures of these π-conjugated systems on their fluorescence were systematically investigated by using fluorescence spectroscopy. In particular, π-extended TPhOHs and TPhOHs containing electron-withdrawing groups exhibited significant solvatochromism, and fluorescence colors varied from blue to red. Detection of ethanol contents in alcohol beverages (detection limit ∼ 5 v/v %) was demonstrated using different TPhOHs revealing the effect of molecular structure on sensing properties. Ethanol contents in alcoholic beverages could be estimated from the intensity of the fluorescence elicited from the TPhOHs. Moreover, when terphenol and Na2CO3 were combined with a water-absorbent polymer, ethanol could be detected at lower concentrations. Detection of ethanol vapor (8 v/v % in air) was also accomplished using a nanofibrous polymer scaffold as the immobilized sensing film.

Recent Evidence on Alcohol and Cancer Epidemiology

This review presents an overview of the impact of alcohol consumption on cancer risk. Results from the 2009 International Agency for Research on Cancer monograph as well as the most recent meta-analyses and epidemiological studies are considered. Alcohol consumption is one of the most important known risk factors for human cancer and potentially one of the most avoidable factors, but it is increasing worldwide. Ethanol in alcoholic beverages is carcinogenic to humans and causes several cancers (oral cavity, pharynx, larynx, esophagus, colorectum, liver and female breast). Cumulative lifetime consumption, frequency and drinking pattern appear to play a role in risk characterization. While the role of heavy drinking has been long recognized, new evidence suggests that light consumption (up to one drink/day) is also associated with adverse effects. In addition, some genetic polymorphisms interact with alcohol metabolism and may modify its impact.

Alcohol and toxicity

Alcohol and toxicity

Sequential injection system for the enzymatic determination of ethanol in alcoholic beverages with in-line dilution

A sequential injection system for the enzymatic determination of ethanol in wines with spectrophotometric detection was developed. The determination was based on the enzymatic reaction involving ethanol and NAD+ in the presence of alcohol dehydrogenase. A gas-diffusion unit was introduced in the system to perform an in-line dilution. It was possible to operate the system with a reagent consumption of 1.3 mg of NAD+, 0.45 U of ADH, and 32 μL of sample, per assay with a determination rate of 21 h−1.The results obtained from the analysis of wine samples were statistically equivalent to those obtained by the reference method, with good repeatability; the relative standard deviation (n = 10) was <1% with a LOQ of 0.3% (v/v).