Nutritional characterization of value-added health mix using germinated barley

ABSTRACT

The present study was designed to explore the nutraceutical potential of orange peel, malted barley, and fenugreek-based value-added/functional health mix. Purposely, orange peel and fenugreek powder have been combined to germinated barley flour with different ratio  . The composite flour was subjected to a nutritional and antioxidant assay. The nutritional analysis revealed that orange peel and malted barley are good sources of protein, fiber, fat, potassium (K), and calcium (Ca). Afterward, the health mix-based functional drink was developed and subjected to physicochemical, functional, and sensory evaluation. The TPC (Total phenolic content), DPPH (1, 1-diphenyl-2-picrylhydrazyl) activity, FRAP (ferric reducing antioxidant power) was found to be increased with the addition of peel flour at various concentrations. The drink (30% peel flour, 60% barley flour and 10% fenugreek flour) is among the other treatments. Sensory scores of formulated value-added drinks decreased with the progression of storage time.

KEYWORDS:

• fenugreek seed (FS)
• orange peel (OP)
• germinated barley
• antioxidant indices

Introduction

The industrialized world of this century tackled to deal with new challenges such as excess healthcare costs, longer life expectancy, and improvements in the way of living. The demand for functional food is increasing due to more health awareness.^[1] The utilization of germinated seeds to create functional flours has been encouraged by the rising demand for healthy food products. It has been shown that germination is a low-cost and long-term process. It enhances the nutritional value and functional component content of grains as well as flavor, digestibility, and bioavailability.^[2] The cereals have been germinated for generations to soften the structure of the kernel, boost nutrient content and availability that reduce the quantity of anti-nutritive chemicals. ^[3] The germ is the center of the grain. The grain starts to germinate after being exposed to light, water, and warmth. The germinated grain is a treasure trove of nutrients packed into the smallest possible space including vitamins, whole grain energy, trace elements, secondary plant compounds, and high-quality protein.^[4]

Cereal grains are a significant source of numerous health-beneficial substances including vitamins, antioxidants, phytochemicals, and micronutrients.^[5] The most significant grain is barley which ranks fourth, globally in terms of cereal production. The major consumer of barley is the United States, where 30% of it is used to make malt and alcoholic beverages, and 65% of it is used as animal feed. ^[6] Due to its nutritional richness and high concentration of physiologically active substances, barley is still a significant grain in various Asian and Northern African societies today. Germination of barley grains compassionate with the action of the enzyme increased total protein and improvements in the profile of amino acids, improves carbohydrates, crude fiber some forms of vitamins and minerals, but low starch levels and loss of total dry matter have been needed to help improve some health conditions.^[7]

Fruit waste byproducts are produced in large quantities during food production and processing in underdeveloped nations which is expensive and has negative environmental implications. Citrus fruits are among the fruit wastes that contribute significantly to a variety of health advantages. Citrus L., a member of the Rutaceae family, is a widespread fruit grown and enjoyed around the world both fresh and as juice.^[8] Citrus fruits, especially orange peel, contain a significant amount of dietary fibers, phenolic compounds, and phytochemical agents that have been shown in studies to promote well-being. These properties include extensive anti-inflammatory, antioxidant, anti-obesity, anti-cancerous, anti-diabetic, and anti-hypercholesterolemia effects.^[9] The culinary and pharmaceutical sectors make extracts from orange peels due to the bioactive components they contain in them. To impart health benefits including antioxidant, anti-inflammatory, and anticancer characteristics, or to create cosmetics and aromas, orange peel extracts have been employed in foods and beverages as flavor enhancers.^[10] Functional components include phytochemicals which are plant-derived, biologically active and non-nutritive chemicals that function in the body to prevent the onset of certain non-contagious diseases.^[11] The facts shed light on a balanced diet and disease prevention strategies as well. Functional and nutraceutical foods must consequently be consumed due to their satiating nature, nutritional value, perishability, and protected status.^[12]

Fenugreek (Trigonella Foenum-graecum) is a plant native to the Leguminosae family that grows annually and is widely grown in the Mediterranean and Asian countries.^[13] Rich sources of gum, fiber, alkaloids, flavonoids, saponin, and volatile material are fenugreek seeds. Fenugreek may be used as a food stabilizer, adhesive and emulsifying agent to alter food texture for some special purposes due to its high fiber content.^[14] The seeds and leaves of this plant are commonly used as anti-diabetic, antimicrobial, anti-inflammatory, anti-cancer, and antioxidant agents for medicinal purposes. Besides, fenugreek seeds have also been reported to have good free radical scavenging operations.^[15]

As a result, it is now preferred for humans to consume ecological and natural products in their meals to stop the oxidation of biomolecules in living things.^[16] Conclusively, Health Mix Using Germinated Barley is helpful for relieving various physiological ailments due to the presence of functional components. Developing citrus peel powder with fenugreek seeds powder and barley powder-based functional beverage is high fiber drink and also beneficial in managing Citrus peel waste to reduce environmental pollution. Functional drinks with more fiber are predicted to boost society’s consumption of fiber. Citrus peel waste is surplus in Pakistan, so the incorporation of citrus peel powder into germinated barley and fenugreek seeds to make a functional drink is a unique idea. As a result, in this study, citrus peel powder is used in barley and fenugreek seeds powder with different proportions for the development of health mix drink and the nutritional and compositional profile of the developed health mix drink are studied.

Material and methods

Procurement of raw material

For the current research work, orange peel was collected as waste material from local market of Faisalabad. Barley and fenugreek seeds were also procured from the local market of Faisalabad

Preparation of germination barley flour

Barley seeds were soaked overnight and then washed with tap water until they reached a neutral pH achieved. Then, the seeds were steeped in tap water at room temperature for 6 hours. After the water was removed and seeds were spread across the moist filter, which were placed in plastic trays containing sterile tap water. Seeds were covered by a filter paper and germination was carried out in times of 2 days. The sprouted seeds were spread in treys and sun-dried for the next 3 to 4 days and then make powder and use for further analysis

Dehydration of sprouting barley

Preparation of barley flour

Preparation of orange peel powder

After the collection of waste orange peel, the procedure of sorting washing and cutting was carried and then spread the material in trays and put for sun drying for 4 to 5 days after the process, the powder was prepared

Preparation of fenugreek powder

Fenugreek was procured from the local market in Faisalabad, and after sorting converted into powder.

Standardization of barley health mix

The health mix powder was formulated in different proportions by using malted barley and orange peel and fenugreek. Information on health mix standardization is given in Table 1.

Table 1. Standardization of malted barley nutritional blend.

Ingredients (gm.)	HM1	HM2	HM3
Malted barley flour	100	60	50
Orange peel	0	30	35
Fenugreek	0	10	15

HM1 *control sample HM2 *peel and fenugreek containing sample HM3 * peel and fenugreek containing sample

Proximate health-mix composition analysis

The proximate analysis of the health mix is evaluated according to the process by following the protocol of AACC to determine moisture content, protein, fat, ash, and dietary fiber.

Mineral analysis

Potassium (K), calcium (Ca), magnesium (Mg), sodium (Na), and iron (Fe) content were determined through an absorption spectrophotometer and flame photometer^[17]The health mix was analyzed for mineral assay following the guidelines of^[17]

In vitro studies

Health mix powder was further analyzed for its in vitro antioxidant potential including total phenolic content (TPC), ABTS, FRAP, CUPRAC and free radical scavenging activity by DPPH (1,1-diphenyl-2-picrylhydrazyl)^[18,19]

Free radical scavenger DPPH

DPPH (1,1-diphenyl-2-picrylhydrazyl) free radical scavenging ability estimation was determined by the DPPH (1,1-diphenyl-2-picrylhydrazyl) assay. ^[20]

ABTS (2, 2’-azino-bis, 3-ethylbenzothiazoline-6-sulfonic acid) assay

ABTS assay of the drink was estimated to the method outlined by^[21]

Ferric Reducing Antioxidant Power (FRAP)

The metal ion chelation ability is another parameter to access the antioxidant capacity of the tested compounds under different challenges and for this ferric reducing antioxidant power is the most adapted test. The FRAP value was calculated by plotting a calibration curve from different concentrations of FeSO4^[22]

Health mix preparation drink

A healthy mix of 20 g of different proportions was added to 200 ml of boiled milk or water and mixed well to get a malted drink.

Physicochemical analysis of composite flour

Functional properties like bulk density, water and oil absorption capacity, the least gelation property, the foaming capacity, foam stability, and swelling property were measured by their respective methods described by Ajatta et al.^[23]

Hunter color

The Hunter color scale is a quality parameter of food products based on their developed color Prepared drinks were analyzed for their color values using CIE-Lab Color Meter following the method of Naufalin et al.^[24,25]

Sensory evaluation

The formulated health mix was subjected to sensory evaluation using a 9-point hedonic scale system (9=like extremely; 1=dislike extremely) as mentioned in Appendix-I following the instructions of Tobin et al.^[26]

Statistical analysis

The data were statistically analyzed to determine the level of significance (p < .05). An analysis of variance (ANOVA) was applied for each parameter. Three times the data were run, and the findings were given as mean ±SD by using method of Steel et al.^[27]

Results and discussion

Proximate and mineral analysis

The nutritional profiling of any food product is provided approximate value for the chemical screening of material. Table 2 indicates the proximate composition of orange peel, germinated barley and fenugreek. The results showed that the higher moisture content was found in fenugreek seeds whereas the lower moisture was noted in orange peel. Similarly, the protein were recorded 4.3 ± 0.4%, 9.7 ± 0.9%, and 14.8 ± 1.3% of orange peel, germinated barley, and fenugreek respectively. The fenugreek seeds were composed of high fat contents as compared to germinated barley and orange peel. Similarly, the ash content was found to be high fenugreek seeds as compared to germinated barley and orange peel. The highest nitrogen free extract (NFE) was measured in orange peel whereas the lower contents was noted in fenugreek. Furthermore, the fiber content was measured to be high in orange peel while the lower value was seen germinated barley.

Table 2. Compositional analysis (%) of germinated barley, orange peel, and fenugreek seeds.

Parameter	Orange peel %	Germinated barley %	Fenugreek %
Moisture	3.16±.12	3.5±0.3	8.9±0.8
Protein	4.3±.4	9.7±0.9	14.8±1.3
Crude fat	1.8±.7	2.2±0.2	5.70±0.5
Ash	2.0±.2	2.5±0.2	3.5±0.3
Fiber	9.1±.7	3.8±0.3	8.97±0.8
NFE	74.1±6.1	69.0±5.8	50.1±4.1

Mineral analysis of germinated barley, orange peel, and fenugreek

High calcium and Mg content were measured in fenugreek while the lower content was found in orange peel. However, sodium (Na) contentswas high in fenugreek seeds while the lower sodium content was found in germinated barley. The iron (Fe) content was high in fenugreek seed whereas lower content was found in orange peel. Similarly, the phosphorous (P) was more in germinated barley as compared to orange peel and fenugreek seeds. The potassium (K) was measured to be fenugreek seeds whereas orange peel contained low quantity of potassium (K) (Table 3).

Table 3. Mineral analysis (mg/100 g) of germinated barley, orange peel, and fenugreek.

Parameters	Orange peel (mg/100 g)	Germinated barley (mg/100 g)	Fenugreek (mg/100 g)
Ca	45.0±.3	69.4±5.9	150±8.5
Mg	41.0±.4	120.5±10.1	134±6.3
Na	19.4±.1	13.4±1.2	23.71±0.24
Fe	7.9±.7	8.9±0.7	15.19±0.15
P	36.3±2.5	320±10.0	215±16.3
K	13.9±1.3	210±4.1	515±26.2

DPPH (trolox mg/g) of health mix flour

DPPH values were changed according to treatments and storage intervals as shown in Table 4. The higher value was found on HM3 at 15 days whereas lower DPPH was measured on HM1 at 0 day of storage. The result showed that DPPH value was observed to be increase with the passage of storage time. However, the high outcome was found at 30 days whereas the lower value was found at 0 day.

Table 4. DPPH (Trolox mg/g) of health mix flour.

Treatments	Duration			Mean
	0 day	15 day	30 day
HM1	38.0±1.2	44.5±1.3	59.5±2.2	47.3±2.3
HM2	47.0±1.1	64.0±2.2	66±2.32	59.0±2.3
HM3	61.0±2.1	67.0±2.1	68±2.23	65.3±0.2
Mean	48.7± 2.4	58.5±1.3	64.5±3.1

FRAP (Gae mg/g) of health mix powder

FRAP outcome changed with respect to different treatments and storage intervals as shown in Table 5. The higher value was found on HM3 at 15 days whereas lower FRAP was measured on HM1 at o day of storage. The outcomes showed that FRAP value was observed to be increase with the passage of storage time. However, the high outcome was found at 30 days whereas the lower value was found at 0 day.

Table 5. FRAP (Gae mg/g) of health mix powder.

Treatments	Duration			Mean
	0 day	15 days	30 days
HM1	17.2± 1.7	22.3±2.2	24.3±2.2	21.3±2.1
HM2	19.3±1.9	26.3±2.6	30.12±3.01	25.2±2.5
HM3	33.3±3.2	39.1±3.9	42.1±4.1	38.2±3.8
Mean	23.2±2.3	29.2±2.9	32.2±3.1

ABTS values of health mix flour

ABTS findings were changed with respect to various treatments and storage periods as shown in Table 6. The higher value was found on HM3 at 15 days while lower ABTS was measured on HM1 at o day of storage. The outcomes showed that ABTS value was observed to be increase with the passage of storage time. However, the high value was found at 30 days whereas the lower value was found at 0 day.

Table 6. ABTS values of health mix flour.

Treatments	Duration			Mean
	0 day	15 days	30 days
HM1	12.1±1.1	14.1±1.4	16.1±1.6	14.1±1.4
HM2	17.3±1.7	22.1±2.2	25.3±2.5	21.6±2.1
HM3	26.3±2.5	30.25±3.01	32.2±3.2	29.6±2.8
Mean	18.6±1.6	22.2±2.1	24.5±2.4

CUPRAC of health mix flour

The content of CUPRAC was high in supplementation flour in HM3. The results of CUPRAC were changed with respect to various treatments and storage periods as shown in Table 7. The higher CUPRAC was found on HM3 at 15 days while lower value was measured on HM1 at 0 day of storage. The outcomes showed that CUPRAC was observed to be increase with the passage of storage time. However, the high CUPRAC was found at 30 days whereas the lower value was found at 0 day.

Table 7. CUPRAC of health mix flour.

Treatments	Duration			Mean
	0 days	15 days	30 days
HM1	7.1±0.5	12.2±1.1	14.1±1.4	11.2±1.1
HM2	9.2±0.7	13.1±1.2	15.3±1.4	12.5±1.1
HM3	12.3±1.1	14.3±1.2	16.4±1.6	14.3±1.4
Mean	9.5±0.8	13.2±1.3	15.3±1.5

Value-added/functional drink analysis

Color value: Color is the main selective visual element in the consumer’s reasonable choice of items. The higher color (a) value was noted in HM1 whereas the lower value was found in HM3. However, the lower color (b) value was observed in the HM1 as compared to HM2 and HM3. The HM3 was composed high color (L) value while the lower value was noted in HM1 as shown in Table 8

Table 8. a, b, and L values for health mix flour blend.

Values	HM1	HM2	HM3	Mean
a	4.6±0.1	4.1±0.2	3.7±0.3	4.2±0.3
b	36.7±1.5	38.5±1.5	38.59±1.58	37.9± 1.8
L	80.7±5.1	84.3±5.8	85.05±6.01	83.3±5.1
Mean	40.7±2.0	42.6±2.5	42.9±2.7

Physicochemical properties of health mix

The physicochemical properties of health mix are measured such as the bulk density, foam stability, gelation capacity, water absorption, swelling capacity and foam capacity as shown in Table 9. However, the higher bulk density was found to be in HM3 and the lover bulk density was noted in HM1. Furthermore, foam stability was more in HM3 while the lower content was found in HM1. However, the gelation capacity was high in HM3 and lower value was calculated in HM1. The water absorption capacity was more in HM3 whereas the lower content was found in HM1. The swelling capacity of HM3 was noted to be extreme whereas least value was counted in HM1. The foam capacity results were high of HM2 whereas the lower value found in HM1.

Table 9. Physicochemical Properties of health mix drink.

Treatments	Bulk density	Foam Stability	Gelatin capacity	Water Absorption capacity	Swelling capacity %	Foaming capacity %
HM1	48.5±1.2	49.0±1.0	11.5±.1	262.0±3.1	1.02±.01	88.1±2.4
HM2	50.0±1.2	50.0±1.1	12.8±.2	265.0±3.2	1.05±.02	9.7±2.7
HM3	51.5±1.3	55.2±1.2	12.95±.2	269.0±3.2	1.06±.01	89.2±1.9
Mean	49.8±1.2	51.4±1.2	12.4±.1	265.0±3.2	1.04±.01	89.3±2.0

Sensory evaluation

Consumer acceptability has the at most importance in achieving the aims of any research focused on dietary intervention. A useful and economical method is to measure customer inclination toward a product’s hedonic response evaluation. Small variations were found according to treatment and storage (Table 10) intervals. The higher appearance was measured on HM2 at 0 day of storage whereas the lower appearance value was found on HM1 at 30 days of storage. The results showed that appearance of all treated sample was reduced with the passage of time. The results of flavor were changed with respect to various treatments and storage periods. The higher flavor was found on HM3 at 0 day while lower value was measured on HM2 at 30 day of storage. The outcomes showed that flavor was observed to be decrease with the passage of storage time. However, the high flavor was found at 0 days whereas the lower value was found at 30 days. Furthermore, the outcomes of taste were varied with respect to various treatments and storage intervals. The higher value of taste was found on HM3 at 0 day while lower value was measured on HM1 at 30 day of storage. The outcomes showed that taste was observed to be decrease with the passage of storage time. However, the high flavor was found at 0 days whereas the lower value was found at 30 days.

Table 10. Sensory evaluation of health mix drink.

Parameters	Treatments	Duration			Mean
		0 days	15 days	30 days
Appearance	HM1	6.90±0.02	6.85±0.04	6.80±0.04	6.85±0.03
	HM2	6.95±0.03	6.92±0.03	6.90±0.05	6.92±0.04
	HM3	6.85±0.05	6.83±0.02	6.80±0.07	6.82±0.06
	Mean	6.90±0.06	6.86±0.03	6.83±0.07
Flavor	HM1	7.1±0.1	6.95±0.02	6.9±0.1	6.95±0.04
	HM2	7.0±0.1	6.98±0.03	6.90±0.03	6.96±0.05
	HM3	7.1±0.1	6.99±0.03	6.92±0.05	6.98±0.03
	Mean	7.04±0.03	6.97±0.02	6.0±0.1
Taste	HM1	7.2±0.1	7.01±0.02	6.91±0.01	7.02±0.07
	HM2	7.1±0.1	7.05±0.04	6.92±0.02	7.03±0.06
	HM3	7.1±0.1	7.1±0.1	6.97±0.03	7.04±0.08
	Mean	7.12± 0.07	7.04±0.03	6.93± 0.02

Discussion

The proximate composition of germinated barley and others ingredients was essential for optimizing the chemical characteristics of flour.^[28] Previous study showed that the barley grain was composed of moisture, protein, crude fat, ash, crude fiber and Carbohydrate. Similarly, the present results are consistent with the previous results.^[29] Another study showed the similar proximate ranges (protein, fiber, fat and ash) for malted barley.^[30] The mineral composition of germinated barley is in agreement to previous results that proved that germinated barley composed of calcium, phosphorus, magnesium, and iron. The previous ranges are similar to current findings in which mineral are present including Ca, Mg, Na, Fe, P, and K in germinated barley powder.

The production of phenolic chemicals for the protection of the seeds from abiotic and biotic challenges during germination may be the cause of the higher TPC and TFC of germinated barley in comparison to non-germinated barley. The current study demonstrated the potential importance of germination in the prevention and mitigation of the risk of type II diabetes mellitus and prostate cancer. Therefore, enhancing the health-promoting qualities of raw barley by germination of the grain could be a tempting technique. The TFC of germinated white, black, and blue barley flours increased.^[31] Tang et al.^[31] studied the phytochemical profile in germinated and raw barley seeds of different varieties and conclude TPC and TFC in all germinated cultivars. The raw seeds as compared to non-germinated seeds, white and blue cultivars gave the richest TPC followed by black cultivar. While the highest TFC was also found in the white cultivar.

Mahfood & Farouk^[32] compared raw grains with soaked grains. The outcomes showed that the mineral and vitamins (A, D, Niacin, and C) are increased in germinated grains compared with the raw grains. However, in the germinated barley, the nutritious components (total phenol, flavonoids, and antioxidant activity (DPPH)) are increased. Additionally, the sensory evaluation of raw and germinated barley flours was also investigated. The results showed that there were no significant differences in variety’s texture or odor but differences in the color, taste, and overall acceptability are measured. The previous findings are similar to Lahouar et al.^[33] who proved that germinated barley the antioxidant activities (AA) using ABTS, DPPH radicals and ferric reducing antioxidant power (FRAP). The concentrations of total phenolic (TPC), total flavonoid (TFC), and condensed tannins (CTC) were measured. The findings showed that the total phenolic content varied according to different treatments.

Nemzer et al.,^[15] studied the physicochemical characterization of germinated barley like water-holding capacity, water-solubility index, fat-absorption capacity, and foaming capacity of flours. Additionally, these barley flour’s higher transition temperature values and lower enthalpy values. However, both significantly increased as a result of germination. Their findings may be useful in determining the impact of germination on the highland barley’s functional characteristics and in selecting the most appropriate uses to encourage the use of germinated highland barley in the food business.

Malomo et al.,^[34] investigated the capacity of yam-soy blend composite flour for improved nutrition for bakery product production. Higher values were observed for functional properties with a higher substitution level, but they were limited to a reasonable range. The pattern of higher water absorption potential of mixed flours is in line with previous results.^[35]

PI and DO^[36] studied the phytochemical combined profile of orange peel and fermented flour and conducted the phytates, oxalates, flavonoids, tannins, alkaloids, and saponins contents of the unfermented sweet orange peel flour. The number of phytates, oxalates, flavonoids, tannins, alkaloids, and saponins in the fermented sweet orange peel flour was 0.1%, 0.10 oxalates, 0.56%, 0.11%, and 0.09% respectively. In comparison to bread made entirely of wheat flour, all bread made with sweet orange peel flour included more phytates, oxalates, flavonoids, tannins, alkaloids, and saponins. The weight and height of the bread made with fermented sweet orange peel flour were not considerably impacted by the fermentation of the sweet orange peel flour.

Grigor et al.,^[37] enhanced awareness of the sensory reactions to fiber fortification that can benefit producers and efforts to promote health. A linear random-effect meta-analysis of 20 eligible studies (869 panelists, 34% male) estimated the effects of fiber fortification (or changed ingredients) on the sensory acceptability of baked cereal foods (bread, cookies and muffins). Fiber fortification of baked cereal foods can lower acceptability, but the magnitude and direction of responses are influenced by product format and base product acceptability. Refining approaches to fiber fortification may enhance market uptake. However, the storage period imparted a substantial decrease in color, flavor, taste, bitterness, and overall acceptability of examined beverages.

Conclusion

In germinated barley-based health mix drinks, orange peel and fenugreek were added due to their maximum anti-oxidative activity based on DPPH radical scavenging activity and others. Three types of value-added/functional beverages were subsequently prepared during the product development process by supplementing different proportions of peel powder with fenugreek powder and barley powder. Physicochemical properties exhibit significant effects due to the addition of peel flour. The health mix blend prepared with 30% germinated barley flour, 15% peel flour, and 5% fenugreek flour was the most preferred drink approved by the hedonic scale.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

The current work was supported by the Higher Education Commission of Pakistan