Screening of Suspected Liquids to Alcoholic Drinks by a Qualitative Chemical Kit

Research Article

Austin J Environ Toxicol. 2021; 7(2): 1038.

Screening of Suspected Liquids to Alcoholic Drinks by a Qualitative Chemical Kit

Hossein Mahdavi SA1, Hassanian-Moghaddam H2,3, Hashemi Nazari SS4, Akhgari M1, Shahbazi F5, Nazari Kangavari H6, Akhavan Tavakoli Ar7, Rafizadeh M8 and Rafizadeh A9*

1Legal Medicine Research Center of Iran, Tehran, Iran

2Social Determinant of Health Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

3Department of Clinical Toxicology, Loghman Hakim Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran

4Safety Promotion and Injury Prevention Research Center, Department of Epidemiology, School of Public Health and Safety, Shahid Beheshti University of Medical Sciences, Tehran, Iran

5Department of Epidemiology & Biostatistics, School of Public Health, Hamadan University of Medical Sciences, Hamadan, Iran

6Department of Epidemiology & Biostatistics, School of Public Health, Iran University of Medical Sciences, Tehran, Iran

7Legal Medicine Research Center, Legal Medicine Organization, Rasht, Iran

8pharmacy faculty, Mazandran University of Medical Sciences, Ramsar, Iran

9Department of Nursing & Midwifery, Islamic Azad University, Rasht Branch, Rasht, Iran

*Corresponding author: Rafizadeh A, Department of Nursing &Midwifery, Rasht Branch of Islamic Azad University, Poletaleshan, Lakan road, Rasht, Iran

Received: April 20, 2021; Accepted: May 20, 2021; Published: May 27, 2021


Alcohol consumption is prohibited in Iran and it has caused many problems in various fields. This research is done to evaluate a new qualitative screening kit (for rapid detection of alcoholic beverages from nonalcoholic liquids) and to avoid more wasting time and money. For this purpose, the ethanol content of 839 suspected liquids prepared from the alcohol lab of legal medicine (Guilan province, Iran) was measured by a gas chromatography apparatus. At the same time, a newly designed kit based on the sulfochromic acid method was also used to separate negative samples (with ethanol equal or less than 3% volume/volume) from the others. Out of 839 samples, the ethanol content of 163 (19.43%) cases was appraised negatively. While most of the samples (676 cases or 80.57%) had ethanol more than 3% (positive results) with a mean of 21.68mg/L. Three samples (0.36%) had high levels of methanol which were interpreted as false positive due to the nonspecificity of the used method. Our results suggest the used kit has suitable efficiency for screening ethanol content of suspected liquids because the negative results of the kit have definite diagnostic value. However, a confirmative test (like gas chromatography) is needed for validation of the positive results.

Keywords: Alcoholic beverages; Alcoholic strength; Ethanol; Ethanol detection


FID: Flame Ionization Detector; GC: Gas Chromatography; GC-Mass: Gas Chromatography-Mass Spectrometry; HPLC: High Performance Liquid Chromatography; LMO: Legal Medicine Organization; NPV: Negative Predictive Value; PPV: Positive Predictive Value; SPSS: Statistical package for social science; V/V: Volume/Volume


Production, trade, and consumption, of alcoholic drinks, are prohibited in Iran and there are severe punishments for them. But, these penalties not only have been unable to decrease alcohol consumption in Iran society but also, have been lead to increase usage of poor-quality alcohol (fake and homemade) that can be accompanied by many dangers including methanol intoxication [1,2]. One of the tasks of the police in Iran is the confiscation of suspected liquids to alcoholic drink and the determination of their owners’ legal status by measuring the ethanol concentration in them. To measure their ethanol content, the mentioned fluids are referred to Legal Medicine Organization (LMO). Based on the Iran Legal protocols, liquids with 3% volume/volume (v/v) ethanol or less are not considered being alcoholic beverages at all, and their owners should be released immediately. But, the owners of solutions contains ethanol more than 3% v/v must be referred to the judicial system for the continuation of the legal proceeding [3,4]. But, due to different reasons, measuring the amount of ethanol of samples is so timeconsuming that put makes their owners waiting for the final court order.

Like methanol, different advanced methods including High- Performance Liquid Chromatography (HPLC), Gas Chromatography- Mass Spectrometry (GC-MS), and usually Gas Chromatography (GC) are currently used to detect or quantify ethanol in different samples including alcoholic beverages [5-11]. Likewise, biological and chemical methods are used for this purpose too [12-18]. GC is usually used as a gold standard method for the quantitative analysis of alcohol in Iran and worldwide [19]. Application of these methods is almost time-consuming and costly and need very expensive devices and high technical knowledge that make them inapplicable in common laboratories with many samples per day [1,2,5-10,19].

For these reasons, now a day rapid and chemical methods have been widely considered for different purposes due to their simplicity, easiness, low cost, short operation time and etc [13]. Accordingly, the accessibility to an easy method for quick screening of these samples and separating negative cases from positive ones (based on a cutoff point) to avoid spending a lot of time and money can be a great advantage.

So, in this study, the ethanol content of 839 samples was measured to evaluate the efficacy of a newly designed kit. For this purpose, the GC method was used as a gold standard. Lastly, the gained results by the kit and GC methods were compared together to confirm kit ability and final conclusion.

Materials and Methods

In this study, 839 suspected liquids to alcoholic beverages were used as samples. They had been discovered by police (Gilan province, Iran) between April 2017 and May 2018 and were referred to the LMO of Rasht, Guilan province, Iran. Their ethanol contents were determined by both kit and GC methods. The obtained results by both techniques were compared together and the kit function was approved by comparison with the GC method. The technicians of each section were blind to the results obtained by another division. Statistical analysis was performed using SPSS software version 24 (IBM Corporations, Chicago, Ill, USA).


A GC apparatus (Shimadzum 148, Japan) was used to determine the ethanol content of the samples. This device contained a packed column and FID detector. To performance of the tests, 0.2μL of the diluted (1:100) samples were manually injected to GC by 1μL Hamilton syringe. The GC injector and detector temperatures were set at 180°C and 210°C, respectively. No internal standard was applied.


The samples (839 suspected fluids) were prepared from alcohol lab of Rasht LMO. To evaluate the ethanol contents of samples by GC, a mix standard solution with 320mg/dL of methanol and ethanol was prepared from with Merck Company (Darmstadt, Germany) trademark. Also, a newly designed kit produced by Arya Mabna Tashkhis Co., Tehran, Iran, was used to qualitatively detect ethanol content of the samples. This kit is designed based on traditional sulfochromic acid method and contains two reactants (a diluent buffer and one color reactant) and an instruction brochure in the pack.

Procedure of the Kit

According to the brochure of the kit, 50μL of each sample was poured into a test tube with 1ml of A reactant (diluent buffer). Then, 1mL of B reactant (color reactant) was added to the test tube and shaken hardly to mix. After one minute, the final result was concluded as positive (turquoise color) or negative (different colors except turquoise or colorless) according to the appeared color.

Study measures

Key indexes in this study were sensitivity, specificity, Positive Predictive Value (PPV), and Negative Predictive Value (NPV). The sensitivity and specificity of a qualitative test refer to its ability to correctly identify the positive and negative results, respectively. In the diagnostic tests, the PPV and NPV should be proportional with the true positive and negative results, respectively. In this research, the accuracy of the newly designed test measured by two area under the ROC curve. The value of this index can be variable between 0 and 1. An area of 1 represents a perfect test; by decreasing the value of this index the test becomes more worthless. Although, positive and negative likelihood, roc area and chance ratio were calculated in the same real and virtual incidences too, the sensitivity, specificity, PPV and NPV are more discussed in continue.


Out of 839 samples, 72 (8.58%) cases had not ethanol and the ethanol content of 91 (10.85%) cases was determined less than 30000mg/L (3% v/v). In other words, 163 (19.43%) cases were determined as negative outcome (nonalcoholic drink). The most of these samples (676 cases or 80.57%) had ethanol more than 3% v/v that was evaluated as positive result (alcoholic drinks). Table 1 shows the analytical quality assurance of the kit including four main parameters (sensitivity, specificity, PPV and NPV).

Citation: Hossein Mahdavi SA, Hassanian-Moghaddam H, Hashemi Nazari SS, Akhgari M, Shahbazi F, Nazari Kangavari H6, et al. Screening of Suspected Liquids to Alcoholic Drinks by a Qualitative Chemical Kit. Austin J Environ Toxicol. 2021; 7(2): 1038.