Determination of Alcohol Content Using Gas Chromatography

Determination of inebriantic beverage cognitive content Using vaunt Chromatography1.0TITLEDetermination of intoxicant beverage content in Tapai by apply mess up chromatography technique2.0INTRODUCTIONAlcohol is an thoroughgoing compound which contain hydroxyl functional group (-OH) that bonded to saturated snow (C) atom. For centuries, inebriant has been utilise for many purposes for examples as fuels, to subscribe to drinks, as a solvent and as an industrial feedstock. Alcohol disregard give us many benefits if we use it in the right carriage but it too stop give bad effects to us if we misuse it.Alcohol quarter be modernized commonly by victimisation excitement process. ferment process is a process which uses yeast as the catalyst to grass down sugar or glucose into alcohol and carbon dioxide. Ethanol is the or so common type of alcohol present from the process of hullabaloo.Yeast + glucose alcohol + carbon dioxideTapai is maven of the most general desse rt or fermented nutriment that can be lay out easily in East- and Southeast Asia such(prenominal) as Malaysia, Ind sensationsia, Singapore and Brunei. Tapai can be made from different primary(prenominal) ingredient such as glutinous sift, white sieve and cassava. During prep of Tapai, the ingredients be cleaned and washed before cooked. Cooked glutinous rice is then leave to cool down at temperature 30oC. To produce a matured and uncorrupted quality Tapai, it has to undergo upheaval for 3-4 days.Gas chromatography technique has been selected in this conduct to jog the summate of fermentation alcohol content in Tapai. A few aspects puddle been considered in choosing the right technique to be apply. Since Tapai contains alcohol, the Halal make love has been rise in Moslem community whether it is allowed to be consumed or veto because of some of the similarities amongst Tapai and boozer beverages.3.0LITERATURE REVIEW3.1 zymolysisFermentation is a process of muta tion of carbohydrates into alcohols and carbon dioxide or organic mordant by using yeast or bacteria as the catalysts. The yeast works to break down the carbohydrates or sugar. Fermentation refers to the chemical conversion of sugars to neutral spirits. It is a process which is commonly apply in found to produce soaker beverages such as beer and booze. anyway that, fermentation process is genuinely profitable for preservation of food. It produces lactic acids which can make the food can be used in a long period of time. For an example, it is found in such sour foods and pickled cucumber, yogurt and kimchi.Yeast + glucose alcohol + carbon dioxideIn Malaysia, fermented glutinous rice likewise known as Tapai is one of the most popular dessert among the Malaysian consumers. Tapai is a agreeable or sour dry taste and can be used as a dessert or traditional recipes. Tapai can be made from various types of carbohydrate sources, but commonly from cassava, glutinous rice and whit e rice. Yeast and sugar are used in the map to make Tapai.Fermented foods generally produce pleasant aroma, texture and create a salutary keeping quality under ambient environment. Fermented foods are fain from raw materials. Yeast fermentation generally is an alcoholic fermentation which involves the take of fermentation alcohol. Southeast Asia produces 150 million tones of paddy annually. Rice booze is one of the most popular sources to make alcoholic beverages in Asian country. In Malaysia and Indonesia, cassava tubers commonly used as fermented foods because of the soaring carbohydrates content in it.To make Tapai, the first look is soaked the rice in a gallon of wet and left overnight. Then, the soaked rice is steamed for 45 minutes. Cool the steamed rice to body temperature. After that, the Tape catechumen motor is sprinkled on the rice and mixed for a minute to distribute uniformly. The starter and rice have to be mixed well to prevent the spoilage. half(prenomin al) cup of sugar is mixed with water. The rice is jammed in layers into a jar and every layer is sprinkle with some sugar water. This step is taken to help the fermentation process to start. The rice is left for fermentation process to take place for 2-4 days. When the rice already produces a maculation of liquid at the bottom of the container or jar and distinctive olfaction is produced, the fermentation process is complete.There are 3 stages of fermentation which are Initial storey (Lag Phase), Middle Stage (Log to Stationary Phases) and Latter Stage (Death Phase). Initial stage is central to make natto bacilli grow sufficiently. The temperature is raft at body temperature 37-40oC. This temperature is an optimum temperature for the bacteria to grow. During the log phase, natto bacilli release a luckive deal of heat produced by the fermentation process. Sufficient oxygen must(prenominal) be supplied during this phase to make the process runs smoothly. After the stationary phase, the natto bacilli ambit the death phase. It undergoes autolysis gradually. 11% of glutamic acid is deliberated 16 hours after the fermentation process started. Organic acids are synthesized in this phase.Theoretically, fermented food is non considered to the full safe since they are prepared under uncontrolled conditions. Neverthe little, natural processes have been related with a few documented cases of food induced disease. inherent processed food has been known to cause serious food tipsiness compared to prepared packed food because the output of prepared packed food is been done in a factory with a regular observe from the quality control unit. look-alike 1Process of making Tapai3.2comparability between Tapai and alcoholic beveragesThe process of Tapai harvest-feastion is more likely to alcoholic beverages production where both of these productions have to go through the fermentation process. both processes use yeast in order to produce alcohol.There powerfulnes s be some differences for alcohol content in wine and Tapai since the new(prenominal) processes except fermentation which are used for alcoholic beverages but not in Tapai production. For Tapai production, the process takes place naturally while for alcoholic beverages production requires temperature control where there are hot fermentation and cool fermentation. blare et al. Went and Geerligs were the first person reported to segregate and identify the essential microorganisms for the fermentation of Tapai in 1895. In order to produce a dangerous Tapai, a mixture of Candida oryzae and Endomycopsis burtonii with 17% of reducing sugar and 2.7% of ethanol are needed. For wine and grapes, the healthy properties can be achieved due to the presence of antioxidants like polyphenols.From a chapter entitled Tapai processing in Malaysia A technology in musical passage by Zahara Marican and Yeoh Quee Lan in a book edited by KH Steinkraus entitle Industrialization of indigenous fermented f oods, the authors write more or less the making of Tapai in Malaysia. There are some points that can be considered as useful tuition from the quote on page 252 Tapai fermentation is not an anaerobic process and this partly accounts for the low alcohol content. Furthermore the fermentation is normally arrested before all of the sugars are fermented, as the desired end product should not be alcoholic if it is to be consumed by those opposed to alcohol consumption for religious or some different reasons. There is some information gained where Tapai fermentation is not an anaerobic. In the beginning, the enzymes are release to breakdown the sugars by the process of enzyme hydrolysis. This explained why Tapai tasted sweet.Besides that, Tapai production might be a little different compared to the wine or beer fermentation even though alcohol is produced and sugar is used. For Tapai fermentation, the theory of solid substrate fermentation is involved. separately of the white rice or g lutinous rice is coated with a thin layer of sticky liquid to make it difficult for oxygen to diffuse or penetrate. The reaction of the sugar molecules and the liquid caused the Tapai sticky or viscous by the interaction of hydrogen bond between the water molecules and sugar molecules. Furthermore, the low alcohol content in Tapai is caused by the somewhat open or not an air tight packing and senior high school room temperature which resulting in the vaporization of alcohol and the alcohols are escaped to the environment. The alcohols as the fermentation product have a short chain and gave a lower evaporation point and boiling point.Alcoholic beverages contain high amount of ethanol compared to Tapai. Beer contains approximately 6% of ethanol, wine contains 7 to 24% v/v of ethanol, whisky contains 35% of ethanol and Tapai contains only about 5% of ethanol. Besides ethanol, alcoholic beverages contain another chemical authorship such as water, glycerine, pestins, acids, polyphenols and some flavours. The least amount of ethanol contained in Tapai is caused by the volatilities of ethanol from the fermented rice due to non-air tight container.Figure 2Production of alcoholic beverages3.3Analysis of ethanol in fragmented rice using many techniquesRecently, many searches have been made in order to determine ethanol content in fragmented food such as wine and Tapai. Various types of techniques can be used for ethanol mark such as enzymatic method, biosensor, high liquid movement liquid chromatography (HPLC), near-infrared emission (NIR) spectroscopy and flow dig summary.3.3.1high-performance liquid chromatography (HPLC)High-performance liquid chromatography (HPLC) analysis was used to quantify the presence of individual phenolic resin compounds using an HPLC system consisting of a Shimadzu Prominence LC 20A, with an LC-20AT quaternary pump, a SIL 20 auto haver A, a DGU-20A5 on-line degasser, a CBM-20A integrator and a SPD-20AV DAD detector.HPLC technique ha s been designed to determine the amount of ethanol produced and amount of unwanted byproducts produced during the fermentation process. HPLC can produce the shortest analysis time and maintaining abounding solving between components for proper quantification and identification. By using this technique, all components can be analysed less than 10 minutes.3.3.2short circuit wave-near infrared (SW-NIR)Short wave-near infrared (SW-NIR) is one of the type of NIR. SW-NIR is one of the techniques that can be used to detect alcohol content in beverages and food products. The region of SW-NIR is between 700 to 1100 nm (Fu et al., 2012). The SW-NIR is suitable for non-destructive or non-invasive analysis of biological and biomedical materials (Ozaki et al., 1992 ai Ozaki, 2001 Sato et al., 2000). The SW-NIR can penetrate more intensely into a hear with less heating effect compared than long-wave NIR and the obstruction arising from the intense water bands can be diminished (Reeves III, 1 994). Tungsten lamps, LEDs or the sunlight can be utilized as the SW-NIR light source and the light can be transmitted through glass optical fibers (Fu et al., 2012). In order to determine the content of alcohol in aqueous samplings, temperature factor needs to be emphasized (Wlfert et al., 1998 Barboza Poppi, 2003). It is because temperature can cause a change in the vibe spectra (Fu et al., 2012). Figure 2 below shows the schematic diagram of the multichannel SW-NIR spectrometer module.Figure 2Schematic diagram of the multichannel SW-NIR spectrometer module3.3.3Fourier transform infrared spectroscopy (FTIR)In determination of alcohol, FTIR had been used for the quality control of alcoholic drinks such as spirit drinks and beer. FTIR with combination of multivariate data analysis can determine the essential compound quantitatively (Lachenmeier, 2007). Apart from that, Vonach et al. (1998) also stated that the combination of HPLC-FTIR can be used to determine the components of win e such as acetic, fructose, succinic, glycerol, ethanol, glucose, citric, lactic, malic and tartaric acid directly when used a real wine as a sample. By using FTIR, to buzz off the information about the product, for example the chemical composition, possibly we can make it within in a short time. Liquid Fourier transform-middle infrared spectroscopy can be used to analyze wine in a 90 s (Patz et al., 2004). Nowadays FTIR can give us advantages in order to determine and analyze the beverages because it can help us to subordinate the time cost, and at the same time, it is also contribute for a good clearcutness and accuracy when dealing with the parameters (Moreira et al., 2004). Furthermore, according to Lachenmeier (2007), a simple sample preparation is needed when dealing with FTIR spectroscopy compared to the other analytical methods.3.3.4 another(prenominal) spectroscopyRaman spectroscopy is not a preferable technique in determining ethanol content in Tapai due to the expensiv e pecker required. As HPLC method is low sensitivity but accurate deliberation process and the sample distillate are required, HPLC is not a good technique in this study. NIR spectroscopy is time consuming in establishing calibration curves and has low accuracy. It can be disturbed by other alcohol in the sample (Wang et al., 2003).3.4Determination of ethanol in Tapai by using Gas ChromatographyGas-liquid chromatography is one of the most sophisticated analytical techniques in spectroscopy. Gas chromatography becomes more preferable because of its utilization of electronic digital data-processing equipment for accurate qualitative and quantitative analysis.Traditional ethanol analysis methods are distillation and mass determination. These methods include grim accuracies because it can be interfere by other component in the sample. In order to evaluate the accuracy and precision of the gas-liquid chromatography, the ethanol standards and typical Tapai samples are necessary to be analyzed (B.Stackler and E.N.Christensen., 1974)To analyze ethanol contents in alcoholic beverages, Bouthilet et al. had made a packed GC method. Yet, the method needs at least 100mL of sample and distillation as the pretreatment process. These official gas liquid chromatography methods can be change with different sample preparation procedures when dealing with different samples. These samples require distillation or dilution as the pretreatment process. By using packed column, lower resolution and interference by other alcohols in alcoholic beverages still exist. When capillary tube vessel tube gas chromatography is used for determining the ethanol contents in alcoholic samples, time-consuming pretreatment procedures are still required. The capillary GC method is one of the most important techniques because of its advantage of high resolution and sensitivity (Wang et al., 2003).Wang et al. (2004) has analyze the gas chromatography coupled with flame ionization undercover wor k (GC-FID) to detect the ethanol content in alcoholic beverages. A selected column used is mega-pore capillary column (CP-Wax 58 CB). Acetonitrile was elect as the internal standard for ethanol and, 2-pentanol is used for methanol. This selection is due to the water solubility of acetonitrile is more or else than 2-pentanol. Direct snapshot GC coupling with dual internal standards was used to determine ethanol and methanol content can be obtained less than 12 min.Due to the success in the previous study, Brill and Wagner (2012) have performed the new method using GC-MS in order to detect the alcohol content that has been adapted by Wang et al.s (2004) experiment that used capillary gas chromatography together with FDI detector. This further study is quite costly, but it is expense since the accuracy is high. The column in this study was replaced by polar capillary column instead of mega-pore column in the previous study. 0.2 L splitless injections were chosen to directly inject t he samples to the GC-MS. Helium set at pressure set at 7.35 psi and 1mL/min of flow rate acted as the carrier gas in GC-MS. The polar VF-WAXms selected was the reliable because the samples could be run in 15 minute intervals.Capillary gas chromatography is a simple and speedy method that developed to determine ethanol content in alcoholic beverages using megapore polar column (CPWax58 CB, 30 m 0.53 mm) with direct injection gas chromatography. Contrary to packed GLC method, distillation or stepwise dilution of samples were not necessary by the method developed in this study. In conclusion, gas chromatography method is the most suitable and quick method for determination of ethanol contents in alcoholic beverages with complex alcohol contents and small sample amount.Solid-phase microextraction (SPME) is a non-harmful environmentally pre-concentration technique which is clean, selective, rapid, efficient, simple and solvent- excuse. The derivation of polyphenols allowed non-volatile compounds to be converted into volatile derivatives, thus improving the separation, selectivity or sensitivity of SPMEGC determination. This step can be performed in the sample solution foregoing to sampling, on the SPME fibre coating or during desorption in the GC injection port. For on-fibre derivation, the chemical agent has to be loaded on the fibre previous to or after analyte extraction. Commonly, the analytes are first extracted and then the fiber is unfastened to the vapour of the derivatizing agent. The coupling of GC and SPME has been used to determine resveratrol isomers and the total free resveratro was also determined by direct-exposure-probe positive-ion chemical ionization mass spectrometry. However, no references have been found to the use of the hyphenated technique SPMEGCMS for the determination of polyphenol mixtures (Pilar Vinas et al., 2009).4.0PROBLEM STATEMENTSThe purpose of this study is to determine the most suitable and countenance method for alcohol de termination in the sample used which is fermented rice or commonly called as Tapai. Tapai is a traditional dessert or fermented food frequently found in East and Southeast Asia. Nowadays, Tapai has been launch and being promoted to many countries. Determination of ethanol perhaps is the most important analysis in the fermentation patience for the quality control. It is necessary for the industry to use the most accurate and easiest method in order to view the alcohol composition in the products manufactured before sending them to the markets.Besides that, this study is fundamental in order to determine the Halal limits for alcohol content in the sample. The consumption of Tapai had raised the Halal issue to Muslim community since it has similar alcohol content with beer which is 5%. So, the study should expand the method that can give the short period for the detection and the easiest way to handle the experiment or the test include the easy sample preparation with the accurate results.5.0OBJECTIVESThese are few objectives that have been identified in order to deal up with the problem statement and carried out the research on determination of alcohol content in Tapai by using gas chromatography spectroscopy.To develop gas chromatography method with the best accuracy compared to other methods for alcohol detection in the sample.To determine the lower limit of detection (LOD) of alcohol.To identify the amount of ethanol content in the sample.