Effect of nitrite substitutes for vegetable sources on meat color fixation on fermented sausage production

다국어 초록 (Multilingual Abstract)

The purpose of this study is to screening vegetable powders can substitute nitrite had used for curing agents in meat products and confirming the preservative effects by apply to fermented sausage.

As known in previous research, vegetables contained much nitrate were selected. Total 14 kinds of vegetables nitrate contents were measured by Ion Chromatography System and nitrite contents were measured by diazotization method of Korean Food Standards Codex. The nitrate contents were measured range to minimum 800 mg/kg to maximum 8223.5 mg/kg. The nitrite contents were measured range to not detected (ND) to maximum 339.4 mg/kg.

Five vegetables (vitamin, young radish, crown daisy, leaves of Brassica juncea L. celery) which had more than 5,000 mg/kg nitrate contents were selected to investigate the nitrite produce with nitrate reductive strains. Samples were cultivated respectively with Staphylococcus carnosus KCTC 3580, Staphylococcus xylosus KCTC 3342 and Commercial fermented sausages starter BITEC Advanced LD-20 (Staphylococcus carnosus and Lactobacillus sakei) in TSB broth at 30℃, 37℃ in 24, 48, 72hr, to confirm the nitrate reductive activities. Optical density (OD) of each samples were observe to probe nitrate reduction and nitrite production.

The portion of vegetable powders were selected according to most nitrate reductive activity of each vegetable powders, vitamin and young radish was 1.0%, celery and leaves of Brassica juncea L. was 1.5% and crown daisy was 2.0% of total curing spices. There were no significant difference in OD of 30℃ and 37℃ (p<0.05).

Fermented sausages were prepared by 3 types, nitrate added (Control-1) and nitrate, nitrite added (Control-2), and nitrite-free and vegetable powder added (Experimental batches). Control-1 was manufactured applying 120 ppm of sodium nitrate and Control-2 was manufactured applying 120 ppm of sodium nitrate and 70 ppm of sodium nitrite. Vegetable powder added fermented sausage was manufactured with each vegetable powder’s most effective nitrate reduction ratio. The commercial fermented sausage starter culture BITEC Advanced LD-20 was added all fermented sausages. Fermented sausages were ripened 30-days fermentation.

The weight loss of drying was reached 40% after 24 days. The pH value of fermented sausages was decreased from average 5.80 to 4.50 for 2 days, and then gradually increased to 4.80 levels. The water activity of fermented sausages was gradually decreased from average 0.970 to 0.860.

The chromatic value CIE a* of batch D and I were 6.24 and 6.89, similar or even better than Control batches, but other Experimental batches were lower than Control batches significantly.

Lipid rancidity of fermented sausages added nitrate was 0.52±0.13 TBARS value at 4th week of fermentation, and the TBARS value of experimental batch D and I was significantly no difference compared with Control batches.



Pathogenic microorganisms such as coliform, E.coli O157:H7 KCCM 40406, Staphylococcus aureus KCTC 1916, Clostridium perfringens KCTC 3269 were not detected in all raw materials and tested samples during fermentation.

Biogenic amines were not detected all of the Control batches and Experimental batches during and after fermentation periods.

The sensory evaluation of Experimental batch D and I were not significantly different between Control batches and Experimental batch D and I on color, flavor and overall preferences.

As the results, high-nitrate containing vegetable powders such as crown daisy and leaves of Brassica juncea L. could substitute the nitrite usages in fermented sausage production applied with nitrate reductive microorganism.

목차 (Table of Contents)

• Contents
• Contents ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ i
• List of Figures ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 4
• List of Tables ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 5
• Abstract ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 7
• Ⅰ. Introduction ·····························································10
• Ⅱ. Materials ··················································· 13
• 2.1 Materials
• 2.1.1 Vegetable powder ····························································· 13
• 2.1.2 Microorganisms ·······························································13
• 2.1.3 Media ·········································································13
• 2.1.4 Preparation of fermented sausage ·······································13
• Ⅲ. Methods ································································· 14
• 3.1 Vegetable powders preparation ················································ 14
• 3.2 Analysis of nitrates contents in vegetable powders ······················ ···14
• 3.3 Incubation conditions for nitrite formation from vegetable sources with microorganism ······································································· 15
• 3.4 Manufacturing of fermented sausages (In situ) ····························· 16
• 3.5 Analysis of physicochemical characteristics ·································19
• 3.5.1 Weight loss on drying of fermented sausages ··························· 19
• 3.5.2 Changes of pH ······························································· 17
• 3.5.3 Changes of aw ································································ 20
• 3.5.4 Analysis of fermented sausages color ···································· 20
• 3.5.5 Thiobarbituric acid reactive substance (TBARS) ······················ 21
• 3.6 Microbiological analysis ······················································ 22
• 3.6.1 Detection of pathogen microorganism ··································· 23
• 3.7 Analysis of residual nitrite contents ·········································· 26
• 3.8 Determination of biogenic amines (BAs) by HPLC ························ 27
• 3.9 Sensory evaluation ······························································ 28
• 3.10 Statistical analysis ····························································· 29
• Ⅳ. Results and Discussion ················································ 44
• Ⅴ. Conclusion······························································· 79
• Ⅵ. Reference ································································ 81
• Acknowledgement ····················································· ·····87