PRESERVATIVE EFFECT ON SHELF STABILITY OF ZOBO WINE USING SODIUM BENZOATE

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ABSTRACT

This study was aimed at determining the shelf stability of zobo wine using sodium benzoate. Zobo wine was produced by fermentation of the must using brewer’s yeast, Saccharomyces cerevisiae at 280C for a total of 28 days with addition of sodium benzoate (0.5% w/v) and stored for 21 days. The sample and a red wine from grape (oster wine) were analyzed for their physicochemical, microbial and sensory qualities. The results obtained revealed that the physicochemical properties ranged from 5.50 to 6.45, 0.99 to 0.99, 114 to 487 mg/l, 67.50 to 78.50 g/100ml, 21.00 to 40.50%, 0.28 to 0.31 nm, 13.85 to 76.45 mg/l, 0.50 to 0.69 g/100 ml and 6.61 to 10.04 % for pH, specific gravity, total dissolved solids, total titratable acidity, dry matter, colour intensity, phenol content, total solids and alcohol content, and total solids. In as much as red wine from grapes was higher in the physicochemical properties pH, phenol content, total solids and alcohol content; zobo wine showed higher values in total dissolved solids, total titratable acidity, dry matter and colour intensity. From microbial analysis, no significant bacteria growth was isolated from the samples, although lower in zobo wine. Also, no fungal or coliform growth was observed. The zobo wine was of comparable microbial quality with the red wine from grapes possibly due to presence of sodium benzoate generally used as preservative in beverages. Sensory evaluation indicated higher acceptance of zobo wine than the red wine from grapes (oster wine). The sensory attributes ranged from 6.90 to 7.65, 5.80 to 7.00, 6.15 to 7.05, 5.85 to 7.05, 5.25 to 7.20 and 6.10 to 7.75 for colour, appearance, clarity, taste, flavour, mouth-feel and general acceptability respectively when evaluated on a 9 - point hedonic scale. Although zobo (Hibiscus sabdariffa) extracts are readily contaminated by spoilage microbes, it can be used in the production of shelf stable wine with addition of sodium benzoate and could favourably compete with imported wines.  
 







TABLE OF CONTENTS 
 
Title Page                                                                                        i
Declaration                                                                             ii
Certification                                                                          iii
Dedication                                                                               iv
Acknowledgements                                                     v
Table of Contents                                                                 vi
List of Tables                                                                           ix
List of Figures                                                                  x
List of Plates                                                                         xi 
Abstract                                                                         xii 

CHAPTER 1: INTRODUCTION 
1.1 Background of study                                              
1.2 Statement of problems                                     
1.3 Justification                                                   
1.4 Objectives of the study                                                 

CHAPTER 2: LITERATURE REVIEW 
2.1 An overview of zobo calyces                                       
2.1.1 Nutritional composition of zobo calyces                 7                
2.1.2 Medicinal benefits of zobo calyces and its use in food   7     
2.1.3 Utilization of zobo calyces in food and beverage production  8    
2.2 Wine production and classification                      9  
2.2.1 Raw materials used in wine production      11 
2.2.2 Cultured yeast and inoculation                                  12 
2.2.3 Fermentation of wine                            12 
2.2.4 Stages involved in zobo wine production                14
2.2.5 Maturation and bottle ageing of wine                 18 
2.3 Physicochemical properties of wines                           20 
2.3.1 Importance of phenolic compounds                 20  
2.3.2 Effect of organic acids and ph on wine quality and stability     21 
2.3.3 Alcohol content of wines                                   21
2.3.4 Development of aroma compounds in wine undergoing maturation        22  
2.3.5 Effect of volatile acidity /acetic acid on wine quality        23 
2.3.6 Residual sugars in wines and their effect on consumer choices of wines  24   
2.4 Sensory properties of wines                                26 
2.5 Standards for wines                                                          27  
2.6 Spoilage of wines                                                                27 

CHAPTER 3: MATERIALS AND METHODS 
3.1 Sources of raw materials                                                   29 
3.2 Sample preparation                                                          29 
3.2.1 Preparation of must                                                29 
3.3.2 Fermentation of must                                               29 
3.2.3 Clarification and storage of young wine                  30 
3.3 Physicochemical analyses of the wine samples         34 
3.3.1 pH                                34 
3.3.2 Specific gravity                                         34 
3.3.3 Total dissolved solid                                                34 
3.3.4 Total titratable acidity                                    35
3.3.5 Dry matter                                                 35  
3.3.6 Colour intensity                                                 35 
3.3.7 Determination of phenols content               36 
3.3.8 Total sugar                                                                 37 
3.3.9 Alcohol content                                                   37 
3.4 Microbial analysis                                                                38 
3.4.1 Total viable microbial count (TVC)                              38 
3.4.2 Total fungi count (TFC)                                                   38 
3.4.3 Total coliform bacteria count                                   38 
3.5 Sensory evaluation of wine                                                     39 
3.6 Experimental design                                                    39 
3.7 Statistical analysis                                                      40 

CHAPTER 4: RESULTS AND DISCUSSION 
4.1 Production of zobo wine                                                 41 
4.2 Physicochemical properties of wine samples               42   
4.3 Microbial characteristics of wine samples             46 
4.4 Sensory evaluation of the test wine samples                                                    
CHAPTER 5: CONCLUSION AND RECOMMENDATIONS       
5.1 Conclusion                                                                              52 
5.3 Recommendations                                                        52 
5.2 Contribution to knowledge                                       53 
REFERENCES            53
 





LIST OF TABLES 

Table 2.1: Classification of wines based on residual sugars        25 

Table 4.2: Physicochemical properties of wine samples        43 

Table 4.3: Microbial quality of test wine samples             47 

Table 4.4: Sensory properties of wine samples                49 
 
 
 



 
LIST OF FIGURES 

Figure 2.1: Flow diagram showing production of red wine      17

Figure 3.1: Flow chart for the production of wine from Zobo wine  31




LIST OF PLATES

Plate 1: Zobo calyces                                                                 32 

Plate 2: Granulated sugar                                               33 

Plate 3: Zobo wine                                                         41 

 



CHAPTER 1
INTRODUCTION

1.1 BACKGROUND OF THE STUDY
Fruit wines are fermented alcoholic beverages made from a variety of base ingredients (other than grapes). They may also have additional flavours taken from fruits, flowers and herbs. Fruit wine can be made from virtually any plant matter that can be fermented. Most fruits and vegetables have the potential to produce wine. Few foods other than grapes have the balanced quantities of sugar, acid, tannin, nutritive salts for yeast feeding, and water to naturally produce a stable, drinkable wine; so most wines are adjusted in one or more respects at fermentation. However, some of these products do require the addition of sugar or honey to make them palatable and to increase the alcoholic content (sugar is converted to alcohol in the fermentation process) (Ihekoronye and Ngoddy, 2015).

In recent times, there has been an increase in the use of several locally grown tropical fruits and vegetables as raw materials for alcoholic beverages (wine) production in Nigeria. Among the assortments of such indigenous tropical fruits are the kolanut (cola acuminata), cocoa (theobroma cacao L.), African star apple (chrysophyllum albidium), pawpaw (carica papaya) and zobo calyces. The temperature restriction of grape to the temperate regions and the very high duty on imported wines has stimulated interests in producing wines from tropical fruits (Odigie et al., 2003).

Roselle, which is zobo calyces (Hibiscus sabdariffa) belongs to the family Malvaceae, and is an important annual crop grown successfully in tropical and sub-tropical climates. The plant is cultivated for its stem, fibre, edible calyces, leaves and seeds which are used in making various foods. The commercially important part of the plant is the fleshy calyx (sepals) surrounding the fruit (capsules). It is used for making wine, juice, jam, jelly, syrup, gelatin, pudding, cakes, ice cream and flavours and also dried and brewed into tea, among other things (IfieIdolo et al., 2012).

The red calyces contain antioxidants including flavonoids, gossypetine, hibiscetine and sabdaretine. Delphinidin 3-sambubioside and cyaniding 3-sambubioside are the major anthocyanins comprising 70 and 29 % of total anthocyanins respectively. Its calyx also contains 4 % citric acid, organic acids, minerals and amino acids. The seeds contain 17.8 to 21 % non-edible oil and 20 per cent protein, and are sometimes used for animal feed (Odigie et al., 2003).

A very widely-used preservative, sodium benzoate is used throughout various industries, including food and beverage, pharmaceutical, cosmetic, and industrial. As a food additive, sodium benzoate was the first preservative allowed in foods by the FDA. Today, it is classified as Generally Recognized as Safe (GRAS), meaning experts consider it safe when used as it’s intended. It is internationally approved as a food preservative and is identified by the number 211 (listed as E211 in Europe). It functions as a preservative by inhibiting the growth of harmful bacteria, mold, and other microbes, preventing product spoilage. It’s also used to preserve over-the-counter medications such as cough syrup and as a lubricant in pill manufacturing (Odigie et al., 2003).

Sodium benzoate is commonly found in soft drinks, it can also be used in winemaking both to prevent spoilage and stop the fermentation process. Like potassium sorbate, sodium benzoate is a yeast inhibitor. Rather than killing off the yeast, it hinders yeast’s ability to multiply and be active. Winemakers usually add small quantities of sodium benzoate to sweet and sparkling wines when active fermentation has ceased. For sweet wines, it can also be added at the same time as the final syrup. Sodium benzoate can also be added to wines that have difficulty maintaining clarity after fining. It can also be used post-fermentation balance the wine’s pH and pressure. When using sodium benzoate as a stabilizer, it must be used in conjunction with an aseptic dose of potassium metabisulfite (Campden tablet).

1.2 STATEMENT OF PROBLEM
Most wines consumed in Nigeria are imported and there is little or no local production. Wines are mostly produced from grapes, which are not available in the tropics hence the need for alternative use of plant materials and other fruits for wine production in the tropics. A survey conducted in Lagos and other states in the Western part of Nigeria to know their choice of wines indicated that the wines used by most respondents were all imported wines from countries like Israel, Spain, Italy, America and South Africa, with alcohol levels below 7%.  

Despite the fact that the popularity of beverage from zobo calyces is increasing, one of its greatest limitation for large scale production is that it has a very short shelf life of 24 hours if not refrigerated (Omemu et al., 2006). Therefore, there is urgent need to explore various preservation methods that could be employed to extend the shelf life of this product.

Most producers depend heavily on the use of preservatives. Electricity supply in which processors depend mainly on for refrigeration is grossly inadequate and often characterized by frequent power outage. On the overall, when electricity supply is low, the drink could easily be invaded by spoilage microorganisms. 

Food safety is an essential concern of both the consumers and the producers (Witkowska et al., 2013). Microorganism in food is not always detrimental, because their growth may result in pleasant taste and texture (Bukar et al., 2010). But microorganisms such as Staphylococcus aureus, Pseudomonas aeruginosa, E. coli, Vibro cholera, Salmonella, Bacillus, Clostridium species etc. could contaminate food and transfer a wide range of disease conditions in food (Bukar et al., 2010).

1.3 JUSTIFICATION 
Low alcohol wine produced from extracts obtained from zobo calyces extract will provide health benefits to its consumers due to its rich source of phenolic compounds and low alcohol. The production of low alcohol wine will help in the alcohol control measures intended to curb alcohol consumption levels and drinking habits and reduce related social and health problems. This move is in line with the World Health Organization’s (WHO) public health objective on alcohol which is to reduce the health burden caused by the harmful use of alcohol and thereby, to save lives, reduce disease and prevent injuries. The product will be a potential red wine which can compete with imported wines on the market and the product would be beneficial to the local churches that rely on imported wines for communion services. This product could be relatively cheap because of the use of locally available raw materials and will save the government some foreign exchange for development.  The near absence and epileptic public electricity power supply used to power home appliances including refrigerators has made the preservation of such beverages in Nigeria difficult limiting the shelf life of many of such beverages to just few days. Research into the shelf stability of wine product without refrigeration has therefore intensified. The aim of this research was to examine the preservative effect of sodium benzoate on wine produced from zobo calyces

1.4 OBJECTIVES OF THE STUDY
The main objective of this study was to examine the preservative effect of sodium benzoate on wine produced from zobo calyces.
The specific objectives include to:

i. produce wine from zobo calyces.

ii. determine the physicochemical properties of the produced wine.

iii. determine the microbial quality of the produced wine. 

iv. determine the sensory characteristics of the wine product after storage.


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