Nutritional Properties of Innovatively Prepared Plant-Based Vegan Snack

Abstract

This research examines the nutritional characteristics of novel-prepared plant-based vegan snacks (PVSs). The proximate composition, mineral content, amino and fatty acid profiles, volatile compounds, phytochemicals, antioxidant activity, and in vitro protein and carbohydrate digestion in prepared snacks were analyzed. The PVSs were mainly prepared by mixing blanched broccoli, pumpkin, chickpeas, whole oat flour, red sweet pepper, fresh onion and garlic, leafy vegetables, and mixed spices, then homogenated, shaped, and freeze-dried. Consequently, sensory evaluation was used to select the most favored PVS; PVS2 contains 25% broccoli and 25% chickpeas, which was the superior model of this study and was analyzed further. The moisture content, crude protein, crude fat, ash, crude fiber, and available carbohydrates were 74.80, 3.40, 28.18, 4.97, 7.86, 3.69, and 51.89 g 100 g⁻¹ of PVS2 (containing 25% of either broccoli or chickpeas), respectively. The results showed that the highest mineral content in macro-elements was sodium, followed by potassium. The PVS2 formula provides 95.24 Kcal 100 g⁻¹ and 54.28 mg 100 g⁻¹ of vitamin C on fresh weight (fw). Consequently, TPC, TCs, TF, TFL, and AOA exhibited valuable content. The HPLC analysis revealed that fifteen phenolics were quantified, with predomination of chlorogenic acid (1741.60 μg g⁻¹), phenolic acid, and naringenin (302.38 μg g⁻¹) as flavonoids, as well as Daidzein (22.27 μg g⁻¹) as an isoflavone. The GC-MS quantification of volatiles exhibited more components; at least 37 displayed concentrations higher than 0.1%. The predominant volatile was cis-13-octadecenoic acid. The percentage of essential amino acids (EAAs) was 25%, and the percentage of non-essential amino acids (NEAAs) was 75%. Among the EAAs and NEAAs, phenylalanine and glutamic acid were the highest, respectively. The fatty acids (FAs) profile exhibited that saturated fatty acids (SFA) and unsaturated fatty acids (USFA) were 20.2% and 79.2%, respectively. The predominant FA in PVS2 was cis-11,14,17-Eicosatrienoic acid, with a percentage of 37.13%, followed by cis-8,11,14-Eicosatrienoic acid, with a percentage of 36.02%. Omega-3 fatty acids reached 39.04%, followed by omega-6 fatty acids at 38.95%. The degree of protein degradation values for the sample was 292.09 mg NH₃ g⁻¹ before digestion and increased to 2106.77 mg NH₃ g⁻¹ after enzymatic digestion. The glycemic index (GI) of PVS2 was estimated to be 21.12, slightly higher than individual vegetables’ GI. Finally, a prepared PVS may be advantageous for recommending the enhancement and further development of diverse snacks to satisfy the nutritional needs of healthy persons and patients across various age demographics.

1. Introduction

Fast and junk food with excessive calories and unhealthy ingredients are harmful when taken in excess and affect eating habits, as well as leading to numerous diseases at different ages [1]. Unhealthy nutrition impairs immunity and hampers physical and mental development in children, in addition to causing obesity, heart disease, and atherosclerosis in adults [2]. Undoubtedly, providing enough nutrients to regulate metabolism is essential. However, if high-calorie foods are our primary energy source, we will be malnourished while feeling full [3]. Indeed, snacking is a prevalent habit among adolescents and children. To enhance the nutritional quality of their diets, it may be beneficial to substitute current snacks with nutrient-rich alternatives [4]. According to recent studies, at least 50% of meals should consist of fruits and vegetables [5,6] due to their nutritional properties. For instance, due to their vitamins and bioactive components, potential health benefits were shown when moringa was added to noodles. Since the contents of both flavonoids and phenols are high enough, it is considered a promising healthy snack for children [7]. Even recent studies have investigated the possibility of making vegetable snacks from vegetable processing residues to reduce caloric intake [2,8].

Innovatively, our research was undertaken to create novel plant-based vegan snacks using locally accessible vegetables, fruits, grains, legumes, seeds, and spices. This research may lead to more nutritious snack options and replace unhealthy snacks with high-nutrient ones. For instance, cruciferous plants like broccoli, cabbage, and cauliflower contain phytochemicals such as glucosinolates, phenolics, antioxidant vitamins, and minerals [9]. Consuming broccoli provides antioxidants, regulates enzymes, and controls apoptosis and the cell cycle [10]. Broccoli’s health advantages are attributed to organosulfur compounds like glucosinolates and S-methyl cysteine sulphoxide; vitamins E, C, and K; minerals like iron, zinc, and selenium; and phenols including kaempferol, quercetin glucosides, and isorhamnetin [11]. These effects are usually caused by breakdown products such as isothiocyanates and indoles from plant myrosinase hydrolysis and/or human microbiota glucosides [12]. Chickpea has been widely consumed for centuries for its protein-rich seeds. It consists of about 18–22% protein. It is a sustainable source of protein, especially for vegetarians, and is used in many traditional food products such as bread, snacks, extruded, weaning, roasted chickpeas, and plant-based beverage items. Chickpea has anti-diabetic, anti-inflammatory, low cholesterol, and cardiovascular disease-preventative advantages [13]. Chickpea protein exhibits inhibitory activities against angiotensin-converting enzyme I (ACE-I) through hydrolysis. The α-galacto-oligosaccharides in chickpeas function as prebiotics due to their indigestibility, and the manufacturing and consumption of fermented chickpeas are significant in legume nutrition [14]. Chickpeas are a considerable source of unsaturated fatty acids, particularly linoleic and oleic acids. Isoflavones and carotenoids are the primary bioactive components found in chickpeas [15]. Chickpea oil contains three significant sterols: β-sitosterol, stigmasterol, and campesterol [16]. Ca, Mg, and P, as well as vitamins such as riboflavin, niacin, thiamin, folic acid, and the precursor of vitamin A, β-carotene, are presented [15,17].

Hence, stress increases unhealthy food consumption in individuals. An online experiment revealed and validated the negative impact of stress on healthy food choices and showed that the Regulation of Craving-Training (ROC-T) intervention promotes healthy food choices under stress and no-stress conditions [18]. Barakat and Rohn [19] indicated that the health-promoting compounds in the prepared vegetative snacks were significantly affected by cooking methods. They suggested the presented data may help to select optimal cooking conditions for innovative plant products. Similar findings were observed when six innovative vegan formulas in ready-to-use and ready-to-eat vegetarian forms were prepared [20]. Vegetables (cauliflower, taro, green zucchini, peas, beans, and spinach) were applied with 20% and 30% chickpeas, as well as 50% additional edible ingredients, and these are considered a promising approach to improve human health and good dietary practices [20,21,22]. The trend towards plant-based foods and vegan diets has recently increased globally [23]. Most recently, Bayindir Gümüş et al. [24] claimed that the glycemic index (GI) and glycemic load (GL) values of vegan analogue food have never been published, and more vegan GI and GL studies are recommended. Consequently, processed vegan meals contain higher carbohydrate contents than animal-based proteins; substituting animal-derived protein sources with plant-based food products in older persons may pose an imminent risk of insufficient protein consumption [25]. Additionally, Atik et al. [23] noted that the edible covering of pomegranate peel extract protects the goods and adds health benefits, providing a novel vegan and eco-friendly food packaging option. Additionally, Butson et al. [26] indicated that probiotic fruit and vegetable snacks are innovative non-dairy foods, and the enriching of snacks with probiotics faces cell viability challenges.

Recently, researchers have hardly struggled to enrich daily meals with healthy snacks containing most of the daily requirements of elements to reduce the oxidative stress in the human body—the presence of such types of snacks may be helpful for all age groups as a source of antioxidants. However, Granato et al. [27] stated that exploring the connections between the dose and the properties of functional substances, delving into the marketing and technical facets of the functional food industry, studying the effects of natural extracts on the characteristics of food, improving extraction methods to increase yield while maintaining stability, and looking into the effects of functional foods on health through interdisciplinary research are the key factors for developing functional products. Therefore, the current study aims to characterize the nutritional properties of PVSs by defining their content of macronutrients, calories, minerals, antioxidants, volatiles, fatty acids, amino acids, and phenolic compounds. In addition, compositional changes, digestibility, and release of food components under simulated digestive system conditions were investigated by estimating the protein digestion coefficient and assessing the glycemic index of prepared snacks.

2. Materials and Methods

2.1. Ingredients

Fresh broccoli florets (Brassica oleracea L.), dried chickpeas (Cicer arietinum L.), fresh pumpkin pulp (Cucurbita pepo L.), whole oat flour (Avena sativa L.), red sweet pepper flesh (Capsicum annuum L.), peeled fresh onion bulb (Allium cepa L.), peeled fresh garlic bulb (Allium sativum L.), mixed spices, green leafy herbs (coriander leaves (Coriandrum sativum L.), dill leaves (Anethum graveolens L.), and parsley leaves (Petroselinum crispum Mill. Fuss)), and edible salt were purchased from Tamimi Markets in Unaizah City [28], Qassim region, SA. Oat fiber was purchased from the iHerb website: https://eg.iherb.com/pr/nunaturals-oat-fiber-powder-1-lb-454-g/46824 (accessed on 17 May 2022) [28].Three formulas of broccoli-based healthy snacks (PVS) were prepared according to

Table 1. Raw ingredients of prepared plant-based vegan snack (PVS) formulas (g 100 g⁻¹).

Ingredients	PVS1	PVS2	PVS2
Blanched broccoli	20	25	30
Soaked and peeled chickpeas	30	25	20
Blanched pumpkin pulp	15	15	15
Whole oat flour	10	10	10
Oat fiber	1	1	1
Red sweet pepper	10	10	10
Fresh onion	5	5	5
Fresh garlic	0.75	0.75	0.75
Edible salt	1.25	1.25	1.25
Green leafy mix	6	6	6
Mixed spices	1	1	1

PVS: plant-based vegan snack.

.

2.2. Manufacturing of Broccoli-Based Healthy Snacks

Broccoli was steamed for 3 to 5 min, while pumpkin was steamed for 8 to 10 min. Chickpeas were immersed in water for 12 to 18 h, changing the water 3 to 4 times over the duration of soaking. The green leafy herbs were mixed after being washed and chopped, then blended as (2:1:1 for green coriander: green parsley: green dill). Spices were combined as follows to formulate 100 g: 25 g of ground black pepper, 20 g of ground cumin, 20 g of ground mixed spices, 10 g of ground dry coriander seeds, 10 g of ground ginger, 10 g of sweet red pepper, and 5 g of hot chili powder. The quantities indicated in Table 1 were weighed and mixed in a benchtop food processor (Santos, VITA-MAX CORP Light Industrial Food Preparation Machine Model VM0122E, Orlando, FL, USA) for 5 min at medium speed until the ingredients were homogeneous, then shaped using a pastry bag with a stainless steel nozzle of 1 cm diameter. The pastry bag was filled with the appropriate amount of PVS mixture and pressed by hand to form approximately 2–3 cm long fingers in a regular shape. The shaped fingers were freeze-dried for 48 h at −52 °C and 0.031 mbar (CHRIST, Alpha 1–2 LD plus, Osterode, Germany), as shown in Figure 1.

2.3. Sensory Evaluation

The sensory evaluation was conducted on the prepared snacks to determine the most palatable options. Twelve trained faculty members and graduate students were invited and followed the method described by Showkat et al. [29]. The snacks were tested for appearance, color, taste, smell, texture, ease of cutting, and general acceptability. The characteristics were recorded on a scale ranging from “9 = very good” to “1 = very dislike”. After sensory evaluation, the best samples were selected according to high palatability and acceptability scores.

2.4. Proximate Chemical Composition and Minerals Content in PVS2

Moisture, crude protein, crude lipids, ash, dietary fiber, available carbohydrates (by difference), and energy value were analyzed in the formulated PVS2 using AOAC methods [30]. According to Nielsen [31], ascorbic acid was measured using 2,6-dichlorophenol indophenol dye titration. Flame photometry (PFP7, Model Jenway 8515, Cole-Parmer Ltd. Beacon Road Stone, Staffordshire, ST15 0SA, United Kingdom), technique 956.01, assessed sodium and potassium. An atomic absorption spectrometer (Perkin-ELMER, 2380, Beaconsfield, Buckinghamshire, England) was used to measure calcium, magnesium, iron, copper, manganese, and zinc according to AOAC method 968.08 [30]. Phosphorus was measured using a conventional colorimetric method, as mentioned by Borah et al. [32].

2.5. Phytochemicals Analysis of PVS2

Using the Folin–Ciocalteau method, the total phenolic compounds (TPC) in PVS2 samples were quantified and expressed as mg gallic acid equivalents (mg GAE 100 g⁻¹ dw) following Bettaieb et al. [33]. The modified method of Khalifa et al. [34] was used to determine the total carotenoids (TCs) content through a colorimetric assay. The concentrations of total flavonoids (TFs) [35] and total flavonols (TFLs) [36] were quantified, and the findings were expressed as mg quercetin equivalent (mg QE g⁻¹).

2.6. Antioxidant Activity of PVS2

The antioxidant activity (AOA) was determined colorimetrically using 2,2-diphenylpicrylhy-drazyl (DPPH) radicals to determine the DPPH radical scavenging activity (DPPH-RSA). The Trolox standard was graphed, and the results were expressed as μmol TE g⁻¹ dw [37].

2.7. Quantification of Phenolic Compounds in PVS2 by HPLC-DAD

The phenolic chemicals in PVS2 were analyzed using the HP1100 HPLC system (Agilent Technologies, Palo Alto, CA, USA), which is outfitted with an autosampler, quaternary pump, and diode array detector (Hewlett Packard 1050). The column (Altima C18, 5 × 150 mm, 4.6 mm I.D.) and a guard column (Altima C18, 5 mm (Alltech)) with a gradient system of 95% eluent (A) 0.1% formic acid in water and 5% eluent (B) 0.1% formic acid in acetonitrile were used by Kim et al. [38]. A flow rate of 1 mL min⁻¹ was set, 10 µL of the produced sample extract was automatically injected, and separation occurred at 25 °C. Chromatograms were acquired at wavelengths of 280, 320, and 360 nm. The detected phenolic compounds’ peaks were measured in µg g⁻¹ by contrasting the results with a built-in library.

2.8. GC-MS Quantification of Volatile Components in PVS2

This GC-MS analysis employed a Thermo Scientific Trace GC Ultra/I.S.Q. Single Quadrupole MS equipped with a TG-5MS fused silica capillary column (30 m, 0.251 mm, 0.1 mm film thickness). The ionization energy of the electron ionization device employed for GC-MS detection was 70 eV. Helium served as the carrier gas, with a 1 mL min⁻¹ flow rate. The MS transfer line and injector temperatures were sustained at 280 °C. The oven is engineered to elevate its temperature from 50 °C to 150 °C at a pace of 7 °C per minute (maintain for 2 min), then to 270 °C at a rate of 5 °C per minute (maintain for 2 min), and finally to 310 °C at a rate of 3.5 °C per minute, sustaining this temperature for 10 min. By determining the relative peak area as a percentage, we could examine the quantification of the designated components. The chemicals were provisionally identified by comparing their relative retention times and mass spectra with the GC-MS instrument’s N.I.S.T. and WILLY library data [39].

2.9. Determination of the Amino Acid Profile in PVS2

The amino acid profile in PVS2 was determined after the acidic hydrolysis in evacuated ampoules at 110 °C for 24 h using HPLC-PICO-TAG (Agilent Technologies, Palo Alto, CA, USA). The quantitative determination of amino acids was conducted following Cohen et al. [40]. According to Blouth et al. [41], tryptophan was colorimetrically measured in the alkaline hydrolysate. The predicted biological value (BV) and amino acid score are contingent upon the WHO [42] and were calculated as cited by Chavan et al. [43].

2.10. Determination of the Fatty Acids Profile in PVS2

According to Petrović et al. [44], total fatty acid fractions were methylated. Gas–liquid chromatography (GLC) equipped with a DFI detector was employed to determine the methyl esters of fatty acids (F.A.s) obtained from PVS2. The GLC conditions were updated using an incremental elevated temperature program from 100 to 200 °C at various times, followed by a 10 min period of cooling at 2 °C per minute to 230 °C and a subsequent 10 min holding period. The injection temperature was set at 250 °C, while the detector temperature was maintained at 300 °C. The GC method for quantifying fatty acid methyl esters was subjected to a validation process. A standard integrator program (Saturn GC Workstation Software version 5.51) was utilized to assess the results. Individual fatty acids were measured using the normalized peak area results without applying a correction factor.

2.11. In Vitro Glycemic Index (GI) and Hydrolysis Index (HI) Analysis

In vitro GI determination in PVS2 was applied using the method described by Aribas et al. [45] and modified by Barakat and Almutairi [46]. Briefly, the sample, weighing 0.1 g, was mixed with 2 mL of HCl (0.05 M) containing pepsin (0.117 g mL⁻¹; Sigma, Louis, MO, USA) and incubated at 37 °C for 30 min with shaking. Afterward, 1 mL of enzyme solution containing 0.243 g pancreatin (Sigma) and 14.45 U (56 μL) amyloglucosidase (260 U mL⁻¹; Sigma) was added. In a horizontal, 37 °C stirring water bath, samples were incubated. At 20 min intervals, 100 μL aliquots from prepared tubes were transferred to Eppendorf tubes for up to 180 min before mixing with 1 mL of 100% ethanol. The glucose content was then measured using GOD-PAP (FDL, UK) following centrifugation at 800× g for 10 min. Shimadzu UV-1800 spectrophotometers (Kyoto, Japan) recorded absorbance at 500 nm. IG and HI were calculated using the mentioned equations:

$$\mathrm{HI}={\displaystyle \frac{\mathrm{The}\mathrm{area}\mathrm{under}\mathrm{the}\mathrm{hydrolysis}\mathrm{curve}\mathrm{of}\mathrm{the}\mathrm{sample}}{\mathrm{The}\mathrm{area}\mathrm{under}\mathrm{the}\mathrm{hydrolysis}\mathrm{curve}\mathrm{of}\mathrm{white}\mathrm{bread}}}$$

Then, the in vitro GI was determined by using the following equation:

GI = 39.71 + 0.549 HI

2.12. Statistical Analysis

The Statistical Package for the Social Sciences (SPSS) program (Ver. 24) was used to analyze sensory arbitration results at a significance level of 0.05 to select the best formula significantly, and multiple Duncan test comparisons were made according to Steel et al. [47].

3. Results and Discussion

3.1. Sensory Evaluation of PVS Formulations

Table 2. Sensory evaluation of plant-based vegan snacks (PVS), (mean ± SE), n = 12.

Organoleptical Characteristics	PVS Formulas
	PVS1	PVS2	PVS3
Appearance	6.18 b ± 0.35	7.65 a ± 0.34	5.22 b ± 0.33
Color	7.66 b ± 0.38	8.28 a ± 0.34	6.25 b ± 0.41
Taste	6.84 c ± 0.25	8.66 a ± 0.35	6.88 b ± 0.35
Smell	7.25 b ± 0.29	7.51 a ± 0.33	7.29 b ± 0.41
Texture	6.15 b ± 0.39	8.51 a ± 0.31	5.82 b ± 0.44
Easy cutting	7.58 c ± 0.35	8.22 a ± 0.44	6.85 b ± 0.39
Overall acceptability	6.95 c ± 0.26	8.13 a ± 0.38	6.39 b ± 0.27

^a,b,c: Column means with identical superscripted letters are not statistically significant (p > 0.05).

illustrates the sensory evaluation of various PVS formulations. The results showed that PVS2 was the favored formula among the panelists, as evidenced by the high scores in all sensory attributes. In contrast, the panelists did not favor the sensory scores of the prepared PVS1 and PVS3. Consequently, panelists chose PVS2 as the most favored formula, and it was subsequently subjected to a comprehensive nutritional characterization.

3.2. Proximate Composition and Minerals of Plant-Based Vegan Snack (PVS2)

The overall acceptability of the prepared snacks with 25% broccoli and 25% chickpeas was the highest, according to the nutritional characteristics. The results in

Table 3. Proximate chemical composition and mineral content in plant-based vegan snack (PVS2) formula containing 25% broccoli and 25% chickpeas (mean ± SE), n = 3.

Composition	PVS2 (g 100 g−1) *	PVS2 (g 100 g−1) **	Minerals	PVS2 (mg 100 g−1)
Moisture	74.80 ± 0.21	3.40 ± 0.12	Sodium	958.87 ± 21.24
Protein	7.35 ± 0.11	28.18 ± 0.11	Calcium	157.89 ± 5.13
Total fat	1.30 ± 0.24	4.97 ± 0.32	Magnesium	97.86 ± 5.27
Ash	2.05 ± 0.05	7.86 ± 0.06	Phosphorus	5.67 ± 148.57
Dietary fiber	0.96 ± 0.27	3.69 ± 0.34	Manganese	4.19 ± 0.98
Total carbohydrates	13.54 ± 0.23	51.89 ± 0.27	Potassium	530.87 ± 8.87
Calories (Kcal)	95.24 ± 1.36	340.17 ± 2.63	Copper	2.68 ± 0.55
Vitamin C mg 100 g−1	54.28 ± 2.67	145.32 ± 5.98	Zinc	5.19 ± 0.64
			Iron	2.35 ± 0.98
			Selenium	0.15 ± 0.08

*: Presented on fresh weight (fw), **: presented on dry weight (dw).

summarize the chemical composition and mineral content of PVS2 calculated according to the dry weight. The moisture, crude protein, fat, ash, crude fiber, and available carbohydrate contents were 3.40, 28.18, 4.97, 7.86, 3.69, and 51.89 g 100 g⁻¹ on dw, respectively. Each 100 g of PVS2 provided 340 kcal and 145.32 mg of vit. C. The results agree with previous studies and are similar to the currently prepared and presented diets [21]. He mentioned that all fresh diets demonstrated appropriate vitamin C content, which depends on the initial ingredients. However, the average levels of vitamin C were 45.75 to 50.95 mg of vit. C in a fresh diet. Barakat [20] indicated that the results of the proximate analysis were 25.02 to 33.96, 1.87 to 2.36, 7.83 to 9.15, 8.14 to 8.84, and 46.79 to 56.16% for crude protein, fat, ash, fiber, and carbohydrate contents of the ready-to-use meals, respectively. In addition, broccoli is rich in various nutrients and biologically active compounds, such as vitamins, minerals, fiber, glucosinolates, and phenolic compounds, and their contents vary according to the parts. Its flowers, leaves, stems, buds, and seeds can be developed into functional foods to prevent and treat some chronic disorders [48].

According to Dietary Reference Intakes [49], the Recommended Dietary Allowances (RDA) of protein range from 34 to 56 g d⁻¹ for ages ranging from 9 to 70 years in both genders, which increases to 71 g d⁻¹ for females during pregnancy and lactation. One hundred grams of PVS2 on a dry basis could provide at least 50% of the RDA for adults and at least 40% of the RDA for pregnant and lactating women daily. In context, Adequate Intake (AI) of dietary fiber could be compensated by at least 13% when consuming 100 g dw of PVS2 daily for an individual consuming 2000 calories. Moreover, the RDA of carbohydrates is 130 g d⁻¹ for ages ranging from 9 to 70 years in both genders, which increases to 210 g d⁻¹ for females during pregnancy and lactation. Consuming about 100 g·dw PVS2 could provide at least 40% of the RDA for adults and at least 25% of the RDA for pregnant and lactating women. Accordingly, as shown, 100 g dw of PVS2 could provide about 340 kcal, which covers the requirements of an adult person (70 kg, 2000 Kcal) for about 3.5–4.0 h [49].

The results in the same table indicate the mineral content (mg 100 g⁻¹) in PVS2 relative to the wet weight. The results show that the highest mineral content among the macro-elements was Na, followed by K, P, and Ca, while Mg was the lowest. Among the micro-elements, the Zn content was the highest, followed by Mn, Cu, and Fe, whereas Se recorded the lowest. The results obtained were consistent with the results reported previously [20,21]. It is worth noting that the snacks provided contain many elements that cover the body’s needs for many age groups, as there are many nutritional benefits when eating such a snack [22].

For human requirements, the presented mineral content in 100 g dw of prepared PVS2 could provide 64–42%, 18–11%, 22–12%, 1–0.5%, 121–23%, 34–13%, 788–297%, 349–182%, 173–47%, and 750–272% of the daily AI and RDA of Na, K, Ca, P, Mg, Fe, Cu, Mn, Zn, and Se. As mentioned in Dietary Reference Intakes [50], the AIs for sodium, potassium, calcium, phosphorus, and magnesium are 1500–2300, 3000–4700, 700–1300, and 80–420 mg d⁻¹, whereas the RDAs for manganese, iron, copper, zinc, and selenium are 1.2–2.3, 7–18, 0.34–0.9, 0.34–1.3, 3–11, and 0.02–0.055 mg d⁻¹.

3.3. Phytochemicals and Antioxidant Activities

The results shown in

Table 4. Total phenolic content, flavonoids, flavonols, and antioxidant activity of plant-based vegan snack (PVS2) formula containing 25% broccoli and 25% chickpeas (mean ± SE), n = 3.

Items	PVS2
TPC [mg GAE 100 g−1]	630.63 ± 13.98
DPPH-RSA [µmol of TE 100 g−1]	978.25 ± 19.28
Carotenoid [mg 100 g−1]	1.03 ± 0.08
TF [mg QE g−1]	1845.57 ± 54.27
TFL [mg QE g−1]	1487.27 ± 27.19

indicate the TPC content and AOA of PVS2 per 100 g of dried sample. The TPC was 630.63 mg GAE 100 g⁻¹, and the AOA was 978.25 µmol of TE 100 g⁻¹. The results were similar to those previously studied by Barakat [21], who prepared six innovative vegetarian meals containing different vegetables (cauliflower, taro, green zucchini, peas, beans, and spinach) at 20%. The prepared meals were supplemented with 30% chickpeas and 50% additional edible ingredients. They were also similar to Barakat and Rohn [19]. The result was very similar to Barakat [20], where the applicability of different vegetables to produce vegetarian diets for ovo-vegetarians was investigated when mixed with 25% chickpeas. It also agreed regarding the high content of phytochemical compounds in vegetable products, as indicated by Ciurzynska et al. [2] in a recent study on the possibility of manufacturing fast vegetable meals from vegetable processing residues. Also, when noodles were fortified with moringa powder [7], potential health benefits were shown due to the high content of phytochemicals.

The TCs content was 1.03 g 100 g⁻¹, the flavonoids content was 1845.57 mmol QE 100 g⁻¹, and the flavonols content was 1487.27 mmol QE 100 g⁻¹. The high phytochemical content of vegetable products was also consistent with a recent study on the potential of vegetable snacks made from vegetable processing residues [2]. Also, noodles fortified with moringa powder [7] have shown potential health benefits due to their nutritional content (vit. A, vit. C, and Ca) and phenolics such as flavonoids. Since the contents of macro- and micronutrients, in addition to both phenolics and flavonoids, are highly valuable compared to similar diets, these are considered as promising healthy snacks for all groups, especially those living in food-insecure areas. The results were identical to those previously studied by Barakat [21], who prepared six innovative vegetarian meals containing different vegetables at 20%. The meals were supplemented with 30% chickpeas and 50% additional edible ingredients. They were also similar to Barakat and Rohn [19] and close to Barakat [20].

3.4. Quantification of Phenolic Compounds in PVS2 by HPLC

The results in

Table 5. Identification and quantitative estimation of some phenolics and flavonoids of plant-based vegan snacks (PVS2).

Item	R.T. (min)	Compound	(µg g−1)
Phenolic acids	3.39	Gallic acid	267.27
	4.17	Chlorogenic acid	1741.60
	5.69	Methyl gallat	44.55
	6.28	Caffeic acid	14.86
	6.63	Syringic acid	46.44
	8.68	Ellagic acid	69.72
	9.04	Coumaric acid	91.57
	14.04	Cinnamic acid	6.39
Flavonoids	8.01	Rutin	172.65
	10.41	Naringenin	302.38
	12.47	Daidzein	22.27
	12.64	Quercetin	18.68
	14.50	Apigenin	27.99
	15.37	Hesperidin	34.20

Phenolic acids, their derivatives, and flavonoids were identified and quantified at 280, 320, and 360 nm.

indicate the quantitative contents of some phenolics and flavonoids in the PVS2 after their separation using HPLC-DAD. Fifteen compounds were quantified, where nine phenolic acids and their derivatives, five flavonoids, and one isoflavones were identified, as shown in Table 5. From phenolics, the highest was chlorogenic acid, with an amount of 1741.60 μg g⁻¹; followed by gallic acid, with an amount of 267.27 μg g⁻¹; then Coumaric acid, with an amount of 91.57 μg g⁻¹, and Ellagic acid, with an amount of 69.72 μg g⁻¹. The highest flavonoid was Naringenin among the five flavonoids identified. In addition, Daidzein as an isoflavone was identified with an amount of 22.27 μg g⁻¹. These results are somewhat consistent with what was reported by Vallejo et al. [51] when they isolated phenolics in broccoli. Jukanti et al. [17] also indicated that chickpeas contain a good percentage of isoflavones, which, for our meals, can be attributed to chickpeas in the formulas [22]. The results also agreed with Barakat and Rohn [19]. At the same time, the content of the current meal is complex due to the difference in the presence of many factors that help us explain this difference, as the components of the formulas, the preparation conditions, and the production conditions of the plant components themselves are among the most important factors affecting this difference. We also agreed on the high content of biochemical compounds in vegetative products, as indicated by Ciurzynska et al. [2] in a recent study on the possibility of manufacturing fast vegetative meals from vegetable processing residues. Also, when noodles were fortified with moringa powder [7], potential health benefits appeared due to the biologically active components presented by adding moringa.

3.5. Quantification of Volatiles in PVS2 by GC-MS

The results in

Table 6. Identification and quantitation of volatile chemicals in PVS22 using GC-MS.

No	R.T. (min)	Compound	(g 100 g−1)
1	5.04	D-Fructose, diethyl mercaptal, pentaacetat	0.78
2	8.78	2-Hydroxytetradecanoic acid	0.41
3	9.72	D-Fructose, diethyl mercaptal, pentaacetat	1.37
4	14.93	10-Heptadecen-8-ynoic acid, methyl ester, (E)-	0.32
5	15.04	Methyl 4,6-tetradecadiynoate	0.16
6	16.25	Methanol, tris(methylenecyclopropyl)-	0.12
7	21.39	Pyrrolizidine-3-one-5-ol, ethyl ether	0.10
8	21.83	9,12,15-Octadecatrienoic acid, (2-phenyl-1,3-dioxolan-4-yl)methyl ester	0.12
9	22.40	Cyclobutanecarboxylic acid, 1-hydroxy-, methyl ester	0.16
10	24.34	1H-Indol-5-ol, 3-(2-aminoethyl)-	0.11
11	24.63	9-Octadecenoic acid (z)-	0.28
12	25.49	1-Heptatriacontanol	0.16
13	26.04	cis-5,8,11,14,17-Eicosapentaenoic acid	0.17
14	26.28	Cyclopropanedodecanoic acid, 2-octyl-, methyl ester	0.22
15	26.46	Hexadecanoic acid, methyl ester	4.14
16	27.93	n-Hexadecanoic acid	4.44
17	28.77	Palmitic Acid, TMS derivative	0.92
18	29.28	9-Hexadecenoic acid	0.34
19	29.58	9,12-Octadecadienoic acid, methyl ester, (E,E)-	4.69
20	29.71	cis-13-Octadecenoic acid, methyl ester	43.89
21	30.17	Heptadecanoic acid, 16-methyl-, methyl ester	1.35
22	30.53	Oxiraneoctanoic acid, 3-octyl-, cis-	0.23
23	31.15	Oleic acid (cis-13-Octadecenoic acid)	22.45
24	31.81	9-Octadecenoic acid	0.17
25	32.27	trans-13-Octadecenoic acid	1.31
26	32.40	cis-13-Octadecenoic acid	0.96
27	32.62	Oxiraneoctanoic acid, 3-octyl-, cis-	0.10
28	32.91	cis-13-Eicosenoic acid	0.12
29	33.10	12-Methyl-E,E-2,13-octadecadienoic-1-ol	0.16
30	33.43	cis-Vaccenic acid	0.26
31	33.57	Oxiraneundecanoic acid, 3-pentyl-,methyl ester, cis-	0.33
32	35.26	9,12-Octadecadienoyl chloride,(Z,Z)-	2.24
33	35.66	9-Octadecenoic acid (Z)-,2-hydroxy-1-(hydroxymethyl)ethyl ester	0.27
34	36.72	Cyclopropanedodecanoic acid, 2-octyl-, methyl ester	0.27
35	37.00	1,2-Benzenedicarboxylic acid	0.55
36	43.76	10,13-Octadecadiynoic acid, methyl ester	0.12
37	47.17	α-Sitosterol	0.12

indicate the quantitative content of volatiles in PVS2 after its quantification using GC-MS. By performing MS analysis, many compounds were obtained; only 37 which were observed displayed concentrations higher than 0.1%. The highest of the obtained compounds was cis-13-octadecenoic acid methyl ester, with a percentage of 43.89%, followed by oleic acid (cis-13-octadecenoic acid), with a concentration of 22.45%. In contrast, 9,12-octadecadienoic acid, methyl ester, (E,E)-, n-hexadecanoic acid, hexadecanoic acid, methyl ester, and 9,12-octadecadienoyl chloride, (Z,Z)- had rates of 4.69, 4.44, 4.14, and 2.24%, respectively. Some compounds were obtained at low concentrations, such as D-fructose, diethyl mercaptal, pentaacetat, heptadecanoic acid, 16-methyl-, methyl ester, and trans-13-Octadecenoic acid, at concentrations of 1.37, 1.35, and 1.31%. The separation also resulted in 28 compounds with concentrations ranging from 0.1 to 0.96%. Our results are consistent with a study [52] performed on several chickpea varieties and with research conducted on broccoli [53]. No chemical characterization was performed on the GC-MS results for the snack developed in our study. Still, this is a foundation for future studies on the nutritional characterization of compounds with nutritional benefits. Our results require further study and scrutiny for confirmation, and we need to draw a path in order to interpret the results of the following studies, which may yield results that are consistent with or different from our results due to the nature of the additive change, the composition, and the circumstances of their results, as mentioned by Munir et al. [54].

3.6. Amino Acids Profile in PVS2

The results in

Table 7. Amino acid composition (mg g⁻¹ protein) * of plant-based vegan snack (PVS22).

Essential Amino Acids	PVS2 (mg g−1 Protein)
Lysine	38.91
Threonine	21.93
Valine	24.41
Methionine	6.01
Isoleucine	19.81
Leucine	37.50
Phenylalanine	39.27
Histidine	26.53
Cystine	15.21
Non-Essential Amino acids
Arginine	53.41
Aspartic	167.32
Serine	50.59
Glutamic	234.88
Proline	37.14
Glycine	63.67
Alanine	48.11
Tyrosine	21.22
Essential Amino Acids	229.58
Non-Essential Amino Acids	676.35
EAA/TAA ratio	0.25
Total Amino Acids	905.93

*: Mean of duplicate analysis.

indicate the quantitative content of amino acids in PVS2 after HPLC analysis. The percentage of essential amino acids (EAAs) was 25%, and the percentage of non-essential amino acids (NEAAs) was 75%. Among the EAAs, the amino acids phenylalanine and lysine were the highest EAAs, followed by a high content of the amino acid leucine, one of the non-polar aliphatic amino acids. Among the NEAAs, glutamic acid was the highest, followed by aspartic acid, glycine, arginine, and serine. Hence, the percentage of protein and its amino acid content changed with the change in maturity. A slight variation was observed in the protein contents of different broccoli types [55]. Our investigations do not describe the amino acid composition and number of such a meal, but the results are used as a foundation for future research. Glutamic acid, seronine, and valine are abundant in broccoli, according to Drabińska [56]. Like our work, broccoli inflorescences have elevated tyrosine, glutamic acid, and aspartic acid [57]. Sulfur amino acids like cysteine and methionine decreased, likely due to chickpeas, limiting amino acids for legumes across the board. Due to its high level, the drop in methionine was less dramatic in broccoli than in cysteine. This is consistent with what was reported by Jukanti et al. [17] regarding chickpeas.

Table 8. Amino acids % and calculated biological value (BV), essential amino acid index (EAAI), and requirement index of different age groups.

Parameters	PVS
Total BCAAs (mg g−1 protein)	81.72
Total aromatic AA (mg g−1 protein)	60.49
Total conditional AA (mg g−1 protein)	412.45
Total basic AAs (mg g−1 protein)	118.85
Total acidic A.A.s (mg g−1 protein)	402.20
Total hydrophobic A.A.s (mg g−1 protein)	275.92
Total polar A.A.s (mg g−1 protein)	343.83
BV	8.82
EAAI	35.70
Requirement index (Infants)	76.47
Requirement index (Preschool child)	83.05
Requirement index (Schoolchild)	90.88
Requirement index (Adult)	95.58

BAAs are basic amino acids, BV is the calculated biological value, and EAAI is the essential amino acid index.

presents the amino acids %, computed BV, essential amino acid index (EAAI), and need index for various age groups. Total branch-chain amino acids (BCAAs) were quantified at 81.72 mg g⁻¹ of protein, whereas total aromatic amino acids were measured at 60.49 mg g⁻¹ of protein. The conditional AA exhibited a protein content of 412.45 mg g⁻¹, with total basic amino acids (BAAs) including lysine, arginine, and histidine at 118.85 mg g⁻¹ protein, total acidic amino acids at 402.20 mg g⁻¹ protein, total hydrophobic amino acids at 275.92 mg g⁻¹ protein, and total polar amino acids at 343.83 mg g⁻¹ protein. The determined biological value (BV) and essential amino acid index (EAAI) were 8.82 and 35.70, respectively. However, the prepared bar was preferred and served to various age groups. The requirement index, which depends on the World Health Organization (WHO, 2007), was calculated, and the results are presented in Table 8. Interestingly, BCAAs comprise three essential amino acids: leucine, isoleucine, and valine, which commonly boost muscle building, enhance exercise performance, and reduce fatigue [58]. The current formula has sufficient BCAA content. Basic amino acids are highly associated with increasing protein bioactivity and possess antioxidant and antimicrobial properties [59]. Total uncharged polar AAs (glycine, serine, threonine, tyrosine, and cysteine) are high, and might be responsible for increasing protein solubility [60]. The calculated BV and EAAI values were slightly low, as this formula has a higher NEAA content. According to WHO’s 2007 amino acid requirements, 100 g of formulated PVS2 covers 76.47–95.58% of the requirement for infants and adults, respectively.

3.7. Fatty Acid Composition of PVS2 by GC-MS

The quantitative content of fatty acids in PVS2, analyzed by GC-MS, is shown in

Table 9. Fatty acid composition (g 100 g⁻¹) of plant-based vegan snack (PVS2).

Fatty Acids	R.T. (min)	Fatty Acids	PVS2 *
Saturated fatty acids
	2.875	Heptanoic acid (C7:0)	0.11
	6.670	Undecanoic acid (C11:0)	0.34
	11.534	Hexadecanoic acid (C16:0)	0.11
	12.242	Heptadecanoic acid (C17:0)	0.11
	13.604	Stearic acid (C18:0)	18.55
	15.819	Arachidic acid (20:0)	0.11
	17.335	Heneicosanoic acid (21:0)	0.51
	19.057	Behenic acid (C22:0)	0.26
Total of saturated fatty acids			20.10
Monounsaturated fatty acids
	13.088	cis-10-heptadecanoic acid (C17:1)	0.20
	14.319	Oleic acid (18:1)	0.49
	16.166	cis-11-Eicosenoic acid (C20:1n9)	0.37
	19.346	Erucic acid (22:1 n-9)	0.63
	19.556	cis-13,16-Docosadienoic acid (C22:2)	0.11
Total of monounsaturated fatty acids			1.80
Polyunsaturated fatty acids
	15.136	Linoleic acid (C18:2n6c)	0.09
	15.346	Linoleliadic acid (C18:2n6t)	0.06
	15.676	α-Linolenic acid (C18:3n3)	0.11
	15.935	γ-Linolenic acid (C18:3n6)	0.23
	16.926	Arachidonic acid (20:4n6)	2.37
	16.696	cis-11,14,17-Eicosatrienoic acid (C20:3n3)	37.13
	16.606	cis-8,11,14-Eicosatrienoic acid (C20:3n6)	36.02
	17.069	cis-5,8,11,14,17-Eicosapentaenoic acid (C22:3n)	0.68
	19.734	cis-4,7,10,13,16,19-Hexaenoic acid (C22:6n3)	0.94
		Total of polyunsaturated fatty acid	77.63
Unknown			0.47
Total of fatty acids			100.00

RT: Retention time of fatty acids, *: Mean of duplicate analysis.

. The percentages of saturated (SFA) and unsaturated fatty acids (USFA) were 20.2% and 79.2%, respectively. The predominant fatty acid in PVS2 was cis-11,14,17-Eicosatrienoic acid, with a percentage of 37.13%, followed by cis-8,11,14-Eicosatrienoic acid, with a percentage of 36.02%, which is a beneficial fatty acid that follows omega-6. This was followed by stearic acid with a percentage of 18.55%, Arachidonic acid with 2.37%, and Undecanoic acid with 0.34%. Eighteen fatty acids were obtained, the percentage of less than 1%. The snack contained a high percentage of monounsaturated and polyunsaturated fatty acids. Omega-3 fatty acids reached 39.04%, followed by omega-6 fatty acids at 38.95%, then omega-9 fatty acids at 1.01%, and then in last place, monounsaturated fatty acids reached 1.81% of the total FA. The PVS2 contained a high percentage of USFA, especially omega-3 and omega-6 fatty acids, which support the health aspect, mainly due to the ingredients containing small amounts of healthy natural fats. Consistent with our finding, Bhandari et al. [61] mentioned that different broccoli parts contain higher quantities of USFA, even when blanched [62], as do chickpeas [63], which supports our data.

3.8. In Vitro Digestion of PVS2

Figure 2a shows the degree of protein degradation in PVS2 in mg NH₃ g⁻¹ on wet weight. The value for the sample was 292.09 mg NH₃ g⁻¹ before digestion, which increased to 2106.77 mg NH₃ g⁻¹ after enzymatic digestion using pepsin and pancreatin in a simulation model of digestion. Previous studies have indicated the presence of many biological properties and benefits of proteins resulting from degradation, and the results are consistent with Burgos-Díaz et al. [64]. Also, the isolation of active compounds occurred, and the antioxidant capacity and activity increased [65].

Figure 2b shows the glycemic index (GI) of PVS2. It was noted that the GI of PVS2 was 21.12. It is well known that vegetables have a very low glycemic index. The results here indicate a slightly high GI, perhaps due to the presence of other ingredients, such as oatmeal and pumpkin.

However, the results are consistent with what was mentioned by Atkinson et al. [66], who indicated a high glycemic index for pumpkin and starchy and sugar-containing foods such as carrots. The PVS given here may help obese or overweight people lose weight. This food is recommended as a healthy snack since it contains various plant components that are helpful to the body, as mentioned in the current study. The limitation of this type of research is that global definitions of functional foods are uncertain. Also, high chemical antioxidant activity does not imply efficacy in vivo, and the interactions between bioactive compounds and their effects are not completely known. Even though this product has many nutritional benefits, the freeze-drying procedure is expensive. The authors of this investigation are optimistic that the sector will grow rapidly and produce nutritious products at lower costs shortly.

4. Conclusions

This study explored the revolutionary nutritional properties of PVS. By using sensory evaluation, this study chose its favorite PVS formula. PVS provided 95.24 Kcal per 100 g and 54.28 mg of vitamin C. The TPC, TCs, TF, TFL, and AOA yielded substantial content. The HPLC study identified 15 phenolics, including chlorogenic acid (1741.60 μg g⁻¹) and phenolic acid, Naringenin (302.38 μg g⁻¹) as a flavonoid, and Daidzein (22.27 μg g⁻¹) as an isoflavone. GC-MS quantified additional volatiles; 37 substances had values over 0.1%. Cis-13-octadecenoic acid was the main volatile. EAAs were 25% and NEAAs 75%. The highest EAAs and NEAAs were phenylalanine and glutamic acid. The FA profile showed 20.2% SFA and 79.2% USFA. PVS2’s major fatty acid was cis-11,14,17-eicosatrienoic acid (37.13%), followed by omega-6-beneficial cis-8,11,14-eicosatrienoic acid (36.02%). Omega-3s comprised 39.04%, and omega-6s comprised 38.95%. Protein degradation levels in the sample ranged from 292.09 mg NH3 g⁻¹ before digestion to 2106.77 mg NH3 g⁻¹ after enzymatic digestion. PVS2 had a GI of 21.12, somewhat higher than veggies. In conclusion, the prepared PVS may help to scale up and prepare distinct PVS to fulfill the nutritional needs of healthy people and patients of different ages; thus, it may be recommended.