Cassava starch extracted from cassava roots (
Cassava (
The composite flour mixture is included with rice flour, maize powder, milk powder, and bread crumbs. Composite flour mixture has been used as binders in comminuted meat products due to their economic benefits. The amount of composite flour mixture used in processed meat varies depending upon the type of product, manufacturing procedure, and government regulations (
Nowadays, starches are used as binders in processed meat products to absorb moisture, which is released from meat protein during heating. A satisfactory binder should have the ability to hold more moisture throughout the processing, cooking, chilling, and storage of meat products. Today scientific principles are employed to improve production procedures, product quality, and product safety.
To maintain the water-holding ability the binder should also assist in binding the fat and maintaining its dispersion throughout the mix. Starches are commonly added to emulsion-type meat products and are popular not only for their functional properties but also to extend the more expensive lean meat portion of products (
Extracted cassava native starch and modified starch were used for the replacement of composite flour mixture as a source of dietary fiber content (
The replacement of composite flour mixture with cassava starch decreases the cost of preparation, improves the organoleptic properties and nutritional composition of fresh chicken sausage.
MU51 variety is one of the most cultivated and recommended cassava varieties in Sri Lanka. Thus, the predetermined quantity of cassava MU51 variety was purchased from a cassava cultivator in Bandaragama, Sri Lanka.
Kjeldahl Tablets (Kjeldahl Tablets, Suphelco, Sigma- Aldrich Chemie GmbH).
Conc. H2SO4 (Concentrated Sulphuric acid, Sigma- Aldrich, Sigma-Aldrich Chemie GmbH, 99.999%).
Conc. NaOH (Sodium Hydroxide, Sigald, Sigma-Aldrich Chemie GmbH, ≥98%)
95% Ethyl Alcohol (Etanol, Sigma-Aldrich, Sigma- Aldrich Chemie GmbH, 95%).
6N HCl (Hydrochloric Acid, Sigma, Sigma-Aldrich Chemie GmbH, 37%).
Diethyl Ether (Diethyl Ether, Sigma-Aldrich, Sigma- Aldrich Chemie GmbH, 99.7%).
Petroleom Ether ( Petroleom Ether, SIGALD, Sigma- Aldrich Chemie GmbH, 99%).
Phenol (Phenol, Sigma-Aldrich, Sigma-Aldrich Chemie GmbH, 99%).
Potassium cyanide (Potassium cyanide, Sigald, Sigma- Aldrich Chemie GmbH, 99%).
Pictric Acid (2,4,6-Trinitrophenol, Sigma-Aldrich, Sigma- Aldrich Chemie GmbH, 99%).
Methyl Red (Methyl Red, Sigma-Aldrich, Sigma-Aldrich Chemie GmbH, 99.5%).
Methyl Blue (Mthyl blue, Sigma-Aldrich, Sigma-Aldrich Chemie GmbH, 70%).
Peptone (Peptone, Sigma-Aldrich, Sigma-Aldrich Chemie GmbH).
Nutrient agar (Nutrient Agar, Sigma-Aldrich, Sigma- Aldrich Chemie GmbH).
Potato Dextrose Agar (Potato Dextrose Agar, Sigma- Aldrich, Sigma-Aldrich Chemie GmbH).
The Brookfield texture analysis (Model: CT3-4500, Brookfield Engineering, USA).
UV–Vis spectrophotometer (Model: UV mini-1240, SHIMADZU CORPORATION, Japan).
Muffle furnace (Model: SHIMADEN SR1, SHIMADEN CO., LTD, Tokyo).
Polarized Zeeman atomic absorption spectrophotometer (Model ZA3000, HITACHI, Japan).
Analysis of storage stability – (
Pre-gelatinized modification (PG) –
Nutrition Analysis of sausage –
Toxicological Analysis – Spectrophotometric Determination of Cyanide (Picric acid method) by
Cassava starch was extracted according to the method described by
Finally, ground starch was packed in a polyethylene package and was stored (-5 °C) in an airtight container.
Pre-gelatinized modification (PG) was done with slight modifications of the method described by
The pre-gelatinized cassava starch was packed in a polyethylene package and stored under cold room conditions at -5 °C.
The basic ingredients used for the chicken sausage were lean meat (minced chicken) fat, ice, isolated soy protein, composite flour mixture, rice flour, maize powder, bread crumbs, milk powder, cassava starch (native and modified), sugar, salt, garlic, black pepper, and chili powder. The amount of ingredients added to sausage preparation is summarized in Table
Ingredients content in sausage formulae.
Ingredient | a0b0 CFM 50%Starch 50% | a0b1 CFM 0%Starch 100% | C CFM 100% Starch 0% | a1b0 CFM 50% Starch 50% | a1b1 CFM 0% Starch 100% |
---|---|---|---|---|---|
|
|||||
Meat % | 57.6 | 57.6 | 57.6 | 57.6 | 57.6 |
Emulsion % | 33.3 | 33.3 | 33.3 | 33.3 | 33.3 |
Composite Flour Mixture (CFM) % | 3.05 | 0 | 6.1 | 3.05 | 0 |
Cassava Native starch % | 3.05 | 6.1 | 0 | 0 | 0 |
Cassava Modified starch % | 0 | 0 | 0 | 3.05 | 3.05 |
Sugar & Salts % | 2 | 2 | 2 | 2 | 2 |
Spices Mixture % | 1 | 1 | 1 | 1 | 1 |
Two cassava starch types (i.e., native cassava starch (a0) and modified cassava starch (a1)) were used in the experimental design. In addition, two replacement levels (i.e., 50% (b0) and 100% (b1)) were included.
Four different sausage batters were prepared by replacing composite flour mixture with cassava native starch in 50% (a0b0) and 100% (a0b1) levels and with cassava modified starch in 50% (a1b0) and 100% (a1b1) levels. A sausage was prepared without replacing the composite flour mixture as a control. Other ingredients and additives were incorporated in the formulated batters (Table
The batter was filled into cellulose casings and cooked in steam for 15 minutes until the core temperature becomes 72 oC. Then the prepared sausage was stored at -5 °C in a freezer for further analysis.
Six sensory attributes (i.e., Appearance, Colour, Aroma, Flavour, Hardness, Overall acceptability) were analyzed by using the five-point hedonic scale (5 = Like extremely, 1= Extremely dislike) and 30 untrained panelists.
The texture profile (TPA) was analyzed according to the method described by
Values for the color dimensions L*, a*, b*, C*, and h* were measured by using the Colorimeter (Lovibond).
Moisture content, crude protein content, crude fat content ash content, and acid insoluble ash contents were analyzed for the top-ranked sausage samples (a0b0 - 50% replacement with Cassava Native starch) following the
The top-ranked sausage sample was subjected to dry ashing (using muffle furnace at 500 oC) and analyzed for the presence/quantity of Na, K, Ca and Mg minerals using Atomic Absorption Spectrophotometric (AAS) method.
Starch content was measured by the method of Dubois (
The cyanide content of the top-ranked sausage sample (a0b0 – 50% replacement with Cassava Native starch) was analyzed according to Spectrophotometric Determination of Cyanide (Picric acid method) by
About 10 – 20g of the grounded top-ranked sausage sample was weighed and the weight was recorded. Then the starch sample was introduced into a triple-necked round bottom flask containing 100 mL of deionized water. About 50 mL of 2N H2SO4 was added to the round bottom flask. The flask was immediately connected to the steam generator and the distillate was collected in 50 mL of Na2CO3 solution till it becomes 200 ml. The solution was transferred into a 250 mL volumetric flask and the volume was made up to 250 mL using deionized water.
Then 10.00 mL of the distillate (final solution should be contained 3 mL of 5% Na2CO3), and 4 mL of 1% Pictric acid were added into a boiling tube and the boiling tube was immersed in a boiling water bath for 12 – 15 minutes. Then the solution was left to cool to room temperature and volume was made up to 25 mL.
Finally, the absorbance was measured at 530 nm in UVVis spectrophotometer (VU mini – 1240) for the developed color of CN- with Picric acid. Reading of absorbance was noted down and CN- concentration was calculated based on the equation (Cyanide Content = (absorbance+0.03)/0.0048) which was been first developed using standard concentration series of KCN solution.
The total plate count and coliforms of the prepared sausage samples were determined during the storage using the methods described by SLS 516 (2013).
The total plate count of the sausage sample was performed using a Colony count at 30 °C by the surface plating technique.
Coliform detection and enumeration of frozen fries samples were carried by the Most Probable Number (MPN) method.
Photographs of sausages with different treatment types and levels. Note: a0b0 – Native 50%, a0b1– Native 100%, a1b0 – Modified 50%, a1b1 – Modified 100%, C- 0% Replacement.
Bar Chart of Friedman overall of Ranks 751 – 50% replacement with Cassava Native starch, 351 – 100% replacement with Cassava Native starch, 451 – 50% replacement with Cassava Modified starch, 651 – 100% replacement with Cassava Modified starch, 551 – Control.
All the parametric and non-parametric data were analyzed by using MINITAB 17 statistical tool. Non-parametric data which are results obtained from the sensory analysis were analyzed using the Friedman ranking test. One-way ANOVA was used to analyze the significant differences of mean values of each sample, followed by Tukey Pairwise comparison test to analyze the samples that had a significant difference in color and texture. Two sample t-test was used to analyze the significant difference in nutritional composition by referring to the reference values obtained from SLS and USDA standards.
As the sensory profile is the most critical factor results of the sensory analysis were used for the selection of the best sample. The top-ranked sample was 551(Control sausage- 0% replacement) according to the average ranking of Friedman analysis. The second best-ranked sample was 751 (a0b0 – 50% replacement with cassava native starch) according to the results of the Friedman ranking analysis.
Results further suggest that the color, appearance, and overall acceptability of all the treatment levels are significantly different from the control sausage sample, (
Bar chart of compares nutritional composition of sausage with referred SLS and USDA standards.
According to the texture profile analysis data a0b0 sample has been given 1475 ±278 (g) for gumminess, 99.5 ±19.7 (mJ) for Chewiness, 0.24 ±0.01 for cohesiveness, and 6.88 ±0.06 (mm) for springiness values.
Since the
According to
In the colorimetric values analysis L*, a*, b*, c*, ho values refer to lightness, redness, yellowness, saturation, and hue, and those dimensions were considered. According to the color values analysis results, there was no significant effect (at 0.05 significant level) by the 50% replacement by cassava native starch (a0b0) on lightness and hue. However, that treatment (a0b0) affected the redness, yellowness, and saturation of sausage.
The 100% replacement by the cassava native starch (a0b1) significantly affected (at 0.05 significant level) lightness, redness, and hue. However, it was not significantly affected by yellowness and saturation. A similar result was observed by
The 50% replacement with Cassava Modified starch (a1b0) was significantly influenced (at 0.05 significant level) on the lightness and redness. Also, it was not significantly affected by yellowness and saturation.
But the treatment of 100% replacement with Cassava Modified starch (a1b1) was significantly affected (at 0.05 significant level) on the lightness, redness, yellowness, and saturation and significantly not affected on hue according to Tukey’s pairwise comparison results.
Since the p values are less than 0.01 (at 1% significant level) for the correlation of a0 vs. L*, a1 vs. a*, a1 vs. L*, a1 vs. a*, a1 vs. b*, a1 vs. C* and a1 vs. ho (
According to the Pearson correlation analysis results, there was a strong negative relationship between a0 vs. L*, a1 vs. L*, and a0 vs. h0. Also, there was a strong positive relationship between a1 vs. a*, a1 vs. b*, and a1 vs. C*.
The top-ranked sample showed average moisture, crude fat, crude protein, total solids, ash, and acid-insoluble ash content of 65.98%, 7.19%, 8.79%, 34.02%, 2.36%, and 0.32% respectively. The starch content of the top-ranked sample was 3.12%. According to
According to the SLS and USDA standards moisture, crude fat, crude protein, total solids, ash, and acid-insoluble ash contents were 74.6%, 33%, 12.1%, 20%, 4 %, 1%, and 0.5% respectively (
Previous research on the effect of substitution of fat with Sorgam and finger-millet showed protein, fat, ash, crude fiber, and carbohydrates contents as 10.4%, 3.1%, 1.6%, 2.0%, and 70.7% respectively (Das et al., 2013).
High fat consumption has been associated with cardiovascular diseases, obesity, cancer, and hypertension, among other illnesses (
According to the AAS analysis results, there was a high Na content in the sausage sample which was 1348.39 mg.100g-1. Also, the K and Mg contents were 232.15 mg.100g-1 and 73.21 mg.100g-1 respectively. According to the analysis the absorbance value for the Fe content was not observed in the standard range. Calculation suggests that the Fe content of the sausage sample should be less than 0.012 mg.100g-1.
Cassava starch has a cyanide content of 7.05 mg.kg-1 on a dry basis (
The detected average pH value of the sausage was 5.64 ±0.11. The standard level for pH 6.6 has been shown by
Microbial analysis was conducted by TPC and coliform test. Total plate count was analyzed with the purpose of shelf life analysis of the best-ranked sausage and coliform analysis was conducted to determine whether the coliform bacteria are present or absent in the sausage sample.
Sausage is usually exposed to high temperatures by heating till the middle temperature becomes greater than 74 °C by using the steaming process. But this temperature might not be enough to inactivate all the microorganisms.
Sausages are also re-contaminated with spoilage bacteria during the processing stages followed after cooking. Since that the initial plate count was around 10 C.F.U.g-1.
The excessive proliferation of present microflora in the sausage content or on the surface during storage causes economic losses because of spoilage and deterioration. Especially, lactic acid bacteria are considered to be a major component of the microbial population found on vacuumpackaged sausages (
Findings from research by
The best composite flour mixture replacement formula was a 50% composite flour mixture with cassava native starch. There was no significant effect on the total solids content by the 50% replacement with cassava native starch and it was significantly affected on moisture content, protein content, fat content, starch content, ash content, acid insoluble ash content, and pH value of the final product. Shelf life analysis of developed sausage suggests that the product was safer to consume up to 14 days of manufacturing under frozen conditions (-5 °C).
The authors express their gratitude to the University of Sri Jayewardenepura, Sri Lanka for the provided facilities to complete the research study successfully and to Ms. Sashie Abeywickrema, Research Candidate (Sensory Neuroscience), Department of Food Science University of Otago, Dunedin, New Zealand, for her generous assistance in language correction.
This research received no external funding.
No conflicts of interest to declare regarding the present study.
Not applicable.