1.1.

THE WINE

Wine is a health drink resulting from complete or partial fermentation of grapes, either exclusively by natural micro-flora of grapes or the added wine yeast culture (Joshi and Attari, 1990). Wine is an alcoholic beverage made from the fermentation of fruit juice (grapes, apple, etc). The natural chemical balance of grapes is such that they can ferment without addition of sugars, acids, enzymes or other nutrient. Although other fruits such as apples and berriers can also be fermented, the resultant wines are normally named after the fruit from which they are produced. Wine is produced by fermenting crushed fruits using various type of yeast which consume the sugars found in the fruits and convert them into alcohol. Various varieties of fruits and strains of yeasts are used depending on the type of wine produced.

1.1.1.

HISTORY OF THE WINE

The latest evidence suggests that wine was being made during the Neolithic period (approximately 6000 B.C.) in the Near East. Many lines of evidence indicate that wine making and grape cultivation began in the Transcaucasian region encompassing the northern portion of Turkey, Iran and Iraq, the southern region of Armenmia and Azerbaijan. This zone corresponds to the closest overlap of natural distribution of the wild Vitis vinifera (spp.silvatica) with the Northern spread of agriculture out of its Near Eastern centre of origin.

Figure 1.1: A 19th century reproduction of the Rigveda Viticulture was believed to have been introduced to India by Persian traders sometime in the 4th millennia BC. Historians believe that these early plantings were used mostly for table

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grapes or grape juice rather than the production of an alcoholic beverage. During the Vedic period of the 2nd and 1st millennia, the Aryans tribes of the region were known for their indulgence of intoxicating drink and it seems probable that wine was a pleasant beverage. The religious text of the Vedas mentions at least one alcoholic drink that may have been wine related-sura which seems to have been a type of rice wine that was fermented with honey. The first known mentioning of grape- based wines was in the late 4th century B.C. writings of Chanakya who was the chief minister of Emperor Chandragupta Maurya. In his writings, Chanakya condemns the use of alcohol while chronicling the emperor and his court's frequent indulgence of a style of grape wine known as Madhu. In the centuries that would follow, wine became the privileged drink of the Kshatriya or noble class while the lower caste typically drank alcohol made from wheat, barley and millet. Under the rule of Muslim Mughal Empire alcohol was prohibited in accordance to Islamic dietary laws. However, there are written reports about at least one Mughal ruler, Jahangir who was fond of brandy wine. In the 16th century, Portuguese colonists at Goa introduced port style wine and the production of fortified wines soon spread to other regions. Under British rule during the Victoria era, viticulture and wine making was strongly encouraged as a domestic source for the British colonists. Vineyards were planted extensively through the Baramati, Kasmir and Surat regions. In 1883, at the Calcutta International Exibition, Indian wines were showcased to a favorable reception. The Indian wine industry was reaching a peak by the time the phylloxera epidemic made its way to country and devastated its vineyards. It was a long road for the Indian wine industry to recover from the devastation at the end of the 19th century. Unfavorable religious and public opinion on alcohol developed and culminated in the 1950s when many of India's states prohibited alcohol. Vineyards were either uprooted or encouraged to convert to table grape and raisin production. Some areas, like Goa, continued to produce wine but the product was normally very sweet and highly alcoholic. The turning part of the modern Indian wine industry occurred in early 1980s with the founding of Chateu Indage in the state of Maharashtra With the assistance of French winemakers, Chateau Indage began to import Vits vinifera grape varieties like Cabernet Sauvignon, Chardonnay, Pinot blanc, Pinot noir and Ugni blanc and started making still

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and sparkling wines. Other wineries soon followed as the emergence of India's growing middle class fueled the growth and development of the Indian wine industry. However, Indian wine yards were totally destroyed by unknown reasons in the 1890.

1.1.2. WINE GROWING REGIONS IN INDIA
Following are the major wine growing regions in the country:<1> Nasik Region (Maharashtra State): It is the biggest wine producing region in India. This region includes Pune, Nasik and Ahmed Nagar. It is above 800 meter sea level. Several top wineries are located in this area including Chateau Indage and Sula Wines. <2> Sangli Region (Maharashtra State): This region includes Solapur, Sangali, Satara and Latur. It is above 800 meter sea level. <3> Bangalore Region (Karnataka State): Nandi Hills located about around 45 kilometer north of Bangalore City. Grover Vineyards is located in Nandi Hills. It is above 800 meter sea level. <4> Himachal Region: It is located at Northern India. It is upcoming state for the wine production. Temperature varies from 20 degree C to 40 degree C. Unique climate of this region attracts the wine makers to produce delicate wine grapes.

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1. Eighty percent of wine consumption in the country is confined in major cities such as Mumbai (39%). Chateau Indage at Narayangaon is a pioneer of French style wines in India. Grover Vineyards and Sula Vineyards make invaluable contribution for production of wines in the country.4 million litres production.2: Wine producing regions of India 1.Figure 1.2million litres annually. There is growing awareness about the wine as a product in the domestic market. Maharashtra is leading among the states with 36 wineries and 5. WINE INDUSTRY: INDIAN PERSPECTIVE Approximately 40 wineries are presently operating in the country with a total production of 6.3. produces exquisite qualities in 4 . Bangalore (9%) and Goa (9%). Delhi (23%). Major wineries such as Champagne Indage.

both still and sparkling wines.4. Tannin is what gives red wines a complexity that is beyond that of most white wines. Wine is classified as dry table wines. The company has the capacity of producing over 3 million bottles annually. 5 .6%) and higher sugar content (atleast 24 Brix).For sweet desert wines. medium acidity (0.9%) and moderate sugar content (19 to 23 Brix) (Bammi. The way that the red wine gets its colour is by letting the skins soak in the juice until the red colour bleeds out. Central governments have announced number of financial incentives and administrative reforms.1. Everyone knows that almost all grapes have colourless juice. 1968). Soaking the skins give red wine its colour and also imparts a substance known as tannin. Thus cultivation of good quality of grapes is very much important.5 to 0. Chenin Blanc is predominantly grown but emphasis should be given to red wine varieties. the must should be of relatively higher pH (3. high acidity (0.35).30 to 3. Red Wines: Red wine is simply wine produced from red (or black) grapes.6 to 0. Some common types of wine are:1. which contain alcohol less than 14% and little or no unfermented sugar and sweet dessert wines are those having 17 to 20% alcohol and some unfermented grape sugars.00 to 3. 1. Quality wines can only be prepared from quality grapes. Sula vineyards at Nasik are new vineyards in India for making wine. Several research workers in India have concentrated their efforts to produce quality wines with improved health benefits.Dry table wines require grapes of fairly low pH (3. Reportedly the world wine market is growing at 14% annually whereas the Indian market at more than 20%. TYPES OF WINES Various research workers specified different compositional requirements of grapes and must for different wines. Sangali is another region for wine production.65).

this firmness can be quite intense. Examples are from California. Higher in alcohol. taste of black cherries and sometimes mint. This is one of the main reasons that red wines usually age better than whites. Nebbiolo Pinot Noir High in tannin and acid and need aging to mellow.2 Other Red Wine (Grape) Varieties 6 . Table: 1. Over time. Ranging from light and fruity to big and spicy depending on the quality. grown in the Piedmont region of Italy. The most finicky of grapes producing the widest range of quality. Table: 1. Syrah/Shiraz Zinfandel Rich and spicy wine with lots of tannin and the sweetness of blackberries. the qualities of the tannin will mellow and blend harmoniously with the other characteristics of the wine.Tannin has a mouth drying quality that causes the wine to feel firm in your mouth. When a red wine is young.1 Major Red Wine (Grape) Varieties Varieties Cabernet Sauvignon Merlot Properties Thick skinned grape with lots of tannin. and blackcurrant flavor.

A good rosé should be delicate and refreshing. Believe it or not. It is made from a blend of grapes which include Pinot Noir and Pinot Meunier (black grapes). 2. It is possible to make white wine out of black grapes by carefully extracting the juice and keeping the skins separated. A rich and spicy wine and is popular in Chile. This wine has medium acidity and tannin. White Wines Most white wine is produced from white grapes. High in alcohol and is usually sweet and peppery. a blush. Wine gets its colour from letting the skins soak in the juice. which is just another term for rosé. Besides colour. Rosés have been given a bad reputation from 7 . Found in France. The best rosés are made from the Grenache grape. Champagne is the most famous example. but not as much as in a typical red wine. is considered a white wine. The oak barrels will impart some tannin to the wine. not allowing the skins and stems to soak in the juice also reduces the amount of tannin in the wine.Varieties Barbera Carmenere Gamay Grenache/Garnacha Malbec Sangiovese Properties Low tannin with high acidity. They are made by allowing the skins to soak for only a short period of time before extracting. Sometimes though. not cloyingly sweet. They actually taste of grapes and are low in tannin. A smooth and plummy variety from Argentina. a white wine will be allowed to ferment or age in oak barrels.

Produces the only wine to actually smell like grapes. white wine with the taste of limes. A faily unexciting grape popular in cooler climates for its early ripening qualities. crisp 8 .some of the extremely sweet and cheap varieties on the market. The most intensely aromatic of all wines. A low alcoholic wine with striking acidity. Table: 1. Famous for making Italy's Soave. It is a highly acidic wine that can range from very dry to very sweet. This wine is fresh and tangy like green apples.3 Major White Wine (Grape) Varieties Varieties Chardonnay Chenin Blanc Gewürztraminer Muscat Reisling Sauvignon Blanc Properties It is usually oak aged and has a buttery flavor. These are the most tangy and pungent of the wine varieties.4 Other White Wine (Grape) Varieties Varieties Verdelho Colombard Garganega Müller Thurgau Pinot Properties A rich. Don't let that stop you from finding some that are truly delicious and worth savoring. Produces a crisp every day wine with tropical fruit aromas. Table: 1. Highly acidic and low sugar levels resulting in a dry. The aromas are of florals and spice.

The scientific community has begun to unveil some of the mysteries surrounding this topic. Their chief danger comes from the damage they can do when they react with important cellular components 9 .Blanc Viognier 3. Port wine (also known as Vinho do Porto. and the media has begun whetting our thirst for knowledge. The wine received its name. "Port". Free radicals are atoms or groups of atoms with an odd (unpaired) number of electrons and can be formed when oxygen interacts with certain molecules. 1. US. Port Wine wine. aging. The purpose of this article is to serve as a beginners guide to what antioxidants are and to briefly review their role in exercise and general health. A low acid wine with floral aromas and an apricot quality. leaving residual sugar in the wine. like dominoes.2. India. What follows is only the tip of the iceberg in this dynamic and interesting subject. Port is produced from grapes grown and processed in the demarcated Douro region. Once formed these highly reactive radicals can start a chain reaction. South Africa. and a variety of diseases. Porto. Canada. and often simply Port) is a portugese style of fortified wine originating from the douro valley in the northern provinces of Portugal. It is also grown in Australia. ANTIOXIDANTS AND FREE RADICALS Antioxidants are intimately involved in the prevention of cellular damage -. The wine produced is then fortified by the addition of a neutral grape spirit known as Aguardente in order to stop the fermentation. in the latter half of the 17th century from the seaport city of Porto at the mouth of the Douro river where much of the product was brought to market or for export to other countries in Europe. and to boost the alcohol content. Athletes have a keen interest because of health concerns and the prospect of enhanced performance and/or recovery from exercise.the common pathway for cancer.

and grains. if this occurs. milk. The RDA is 60 mg per day. Intake above 2000 mg may be associated with adverse side effects in some individuals. Current recommended daily allowance (RDA) is 15 IU per day for men and 12 IU per day for women.) 1. squash. and apricots.2. Beta-carotene is a precursor to vitamin A (retinol) and is present in liver. broccoli. cantaloupe. yams. Although there are several enzyme systems within the body that scavenge free radicals. Because beta-carotene is converted to vitamin A by the body there is no set requirement. cantaloupe. Types of antioxidant Table 1. cabbage. Instead. is sometimes included in this category. (NOTE: Vitamin A has no antioxidant properties and can be quite toxic when taken in excess. the body has a defense system of antioxidants. and vitamin C. egg yolk. Vitamin E: d-alpha tocopherol. green peppers. The body cannot manufacture these micronutrients so they must be supplied in the diet. or the cell membrane.such as DNA. seeds. Additionally. whole grains (esp. spinach. butter.5: Some important enzymatic and non-enzymatic physiological antioxidants 10 . carrots. tomato. Vitamin C: Ascorbic acid is a water soluble vitamin present in citrus fruits and juices. fortified cereals. to clarify the relationship. the RDA is expressed as retinol equivalents (RE). the principle antioxidants are vitamin E. and strawberries. kiwi. a trace metal that is required for proper function of one of the body's antioxidant enzyme systems. vegetable and fish oils. A fat soluble vitamin present in nuts. selenium. To prevent free radical damage.1. Antioxidants are molecules which can safely interact with free radicals and terminate the chain reaction before vital molecules are damaged. peaches. spinach. Cells may function poorly or die. broccoli. beta-carotene. wheat germ). kale.

Glutathione Non-protein thiol in cell Cellular defense Serves oxidant as for 4. Catalase (CAT) Mitochondria & Cytosol Removal of H2O2 B. Bilirubin Product of heme Extracellular antioxidants metabolism in blood 3. Vitamin-C Aqueous phase of cell Free scavenger. Vitamin-E Cell breaking antioxidants 11 .Sr. ENZYMATIC OXIDANT 1. α-lipoic acid Endogenous thiol substitute Glutathione. Glutathione peroxidase (GSH) Mitochondria & Cytosol Removal of H2O2 &Organic hydroperoxide 2. recycle Vit-E Chain 6. Carotenoids Lipid soluble antioxidants Removal in membrane tissue reactive species of Oxygen 2. Recycling Vit-C 5. NON ENZYMATIC OXIDANT 1.no Antioxidant Location Function/property A.

complete and efficient conversion of grape sugars to ethanol. It is not surprising that the finest wines are made using only the finest ingredients and also the finest winemakers use Lalvin dried wine yeast. Using Lalvin wine yeast made by Lallemand is recommended.2. Eg: Vitamin-C & Carotenoids. Lallemand is the world's largest producer of specialty yeast for wine making and produce over 10 different wine yeast strains for the professional winemaker in active dried form. C. Radical scavenging antioxidants . Enzyme inhibitor antioxidants . Yeast is the workhorse that converts the initial sweet syrupy must into great-tasting wine. D.These suppress the free radical formation.These suppress the chain initiation reaction. Uric Acid Product metabolism of Purine Scavenging of OH radical 1. enzymes such as peroxidase.These induce production and reaction of free radicals and the transport of appropriate antioxidants to appropriate active site. lactoferrin. 2. THE YEAST It is very important to use high quality yeast in all winemaking.It comprises of proteolytic enzymes and repair enzymes of DNA and genetic materials.2. Preventive antioxidants . but important metabolites without the development of off-flavours. The primary role of wine yeast is to catalyse the rapid.7. catalase. carbon dioxide and other minor.1. Antioxidants based on defense mechanism Antioxidants based on defense mechanism are of four types: A. Ex. Repair and de novo antioxidant . 12 . etc B. carotenoids.

Kluyveromyces.1. 2. saccharomyces and Zygosaccharomyces.2 STRAIN NAME WINE ALCOHOL CONTENT TEMPERATURE OTHER RANGE INFORMATION 13 . Debaromyces. Kurtzmann suggested that 15 are associated with winemaking Brettanomyces. body.2.A lot of research has been done that confirm that the yeast play a critical role in determining the flavour.1 TAXONOMY Out of the 100 yeast genera representing over 700 species described in the latest edition of the monographic series. Metschnikowia. Cryptococcus. Rhodotorula. viscosity and colour of wines. aroma.1. STRAINS OF WINE YEAST i) Red Star Active Dry Yeasts – Table 2. Hanseniaspora and its asexual counterpart Kloeckera.1. The scientific classification of Saccharomyces cerevisiae Kingdom Phylum Subphylum Class Order Family Genus Species Fungii Ascomycota Saccharomycotina Saccharomycetes Saccharomycetals Saccharomycetaceae Saccharomyces cerevisiae 2. A taxonomic study. Candida. Pichia. Table 2.

White 18% 45-95°F Sparkling wine 13-15%.Assmannshausen Red wines Adds is a a spicy its aroma.Table 2. wine White 12-14% 18-20%.3. fast fermenter. drawback ineffectiveness in high solid content. but does well Pasteur Champagne Premier Curvee Red. 59-86°F not with work high of Epernay Flor Sherry Montrachet White wines Port. 13% low temperatures 59-86°F 59-86°F sugar levels tolerance medium-high alcohol conditions qualified for barrel fermentations it is tolerant to heat and sulfur dioxide Pasteur Red Red wine 16% 64-86°F ii) Lalvin Active Dry Yeasts Many of Lalvin's yeasts are intended for commercial rather than home use. 14 . Madeira Red. Some are as follows: . slow fermenting anaerobic fermentations It tolerates sulfur dioxide well.Lalvin active dry yeast STRAIN NAME ALCOHOL CONTENT Lalvin 43 18% TEMPERATURE RANGE 55-95°F OTHER INFORMATION Restart abilities.

Lalvin 71B-1122 14% 60-85°F Metabolizes more of the malic acid during fermentation. a It requires high- nitrogen nutrient. Lalvin AC 14% 85° F produce low levels of SO2 and H2S Lalvin AMH 15% 68-86° F enhances spicy and fruity flavors. Lalvin BGY 15% 75-86°F It is used in reds. aroma Lalvin BA11 16% 68-86°F It requires a highnitrogen nutrient Lalvin BDX 16% 64-86° F It is highly recommended for the production of quality dry red wines. particularly Noir. for fermentations. Lalvin MO5 14% 59-90° F 15 . Pinot slow Lalvin CSM 14% 59-89° F This strain requires high levels of nitrogen and nutrients but will promote malolactic fermentation. It works particularly well in low maturity white grapes from cool regions.

clean wines. Lalvin W15 16% 50-81° F It was developed to ferment dry whites and rosés at moderate speeds where bright fruit and heavy mouth feel are desired.Lalvin M1 16% down to 54° F This strain is used to produce aromatic rosé and white wines. Lalvin R2 16% 42-86° F It may be used for fruit wines whenever a Sauternes wine yeast is specified. Lalvin QA23 16% 50-90° F Used for Sauvignon Blanc. they have a four-to-six-month shelf-life. especially wines with residual sugar." 16 . Refrigerated. They should be attemperated to within 100F of the must for several hours before "pitching. Colombard. Semillon fruity. in theory they impart better flavours to the must. Chenin Blanc. iii) White Labs Yeasts White Labs wine yeasts are "pitchable" yeast cultures. meaning they contain sufficient living yeast to inoculate five gallons of must without rehydration or starter propagation. and for production of fresh. Because they are fresh.

17 . WLP720 Sweet Wine 15% 70-75° F A good choice for sweet mead and cider.white labs yeast STRAIN NAME ALCOHOL CONTENT WLP715 Champagne 17% TEMPERATURE RANGE 70-75° F OTHER INFORMATION Classic yeast. Semillon. used to produce wines. including Chenin Blanc. dry attenuate wines/ strong ales. as well as Blush wines. and Sauvignon Blanc. and Riesling WLP730 Chardonnay White Wine 14% 50-90° F This is a good choice for all white and blush wines.4. Chablis. champagne. or to fully barley cider. Pinot Noir.Table 2. WLP740 Merlot Red Wine 18% 60-90° F This is a vigorous fermenter well suited for Cabernet. Gewurztraminer. dry meads. Shiraz.

WLP760 Red Wine Cabernet 16% 60-90° F High tolerance. Extensive research has demonstrated the protective properties of antioxidants. Blanc. This is largely due to the scientific development.2. independent of its alcohol content. which scavenge reactive oxygen species (ROS) and their precursors. Wine contains a naturally rich source of antioxidants. the wine industry has seen a revolution in the wine production technology. Studies have shown that despite the high intake of saturated fatty acids within the diets of some populations. fermentation Chardonnay. namely the phenolic acids and polyphenols. a determinant of age-related disease.Chardonnay. which may protect the body from oxidative stress. which provide it with its protective redox potential. Red wine contains a rich source of a large number of antioxidants. a reduced mortality rate from cardiovascular disease is attributed to the high consumption of red wine. as well as up-regulate enzymes involved in the repair of cellular damage. Annually. Chenin suitable for Merlot. Sauvignon Blanc. The current study set out to determine the in vivo effects of moderate red wine consumption on antioxidant status and oxidative stress in the circulation. Moderate 18 . and Semillon. The role of nutrition in health has captured researcher's interest in antioxidants and their capacity to protect the body from damage induced by oxidative stress. temperature moderate speed. Blanc. about 26 billion litres of wine are produced from about 8 million hectares of vineyards across the world. WINE In the last 90 years. and Sauvignon 2.

73± 0.consumption of red wine has also been shown to retard or slow the plasma clearance of high density lipoproteins (HDL). It is also observed that percent body fat. Unmodified tartaric acid esters of hydroxycinnamic acids from wine are 19 . other safety parameters were unchanged ( Flechtner – Mors. 2004). the aim of this study was firstly to understand how moderate red wine consumption (400 ml/day) for two weeks effected circulating lipids. waist circumference. These results indicate that red wine inhibits cell mediated LDL oxidation more efficiently than white wine and at much lower concentrations. blood glucose.1999). antioxidant level and total antioxidant capacity in the circulation and secondly assess the differences in bioefficacy of red wine in young and older populations(Cui. 2007). The absorption in human body of phenolic acids from white wine. whereas white wine lacks polyphenols.7 mg/ethanol/mL respectively. particularly hydroxycinnamic acids from white wine are absorbed from the gastrointestinal tract and being circulated in the blood after being largely metabolized to the form of glucornide and sulfate conjugates. up to a concentration of 1. These compounds possess antioxidant activity and are able to reduce the release of pro-inflammatory cytokines.. It is also being observed that consumption of white wine in moderate quantity help in reducing weight and other problem (Lamuela-Raventos et al.53kg. It is being observed that there is redution in weight upto 4. triglycerides. A large body of evidence which indicates that free radical production can directly or indirectly play a major role in cellular processes implicated in atherosclerosis and CVD.04 and 0. but it contains other compound such as hydroxycinnamic acids (caffeic acids) and monophenols (tyrosol). insulin. The incubation of low density lipoproteins LDL in varying concentrations of red and white wine showed a 50% decline in oxidation at concentrations of 0.J.2002) Recent study on wine against cardiovascular diseases have shown that red wine have high content of antioxidant especially polyphenols. The experimental rats were given white wine and it also showed protection against cardiovascular disease (Mahesh et al.. A group of obese patients were taken and till 3 months they were given moderate quantity of wine. and cholesterol were reduced. a negative risk factor for the development of cardio vascular disease (CVD). Therefore..0 mg/mL. blood pressure. which are known to have antioxidant properties. et al.

2007). et al. there was also a clear increase in homocysteine after a month of wine consumption. contains all the nutrients present in the fruit juice (Bhutani et al. it is being seen that when HCT-15 cells. reduced glutathione. However. CHEMICAL COMPOSITION AND SENSORY PROPERTIES OF WINE Wine is produced by fermentation of fruit juice and therefore. TBARS. 1989).1. Whereas white wine did not contain anthrocyanins. a decrease in superoxide dismutase activities and TBARS concentrations. vitamins. After that levels of glutathione peroxidase. Each participant provided their venous blood samples. A study was carried out on daily consumption of wine on oxidative stress.. In addition it contains alcohol. The anthrocyanin fraction from the red wine and the non-anthrocyanin substances extracted from red and white wine suppressed the growth of the cells. HDL cholesterol. Marco et al. Crude methanol extracts of red and white wines were added to diethyl ether in order to divide them into the anthrocyanin fraction and fractions containing other flavonoids and their derivatives. which is derived from human colon cancer or AGS cells.2007). total antioxidant capacity. are bioavailable to humans (Nardini. glutathione peroxidase and reduced glutathione levels. 2009). were cultured with these fractions..human plasma at low levels. and the suppression rate by the anthrocyanin fraction was significantly higher than that of other fractions (Hashimoto. paraoxonase. Volunteers were taken and each of them was supplied 375ml of white wine daily. Wine hydroxycinnamic acids although present in wine as conjugate form.2. This is clear that daily consumption of white wine is not only associated with both oxidative and antiatherogenic effects but also with a potentially proatherogenic increase of homocysteine concentration (Rajdl. Mirella et al. and other compounds synthesized by fermenting yeasts. 2. Yoko 1998). A typical wine contains ethyl 20 . total cholesterol. superoxide dismutase is being measured and after a month there is significant increase in HDL..D..cholesterol. derived from human gastric cancer. Lipofuschin is an end product of lipid peroxidation which dramatically increases ethanol consumption (Assuncao.

sugars. higher alcohols.1: Wine composition Component Water Alcohol Everything Else Percentage 85 12 3 Principal components of a litre of wine (1000 ml) Figure 2. amino acids. tannins.2: Water glycerol tartaric acid malic acid lactic acid 21 . 1998). esters.alcohol. fatty acids and minor constituents like flavouring compounds (Joshi. aldehydes. Figure 2. acids. anthocyanins.

0. It is reported that 0.248 – 0. The percentage of alcohol in wine is approximately equal to the brix multiplied by the factor 0. 1980 and Joslyn.57 (Cruess 1958).586% tartaric acid and 0. Sucrose is rapidly hydrolysed during fermentation and is present in wine in very small amounts (0. sweet and very sweet wines contain 2.0 to 3.144% 22 . 1977).2 to 15. 1954).5% (Suresh et al. rhamnose. 1985). reported production of 50% alcohol from the total sugar content of the berries (Pandey and Pandey 1990). The alcohol content of the wine prepared from four cultivars of grape was in the range of 11.. sucrose.2 pH (Suresh et al.058 – 0. xylose. The dry wines are practically free of sugars and contain 0 to 1. arabinose.. Wine may contain 85% to 88% and 12 to 15% dry matter (Jarczyk and Wzorek. respectively (Jarczyk and Wzorek. d) Organic acid The titrable acidity in the wines prepared from four cultivars ranged from 3.6 to 4.. 8 to 11 and 7 to 12% sugars. b) Alcohol Wine contains mainly ethyl alcohol and traces of methyl and higher alcohols (Amerine. 1961).06%) (Amerine et al.potassium succinic acid a) Water reducing sugar phenolic compound azoted substances It is the major constituent of wine. 1985). While semidry. 1977).01 to 0.0% sugars. c) Sugars The sugars present in the wine include glucose. fructose.

2001) e) Proteins The nitrogenous compounds present in wine include protein. gallic acid. most wine drinkers stopped swirling for a long minute and took notice. peptones. According to Indian standards for wine. About half of the wine proteins are bound to phenolics. 1960). and wine drinkers across the world swirled and sipped and rejoiced.mallic acid is present in Thompson Seedless wines (Chikkasubbana 1990). amino acid and ammonia. iron. THE HEALTH BENEFITS OF WINE When word got out about the health benefits of wine. 1985). a wine should contain total acids in the range of 4 to 15g/l (as tartaric acid) (Bhalerao. (Ribereau Gayon et al. New wines contain nearly 200mg/l anthocyanin. copper.. polypeptides. 1983).063% in wine (Patil 1994). It is being analysed that the total nitrogen of 0.09% and proteins of 0.01 to 0. cobalt. zinc. iodine. phosphorous. f) Minerals The mineral elements present in wine include magnesium. aluminium and lead. amides. bromine. A wide range of tannin content in white wine (46 to 143mg/ 100ml) and red wines (28 to 328 mg/100ml) was reported (Suresh et al. 1epigallocatechin.. caffeic acid and ellagic acid (Henning and Brukhardt. g) Phenolic compounds The phenolic compounds isolated from wines include catechin. whereas old wines contain 20mg/l anthocyanin (Joshi 1998) and 273 mg/l anthocyanin content in red wines. 1-epictechin. potassium. 23 . 1998) 2. sodium.56% is present in 16 young wines (Somer and Ziemelis 1973). calcium.050 to 0.07 to 0.2. silica. A wide range of total phenols in white wines ranging from 50 to 6500 mg/l and red wines from 1000 to 4000 mg/l were reported (Joseph et al.2. It was the mid 90s when story of the "French Paradox" poured from all media sources. They are present in varying proportion in different type of wine.. Protein content is in the range of 0.

Thomas’ Hospital in London. 2004). • Stronger bones (Twin Research and Genetic Epidemiology Unit. g) Women’s Health Benefits: • Decreased ovarian cancer risk (The Queensland Institute of Medical Research in Australia. f) Stroke Prevention: • Arteries kept clean by polyphenols in red grape skins (William Harvey Research Institute. 2004). 2002). 2004). d) Ulcer Prevention: • Ulcer-causing bacteria reduced (American Journal of Gastroenterology. 2004). 2004). b) Lung Health Benefits: • Improved lung function from antioxidants in white wine (American Thoracic Society. St. 2002). c) Heart Health Benefits: • Coronary heart disease reduced (University of California. 24 . e) Cancer Prevention: • Cancer cells killed by protein in red grape skins (University of Virginia Health System.a) Overall Health Benefits: • Anti-aging effects in red grape skins (Harvard Medical School in Boston. 2003). 1995). • Healthier blood vessels in elderly (University of Ferrara in Italy. Davis.

the nomenclature ‘Alcohol’ denotes the entire family of organic compounds like methanol. Ethanol (C2H5OH) in specific. 25 . In fact. h) Men’s Health Benefits: • Lower risk of heart attack for men with high blood pressure (Worcester Medical Center in Massachusetts. the most commonly ingested of these chemicals. It is highly volatile liquid with distinct odour having great affinity towards water. one must get it in larger quantities. So with all of the benefits of wine discussed. 2. resveratrol is getting all the praise from the medical field.3. The amount to which extent the central nervous system is affected depends upon the concentration of alcohol in blood. its high solubility in water makes it one of the most potent depressant of human central nervous system. 2004). The way to do that is in pill form.2. Alcohol. Hydrogen. In fact. is an organic compound made up of molecules of Carbon. But the best of all is Concentrated Resveratrol. To really benefit. HARMFUL EFFECTS OF WINE i) Alcohol Although. It is a powerful hypnotic sedative with an array of side effects. and Oxygen. too much wine per day or per week can lead to negative health effects in many cases and rather than prevent disease. in more general terms we denote ‘Ethanol’ when we discuss alcohol. isopropanol.• Lower risk of stroke (Centers for Disease Control and Prevention. The problem with getting it naturally from wine is that the quantity is very low. 2001). ethanol. usually denoted by BAC (Blood Alcohol Content). one will actually up your odds of acquiring such diseases.

the increase in fatty acid levels results in forming of plaque in the hepatic capillaries. This stage is often termed as Euphoria where BAC is around 0. hippocampus of brain helps to shed off inhibitions. it can penetrate almost all cellular membranes resulting in absorption by all organs.03 to 0. This situation leads to Liver Cirrhosis. Although. the enzyme secreted by hepatic cells converts alcohol to acetaldehyde. person feels more relaxed and cheerful which is followed by more stumbling movements and animated speech. They become more confident. As liver performs vital role in filtration mechanism of body. iv) Metabolism and its effects Alcohol dehydrogenase. As mentioned earlier. previous studies have failed to establish direct connection between alcohol and its effect on cancer. While. shying off their inhibitions. This acetaldehyde further gets converted into acetic acid and then acetate by acetaldehyde dehydrogenase. increased alpha waves from brain makes person more cheerful. At initial stages. there is a strong indication to suggest that alcohol enhances the effects of other 26 . Rest 80% is then absorbed by small intestine from where it gets distributed throughout the human body. relaxed. the effects of alcohol on human body changes gradually over the time. Then it goes to stomach where only 20% of the total quantity is absorbed by it.12%. Acetate is a compound of fats which gets deposited locally. stimulation by alcohol to cortex. iii) Absorption and Distribution Mechanism When consumed. As alcohol enhances antidiuretic hormone secretion. more urine is formed which results in dehydration. Because of the chronic and continuous consumption of alcohol. This happens because of the increased metabolism in nigrostriatal pathway of brain which is associated with body movements. due to its high affinity towards water. v) Carcinogenic Effects Alcohol comes under Group1 carcinogens as classified by WHO.ii) Effects at Initial Stages When ingested. alcohol first irritates the mucous lining of mouth and then esophagus causing an anesthetic effect. malfunctioning of liver often leads to jaundice (Hepatitis). Alcohol travels through blood and come into the vicinity of cells of almost every organ.

the condition is known as Confusion. Dizziness continues.39%. the condition is known as Lethargy. esophagus more prone to cancer. When BAC crosses the mark of 0. the condition is known as Stupor. its withdrawal also develops symptoms like delirium tremens which has high percentage of mortality rate (35%). 5. It is characterized by unconsciousness when body reflexes are low. overly aggressive. Their reaction with polyamines can end up in forming mutagenic DNA.28%. When BAC count ranges from 0. if gets concentrated in high amounts then it can damage the DNA of cells. They are not certain of what they are doing. When BAC count ranges from 0.carcinogenic chemicals like tobacco. It is characterized by aggravated emotional state where people try to be sentimental. When BAC count ranges from 0.35 to 0. body movements are severely affected and patients lose and regain consciousness intermittently. 4. While consumption of alcohol develops an array of diseases. The excessive consumption of ethanol also makes mouth. alcohol consumption can lead to fetal alcohol spectrum disorder. In this condition. the condition is known as Coma. pharynx. Alcoholism is a major public health problem.23%.50%. 3. their movements’ loose coordination and start losing their body balance. it results in failure of CNS (Central Nervous System) ultimately resulting in Death. It is also characterized by blurred vision. Nausea is the common symptom of this phase.09 to 0. Acetaldehyde. When BAC count ranges from 0. larynx.25 to 0. They are generally classified as follows1. if not treated. breathing rate declines resulting in dropping of heart beat rate. people become sleepy.17 to 0. vi) General Effects on Human Body There are different effects of alcohol on human body subject to their concentrations in blood. During Pregnancy. 2. 27 . They have high risk of coma or even death. In this stage.5%. the byproduct of metabolism of alcohol.

Black grape is 28 .5trihydroxystilbene) and then preserves it in an airtight bottle. peanuts. 2004. 1995) Resveratrol is synthesized in the skin but not in the fleshy part of the berries of Vitis vinifera and Vitis labrusca grapes. worms and mice—has been dramatically lengthened by minute amounts of a red wine extract. fruit flies. Man’s pursuit of long life. Japanese knotweed was known to comprise in Resveratrol. Lead researcher David Sinclair. a unique antioxidant that red grapes produce in great amounts as a defence against fungi. and perhaps as a dietary supplement.. Though red wine had been known to have health benefits since long but in 1992. it was recommended that probably the secret of these benefits lies in increased Resveratrol content in red wine. whose skin consists of Resveratrol that serves as the protective shield against diseases. RESVERATROL Resveratrol is an antioxidant that is commonly found in many plants.2. resveratrol. otherwise it would vanish in days. edged closer to fruition with the recent announcement that a dietary component may increase the human lifespan to the point where it would be common to live 125 years. polygonum cuspidatum extracts and red wine extracts. This redcoloured youth potion can be obtained from a bottle of vino. says the lifespan of all life forms tested so far—yeast cells. extends the life of yeast cells by 70 per cent.. The process of winemaking utilises alcohol to extract resveratrol (or 3. the so-called fountain of youth. Recent research indicates that resveratrol. This substance is usually created in the outer layers of fruits and plants and defends us against diseases caused by pathogens along with other benefits. PhD.3. assistant professor of pathology at Harvard University Medical School. Food supplements often comprise in Resveratrol in pills that contain grape seed extract. Jeandet et al. found in red grapes. Data in the literature indicates that its quantity is very variable (Lamuela-Reventros et al.4. The skin of red wine is the most abundant source of resveratrol. Red wine. Muscadine grapes are utilized in making red wine. it was isolated from white hellebore roots by Takaoka. Originally. grapes and blueberries are some of its rich and popular sources. Later.

internucleosomal DNA fragmentation. 2003). 1999). the sirtuin gene upregulates the activity of an enzyme (histone deacetylase) that prolongs the time cells have in which to repair 29 . Resveratrol can reduce serum lipid levels and can prevent or inhibit cellular events associated with tumor initiation. K 2009).. V. Both cis. Resveratrol treatment resulted in a gradual decrease in the expression of anti-apoptotic Bcl-2 (Young-Joon et al. animals and probably humans (Heilbronn LK.and trans-isomers of resveratrol have been analyzed in skin. starting from must to skin separation and in wine after malolactic fermentation. thus may help prevent cardiovascular disease and cancer (Joshi. trypan blue dye exclusion assay. (Bruno. 2. and cell cycle analysis showed that resveratrol has a dose-dependent antiproliferative and antiapoptotic action on DHL-4 cells. et al. The addition of polyvinylpolypyrrolidone (PVPP) significantly lowers the resveratrol content. researchers have recognised that severe restriction of calories can significantly broaden the lifespan of insects.17 mg/l. Resveratrol concentration increases during fermentation on the skins but the amount extracted depends on the variety and enological conditions. which can damage DNA and may lead to cancer onset. In this study we have indagated resveratrol anticancer action.. Ravussin E. it is being seen that resveratrol is able to neutralize free radicals. seeds and must of grape berries of Gama. and increased proportion of the sub diploid cell population. analyzing its effects on both cell cycle and growing of human lymphoma B (DHL-4) cells.. MECHANISM OF ACTION Since 1935.33 mg/l to 7. The mechanism behind calorie restriction appears to be a survival factor that is turned on when living organisms are exposed to harsh conditions.nature's best source of resveratrol. The growth inhibitory and antiproliferative properties of resveratrol appear to be attributable to its induction of apoptotic cell death as determined by morphological and ultrastructural changes. When a living organism is deprived of calories. These results confirm resveratrol's potential therapeutic role on tumors. 2003).3. Resveratrol reduces viability and DNA synthesis capability of cultured human promyelocytic leukemia (HL-60) cells. promotion and progression. In recent studies.P. MTT colorimetric test.1. Trans-resveratrol concentration in red wines ranges from 1.

The enzymatic activity also ‘silences’ the genes responsible for protein production (ribosomal DNA). In human terms. What Sinclair discovered was the dietary switch to turn on this mechanism. Slowing down the rate at which proteins are produced slows the rate of ageing itself. did indeed turn on the survival switch and extend the life of yeast cells by 70 per cent. Surprisingly. Therefore. the extract obtained from red wine. et al. What grapes use to turn on this survival mechanism. Cells have machinery to increase enzyme activity on their own. can be transferred to humans in a glass of wine. their strands of DNA. The enzyme itself cannot be bottled because to work. Aged cells typically produce hundreds of thousands of extra copies of ribosomal DNA. From a library of thousands of molecules.their damaged genetic material. resveratrol inhibits the over-production of proteins within cells that leads to accelerated ageing. the calorie restriction mimic. that would be equivalent to 30 to 50 years of added life! Resveratrol was superior to the 16 other molecules tested (Howitz KT. The uniqueness of resveratrol may be partly explained by the fact it is utilised by cells and orally absorbed by humans better than other antioxidants found in grapes (Soleas GJ. 2002). Bitterman KJ et al. 2003). Resveratrol. 17 activated the human survival/longevity gene. it has to be delivered to cells at the right place and time. A family of ironcontrolling antioxidant molecules was screened for its ability to increase the activity of the enzyme.. SUPPLEMENTS FALL SHORT Dietary supplements providing resveratrol from red wine or knotweed are available. tests conducted at Harvard University by Sinclair have failed to find any significant biological activity in resveratrol dietary supplements in tablets.2. 2. responsible for activating the same enzyme. Humans have a similar gene.3. The accumulation of these proteins in living cells has been likened to an ageing clock. 2003). cross-species transfer process scientists now call xenohormesis (Hall SS. SIRT1. Sinclair began to research the dynamics of this survival mechanism.. capsules or as 30 .

However. fills gelatin capsules in nitrogen rather than an oxygen environment and seals dietary supplements ingredients in an airtight pill. Mega doses of resveratrol do not produce greater longevity and in fact may work in an opposite manner and become problematic to genes. a relatively new technology called Licaps. resveratrol appears to work in wine. Leroy Creasy. showcasing the need for special measures that should be undertaken in the manufacture of resveratrol supplements.3. Encapsulation fails to duplicate the airtight environment found in a wine bottle. Licaps technology is being utilised to produce the first stabilised red wine extract. 2002). (Kimura Y. produced under nitrogen and preserved in part by refrigeration. which has demonstrated biological activity. and also under laboratory conditions as a pure 100 per cent molecule. Creasy claims it would take thousands of capsules of resveratrol to provide the equivalent amount of resveratrol found in a glass of red pinot noir wine (Leroy Creasy 2003). reported that resveratrol supplements failed to exhibit much biological activity—evidenced by the ability to activate an enzyme that promotes DNA repair and lengthen the life of yeast cells—compared to wine. developed by Capsugel specifically for liquid ingredients. would be sufficient for humans to achieve enzyme activity levels equivalent to those achieved in the laboratory.. a professor of plant science at Cornell University in Ithaca. REQUIRED AMOUNTS A small amount of resveratrol was found to increase the survival of yeast cells by threefold even when the cells were exposed to ionizing radiation (Zoberi I et al. PhD. which provide about 3mg resveratrol.liquid herbal extracts (David Sinclair 2003). much higher amounts of resveratrol have been used successfully in animal tests for treating cancer. Three five-ounce glasses of red wine per day. but apparently his report went unnoticed by manufacturers. Although studies are lacking that show resveratrol in pills work. A nitrogen bubble inside the capsule also retards any spoilage. However. 2. Okuda 2001) 31 . New York.3. which preserves the resveratrol.

This would also slow the ageing process if you start taking in small quantities in your middle age. the principle micronutrient (vitamin) antioxidants are vitamin E. selenium. ANTIOXIDANTS Antioxidants are a type of complex compounds found in our diet that act as a protective shield for our body against certain disastrous enemies (diseases) such as arterial and cardiac diseases.4. Although there are several enzyme systems within the body that scavenge free radicals. beta-carotene. Last but not least. BENEFITS OF RESVERATROL Years of researches have shown several benefits of this natural compound: • It helps you to reverse diseases that come handy as you grow old. • 2. The body cannot manufacture these micronutrients so they must be supplied in the diet. 32 .2.4. and vitamin C. • • • Resveratrol contain in one red wine glass can suppress carcinogenesis that otherwise cause breast cancer. is sometimes included in this category. antioxidant and inflammation damper. Additionally. This can be done with regular intake of Resveratrol while you are still quite young.3. cataracts and also premature ageing along with several chronic diseases Antioxidants are molecules which can safely interact with free radicals and terminate the chain reaction before vital molecules are damaged. such as diabetes. a trace metal that is required for proper function of one of the body's antioxidant enzyme systems. it ensures sound cardiovascular system. arthritis. • It increases your life span as high resveratrol levels counteract with cell death and damage in brain and heart. It is an effective artery protector. Low Resveratrol doses provide the same effect as that of a calorie-reduced diet. cancer and heart diseases.

New data from ongoing studies will be available in the next few years and will shed more light on this constantly evolving area. Antioxidants are also thought to have a role in slowing the aging process and preventing heart disease and strokes. This has lead to the theory that these diets contain substances. It is not possible to directly measure free radicals in the body. possibly antioxidants. Thus far. This is an important finding because it shows how 33 . If the generation of free radicals exceeds the antioxidant defenses then one would expect to see more of these by-products. male smoker who did not supplement. These measurements have been performed in athletes under a variety of conditions.1.Antioxidant help in many ways:Epidemiologic observations show lower cancer rates in people whose diets are rich in fruits and vegetables. is to eat 5 servings of fruit or vegetables per day. scientists have approached this question by measuring the byproducts that result from free radical reactions. which protect against the development of cancer. but the data is still inconclusive. Therefore from a public health perspective it is premature to make recommendations regarding antioxidant supplements and disease prevention. Regular physical exercise enhances the antioxidant defense system and protects against exercise induced free radical damage. EXERCISE AND OXIDATIVE DAMAGE Endurance exercise can increase oxygen utilization from 10 to 20 times over the resting state. In fact one study demonstrated an increased risk of lung cancer in male smokers who took antioxidants vs. none of the large. prompting concern about enhanced damage to muscles and other tissues. 2. Several interesting concepts have emerged from these types of experimental studies. which comes from several authorities in cancer prevention. Whether this effect was from the antioxidants is unknown but it does raise the issue that antioxidants may be harmful under certain conditions. There is currently intense scientific investigation into this topic. Perhaps the best advice. This greatly increases the generation of free radicals.4. well designed studies have shown that dietary supplementation with extra antioxidants reduces the risk of developing cancer.

2. A placebo controlled study done on mountaineers demonstrated less free radical damage and decline in anaerobic threshold in those athletes supplemented with vitamin E. The experimental studies are often conflicting and conclusions are difficult to reach. It is hypothesized that vitamin E is also involved in the recovery process following exercise. 2. These changes occur slowly over time and appear to parallel other adaptations to exercise. the "weekend warrior" who is predominantly sedentary during the week but engages in vigorous bouts of exercise during the weekend may be doing more harm than good. 34 . Thus. Nevertheless. the amount of vitamin E needed to produce these effects is unknown.4. this finding suggests that supplementation with vitamin E might be beneficial in those triathletes who are adapting to higher elevations. including degree of conditioning of the athlete. Antioxidant supplements prevent exercise induced damage or enhance recovery from exercise:Although it is well known that vitamin deficiencies can create difficulties in training and recovery. PERFORMANCE In general. the role of antioxidant supplementation in a well nourished athlete is controversial. The one exception to this is vitamin E which has been shown to be useful in athletes exercising at high altitudes. but some may require supplementation. intensity of exercise. antioxidant supplements have not been shown to be useful as performance enhancers. intense exercise in untrained individuals overwhelms defenses resulting in increased free radical damage. most of the data suggest that increased intake of vitamin E is protective against exercise induced oxidative damage. Currently. The diet may supply enough vitamin E in most athletes. and diet. On the other hand. Although difficult to generalize.smart the body is about adapting to the demands of exercise. To this end there are many factors which may determine whether exercise induced free radical damage occurs. There is no firm data to support the use of increased amounts of the other antioxidants.

Taking chemicals without a complete understanding of all of their effects may disrupt this balance. harmful.3. Antioxidants supplements were once thought to be harmless but increasingly we are becoming aware of interactions and potential toxicity. thus. no one knows if supplements should be taken and. very little is known about the long term consequences of mega doses of antioxidants. in the normal concentrations found in the body. Also. but at higher concentrations they are pro-oxidants and. AMOUNT REQUIRED BY BODY Although there is little doubt that antioxidants are a necessary component for good health. 35 . if so. It is interesting to note that. how much. vitamin C and beta-carotene are antioxidants.2.4. The body's finely tuned mechanisms are carefully balanced to withstand a variety of insults.

OTHER IMPORTANT MATERIALS:Autoclaved flask and beaker. potassium iodide.4. Mumbai. 450ml of sulphuric acid. 4 of port wine and 4 of white wine. HPLC grade water etc according to different biochemical test. gloves.1N NaOH). pH meter. ethanol were taken.1.3. TBA reagents. ascorbic acid. measuring cylinder. momo pan balance. 0.96gm of potassium dichromate. sodium thiosulphate (0. forceps. blade.1N sodium thiosulphate.1. 1. All 12 samples are stored in air tight centrifuge tubes.4.3. 3.2. 36 . pH ESTIMATION OF THE INDIAN WINES – pH of the wine samples was measured by using Toshniwal pH meter. pipettes. 3. PROTOCOL :– The estimation of total alcohol content is being measured by dichromate method. dichromate reagents.4. 3. Sigma. WINE SAMPLE COLLECTION:Different samples of wine are purchased from different wine shops and restuarent.2. Mumbai and Hi-media.4 of red wine. For estimation of alcohol content we need 1% starch. ALCOHOL ESTIMATIONA. Mumbai. B. They were obtained from Qualigens. CHEMICAL AND REAGENTS:Most of the chemicals used in this investigation were of analytical grades. REAGENTS AND STANDARDS: – 1% starch.12 wine samples were collected. They were DPPH. distilled water. BIOCHEMICAL STUDIES ON WINE:3. BSA. 3.

4.e. 50mg sodium sulphite. Then few drops of starch are added as an indicator.2mg/ml stock solution is being taken.5ml and add distilled water to make the volume upto 3. 37 . 0. 0. Titration is carried out and result is being noted.8 ml (3gm absolute alcohol) in flask make volume upto 100 ml with distilled water. For making this we add 1. 100ml 1%NaOH.5 ml.5ml of d/w is added to make the total volume upto 2ml.96gm of K2Cr2O7 add 550ml of water and add 450ml of conc.3. 3.2 ml to 1ml and after that distilled water is added to make the volume upto 2ml.05gm/100ml is taken. After that 10ml of distilled water is added and then reading is taken at 520nm by spectrophotometer. Different concentration of standard is taken which range between 0. REAGENTS:- Crystalline phenol. After that 2 ml of DNSA is added and kept for incubation for 5 min in water bath.5 mg/ml of maltose is taken and stock solution of 100ml is prepared. 1. alcohol is poured which range in between 1ml to 3.5 ml of sample is taken and 1. Six flasks were taken and in each flask different amount of std. after that 100ml of distilled water is added then 4ml of KI is added. Maltose is taken as standard. PROTOCOL:- DNSA is prepared.25% KI in distilled water and K2Cr2O7. (B) Mix thoroughly and remove 4 ml and add 96 ml water and make volume upto 100 ml. B. ESTIMATION OF REDUCING SUGAR BY DNSA METHODA. H2SO4. After that 10 ml of dichromate is added to each flask and kept at room temperature for 30 min. From this 50mg/100 ml i. Distilled water is taken as blank. (A) Take 3. 40% Rochelle salt solution (Potassium sodium tartarate). Ethanol is taken as standard alcohol. whereas in case of wine sample 0.

After 30 minutes of preparation in dark. which disappears on reduction by an antioxidant compound. DPPH has an absorption maximum at 517 nm. The absorbance was 38 .4. REAGENTS:- Tannic acid.1 mmoles/liter solution of DPPH radical in methanol was prepared and 2 ml of this solution was added to 2 ml of water solution containing 100µl of wine samples.4. Lower absorbance of the reaction mixture indicates higher free radical scavenging activity. ascorbic acid B. PROTOCOL:- The principle for the reduction of 1. . The 0.1999).4. the antioxidant reacts with stable free radical. 3. absorbance was measured at 517nm. methanol. 1 ml of Folin Ciocalteu was added and the content in the flask was mixed thoroughly. ANTIOXIDANT ACTIVITY BY DPPH ASSAY A. After 3 minute of incubation 3ml of 2g/100ml Na2CO3 was added then the mixture was allowed to stand for 2hr with intermittent shaking. DETERMINATION OF TOTAL PHENOLIC COMPOUNDSA.1-diphenyl2-picryl hydrazine. Different concentrations were tested using ascorbic acid as standard for calibration and expressed as mg ascorbic acid equivalents per litre of sample.1-diphenyl-2-picryl hydrazyl (DPPH) free radical is that.5.3. DPPH and converts it to 1. Folin Ciocalteu B. PROTOCOL:- 1ml of wine samples in a volumetric flask was taken and 46 ml of distilled water is added to it. REAGENTS:DPPH. The calibration curve was plotted with % DPPH scavenged versus concentration of standard oxidants (Sanchez-Moreno et al.

Fe2+ B. 500ml KH2PO4. samples were boiled with TBA reagent for 30 mins. The mitochondrial pellet was washed thrice with 50mM KPO 4 buffer. 3. LIPID PEROXIDATION ASSAY BY TBARSA.4. homogenizer. sterile forceps and blade. PROTOCOL:i) Isolation of Mitochondrial fraction from Rat Liver Wistar rats (weighing about 240±20 g) were used for the preparation of mitochondria. OTHER MATERIAL:- Wistar rats. to remove cell debris and the nuclear fractions. TBA reagent. to remove sucrose.000 × g for 10 mins to sediment mitochondria.25Msucrose containing 1mM EDTA. REAGENTS:- 500ml K2HPO4. After the incubation.Fe2+ . The concentration of total phenolic compounds was determined with microgram of tannic acid. saline.4.A. Rat liver were excised and homogenized in 0. 1986). The incubations were carried out at 37⁰C in a water bath.6.25 M sucrose.system as described by (Devasagayan..measured at 760nm using spectrophotometer. pH 7. Protein was estimated and pellets were suspended in the above buffer at the concentration of 5 mg protein/ml. 0. gloves.The homogenate was centrifuged at 3000 × g for 10 mins. 1Mm EDTA.P. C. T. The supernatant was centrifuged at 10. ii) Exposure of rat liver mitochondria to oxidative stress Oxidative damage was induced by ascorbate. The pink colour of thiobarbituric acid reactive substances (TBARS) formed were estimated at 532nm spectrophotometrically as malondialdehyde equivalents after accounting for appropriate 39 . All the experiments were carried out thrice at 4⁰C. ascorbic acid.

The qualitative analysis of the polyphenol contents was performed using a HPLC system which consists of a model 515 pump. catechol and vanillin.22 micron filter. Flow was set to 0. HPLC ANALYSIS OF WINESA.22 micron filter. ferulic acid. a model 2487 dual wavelength absorbance detector and a manual injector. b) 75% HPLC water in 1% acetic acid 40 . HPLC unit C. The filtered extracts were used for HPLC analysis. ferulic acid.7. PROTOCOL:Different wine samples (red. 3.75 ml/min.2007). The standard phenolic acids and flavonoids used for HPLC analysis were tannic acid. catechol and vanillin. the two solvents used to make the gradient were as followsa) 25% methanol in 1% acetic acid. The separation of the polyphenols was conducted in a column. The samples were filtered through 0. REAGENTS:- Tannic acid. . white and port wine) were used for the determination of different polyphenolics and flavonoid compounds. caffeic acid. caffeic acid. OTHER MATERIAL:- 0. Malondialdehyde standard was prepared by the acid hydrolysis of tetraetoxypropane (Vinayak V et al.blanks.4. 25% methanol and 75% HPLC water B.

81 3.68 3.05 41 .22 3.56 3.36 3.78 3.4. pH ESTIMATION OF THE INDIAN WINES Table 4.1.39 3.77 3.4 3.72 3.1: pH readings SAMPLE Red 1 Red 2 Red 3 Red 4 Port 1 Port 2 Port 3 Port 4 White 1 White 2 White 3 White 4 pH readings 3.68 3.

68 0.1. Table 4.: Standard readings for alcohol tolerance by dichromate method.85 1.4 1.34 0.: Standard graph for alcohol tolerance by dichromate method.1 1.9 42 .4.9 2.028 Y – axis differences (B – E) 0 1.51 0. X – axis conc. ALCOHOL ESTIMATION ALCOHOL TOLERANCE BY DICROMATE METHOD STANDARD Figure 4. (mg/ml) 0 0.2.2.4 2.

20 13.3 Dilution: 1:700 Table 4.64 16.36 104.20 13.5 0.5 104.48 16.22 131.5 0.48 13.64 16.22 90.22 104.36 104.22 43 .65 131.64 16.36 131.2 3 ALCOHOL ESTIMATION BY DICHROMATE REAGENT Blank: 4.22 104.3.5 0.6 0.20 11.5 0.6 0.6 0.6 0.20 11.45 0.64 13.22 90. Samples Readings (B-D) Concentration (mg/ml) Alcohol estimation of wine (%v/v) 13.45 0.: Readings for alcohol estimation.20 Red 1 Red 2 Red 3 Red 4 Port 1 Port 2 Port 3 Port 4 White 1 White 2 White 3 White 4 0.65 104.1.5 0.20 13.22 104.

4 (0.4.: Standard readings for reducing sugar assay.31 0.1 0. ESTIMATION OF REDUCING SUGAR BY DNSA METHOD REDUCING SUGAR ASSAY STANDARD Figure 4.228 0.3 0. Table 4.2 0.2.2ml) (0.8ml) Y-axis (O.3. X-axis(mg/ml) 0 0.6ml) (0.4.282 0.: Standard graph for reducing sugar assay.D Reading in nm) 0 0.398 44 .4ml) (0.

5.234 ± 0.0ml) 0.254333 0.248667 0.135 ± 0.281 0.580 ± 0.469 ± 0. Samples Red 1 Red 2 Red 3 Red 4 Port 1 Port 2 Port 3 Port 4 White 1 White 2 White 3 White 4 0.001528 16.003464 3.343333 0.: Readings for reducing sugar assay.D Reading at 520 nm 0.0.617 ± 0.290333 0.293333 1.069 ± 0.738 0.617 ± 0.000577 2.987 ± 0.839 ± 0 3.58 ± 0.001732 3.001155 9.002887 2.001155 45 .5 (1.001732 3.586667 0.242 0.003512 3.11 ± 0.271333 Reducing sugar for wine (mg/ml) 3.001155 7.345 ± 0.48 READINGS OF REDUCING SUGAR IN WINE SAMPLES Table 4.23 0.212333 0.000577 2.

193 0.37 90.047 0.162 0.60 100 90.43 87.093 0.: Readings for DPPH assay.4.088 0.89 81.6.17 DPPH assay (% scavenging activity) 80.24 92.59 81. Samples Red 1 Red 2 Red 3 Red 4 Port 1 Port 2 Port 3 Port 4 White 1 White 2 White 3 White 4 Readings at 517 nm 0.06 87. ANTIOXIDANT ACTIVITY BY DPPH ASSAY Table 4.111 0.00 46 .177 0.22 79.071 0.4.18 0.16 78.113 0.88 89.087 0.

11 0.04 0.2 0.16 0.: Standard graph for determination of total phenolic compounds.03 0. Table 4.24 47 .05 0.4. DETERMINATION OF TOTAL PHENOLIC COMPOUNDS STANDARD FOR TOTAL PHENOLIC COMPOUNDS Standard: Tannic acid Stock: 10 mg in 10 ml Figure 4.01 0.07 0.02 0.06 0.3.08 0.axis 0.13 0. Sr.5.054 0.axis 0.08 Y .02 0.no 1 2 3 4 5 6 7 8 X .7.: Standard readings for determination of total phenolic compounds.

1 ± 0 6.8.137 0.18 0. SAMPLES Red 1 Red 2 Red 3 Red 4 Port 1 Port 2 Port 3 Port 4 White 1 White 2 White 3 White 4 Readings at 760 nm 0.3 ± 0 3.18 ± 0.07 ± 0 5.6 ± 0.96 ± 0 0.164 0.5 ± 0.0057735 6.26 0.0057735 0.026 0.177 0.07 ± 0.9 10 0.144 0.09 0.25 READINGS FOR TOTAL PHENOLIC CONTENT Dilution = 100 microliters of the sample + 990 microliters of the distilled water Table 4.07 ± 0 0.6 ± 0 5.00057735 48 .92 ± 0.152 0.029 0.167 0.0057735 6.74 ± 0 1.02 0.: Readings for determination of total phenolic compounds.1 0.00057735 6.086 0.025 Total phenolic content (mg/ml) 5.

Table 4.07 0 0 49 .5 Y .: Standard readings for lipid peroxidation.55 0.4.26 0.6.axis 0.4.145 0. LIPIDS PEROXIDATION ASSAY BY TBARS STANDARD ASSAY Standard : Malondialdehyde (MDA) Figure 4. X – axis 100 50 25 12.: Standard graph for lipid peroxidation.9.

6.92±0.47666667 58.7±0 16.28 % avg protection 44.82±0.2 8. SAMPLES RED 1 RED 2 PORT 1 PORT 2 WHITE 1 WHITE 3 Readings at 532 nm 0.23333333 46.58 9.READINGS FOR TBARS CONTROL.27 63.35nmol/ml DAMAGE.093 0.34± 0. Isolation of mitochondria fraction from rat liver and protein estimation by Bradford reagent 50 .10.23.8±0 17.098 0.18 12.73±0.01 29.: Readings for lipid peroxidation.1.071 0.91666667 4.49 26.486 0.1.074 0.89nmol/ml Table 4.54 Lipid peroxidation of wine (nmole/ml) 13.

43 micrograms/10 microlitres (correspondence to 2.966 1.: Readings for protein estimation.: Standard graph for protein estimation.674 0.06 micrograms/10 microlitres (correspondence to 1.802 Reading at 595nm 10 microliter of mitochondria sample = 13. Table 4.333 0.176 0.119 1.axis 0 0.11.343 mg/ml) 51 .axis 0 10 20 40 60 80 100 Y .306 mg/ml) 50 microliter of mitochondria sample = 23. X .Figure 4.5.

HPLC ANALYSIS OF WINES 1) Sample Name: std-25 52 .100 microliter of mitochondria sample = 32.287 mg/ml) 4.87 micrograms/10 microlitres (correspondence to 3.7.

2) Sample Name: std-50 53 .

3) Sample Name: std-100 54 .

4) Sample Name: Sample.Red wine R1 55 .

Red wine R2 56 .5) Sample Name: Sample.

Red wine R3 57 .6) Sample Name: Sample.

Red wine R4 58 .7) Sample Name: Sample.

59 .

8) Sample Name: sample-port wine 1000 60 .

9) Sample Name: sample-port wine 9 61 .

10) Sample Name: sample-port wine 99 62 .

11) Sample Name: sample-port wine 4 63 .

12) Sample Name: Sample.White wine W1 64 .

13) Sample Name: Sample- White wine W2

65

66

14) Sample Name: sample- White wine W3

67

White wine W4 68 .15) Sample Name: sample.

294 STANDARD 25 STANDARD 50 STANDARD 100 RED 1 RED 2 RED 3 RED 4 PORT 1 PORT 2 PORT 3 PORT 4 WHITE 1 WHITE 2 WHITE 3 WHITE 4 69 . HPLC ANALYSIS Table 4.013 47.000 100.12.: Concentration of tannic acid for the samples.4.990 54.000 56.852 73.099 64.544 72.7. SAMPLES Concentration of Tannic acid (ppm) 25.000 50.403 48.659 65.430 46.438 36.137 46.038 83.1.

069 ± 0.00057735 6.01 46.0034 64 3. 2 29.038 Port 2 3.0011 9.18 ± 0.22 13.Table 4.580 ± 0.0057735 6.430 White 1 3.58 ± 0.0057735 6.18 12.72 16.135 ± 0.1 ± 0 100 36.0015 3.16 65.0028 2.64 6.27 83.013 70 .34± 0.8±0 44.23333 56.659 Port 4 3.64 92.438 Red 4 3.20 79.60 64.469 ± 0.37 73.20 0.24 17.49 72.544 Red 2 3.0011 7.48 3.77 16.56 16.7±0 26.88 46.47666 67 48.68 13.0035 3.6 ± 0.92±0.234 ± 0.5 8 63.81 13.0017 3.07 ± 0 90.3 ± 0 89.403 Red 1 3.64 90.13.617 ± 0. of tannic acid(ppm) Port 1 3.6 ± 0 87.74 ± 0 78.0017 3. Combined data for wines Types of wine pH of Wine Reducing sugar for wine (mg/ml) Alcohol estimation of wine (%v/v) Total phenolic content (mg/ml) DPPH assay (% scavenging activity) Lipid peroxidation of wine (nmole/ml) Lipid peroxidatio n of wine (% avg protection) Conc.5 ± 0.0057735 5.39 11.68 13.20 5.852 Port 3 3.73±0.78 16.987 ± 0.20 80.099 Red 3 3.0005 16.43 8.22 13.64 5.07 ± 0.06 16.11 ± 0.

0005 2.20 0.82±0.990 3.00057735 81.137 3.89 9.07 ± 0 87.345 ± 0.48 1.00 54.0011 11.617 ± 0.2 8 58.96 ± 0 81.4 13.91666 67 46.92 ± 0.839 ± 0 3.White 2 White 3 White 4 3.20 0.59 47.36 2.05 13.294 71 .

Hence we have estimated antioxidant abilities of Indian wine varieties. serum triglycerides. with a gradual increase in the number of patients with acute myocardial infarction (Krishnaswami. The pH of Wine sample is carried out and from the study it is being clear that white wine is more acidic in comparison of red and port wine. Their intake was significantly and inversely associated with mortality from CHD and also showed an inversely associated with mortality from CHD and also showed an inverse correlation with incidence of myocardial infarction (Hertog. Natural product especially derived from dietary components such as fruits and vegetables yield rich divided in terms of potential benefits in controlling diseases. low levels of HDL cholesterol etc. Biochemical test of wine is successfully carried out by using different sample of red.1998). white. Various antioxidants may prevent and/or improve different diseased states. we know that port wine consist of more alcohol content in comparison of white and red wine.4 to 3. fruits and beverages such as tea and wine. we have used different methods such as radical scavenging by using DPPH and inhibition of lipid peroxidation by measuring TBARS in rat liver mitochondria. Reducing sugar was found more in comparison of red and white wine. and port wine. Wine sample were collected from different wineries shop. According to this result port wine is much better after that red wine followed by white wine.It is being reported that there is increase in prevalence of coronary heart disease (CHD) in India and other developing countries. M.65 and average reading of port wine is 3. 1998). There is slight difference between red and port wine. D. For measuring antioxidant activity. After doing alcohol estimation.. Its acidity lies below 3.05 where as average reading of red wine is 3. (Bhatnagar. Polyphenolic antioxidants such as flavonoids occur naturally in vegetables.G. It has been shown that people of Indo-origin may be more prone to CHD due to metabolic syndrome comprising of resistance to insulinmediated glucose uptake.7.L et al. S.. Reducing sugar for wine is being carried out taking maltose as standard. Red wine consists of less alcohol in comparison of other wines. According to the result red wine is good for drinking. The model of scavenging the stable DPPH radical is a widely used method to 72 . Consumption of wine also can help. 1993).

DPPH activity was found maximum in red wine. followed by port and white wine for health and also for antioxidant activity. The high total phenol and flavonoid contents of wine may lead to its good DPPH-scavenging activity.1 and 1. Inhibitory effects of the six samples of wine were carried against lipid peroxidation induced by ascorbate-Fe2+ in rat liver mitochondria. DPPH is a stable nitrogen-centered free radical..or electron..07 to 0. The Comparative studies were made considering tannic acid as standard. vanillin. It was found that the radical – scavenging activity of the extracts increased with increasing concentration. The content of phenolic compounds was determined by the Folin-Ciocalteu for the different wine samples (V.5 to 5. All the 12 varieties of wines were analyzed for total phenol and total flavonoid contents and some select varieties by HPLC to identify different phenolics and flavonoids such as tannic acid. In this we found that white wine show greater percent of protein protection followed by red and port wine.6 to 3. Substances which are able to perform this reaction can be considered as antioxidants and therefore radical scavengers (Brand-Williams et al. From all the above data it is clear that red wine is better.07. than in red wine and very less in white wine (range between-6.donation. 1995). The result showed that the increase of phenolic content is higher in port wine. It was found that port wine has greater amount of tannic acid in comparison of red and white wine. the colour of which changes from violet to yellow upon reduction by either the process of hydrogen. the red wines had significantly higher amounts of total phenolic content. cathachol. As expected.evaluate the free radical scavenging ability of various sample (Lee et al. 6.74 respectively).. 73 . caffic and ferulic acid. 2003).Katalinic et al. 2003).

the pigments molecules. These varieties of wine. Qualitatively there is little difference between port. may confer health benefits. our studies show that among the different varieties of wines examined red wine have the most potent antioxidant activities and after that port wine followed by white wine. In many cases it is being observed that red wine and port wine are similar.In conclusion. "Nothing more excellent or valuable than wine was ever granted by the Gods to man. This prolonged contact during fermentation produces significant levels of resveratrol in the finished red wine. Red wine has a high concentration of resveratrol because the skins and seeds ferment in the grapes' juices during the red wine-making process. Their antioxidant activities. especially in population prone to CVD. red and white wine. if consumed in adequate amounts. assayed at different levels correlate with their chemical composition in terms of total phenolics and flavonoids. Wines also help from cancer and many heart disease when it is taken in moderate amount. 74 ." Those of us who have come to enjoy the variety and tastes that wine have to offer can now look to red wines for greater health benefits. large amounts of catechins and phenolic content. The difference between red and white wines is that red wines contain anthocyanins. Plato may have been wiser than he knew when he said.

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