Puttalingamma V et al.

, IJSID, 2013, 3 (1), 117-125

ISSN:2249-5347

IJSID

International Journal of Science Innovations and Discoveries

Research Article

An International peer Review Journal for Science

Available online through www.ijsidonline.info

SENSORY QUALITIES

EFFICACY OF CARNAUBA WAX WITH NISIN EMULSION ON TOMATOES TO EXTEND THEIR SHELF LIFE AND
1Defence

Puttalingamma .V1* and Khyrunnisa Begum2 Food Research Laboratory, Mysore, Karnataka, India; 2Department of Studies in Food Science and Nutrition, University of Mysore, Manasagongotri, Mysore, India ABSTRACT statistics is nearly Rs.154800 crore (Statistical abstract. India 2003, discussed. With growing demand from changes in legislation, consumer trends and increasing isolation of antibiotic resistant pathogens, alternatives to these non-chemical based bactericides need coatings. These coatings have been used traditionally as a barrier and to minimize water to be found. Research efforts have focused on the eco-friendlily, nontoxic and edible Fruits and Vegetables constitute the fourth largest items in the national

Received: 03-01-2013 Accepted: 14-02-2013

*Corresponding Author

loss and delay the ripening. The role of bio coating substance like Lactic acid bacteria and Address: Name: Dr. Puttalingamma.V Place: Karnataka, India E-mail: puttu_v2005@yahoo.com their metabolites with carnauba wax emulsion is studied. Tomatoes treated with different temp (51oC) were assessed for their sensory attributes as well as microbial spoilage. KMS+ nisin and nisin (1000) ppm treated samples stayed up to 40 days at low temp and the shelf life of tomatoes without much altering texture, flavor and moisture.

biocoating and natural preservatives, studied at two different temp, RT (283 oC) and Low carnauba wax with nisin treated samples stayed up to 48 days, L.lactis samples upto 45 INTRODUCTION unto 15 days at low temp and 8 days at RT. Storage at low temperature (5 10C) prolongs Key words: Pathogens, Nisin, carnauba wax, lactic acid bacteria, tomatoes and shelf life. days and lowest among all is samples treated with L. plantarum. Whereas control stayed

INTRODUCTION

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Puttalingamma V et al., IJSID, 2013, 3 (1), 117-125 (FAO) statistics, India contributes 15% vegetables to world production of 6.91894X10 3MT (FAO-1995., Singh Uadal et al.2008). Among the various vegetables, tomatoes (Lycopersicon esculentum) stand the highest in world production figures. It been reported. Marketing of fruits and vegetables is the major bottleneck, 30 to 40 % will get perished because of poor marketing infrastructure (Ram P Aneja et al., 2007). Recently consumers have increased interest in healthy and convenient perishability (Cantwell, 2000). Among the oldest methods of preservation are drying, refrigeration, and fermentation. packaging materials have played an important cause of vegetable spoilage by microorganisms. And many enteric pathogens Rungslwee sothornvit & Pecriya kiatehanapaibul,2009). The source of microbes includes fecal maters, during transport insect, is well known that vegetables have shorts shelf life and therefore incurs highest post-harvest losses. In India processing of foods. Whereas most food processing techniques lengthen the shelf life of products, minimal processing increases their vegetables is still under development 2% of the vegetables are reported to undergo processing mean waste of 35-40 % has India is the second largest country to produce horticulture food. INTRODUCTION According to Food and Agriculture Organization

cross contaminated eg, salmonella, spp, Pseudomonas, Listeria spp, B.cereus and pathogenic E.coli.etc.(Shi,et al, 2007., dust and processing conditions, where in storage temperature acts like a booster. Research report (Puttalingamma et al,2008) indicates a predominance of bacterial soft rot, ie, Erwinia, Pseudomonas, water and soil pathogens, yeast and moulds are vegetables with suitable technologies is worthwhile. Such technology would lead to tremendous economic contribution and increased availability of vegetables to the consumers thought the year. present on vegetables. Gram negative bacterial flora causes major spoilage vegetables. They thrive and multiply faster at Novelty and convenience can be added to merchandise vegetables products, like ready to use vegetables, minimally

Modern methods include canning, pasteurization, freezing, irradiation, and the addition of chemicals. Advances in

ambient temperatures and high humidity. An attempt to extend the shelf life by reducing this surface microflora from processed vegetables etc. Consumers awareness has created a demand for safe bio preservatives in food preservation. Application of nisin, which is declared as GRAS preservative could be one effective technology. A few studies have reported worthwhile to use nisin as surface applicant on tomatoes to study its storage behaviors. Vegetables are chief sources of antioxidants and minerals (Gopalan et al., 1993).They also add variety, flavour and

encouraging results in extending shelf life of foods like dairy products, meat and meat products. It was therefore felt colour to the diet. Vegetables like tomatoes are rich source of lycopene and vitamin C. Amir et al., (1999) reported that reductase homocystene and platelet aggregation.(Willcox et al 2003). Hence foods containing lycopenes are referred as Tomatoes by virtue of being rich in lycopene and certain other protective phytochemicals are considered as a health Nisin a class I, bacteriocin, has got wide applications as food preservative (Thomas et al 2000). lycopene treatment could result in a concentration dependant reduction in growth of HL-60 pro myelocytic leukemia cells.

Tomatoes and tomato products (Vit-C, E, flavonoids, lycopenes) control cardiovascular problems through altering HMG-CoA functional foods that reduce major age related conditions such as cancer, osteoporosis, mental aging, etc. (Willcox et al., 2003). promoting food that has the ability to reduce low-density lipoprotein (LDL) cholesterol, homocysteine, platelet aggregation and blood pressure. Tomatoes can be treated with carnauba wax (Copernica prunifera, palm family) emulsion containing nisin by dip coating method to improve appearance, retain their colour, lower moisture loss and improve post harvest storage life.

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Puttalingamma V et al., IJSID, 2013, 3 (1), 117-125 MATERIALS AND METHODS Procurement of materials: tomatoes with uniform size without blemishes or cuts were selected. Preliminary treatments: Selection of tomatoes was done to obtain fruits with uniform characteristics, from local market. Semi ripened They were washed under running tap water for 1-3 minutes to remove surface soil and dirt. The cleaned tomatoes

were transferred to a tub containing chlorine water (100 ppm) with a ratio of 1:5 W/V ratio and kept for 1 minutes. The method, for Total plate counts., H2 S producing organisms, Yeast and moulds and food borne pathogens. Preparation of carnauba wax emulsion:

tomatoes were than drained and dried by spreading them over filter paper for 10 - 15 min. Microbial load including yeast and moulds were present on the surface of tomatoes were determined before and after the treatment according to pour plate was stirred continuously using a motor attached to round bottom flask. Gum Arabica was dissolved in hot water and added to 10 min at a speed of 10,000 rpm. pH was then adjusted to 4 and stored at refrigerated temperature for further use. Method of Application. without treatment served as control. ppm level. Carnauba wax was melted at 95oC in presence of propylene glycol followed by addition of distilled water. The mixture

the melted wax. The mixture was heated to 85-90oC with addition of potassium sorbate (the proportion, the ingredients used

in the preparation is presented in Table-1). Small portion of hot emulsion was poured in to vortex, homogenize and mixed for

Tomatoes were divided into 5 batches; four batches were treated with different surface coating mixtures, while the one ATCC 11454, L. plantarum NCIM 2084). Batch II: Coated with nisin (Nisapline, 2.5% activity ingredients, obtained from Applin & Barrat Co, London) solution at 1000 Batch III: Coated with carnauba wax emulsion containing Nisin (1000 ppm). Batch IV: Coated with nisin and Potassium Metabisulphite (KMS) solution. 51oC, for varying periods. Sensory profile. After the treatment, each batch consisting of 500g tomatoes were air dried and packed in polyethylene

Batch I: Coated with Lactic Acid Bacteria (LAB) live cells suspended in (de Man, Rogosa and Sharpe) selective broth. (L. lactis,

Dip coating method was employed for the surface coating of the tomatoes using various suspension mixtures.

pouches [200 gauge] with perforations (12 holes, 5 mm dia and at a distance of 2) and stored.

Storage studies: Each batch of the treated and control samples was stored at two different temperatures i.e., 283 oC and Assessment of stored vegetables: The stored tomatoes were analyzed for physical quality and physicochemical properties. spoilage. Based on these observations, the storage periods were decided. Determination of physicochemical characteristics: The selected tomatoes were assessed for appearance, colour, shriveling, texture, fungal growth as well as bacterial

Solids (TSS) and texture and compared with fresh vegetables. TSS was assessed by hand refractrometer (Erma No. 83140 0International Journal of Science Innovations and Discoveries, Volume 3, Issue 1, January-February 2013

Tomatoes (treated and control) stored under two different temperature conditions were tested for Total Soluble

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32) and texture was analyzed using a textrometer (CHTILLON-66502, made in USA). The methods adopted for both the experiments were standard techniques (AOAC, 1995). Sensory Evaluation:

Puttalingamma V et al., IJSID, 2013, 3 (1), 117-125

assessed for their sensory attribute. Since tomatoes are used as salad vegetable they were evaluated in raw form as salad for sensory analysis. Ten-trained panelists were requested to assess the samples using a 9-point Hedonic scale. Statistical analysis: Duncans new multiple range test at 5% significance. analysis of variance were employed wherever comparisons among the means were made. Differences were noted according to Knowledge about vegetable storage practices followed from pre historic period indicates application of a variety of RESULTS AND DISCUSSION

At the end of storage period when the physical characteristics were found ideal for consumption of tomatoes were

The observations are presented as arithmetic means along with standard deviations. Students t test and one-way

edible gums and waxes on the surface to extend the shelf life (Hardenburg, 1967; Coma et al., 2001; Ko et al., 2001). There synthetic materials as coatings (Table-1). Suitable explanations regarding the bio-chemical aspects of such treatments have been hypothesized. One of such substances known to be used in 12 and 13 th century is carnauba wax; it is a plant wax from Brazil origin (Kaplan, 1986; Sebtig et al., 2002; Jennifer et al., 2004). It was proposed to use carnauba wax as a base for holding crude bacteriocin and nisin. Surface flora and pathogens on the surface of the vegetables is evident from fig 1,2,and 3

have been modifications in these techniques through the addition of chemical preservatives, in such coatings as well as use of

Surface flora and pathogens were also analysed by pour plate method and results were given in the table-2. It Table 1: Composition of Carnauba wax Emulsion Carnauba wax 100g Gum Arabica 110g Distilled water 250ml Propylene Glycol 50ml Glycerol monostearate 10-15g

Title: Total plates ,Yeast and m oulds counts of tom ato (log)
6 4 2 0 TPC Y &M sur f ace mi cr o f l o r a Series1

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Fig.1..TPC and Y & M counts on the surface of the tomatoes.

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Puttalingamma V et al., IJSID, 2013, 3 (1), 117-125

T i t l e: P at hogens pr esent on t he sur f ace of T o mo a t o e s .

40 20 0 1 Sa l , A h, 2 3 V i b, L mo n o 4

Series1

In most cases nisin exhibited superior effect to that seen with intact cells (Thomas et al 2000). Therefore it was felt interesting Table- 3, that use of carnauba wax as a fixing agent altered the shelf life of the tomatoes.
35

to investigate whether fixing agent such as carnauba wax or adding antifungal agent to nisin is beneficial. It can be seen from

Our observations clearly demonstrated that LAB as intact viable cells or bacteriocins are potent antimicrobial agents.

Fig.2..Pathogens present on the surface of the tomatoes.

Title: Surece flora and H2S producing oganisns inTomatoes

Log no of organisns

30 25 20 15 10 5 0 1 TPC 2 Y& M 3 H2S Series1

Vegetables Tomato

TPC (log) 5.61

Fig. 3. H2 S producing organisms on the surface of the tomatoes. (%) Table -2: Different microflora on the surface of the vegetables. Yeast & Pathogens (%) Mould (log) 3.66 H2S producing organisms 33.3 Salmonella 31.2 Vibrio 18.7 Aeromonas hydrophila 37.5 Listeria monocytog enes 18.7

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with nisin coating on the surface at low temperature (+50C).(Fig-4). A marked difference in the physical appearance of tomatoes preserved with carnauba wax containing nisin was evident. The reason for such a change in the texture and provided sufficient protection from spoilage micro flora as well as pathogenic microorganisms. Table- 3: Shelf life of bio-coated tomatoes stored under different temperature conditions [n=] Coatings Nisin [1000 Carnauba wax + KMS + nisin Temperature Control L. lactis L. plantarum ppm] nisin emulsion [1000 ppm] Control - RT 8 0.5 Treated- RT 15 0.95 13 0.57 16 1 22 0.1 210.5 Control 15 1.28 +5 1oC Treated 45 1.5 28 1.15 40 0.57 480.1 400.1 + 5 1oC*Mean SD Sensory profile of stored vegetables:-

appearance of tomatoes coated with carnauba wax is not clear. Application of nisin suspension as a dip coating method

Tomatoes stored for thirty days appeared to be fresh. It can be stored up to 48 days using carnauba wax emulsion

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shelf life of tomatoes. The success of such an attempt would depend on the acceptability profile of treated and stored vegetables. Improvement of sensory attributes due to treatment and storage can be considered as a gainful attempt. On the

other hand a small decrease in acceptance may be indicative of a scope for improvement in the techniques or treatment or storage was decided based on the shelf life at ambient / room temperature conditions. Such a comparison was made to 6, presents data about shelf life behavior of tomatoes. It is important to mention here that scores for fresh tomatoes were the reasons for low score. The scores increased significantly (5% level) after one week of storage. The comparison between the scores for colour.

Sensory attributes are essential for acceptance of any food. In the present study an attempt was made to increase the

storage conditions so as to effectively improve shelf life without much alteration in the sensory profile of stored tomatoes. In the present study sensory profile of tomatoes treated and stored at two different temperatures were assessed. Period of identify the changes in sensory attributes of treated and untreated vegetables due to storage temperature. Tables .4 and 5 and significantly (5% level) lower to those of stored tomatoes, since semi ripe, tomatoes were procured, and this could be one of

the sensory attributes therefore was made among the control and treated samples stored under two different temperature condition. However colour, taste and texture indicated a significant (p<0.05) difference, with a small but significant increase in of low temperature. This difference could be due to tissue profile, since the tomatoes stored in low temperature had firm and intact tissues as against those stored at R.T (Table 5 and Table 6).

conditions. It is interesting to point out that aroma and taste of tomatoes did not differ due to either the treatment or storage

The difference in texture was large, a mean difference in score of 1.4 occurred between tomatoes stored at RT to that

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Puttalingamma V et al., IJSID, 2013, 3 (1), 117-125

Title : Biocoating and s he lf life of Tom atoe s 60 50

days of shelf life

40 30 20 10 0

Series 1 Series 2

Table 4: Physicochemical characteristics of treated and untreated vegetables Stored at two different temperatures Variations Storage period Moisture [%] Texture [kg] pH TSS Fresh Tomato 8 days 92.7 1.12 5.2 0.10 4.0 0.107 5.60.17 Control 91.01.59 4.00.05 4.80.08 5.00.05 Room Temperature Treated 92.8 0.05 4.91.59 4.60.01 4.90.17 Control 88.4 0.43 4.31.59 4.9 0.10 4.60.15 Low temperature Treated 88.7 0.75 4.41.59 4.00.10 4.80.15 SEM 0.58 0.045 0.086 F ratio 0.6866 NS 76.692 19.239 *Mean SD Table 5: Comparison of sensory profile * between treated and untreated Tomatoes stored under two different temperature conditions Stored for 8 days Room Temperature Low Temperature [51oC] F ratio SEM df [283oC] Fresh Control Treated Control Treated Mean 4.8c 6.7b 6.8b 7.3a 7.5a 0.789 0.675 0.919 0.823 0.972 Colour 16.180 0.079 Aroma Taste OQ Mean 4.8b 6.9a 7.3a 7.2a 7.2a 2.647 0.919 0.875 0.675 0.789 0.919 Mean 5.2b 7.2a 7.1a 7.3a 7.5a 11.451 0.632 0.632 0.994 0.999 1.080 Mean 5.2c 6.6b 6.7b 8.0a 8.1a 25.506 0.789 0.516 0.675 0.942 0.738 Mean 7.7a 4.9c 5.6c 6.5v 7.5a 42.25 0.483 0.316 0.699 0.707 0.527 *Mean SD; Values with different superscripts in rows are significantly different [P <0.05]. 0.426 0.086 0.062 0.179

Fig: 4. Shelf life of tomatoes with different treatments Ser (1) Rt samples , Ser (2) Low temp samples. (1) L. lactis , ( 2) L. plantarum (3) Nisin [1000 ppm, (4) Carnauba wax + nisin emulsion, (5) KMS+nisin [1000 ppm.

1 2 3 4 5 6 Diffe re nt biocoating tre atm e nts

45

Texture

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Table 6: Comparison of sensory profile *of treated and untreated Tomato stored at low temperature [5 1oC] [n=] Period of storage F Fre 8th day 16th day 25th day 32nd day 40th day rat SEM sh Contr Treat Contro Treat Contr Treate Contr Treat Control Treate io ol ed l ed ol d ol ed d Mea 4.8d 7.3c 7.5bc 7.9ab 8.1ab 7.7b 8.2a 7.7 d b 7.8b 7.7b 7.8b 21. n Colo 0.2 87 ur 09 0.7 0.823 0.972 0.568 0.568 0.483 0.632 0.483 0.422 0.483 0.422 6 89 Mea 4.8c 7.2b 7.2b 7.6a 7.2b 7.6a 7.2b 7.9a 7.2b 7.9a 7.2b 10. n Aro 0.2 14 ma 74 0.9 0.789 0.919 0.699 0.919 0.699 0.919 0.568 0.919 0.568 0.919 7 19 Mea 5.2d 7.1c 7.3c 7.5 d bc 7.8a 8.1a 7.8a 7.8a 7.8a 7.8a 7.8a 14. n Tast 0.2 78 e 18 0.6 0.994 1.080 0.422 0.568 0.422 0.632 0.422 0.632 0.422 0.632 3 32 Mea 5.2c 8a 8.1a 7.8ab 8.1a 7.6b 8.1a 7.6b 8.1a 7.6b 8.1a 18. n Text 0.2 23 ure 09 0.7 0.942 0.738 0.422 0.567 0.516 0.567 0.516 0.568 0.516 0.568 6 89 Mea 7.7a 6.5c 7.5a 6.1c 7.0b 5.5d 6.2c 4.2f 5.6d 3.0g 5.0e 54. n 0.1 OQ 37 91 0.4 0.707 0.527 0.738 0.441 0.707 0.422 0.919 0.516 0 0.667 3 83 *Mean SD; Values with different superscripts in rows are significantly different [P <0.05]. CONCLUSION

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df

99

10

coatings are protective coatings, which help to slow dehydration and decay while retaining moisture and increasing shelf life. The protective coatings also offer an attractive glossy sheen to some produce and may protect from brushing, injury. Particularly in the case of tomatoes it retains the important nutritional quality like minerals and lycopen pigments. As a result, low temperature storage was the most effective solution for both microbial spoilage and delay in the ripening of the tomatoes (Yaman, L. Bayndrl , 2001). Storage at low temperature (5 10C) prolongs the shelf life of many foods. In general, low foods. The antimicrobial properties of lactic acid bacteria and their metabolites and commercially available pure bacteriocin eg nisin have been recognized for decades. The vegetables were shredded and used for sensory analysis. Tomatoes were analyzed for sensory attributes. The temperatures reduce the growth rates of microorganisms and slow many of the physical and chemical reactions that occur in

Edible coating technique seems to be very promising as long as consumers accept this technique as safe. Surface

visual appearance and quality was used as a key factor to decide the shelf life of tomatoes. Ten trained panelist were requested to assess the samples using a 9 point Hedonic scale score card. It is conclusive therefore that Nisin+KMS and carnuba wax spoilage 1. +nisin emulsion are effective in controlling growth retaining the freshness of the carrot and beans at low temperature and also extending their shelf life considerably. However, this approach has been suggested for some nutritious perishable vegetables REFERENCES such as carrot, beans. This is thought to help in channeling resources to steps that provide effective protection from microbial Amir, H., Karas, M., Giat, J., Damilenko, M., (1999). Lycopene and 1,25-hydroxy vitamin D, cooperate in the inhibition of cell cycle progression and induction of differentiation in Hl-60 leukemic cells. Nutr. Cancer. 33 : 105. International Journal of Science Innovations and Discoveries, Volume 3, Issue 1, January-February 2013

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ether, fatty acids and nisin incorporation to inhibit Listeria innocua and Staphylococcus aureus. J. Food Protection. 64(4): Hardenburg, R.E. (1967). Wax and related coating for horticultural products A bibliography. Agricultural Research Jennifer, L, Grower., Kay Cooksey., Kelly, J.K., & Getty. (2004). Development and characterization of an antimicrobial packaging film coating containing nisin for inhibition of Listeria monocytogenes. J. of Food Protect., 67(3): 475-479. containing nisin and their action against Listeria monocytogenes. J. of Food Sci., 66(7): 1006-1011. Potential and perspective. Tech Reviews, Indian food Industry, May-June26,(3) p 37-42. modified atmosphere packaging.LWT- Food Science Tech, 42:1484-1490. Packaging applications. J. of Agricultural and Food Chemistry., 50(15): 4290-4294. leafy vegetables. J of Food Sci Techonlogy,45(5), 450-453. Press, London, W.D.C. Chapter 18 [Nisin], 463-524. 1-18. Kaplan, H.J. (1986). Washing waxing and colour adding in fresh citrus fruits. WF Wardowdki S Nagy and W. Grierson. Puttalingamma .V and . Khyrunnisa Begum.(2008). Occurrence of pathogens on surface of few selected vegetables procured from urban marketed during the year.Int.J. of Food Science , tech and Nutrition.Vol 2,no 1 p,129-136.

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