Effect of saffron (Crocus Sativus L) petal as preservative during refrigerated storage of mutton nuggets

Introduction

The presence of high quality nutrients especially the presence of essential amino acids and fatty acids make meat products an indispensable part of human diet (Dominguez et al., 2019). Meat products as an exceptional source of premium quality proteins with high biological value, is valued by developing various comminuted value-added meat products (Madane et al., 2019) [19] . However, the comminution of meat by mincing and chopping increase the tendency of pigment and lipid oxidation. This lipid oxidation along with microbial growth are responsible for limiting the shelf-life and consumer acceptance by causing rancidity that affects flavor, color, and even nutrient value of meat products (Ahmed et al., [2,12,20,21] . To prevent this oxidation problem in meat products, researches were using antioxidants from so many years. Synthetic antioxidants like Butylated hydroxyanisole (BHA) and Butylated hydroxytoluene (BHT) can generate health disease and carcinogenic if consumed frequently (Lishianawati et al., 2021) [18] . Thus, alternative antioxidants derived from safe natural ingredients are being advocated now a day for use in meat products. Kashmir is known to produce world's best saffron. During the harvesting of saffron, the petals are generally thrown away as waste. However, these petals are rich in biologically active compounds like phenolics, flavonoides and anthocyanin; hence can be exploited as functional ingredients in different meat products especially for their antioxidant potential (Hadizadeh et al., 2003) [13] , thereby developing novel meat products with improved shelf life, quality and enhanced health benefits. Although enough literature is available regarding the use of natural antioxidants such as rosemary, sea buckthorn, sage, oregano etc. in different meat products, but there was no data available with this regards to the use of saffron petals in mutton nuggets. Therefore, the current investigation was carried out with an intention to explore saffron petals as a potential natural antioxidant source during refrigerated storage of mutton nuggets.

Material and Methods

Procurement of materials for the preparation of mutton nuggets

Mutton was procured from local market of Shuhama, Kashmir and deboned manually. The lean meat obtained after deboning and trimming the fat manually was packed in low density polyethylene (LDPE) pouches (50-100) micron and stored at standard freezing temperature (-18 ± 2 0 C) until used. Condiment paste used contained fresh onion, garlic and ginger in the ratio of 3:2:1. The spice mix formula was standardized in the laboratory. Saffron petals were dried in an oven at 50-60 o C (overnight) to a moisture content of less than 8% followed by grinding to form saffron petal powder (SPP) and stored in air tight container for further use. SPP was incorporated in mutton nuggets at three different levels replacing lean meat in the formulation (Table-1).

Preparation of mutton nuggets

Thawed mutton was cut into pieces and minced in a mincer (Sirman, Italy) after passing through 8mm plate (2 times) and 4mm plate (once), salt was added into it along with half of chilled water and mixed for 1 minute. Vegetable oil was gradually added and thoroughly mixed for 1 minute followed by addition of whole egg liquid, dry spice mix and condiment paste for 1-2 minutes till proper emulsion was formed. This formulation served as control. Throughout the preparation, the SPP in different levels (1%, 2% and 3%) was added by replacing lean meat from the basic formulation of the product to produce the emulsion containing SPP. The emulsion prepared was then weighed and filled in stainless steel moulds. Moulds were covered with lid and tied with thread and cooked in hot water (90 o C) for 35 minutes to an internal temperature of around 80 o C. Mutton loaves so obtained were cooled to room temperature and cut into nuggets of approx 1cm 3 size. Samples were evaluated for various quality parameters to select the optimum level of SPP. Mutton nuggets containing optimum level of SPP (2%) along with control were packed aerobically in LDPE pouches and stored at refrigerated temperature (4±1 o C) for 21 days. During the refrigerated storage, nuggets were evaluated at an interval of 7 days on 0 th , 7 th , 14 th , and 21 th day for physio-chemical, antioxidant, microbiological and sensory parameters.

Analytical procedures

The emulsion stability was determined as per the method of Anjaneyulu et al. (1989) [3] . The pH of mutton nuggets was estimated by using digital pH meter (Model CP 901, Century Instrument Ltd. India). The proximate composition in both treatment sample and control was determined by using standard procedures of AOAC (2000). Tyrosine value was measured by the procedure put forward by Strange et al., (1977) [22] . with slight modifications. Thiobarbituric acid reactive substances (TBARS) value of cooked products was determined using standard procedure of Witte et al., (1970) [24] . The DPPH assay was done according to the method of Brand et al., (1995) [10] . with some modifications. Total plate count, coliform count and yeast and mould count were determined by methods of APHA (1984). The sensory evaluation of all sample products was performed by a panel of seven semi-trained members consisting of scientists and post-graduate scholars of the Division of Livestock Products Technology based on 8-point hedonic scale, wherein 8 denoted "extremely desirable" and 1 denoted "extremely undesirable" (Keeton et al., 1984) [15] . The data generated was analyzed by analysis of variances using SPPS software and the significance was checked at 5% level of confidence.

Results and Discussion

Optimisation of concentration of SPP

The physico-chemical properties like pH, cooking yield, emulsion stability and moisture content of SPP treated mutton nuggets was non-significantly (P>0.05) affected as compared to control (Table -2). Non-significant (P>0.05) effects on physio-chemical parameters were also seen by various workers in different meat products (Verma et al., (2013) [23] . in mutton nuggets with the incorporation of guava powder; Hazra et al. (2012) [14] . in buffalo meat treated with the crude extracts of Moringaoleifera (Lam.) leaves. There was significant (P <0.05) increase of ash content with increase the level of saffron petal powder.

Overall, slightly increased ash percentage of SPP treated mutton nuggets might be attributed to higher mineral content of SPP. Fat content SPP treated mutton nuggets was significantly (P <0.05) higher than control. Protein content of SPP treated mutton nuggets was significantly (P <0.05) lower than control. Similar type of results were reported by Bhat et al. (2014) [6] . in chicken nuggets treated with Aloevera pulp. The sensory evaluation revealed that upto 2% addition of SPP, the product remained acceptable for all parameters included overall acceptability (Table -3). However, the panelists rejected the product containing 3% level of SPP. Based on the physico-chemical properties and sensory evaluation of the products, 2% of SPP was selected as optimum level for use as antioxidant source in mutton nuggets. Therefore, the 2% SPP treated product along with control were stored in low density polyethylene (LDPE) pouches and kept at refrigerated temperature for a period of 21 days to ascertain their storage stability.

Storage studies

During storage, an overall increase in pH of products could be due to production of basic amines from protein breakdown by micro-organisms. The loss of moisture might be due to degradation of proteins responsible for water holding (Das et al. 2008) [11] . Ahmad (2014) [1] . found similar trend for moisture in aerobically packaged restructured buffalo meat fillets stored at refrigeration temperature. Significantly lower (P <0.05) TBARS and tyrosine values for treatment products could be due to the presence of phenolic compounds in SPP. Kumar and Tanwar (2011) [16] . reported that chicken nuggets containing clove powder (optimized product) maintained significantly (P <0.05) lower TBA values throughout the storage period. DPPH radical scavenging activity (DPPH-RSA) values of SPP treated mutton nuggets were significantly (P <0.05) higher than control. The significantly (P <0.05) higher DPPH-RSA value might be due to the addition of antioxidant rich SPP. These results (increased DPPH-RSA and reduced TBARS and tyrosine values) indicated that saffron petal powder is an excellent natural antioxidant source. During storage, an overall increase in microbial counts (TPC, CC and YMC) were observed, however a comparatively slow increase in SPP treated mutton nuggets might be attributed to the possible antimicrobial activity of the SPP. Similar observation was reported by Ahmad (2014) [1] . in buffalo meat fillets, Bhat et al. (2013a) [8,9] . in chicken seekhkababs, Bhat et al. (2013b) [8,9] . in chicken meat balls; Kumar and Tanwar (2011) [16] . in chicken nuggets treated with different natural ingredients. During storage, coliform counts were not detected during initial period upto 7 th day and yeast and mould count were not detected up to 14 th day of storage. YMC were not detected during early period of storage, which could be due to thorough cooking, good hygiene and absence of post processing contamination. [8,9] , Bhat and Pathak (2009) and Kumar and Sharma (2004) [17] . The products were however, acceptable even on 21 st day of storage.

Conclusion

The outcome of the current investigation pointed out that saffron petal powder is an excellent source of antioxidants as was confirmed by the results during refrigerated storage of mutton nuggets. The saffron petals also showed antimicrobial effects. It was thus concluded that the waste product i.e., saffron petals, if exploited properly can add value and storage stability to meat products.

Acknowledgement

AcknowledgementThe authors are thankful to the Dean, faculty of veterinary International Journal of Advanced Biochemistry Research https://www.biochemjournal.com sciences and animal husbandry SKUAST Kashmir for providing the necessary facilities.

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