Topic for Presentation:

Analysis of Beverages, Analysis of Alcoholic Beverages as per BIS and PFA Act, Detection & Determination of ethanol, furfural, organic acids, aldehydes, chloral hydrate, methanol and ethylene glycol in liquors by colour tests, TLC, GC, & GCMS methods, Distinction between licit & illicit liquors.
By, Praachi Deshmukh MSc I Institute of Forensic Science, Mumbai.

Analysis of Beverages:
Beverages are nothing but Drink / Liquid for human consumption. There are main two types: Alcoholic & Non-Alcoholic Non-Alcoholic Beverage is also called as Carbonated Water means Water (as per PFA std.) impregnated with carbon dioxide under pressure and may contain any of the following singly or in combination: Sugar, liquid glucose, dextrose monohydrate, invert sugar, fructose, honey, fruits and vegetable extractives, permitted flavouring, colouring matter, preservatives, emulsifying and stabilizing agents, citric acid, fumaric acid and sorbitol, tartaric acid, phosphoric acid, lactic acid, ascorbic acid, malic acid, edible gums, edible gelatin, salts of sodium, calcium, magnesium, vitamins, caffeine not exceeding 145 parts per million, ester-gum (glycerol ester of wood rosin) not exceeding 100parts per million, quinine salts (sulphate)<100ppm. It may also contain saccharine sodium,100ppm

Analysis of Nonalcoholic drink:

Tests for presence of Glycerol esters of wood rosins commonly known as ester-gum: 1. Identification: Insoluble in water, soluble in acetone and in benzene 2. IR spectrum: obtain IR spectra of a sample deposited on potassium bromide plate scan between 600 and 4000 wave numbers. Compare with pure ester gum. Tests for Absence of toll oil rosin (Sulphur test): When sulphur containing organic compounds are heated in the presence of sodium formate, sulphur is converted into hydrogen sulphide which is readily detected by use of lead acetate paper. A positive test indicates use of tail oil rosin instead of wood rosin.

 Arsenic Not more than 3ppm Lead - Not more than 10ppm Heavy metals - Not more than 40ppm Acid value Between 3 and 9 Hydroxyl number Between 15 & 45. Analysis of Alcoholic Beverages An alcoholic beverage is a drink containing ethanol, commonly known as alcohol. Toddy: Toddy means the sap from coconut, date, toddy palmtree or any other kind of palm tree which has undergone alcoholic fermantation. 1. White cloudy in appearance 2. with sediments on storage, 3. possess characteristic flavour derived from sap 4. fermentaion without adding extraneous alcohol. 5. Also be free from: chloral hydrate, paraldehyde sedative, tranquilizer & artificial sweetener. Standards to be confirmed: Alcohol content Not less than 5%v/v Total acid as tartaric acid - Not more than 400gms Volatile acid as acetic acid Not more than 100gms Analysis of Alcoholic Beverages The BIS (Bureau of Indian Standards) prescribes methods of test for alcoholic drinks. For estimation of esters, higher alcohols, aldehydes, furfural, & methanol GC (gas chromatographic) method is given in Annex A as alternative method. Purpose: To follow 1. Prevention of Food Adultration Act 1954 and rules framed thereunder; 2. Standards of Weights & measures Rules, 1977; & 3. State Excise Duty Rules Alcoholic Drinks : Methods of Test Colour Tests: Qualitative Analysis of Liquor Test for Ethyl Alcohol: 1. Iodoform Test: 1ml sample + 1ml 5% sodium hydroxide + iodine soln dropwise with shaking until liquid becomes persistent dark brown in colour. Keep for 2-3 min. If iodine colour disappears, add more drops of iodine until persistant brown colour. Add few drops of dil sod

hydroxide to remove extra iodine. Add equal volume of water, keep for 10min. Yellow crystalline ppt = positive test for presence of ethanol. 2. Dichromate test: 1ml sample + 0.2ml2%pot dichromate sol + 1ml concn.sulphuric acid = Yellow colour of dichromate changes to green or blue = presence of ethanol. Test for Methanol: 1. Chromtropic test: 1ml sample in tt + 2ml pot permangnate sol = shake well + few crystals of sod bisulphate till disappearance of colour + 1ml chromotropic acid + concn sulpuric acid slowly with inner side wall of the tt upto 15ml. Appearance of violet colour = presence methanol. 2. Schiffs Reagent Test: 4.5ml sample + 0.5ml ethanol + 2ml 3% pot permangnate sol + 2ml phosphoric acid. Keep for 10min. Add 1ml 10%oxalic acid followed by 1ml conc sulpuric acid. + 5ml Schiffs reagent. Keep for 30min. Observe colou, if purple = methanol present. Parallel expt. To be carried out with control and blank. Test for Furfural: 1. 5ml sample + 1ml aniline + 0.5ml hydrochloric acid + Keep for 15 min. = Red colour = Furfural is present. 2. 2ml sample + 0.2ml aniline + 0.4ml glacial acetic acid. Red colour develops in few seconds & reaches its max. intensity in 5-10 minutes, if furfural is present. 2ml sample + 1ml aniline acetate sol. (10%v/v) = Red colour develops at RT 25-30C & reaches its max in 1-5 minutes. = furfural is present. Quantitative Determination Determination of Furfural: If red colour develops in the colour test of furfural, proceed for quantitative estimation. Dilute measured portion of distillate (V1) with 50% furfural free alcohol to 50ml. Add 2ml colourless aniline & then 0.5ml hydrochloric acid. Mix and keep at 15C for 15min. Compare colour developed with std furfural solution. Calculation:

Furfural, gms/100litres of absolute alcohol = W * 1000 * 100*100 / V1 * V2 where, V1 = Volume of experimental solution used for estimation, V2 = Alcohol % by volume. Determination of Ethyl Alcohol Content: By finding out specific gravity of the distillate at a particular temperature with the help of Pyknometer .

 Obtain corresponding alcohol percent by volume from the tables given in IS 3506. Determination of Aldehydes: 50ml distillate of liquor + 10ml bisulphite sol = Keep in dark fo 30min with occasional shaking. Add 25ml std iodine sol & back titrate excess against std sodium thiosulphate sol using starch indicator. Run a blank taking 50ml distilled water. The difference in titration value, in ml of sodium thiosulphate sol (V), gives equivalent aldehyde. Calculation: V * 0.0011 * 100 *1000* 2 / V1 V1 = alcohol % by volume Note: 1ml of sodium thiosulphate sol = 0.0011 g of acetaldehyde. Determination of Methanol: Two methods, viz., 1. spectrophotometric method & 2. gas chromatography method are employed. The spectrophotometric method is sufficiently sensitive for routine type of analysis. The gas chromatography method is more sensitive and shall serve as an alternative method. Spectrophotometric Method Spectrophotometer of any make with wavelength range from 350 to 700nm & maximum band width of 5nm. Has to prepare Methanol std sol from stock solution, sod salt of chromtropic acid sol, pot permangnate sol. Distill 50ml of sample, take 40 ml of distillate. Dilute 1ml of distillate to 5ml with dist. Water, shake well, take 1ml of this solution, 1ml dist. Water (for blank), & 1ml of methanol std sol in tt and keep in ice-cold water bath. Add to each tt 2ml pot permangnate, keep for 30min. Decolourise by adding a little sod bisulphite & 1ml chromotropic acid. Mix well, add 15ml sulphuric acid slowly with swirling, place in hot wterbath at 80C for 20min. Observe colour development from violet to red. Cool reaction mixture and measure the absorbance at 575nm using 1cm path length cell. Calculation: Methanol content in g/100litres of absolute ethanol = A2*C*D*1000*100*100 / A1*S where,

A2= Absorbance for sample solution, C= Concentration of methanol std solution, g/ml D= Dilution factor for sample solutuio, A1= Absorbance for methanol std solution, & S= Ethanol content of liquor sample in %v/v. Gas Chromatography Method GC equipped with Flame ionization detector & split injection port with tempretures at 250C. Inject 2microliter of methanol std sol into chromatograph and record chromatogram. Determine the retention time of methanol and n-pentanol. Inject 2 microlire of sample solution into chromatograph and record chromatogram. Calculation: Methanol content in g / 100l absolute alcohol = R2*C*D*1000*100*100 / R1*S where, R2=peak ratio of methanol to n-pentanol for sample solution, C=Concentration of methanol in std solution, D= dilution factor for sample solution, R1= peak ratio of methanol to n-pentanol for std solution, S= ethanol content of liquor sample in percent v/v. Distinction between licit & illicit liquors