View of Utilization of Spirulina platensis Algae as A Functional Ingredient for Improving the Nutritional value of ketchup Product

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Utilization of Spirulina platensis Algae as A Functional Ingredient for Improving the Nutritional value of ketchup Product

Shaden, H. Foudah and Nisreen M. Abdulsalam

Food and Nutrition Department, Faculty of Human Sciences and Design, King Abdulaziz, Jeddah-Saudi Arabia.

ABSTRACT

The Spirulina platensis algae also called blue green algae is a class of gram-negative bacteria which possesses a wide range of bioactive colored components such as phycocyanin, carotenoids, and chlorophyll. Spirulina algae is one type of microalgae containing nutrients that have been used as a functional ingredient. In addition to its therapeutic and pharmaceutical applications. This study is aimed to examine the effect of Spirulina algae to improve the nutritional value of ketchup as it is a commonly used product.

Objective: This study aims to assess the effect of adding Spirulina platensis algae powder as a functional ingredient to ketchup, in increasing minerals (Ca, Fe), and improving antioxidant activity (polyphenols, lycopene).

Method: A homogenous mixture of Spirulina algae powder cultured by the open commercial farms was used in this study. Three treatments (1,2,3 %) that applied to ketchup were compared to a control. Chemical compositions of Spirulina algae, as well as minerals, determined the antioxidant activity using (DPPH), and sensory evaluation were determined.

Results: The results revealed that adding Spirulina platensis algae to ketchup increased its minerals content (Ca, Fe). Antioxidant activity (polyphenols and lycopene) was also accepted. Regarding the sensory evaluation and consumers acceptability, tests show no significant difference between the three treatments and control .

Conclusion: The study data regarding Spirulina platensis algae as a functional ingredient indicated that Spirulina algae was a good selection when fortifying tomato products to improve the nutritional value, it is similarly a powerful antioxidant that can be used in alternative treatments. Furthermore, all ketchup samples had good organoleptic properties. The sensory evaluation of the consumers acceptability was good in general, but the most preferable one was the ketchup fortified with 1%S.

platensis.

Key Words: Spirulina Algae, nutritional value, antioxidants, minerals, ketchup .

INTRODUCTION

Ketchup is a commonly consumed and popular foodstuff that has been more often discussed in recent years, particularly because of its lycopene content and related beneﬁcial properties. In the European market, products of this type are abundant varying from each other in nutritional and sensory quality. With regard to the fact that the food market is generally saturated, the assertion of food products including ketchups is possible only in case of their systematic innovation. For the desirable innovation, sensory quality is of the primary importance as the majority of consumers are most aware of this quality aspect (Tauferova, et al., 2015).

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Food fortification (FF) is defined as the addition of one or more essential nutrients to a food, whether or not it is normally contained in the food, for the purpose of preventing or correcting a demonstrated deficiency of one or more nutrients in the population or specific population groups. Food fortification includes biofortification, microbial biofortification and synthetic biology; commercial and industrial fortification, and home fortification. The several types of FF are distinct because different techniques and procedures are used to fortify the target foods.

Biofortification involves creating micronutrient-dense staple crops using traditional breeding techniques and/or biotechnology (Hettiarachchi, 2011).

Compared to other foods or by weight, Spirulina is recognized as one of the most nutritious foods on the planet: high in proteins, containing all essential amino acids, also high in B vitamins, iron, magnesium, potassium and many other vitamins and minerals, as well as antioxidants (Junga, et al., 2019).

Spirulina algae are an important source of nutrients in the traditional diet of some populations of Africa and Mexico. These algae can be extensively grown to obtain a protein-rich material of alimentary use (foodstuff for diet complementation) or industrial use (blue pigments, emulsifiers, thickening and gelling agent). The chemical composition of Spirulina indicates that it has a high nutritional value due to a wide range of essential nutrients, such as vitamins, minerals and proteins (Miranda et al., 1998).

It is well known that microbial pigments are highly responsible for the health benefits and plays a key role as antioxidant due to the presence of hydroxyl substituent and their aromatic structures which enable them to scavenge free radicals. Antioxidants are substances that protect the cells from harm caused by instable molecules known as free radicals. Antioxidants include carotenoids: beta carotene, phenolic compounds:

gallic acid, flavanoids, quercetin, alkanoids, capsaicin, hydroxytoluene (Safari et al., 2019).

This study aims to assess the effect of adding Spirulina platensis algae powder as a functional ingredient to ketchup, in increasing minerals (Ca, Fe), and improving antioxidant activity (polyphenols, lycopene).

Methodology

Preparation of ketchup fortified Spirulina algae: Tomatoes were sorted based on color, firmness and freedom from diseases, lemishes, and rotting. Washing of the tomatoes was done thoroughly under running tap water to reduce initial microbial load which was present on the surface of tomatoes along with dirt, dust, and other extraneous matter. Chop the tomatoes into four pieces, then put them on the stove and cover the pot until the tomatoes are soft and the liquid comes out and Ladle the tomatoes into a fine strainer and press mixture with the back of a ladle to strain out any skins and seeds. The tomato juice was put in an open pan and the spices were wrapped in a muslin cloth and dipped into the juice. Onion, ginger, and garlic were also pulped and added to the juice. The sauce was heated on a low flame with constant stirring. The sauce was filled hot in glass bottles, closed and stored at 4°C in refrigerator for no more than 48 hours (Gujral, et al., 2002). (Ketchup ingredients include whole tomato, sugar, salt, onions, ginger, garlic, mace, cloves (headless), cinnamon, red chili powder and black pepper, acetic acid).

Mineral’s content: The method described by A.O.A.C. (2016) was used for mineral analysis. The ash was digested with 3 cm³ of 3 Ml HCl and made up to the mark in a

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100 Cm³ standard flask with 0.36 Ml HCl before the mineral elements were determined by atomic absorption spectrophotometer.

Determination for lycopene: Determination for lycopene content was by using Spectrophotometer UV-VIS SPECORD 205 by Analytic Jena. Lycopene in the sauce samples was extracted with hexane: ethanol: acetone (2: 1: 1) (v / v) mixture following the method of Sharma and Le Maquer. One gram of the homogenized samples and 25 ml of hexane: ethanol: acetone, which were then placed on the rotary mixer for 30 min., Adding 10 ml distilled water and was continued agitation for another 2 min. The solution was then left to separate into distinct polar and non-polar layers. The absorbance was measured at 472 nm and 502 nm, using hexane as a blank.

The lycopene concentration was calculated using its specific extinction coefficient (E 1%, 1 cm) of 3450 in hexane at 472 nm [8] and 3150 at 502nm. The lycopene concentration was expressed as mg / 100g product. All determinations were repeated for Next; an 8-mm Gaussian smoothing kernel was created around each peak such that voxels closest to the peak were given a value of 1 with those further away receiving values closer to 0. This allowed us to create graded contrast-indicator maps (CIMs) for each peak reported. These CIMs were then combined to form a single nested map for each contrast, thus ensuring each contrast could contribute no more than a value of 1 for a given voxel when overall proportion statistics were calculated.

These combined CIMs were then weighted by the square root of the sample size of the contrast, so that data based on many participants would exert greater influence on the meta-analyses than data based on just a few participants. The 8-mm kernel used in the present analyses is slightly smaller than what has been used in other MKDA meta- analyses (typically 10–15 mm), making this approach somewhat more conservative.

When identical analyses were performed using a 10-mm kernel, all peaks identified using the 8-mm kernel were found to be included within the clusters identified using the 10-mm kernel. Yet, the clusters identified using the 10-mm kernel contained more voxels. One additional cluster was identified in the 10-mm kernel results that was located in left middle frontal gyrus (−27 51 30). This cluster did not meet FWE correction for the 8-mm kernel analysis (k = 38) and thus was not reported in the present results (Buhle et al., 2014).

Determination of total phenolic content: The total phenolic content in the extracts was determined with Folin-Ciocalteu reagent using the method of Sidduraju and Becker (Siddhuraju & Becker, 2003).100 µL of the extracts (10 mg/ml) were taken in test tubes and made up to the volume of 1 ml with distilled water. Then 0.5 ml of Folin-Ciocalteu phenol reagent (1:1 with water) and 2.5 ml of sodium carbonate solution (20%) were added sequentially in each tube. Soon after vertexing the reaction mixture, the test tubes were placed in dark for 40 min and the absorbance was recorded at 725 nm using a spectrophotometer (Shimadzu 1650, Japan) against the reagent blank. The analysis was performed in triplicate and the results were expressed as the gallic acid equivalents (Nagmoti, et al., 2012).

Antioxidant activity: (Free radical scavenging activity) 2, 2-diphenyl-1-picryl- hydrazyl (DPPH) of ketchup was determined following the method of (Rajan et al., 2011). Samples of different concentrations were mixed with an aliquot of DPPH (1 ml, 0.004% w/v). The mixture was vigorously shaken and left to stand for 30 min in the dark at room temperature. The absorbance at 517 nm was recorded to determine the concentration of remaining DPPH. The radical- scavenging activity was calculated as % inhibition by the following formula:

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Inhibition (%) = (A control – A test) / A control X 100. Where; A control = the absorbance of the control reaction. A test = the absorbance of the ketchup. Ascorbic acid was used as a reference compound. Effective concentrations at 50% (EC50) were calculated from regression equations of calibration plots (Y= 1.0445x + 28.826, R² = 0.9658 and Y = 2.8301x + 41.276, R² = 0.9667 for ketchup, respectively) to denote the effective concentration of a sample required to decrease the absorbance at 517 nm by 50%.

Sensory evaluation: Sensory panel was composed of 60 trained assessors (from Female students of King Abdulaziz University) who do not have enough experience with sensory analysis of foodstuffs. The panel composition was chosen on purpose in response to the fact that women go shopping for food in most households. The assessors had not used any perfume, smoked, or consumed any food or beverages that might influence their perceptions for a period of one hour before the analysis. The analysis was carried out for assessors based on the prepared forms for sensory evaluation of ketchups. The samples were evaluated for the overall pleasantness, partial textural properties which included viscosity and density as well as texture acceptability, using a nine-point scale, where nine corresponded to the highest overall pleasantness /viscosity /density /texture acceptability. All panelists classified of five groups that included individual classes of texture acceptability, by which it was possible to detect even subtle differences between samples. In order to create groups of samples evaluated by individual assessors, a statistical model of balanced incomplete block was applied, which allowed reducing the number of samples evaluated by one assessor. Each assessor evaluated five samples, whereas, each sample was evaluated in triplicate by three different assessors. The order of samples in the evaluation of individual assessors differed for reducing its impact on the evaluation. Samples were served in plastic cups of 50 ml volume identified by numerical codes. Stainless spoons were available for manipulation with the samples.

Clean still water was available as a neutralizer (Forde et al., 2007).

Ethical approval: The study protocol was approved by Research Ethics Committee, Faculty of Medicine, King Abdul-Aziz University

Statistical analyses: All obtained data was analysed using Statistical Package for the Social Sciences (SPSS) for Windows, version 20 (SPSS Inc., Chicago, IL, USA).

Data will be presented as mean± standard deviation (SD). Analysis of Variance (ANOVA) test will be used for determining the significances among different groups according to (Armitage et al., 2001). All differences will consider significant if P- values were ˂0.05.

RESULTS AND DISCUSSION

Minerals contents of sauce supplemented with spirulina algae: The results listed in Fig. 1 show the content of mineral elements (calcium, iron and sodium) in spirulina-fortified ketchup samples, as well as standard samples. Samples of ketchup supported with spirulina at concentrations of 1, 2 and 3% showed an increase in the content of calcium with an increase in the concentration of spirulina compared to laboratory and standard control subjects, where the values increased from 21.67±0.98 to 23.65±1.04%.

The same thing was the content of iron and sodium, as the content of iron and sodium increased with the increase in the concentration of added spirulina. The increase in the

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content of calcium, iron and sodium may be due to the increase in the percentage of

spirulina added to the ketchup samples.

Total polyphenols and lycopene content (mg/kg) of ketchup supplemented with spirulina algae: Spirulina is characterized by its high content of phenols in addition to carotenoids. Tomatoes also contain lycopene, which is one of the types of carotenoids in addition to phenols. Results for the estimation of both phenols and lycopene in ketchup and control samples are shown in Fig 2. The samples showed an increase in total phenolate values in ketchup samples fortified with spirulina at concentrations of 1%, 2% and 3%, which increased from 892.00±10.52% to 1011.00±11.21% compared to commercial control samples. On the other hand, we found a higher lycopene content in spirulina-fortified ketchup samples compared to commercial control samples.

From the previous results, the higher content of phenols and lycopene in ketchup samples supported with spirulina compared to control may be due to the higher content of phenols in spirulina in addition to carotenoids.

Fig 1. Minerals contents of ketchup supplemented with spirulina algae. Data are expressed as mean ± SD values given represent means of three determinations.

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DPPH radical-scavenging activity: There are different methods for estimation of antioxidant activity but the most widely methods are those that involve generation of free radical species which are then neutralized by antioxidant compounds. DPPH radical is commonly used as substrate to evaluate antioxidant activity; it is useful and stable free radical that can accept on electron or hydrogen radical to become a stable molecule. The reduction of DPPH free radical was determined by the decrease in its absorbance at 517 nm induced by different antioxidants. DPPH free radical reacts with antioxidant, consequentially, absorbance decreases and the DPPH free radical is converted into the DPPH form. The degree of discoloration indicates the scavenging potential of antioxidant compounds of extracts in terms of hydrogen donating ability (Jiao et al., 2012). Fig. 3 three levels of the concentrations of spirulina algae (1%,2%and 3%) compared with BHT (200ppm) were used with a very high scavenging capacity of 40.00 after only 10 min. in all cases the scavenging capacity did not increase after the first 10 min of incubation. The reactions of BHT with DPPH were similar to olive and pomegranate leaves juice with DPPH; the scavenging capacities were similar.

The overall DPPH radical scavenging effect of the spirulina algae were to donate electron, which could then react with free radicals to convert them to more stable products, terminating radical chain reactions.

Fig. 2.

Total polyphenols and lycopene content (ppm) of ketchup supplemented with spirulina algae. Data are expressed as mean ± SD values given represent means of three determinations.

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Organoleptic properties of ketchup fortified by Spirulina algae: Data presented in Fig. 4 shows that the organoleptic properties of sauce fortified with 1.00%. 2.00%

and 3.00% small grains of S. platensis. The color and appearance observe difference (p< 0.05) among of sauce samples fortified with S. platensis. The concentration 1.0% S. platensis recorded the highest score among the sauce fortified S. platensis samples that may due to increasing microalgae causes darken of ketchup and decreasing causes light green color. Furthermore, the manufacture ketchup by using small grains of S. platensis was preferred by panelists because the color of ketchup fortified with microalgae. Increasing microalgae than 3.00% causes darken of sauce and consumers rejected of primary experiment. The flavor of sauce fortified with S.

platensis have significantly (p< 0.05) different among the sauce samples except between control sample and sample containing 1.00% S. platensis there is no significantly (p>0.05) differences between them. The body and texture of ketchup fortified with S. platensis have significantly (p< 0.05) different among the ketchup samples. ketchup fortified with%3.00 S. platensis recorded the highest score of body and texture that due the reduction of water and increase of dry matter by S. platensis.

Overall acceptability of ketchup fortified with 2.00% S. platensis gained the highest ketchup than fortified with 2.00% and 3-00% S. platensis that due to 1.00% light green color and 1.5% dark green. Generally, all ketchup samples had good organoleptic properties, but the most preferable one was the ketchup fortified with 1%S. platensis.

Fig. 3. Antioxidant activity of ketchup by DPPH caving assay. Data are expressed as mean ± SD values given represent means of three determinations.

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Fig. 4 Organoleptic Properties of ketchup supplemented with spirulina algae. Cc = commercial control, A sauce + 1%

spirulina, B= sauce with 2% spirulina and C= sauce with 3%

spirulina. Data are expressed as mean ± SD values given represent means of three determinations.

0 2 4 6 8 10

Color Texture Odour Taste Overall

acceptability

Sensory attributes

Treatments

Commercial control supplemented with spirulina algae 1%

supplemented with spirulina algae 2% supplemented with spirulina algae 3%

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