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Journal of Food Science and Technology logoLink to Journal of Food Science and Technology
. 2021 Aug 1;59(5):2013–2024. doi: 10.1007/s13197-021-05216-z

Optimization of apricot (Prunus armeniaca L.) blended Aloe vera (Aloe barbadensis M.) based low-calorie beverage functionally enriched with aonla juice (Phyllanthus emblica L.)

Rakesh Sharma 1,✉, Geney Burang 1, Satish Kumar 1, Y P Sharma 2, Vikas Kumar 3
PMCID: PMC9046520  PMID: 35531412

Abstract

The shift in food consumption pattern to accommodate an array of bioactive compounds in foods with minimum calorie intake has resulted in the development of many novel beverages in the last decade. In the present study, optimization of apricot (AP; 0–50%) blended Aloe vera (AV; 50–100%) based low-calorie beverage functionally enriched with aonla juice (AJ; 3,5,7,10%) using steviol glycoside (SG; 0–100%) was performed. The preliminary screening suggested that 40% addition of AP (AV-AP; 60:40) had an acceptable taste. While, for functional enrichment of squash, incorporation of 7% AJ with 40ºB TSS was optimized based on color, consistency, taste, and overall acceptability scores. The developed squash had higher ascorbic acid (26.83 mg/100 g), total phenols (68.77 mg/100 g), antioxidant potential (36.50%) and antimicrobial activity against Staphylococcus aureus (18.00 mm). The aloin content was recorded 0.9 ppm, well within the maximum permissible limits. Additionally, replacement of sucrose with stevioside up to 50 per cent resulted in equi-sweetness level without characteristic bitter aftertaste and a significant reduction in energy value from 142 kcal/100 g to 85.26 kcal/100 g. The developed beverage recorded a high amount of functional compounds with reduced energy values. The beverage can be a recommendation for general consumption for health- conscious people to cover their functional food desires.

Keywords: Functional beverage, Low-calorie, Phenols, Antioxidants, Aloin, HPLC

Introduction

The health-aware people seeking nutritionally enriched functional food products have always created a new vacuum for novel innovations in the food industry. This has continuously shaped the food industry to come up with new food products to meet the specific product- oriented desire of the masses. This has resulted in hike in the demand for fruit beverages especially those loaded with health- promoting and functional incorporations. Such products are designed keeping in view the desire of the general public or the need of specific consumer groups. The novel beverages, now-a-days not only are used for quenching thirst, but also to prevent nutrition-related diseases while improving physical and mental well-being at the same time (Palamthodi et al. 2019). Functional beverage market continues to be the fastest growing segments in the food market which are highly appreciated especially for their nutritional and pharmaceutical properties (Chavan et al., 2018). In the last decade, the beverage market has seen the entry of some big multinational companies. This has resulted in a rapid inflow of various beverages like isotonic and hypertonic beverages, sports and energy drinks, reduced energy/ zero-calorie beverages as well as fruit based functional beverages in the market with expected growth in the near future (Kaur et al. 2021).

Aloe vera (Aloe barbadensis M.), is one of the oldest known medicinal plant and is regarded as a gift of nature. Aloe vera contains a wide range of nutraceutical compounds in the form of polysaccharides (mannan, acetylated glucomannan, glucogalactomannan, galactan, galactogalacturan, arabinogalactan, xylan, cellulose), vitamins (vitamins A, C and E, B12, folic acid), minerals (Ca, Cu, Fe, Mg, K,P, Na, Zn), enzymes (alkaline phosphatase, amylase, carboxypeptidase, catalase, cyclooxygenase, lipase, oxidase, superoxide dismutase) and phenolic compounds (anthrones like aloenin, aloenin B, isobarbaloin, barbaloin and other aloin derivatives) (Ahlawat and Khatkar 2011; Ramachandran and Nagarajan 2014). Different health benefits of Aloe vera (AV) and its derived products have been well documented in the traditional healthcare systems. However, bitter taste, gel-like consistency, poor appearance (color), and stability of Aloe vera gel/juice are the major factors repelling the consumer for its direct consumption and beverage manufacturers for its utilization as a raw ingredient for functional beverage development(Mishra and Sangma 2017; Sharma et al., 2018). The potential nutraceutical and functional properties of Aloe vera are recently being explored for its potential utilization as a blending agent (Sharma et al., 2015). One of the simplest and most economical ideas to overcome these problems may be blending Aloe vera with fruit juice/pulp to make it more palatable and acceptable, while still counting on its functional attributes.

Apricot (Prunus armeniaca L.) is an important fruit with a bright color and strong characteristic aroma with a sumptuousness quality of minerals (Ca, Fe, K, Mg, Na and S), organic acids (oxalic acid, quinic acid, malic acid, shikimic acid, citric acid, and quinic acid), and phenolic compounds (gallic acid, chlorogenic acid, ferulic acid, procatechin, salicylic acid, p-coumaric acid, quercetin and quercitrin and the isomers epicatechin, (-)- and ( +)- catechin) making it suitable option for blending with Aloe vera based beverages (Wani et al., 2015). Aonla, also known as Indian gooseberry (Emblica officinalis Gaertn) is amongst the highest utilized medicinal plant with enormous health benefits. The potential functional properties of aonla fruit have been largely explained in various traditional health care systems. It is rich in various biologically active compounds including polyphenols (gallic acid, ellagic acid, quercetin), minerals (Fe, Na, Zn, K, Ca), vitamins (C, A and B complex) and many enzymes signifying its functional importance in the general wellbeing (Kumari and Khatkar 2018). The aonla fruits are consumed fresh or processed into various value-added products like powder, candy and murabba. Harsh processing conditions, on one hand, result in losses to various sensitive phytochemicals like vitamin C, while, the fruit (fresh or processed) on the other hand has an acidic and astringent taste which makes value addition a necessary requirement. The beverages prepared from the fruits generally carry these features resulting in poor acceptance despite its enormous health benefits. The juice can however be utilized for vitamin C enrichment of various beverages which can mask the acidic and astringent taste of aonla juice while increasing the functional importance of the beverages (Jain and Khurdiya, 2004). The potential of blending as an efficient and economic technology for the nutritional and functional enhancement of novel food products has been long recognized (Bhardwaj and Pandey 2011; Sharma et al., 2018).

Sugar has a very important role to play in our food and nutrition while the sweet taste has been exploited over the centuries for the development of a variety of food products. On one side it is an important ingredient of the food and beverage industry while, on the other side its excessive consumption is reported to be associated with critical disease severity especially in diabetes, obesity and cardiovascular diseases (Twarog et al. 2020). The sugar-sweetened beverages (SSB) are special category of food products which are successful only due to a sweet taste, an unavoidable necessary component of the fast-food industry. Hence, from the last decade, the major focus of the food scientists has been on effective replacement of sugar with non-nutritive zero-calorie sweeteners for the development of low-calorie beverages (LCB). These LCB can be recommended as effective alternates over the SSB with the potential to satisfy both thirst and an innate desire for sweetness with minimal calorific intake (Patel et al. 2018). Stevia (Stevia rebaudiana L.) is popularly known as sweet leaf, sugar leaf and honey leaf which contains a mixture of sweet diterpene glycosides viz., stevioside, steviolbioside, rebaudiosides (A, B, C, D, E, F) and dulcoside A (Gupta et al., 2014). Its high stability at broad pH range (2 to 10), temperature (< 140 °C) and non-fermentable nature have favored its usage in various food applications (Balaswamy et al., 2014). Stevioside is 150 to 300 times sweeter than sucrose and its applicability has been studied by various researchers in various segments of the food industry including bakery, soft drinks, beverages and home foods with diverse success (Adesh et al., 2012; Singh and Singh 2014; Sharma et al., 2018).

Keeping in view the functional importance of Aloe vera and related concerns associated with its direct consumption and industrial applications for the development of various beverages, the present investigation was undertaken to develop a functionally enriched, low-calorie therapeutic beverage by blending Aloe vera juice with apricot pulp and aonla juice and replacing sucrose with equi-sweetness levels of stevioside (steviol glycoside).

Material and methods

Procurement of raw material and preparation

Healthy disease-free Aloe vera leaves were procured from the Botanical Garden of Dr. YS Parmar University of Horticulture and Forestry, Solan, India during 2018–19. Fresh apricot fruits were procured from the local fruit grower which were harvested at optimum maturity (high i.e. > 10% TSS and moderate acidity i.e. 0.7–1.0%). The stevioside powder (steviol glycoside, 80% pure) was purchased from the Council of Scientific and Industrial Research-Institute of Himalayan Bioresource Technology (CSIR –IHBT), Palampur, India. The Aloe vera gel/pulp was extracted using the cold extraction method as described by Sharma et al. (2015). The gel was scrapped with a knife and was then passed through the fruit pulper and pasteurized at 80ºC, followed by pH adjustment to 3.5 by adding citric acid to improve its flow properties. The processed gel/pulp was then packed in pre sterilized glass bottles and processed at 90 ± 2 °C for 25 min, followed by storage at low temperature (7-10ºC) for later use in product development. Whereas, apricot pulp was prepared by hot break/ hot pulping method and aonla juice was extracted through the hydraulic press after grating the de-stoned fruits. All chemicals and reagents used in the present study were of analytical grade (MERK and Sigma Aldrich).

Optimization of Aloe-apricot based vitamin C enriched beverage

Different beverage formulations were prepared by blending Aloe vera (100, 90, 80, 70, 60 and 50%) and apricot (0, 10, 20, 30, 40 and 50%) in varied proportions targeting the highest possible utilization of Aloe vera without any significant decline in the overall acceptability of the product. The control sample consisted of pure Aloe vera pulp (100%) based beverage with standard specifications of 25 per cent juice content, 40oB TSS and1.2% acidity. Whereas, other beverages prepared from different blends contained two different fruit parts (25 and 30%) with constant TSS (40 oB) and acidity (1.2%) in each consecutive formulation viz. T2-T3 (90:10), T4-T5 (80:20), T6-T7 (70:30), T8-T9 (60:40) and T10-T11 (50:50). The beverages were prepared as per the standard method and specifications proposed by FSSA-2006. The selection of the most acceptable blend was made based on the observation of the sensory panel members. The selected blend was then functionally enriched by incorporating aonla juice at different levels (viz. 3, 5, 7 and 10%) in the pre-optimized blend ratio with a constant acidity level of 1.2 per cent.

Optimization of stevioside sweetened low-calorie Aloe vera-apricot beverage

Once the protocol for aonla juice fortified Aloe-apricot based beverage was standardized, attempt was made to reduce the calorific value of the developed product by replacing sugar (sucrose) sweetness with equi-sweetness of stevioside at different proportions of sucrose: stevioside viz. 100:0, 75:25, 50:50, 25:75 and 0:100. The relative sweetness of stevioside was calculated as 270 times the sweetness of sucrose. The resultant quantity of stevioside was added to the beverages to replace the calculated volume of sucrose. The prepared beverages were filled into pre-sterilized PET bottles (250 mL capacity) after adding suitable and recommended preservative (KMS @350 ppm) and kept under ambient storage (21 ± 2 °C & 65 ± 5% RH) for periodic analysis to track important physico-chemical and sensorial changes in the product.

Physico-chemical, nutritional and sensory quality analysis

Total Soluble solids

The total soluble solids (TSS) content was determined by using a digital hand held refractometer of 0–100% range at room temperature and expressed as degree Brix (ºB). The values obtained were corrected for temperature variation to 20 oC.

Titratable acidity and pH

Titratable acidity was estimated as per the standard procedure (AOAC, 2012) by titrating a known volume of sample with NaOH solution against phenolphthalein as an indicator after thorough gelatinization to free acids which are otherwise not titratable. Whereas, the pH of the samples was determined by using a digital pH meter (CRISON Instrument, Ltd. Spain). Before estimating the pH of the sample, pH meter was standardized with standard buffers of 4, 7 and 9.

Ascorbic acid

Ascorbic acid was determined with 2, 6-dichlorophenol-indophenol dye as per the method described by Ranganna (2009) in which aliquot was prepared in 3 per cent meta-phosphoric acid which was titrated with the dye to an endpoint indicated by the development pink color.

Sugars

The sugar content of the developed beverages was estimated by Lane and Eynon method given by Ranganna (2009). A known volume of sample was neutralized with NaOH and then kept as such for ten minutes followed by the addition of 2 mL of lead acetate and 2 mL of potassium oxalate. After diluting it, the filtrate was taken to estimate reducing sugars by titrating against a known quantity of Fehling’s solutions using methylene blue as an indicator. Similarly, the total sugar content was estimated by titrating the prepared sample, after hydrolysis with citric acid, against the known quantity of Fehling’s solution using methylene blue as an indicator. The endpoint was attained when a brick red precipitate appeared in the solution.

Total phenolic content

Total phenolic content (TPC) was estimated according to the method of Jiang et al., (2016). Phenols were extracted in 80 per cent ethanol and estimated based on their reaction with an oxidizing agent phosphomolybdate in Folin-Ciocalteu reagent under alkaline conditions. The absorbance of the mixture was analyzed at 765 nm by a UV spectrophotometer. Gallic acid was used as a standard and results were expressed as mg (GAE)/100 g fresh weight.

Antioxidant activity

The antioxidant activity was determined by using 2, 2-diphenyl-2-picryl hydrazine inhibition method (Brand-Williams et al., 1995). A quantity of 3.9 mL of 6 × 10–5 mol/L. DPPH in methanol was put into a cuvette with 0.1 mL of sample extract and the decrease in absorbance was measured at 515 nm for 30 min or until the absorbance becomes steady.

Antimicrobial activity

Antimicrobial activity was assessed by the well diffusion method against human pathogen Staphylococcus aureus (Schillinger and Lucke 1989). The inoculum was spread with the help of a swab uniformly on the plate and a standard cork borer of 7 mm diameter was used to cut uniform wells on the surface of the solid medium. In each well 100 µl of sample was loaded and the plates were then incubated at 37 ºC for 24 h. The antimicrobial activity was expressed in terms of the mean diameter of the zones of inhibition measured.

Aloin content

The quantitative estimation of aloin content in different samples was done on Binary Waters HPLC unit using Waters HPLC pump 515 with Sunfire TM C-18 analytical column (4.6 × 250 mm, 5 µm) and dual λ absorbance detector 2487 as per the procedures described by Sharma et al., (2014).

Stevioside (steviol glycoside)

The sample was extracted with methanol (sonicated in methanol for 10 min in triplicate at 50ºC) and the methanol fraction was collected, filtered and then evaporated by drying on rotavapour and then lyophilized. A known amount of sample was weighed and dissolved prefixed volume of the mobile phase. The filtrate was re-extracted with methanol and the process was repeated four times. The prepared sample was then passed through a membrane and put in a glass vial. The HPLC was performed using sample and standard stevioside as per the standard method (AOAC, 2012).

Energy value

The energy value of the developed products was measured in a bomb calorimeter (Model Toshiwal DT-100). The energy value of the products in the study was expressed as Kcal/100 g (AOAC, 2012).

Sensory evaluation

Consumer preferences for the experimental beverage samples were found out through sensory evaluation performed immediately after blending was carried out and also during the storage at periodic intervals. All the treatments combinations i.e. Aloe vera + apricot; Aloe vera + apricot + aonla juice and Aloe vera + apricot + aonla juice + stevioside were sequentially evaluated to their sensorial acceptability. At least 10 panelists consisting of trained and semi-trained staff members were given coded samples for three successive sessions consisting of all the blended treatments individually for giving their views on overall acceptability based on color (appearance), consistency (body), flavor and overall acceptability. The evaluation was performed on the 9-point hedonic scale (Amerine et al., 1965).

Statistical analysis

Results are presented as the mean of at least three independent determinations. Statistical evaluation was performed by the two-way analysis of variance in the IBM SPSS statistics program with a significance level α = 0.05 to evaluate the effect of different variables on the quality (physico- chemical and sensorial) and shelf life of the developed beverage. Tukey's HSD (honestly significant difference at p < 0.05) was used for significant mean separation (ranking sample means, largest to smallest) of the standardized combinations and shelf-life periods under refrigerated storage conditions.

Results and discussion

Optimization of Aloe vera-apricot beverage

The sensorial attributes of the apricot (10–50%) incorporated Aloe vera squash (100–50%) with two levels of fruit part (25 and 30%) are presented in Table 1 and Fig. 1. The post hoc analysis using Tukey's HSD test p < 0.05% revealed that the scores for color, consistency, flavor, bitterness and overall acceptability increased significantly (p < 0.05) up to 40 per cent incorporation of apricot juice in the Aloe vera pulp, while further addition didn’t have any positive effect on the sensorial attributes of the developed squash. Minimum values for the sensorial attributes i.e. color (6.41), consistency (6.75), flavor (6.56), bitterness acceptability (6.40) and overall acceptability (6.50) were recorded in the control beverage containing 100% Aloe vera with a non-significant difference amongst the treatments. The highest sensorial scores were observed in the treatments T8-T11 with a non significant differences and the post hoc analysis revealed that treatment T8 with 40 per cent addition of AP had significantly highest sensory scores viz. color (7.90), consistency (7.89), flavor (7.90), bitterness acceptability (7.88) and overall acceptability (7.93). Further, blending of AP up to 40% had some notable changes in the sensorial attributes of the product while, there was no significant change in the sensory attributes beyond this level as can be seen in Table 1. Taking into consideration the maximum utilization of AV pulp and based on the observations of Table 1, the treatment combination containing 60% AV and 40% AP was standardized as the best treatment combination for further studies. To get an overview of the effect of different blending ratios and fruit part incorporation on the sensory attributes (color, consistency, flavor, bitterness acceptability and overall acceptability) of the apricot blended Aloe vera based functional beverage, the overall data was analyzed using rescaled distance cluster analysis (Fig. 1). It is evident from the figure that major clustering was on the basis of blending ratio rather than the fruit part consumed. Two major clusters were formed at a rescaled distance of 5 and were grouped between AL50:AP50 to AL70:AP30 and Al100 to Al90:AP10 than the other treatments indicating that blending as such had a significant effect on the sensory quality of the product. The improved acceptability of various Aloe vera blended beverages has also been reported in earlier studies by Boghani et al. (2012) in blended papaya-Aloe vera RTS beverages, Elbandy et al. (2014) in mango nectar supplemented with Aloe vera gel, Wijesundara and Adikari (2017) in Aloe vera incorporated yogurt and Sharma et al. (2018) in Aloe vera-aonla blended squash.

Table 1.

Sensory evaluation of different Aloe vera- apricot squash

Details of treatments Sensory score
Treatment % Fruit part of blended pulp Color Consistency Flavor Bitterness acceptability Overall acceptability
T1 (Al100) Control 25 6.41 ± 0.11Aa* 6.75 ± 0.13Ba 6.56 ± 0.07Aa 6.40 ± 0.06Aa 6.50 ± 0.10Aa
T2 (Al90:Ap10) 25 6.56 ± 0.16Ab 6.85 ± 0.13Bba 6.78 ± 0.10Bb 6.42 ± 0.10Aa 6.72 ± 0.07Ba
T3 (Al90:Ap10) 30 7.15 ± 0.06Ac 7.31 ± 0.11Bc 7.15 ± 0.08Ac 6.50 ± 0.10Bb 7.23 ± 0.13Ab
T4 (Al80: Ap20) 25 7.51 ± 0.05Cd 7.40 ± 0.06Bc 7.40 ± 0.08Bd 7.15 ± 0.09Ac 7.50 ± 0.11Cc
T5 (Al80:Ap20) 30 7.56 ± 0.08Cd 7.56 ± 0.09Cd 7.44 ± 0.07Bd 7.20 ± 0.06Ac 7.54 ± 0.09Cc
T6 (Al70:Ap30) 25 7.72 ± 0.10De 7.60 ± 0.07Cd 7.51 ± 0.05Ae 7.52 ± 0.07Ad 7.60 ± 0.03Bc
T7 (Al70:Ap30) 30 7.80 ± 0.02Ae 7.86 ± 0.04Bf 7.78 ± 0.04Af 7.76 ± 0.06Ae 7.84 ± 0.11Bd
T8 (Al60:Ap40) 25 7.90 ± 0.04Afg 7.89 ± 0.01Af 7.90 ± 0.03Ag 7.88 ± 0.02Af 7.93 ± 0.06Ad
T9 (Al60:Ap40) 30 7.90 ± 0.01Afg 7.87 ± 0.03Af 7.92 ± 0.04Ag 7.90 ± 0.07Af 7.92 ± 0.06Ad
T10 (Al50:Ap50) 25 7.95 ± 0.01Ahg 7.86 ± 0.01ABf 7.90 ± 0.06Ag 7.90 ± 0.01Af 7.92 ± 0.03Ad
T11 (Al50:Ap50) 30 7.92 ± 0.11Cg 7.80 ± 0.05Ae 7.88 ± 0.03Bg 7.86 ± 0.04Bf 7.88 ± 0.01Bd
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AL Aloe vera, AP apricot pulp

*The mean values ± the standard deviation at p < 0.05%

The values followed by the same lower case letter, in the same column and parameter and by the same upper case letter in the same row are not significantly different at 95% confidence level. (Tukey's HSD test at P < 0.05)

Fig. 1.

Fig. 1

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Cluster analysis showing effect of the blending combinations and fruit part levels on the sensorial attributes of developed beverage

Optimization of Vitamin C enriched Aloe vera-apricot beverage

The incorporation of AJ resulted in minor changes in the sensorial attributes of the developed squash apart from the increase in vitamin-C content of the developed beverage as presented in Table 2. The effect of various levels of AJ incorporation on the sensory attributes of the squash has been presented in Fig. 2 which articulates that the squash fortified with 7 per cent AJ beyond having highest vitamin-C content also had an acceptable appearance, flavor and overall acceptability. With the addition of AJ up to a level of 7 per cent, overall acceptability decreased marginally and non-significantly while, it decreased significantly above 7 per cent addition of AJ. Thereby, AJ fortification up to a level of 7 per cent did not adversely affect the sensory quality of the prepared beverage and contained higher vitamin-C also and hence was optimized. Further, compared to control AV squash (3.89 ± 0.02 mg/100 g Vit. C) and apricot blended AV squash (9.35 ± 0.10 mg/100 g Vit. C), the AJ fortified squash contained 26.83 ± 0.13 mg/100 g Vit. C which was 6.89 and 2.87 times higher compared AV based (control) and apricot blended AV based squash, respectively. Our findings conform to the results of Balaji and Prasad (2014) in Kinnow-aonla blended RTS beverage and Sharma and Thakur (2017) in bitter gourd-aonla blended squash.

Table 2.

Physico-chemical and sensory attributes of fresh fruits and the developed squash

Parameter* Fresh fruits Squash
Aloe vera leaves Aonla fruits Apricot fruits Aloe vera (100) Aloe vera- apricot (60:40) Aloe vera-apricot-aonla (60:40:7)
TSS (ºB) 2.02 ± 0.14a 12.30 ± 0.50b 15.50 ± 1.00c 40.00 ± 0.02a 40.00 ± 0.04a 40.00 ± 0.10a
Titratable acidity (%) 0.25 ± 0.06a 2.65 ± 0.05b 0.95 ± 0.04c 1.21 ± 0.02a 1.22 ± 0.03a 1.20 ± 0.03a
pH 4.5 ± 0.12a 3.20 ± 0.06b 3.81 ± 0.04c 3.80 ± 0.01a 3.82 ± 0.02a 3.79 ± 0.04a
Reducing sugars (%) 0.58 ± 0.12a 4.70 ± 0.65b 6.84 ± 0.13c 12.24 ± 0.13a 13.66 ± 0.28b 13.48 ± 0.21b
Total sugars (%) 1.02 ± 0.05a 8.98 ± 0.30b 12.38 ± 0.22c 33.12 ± 0.05a 33.61 ± 0.36b 33.86 ± 0.14b
Ascorbic acid (mg/100 g) 7.89 ± 0.15a 446.62 ± 6.15b 14.18 ± 3.10c 3.89 ± 0.02a 9.35 ± 0.10b 26.83 ± 0.13c
Total phenolics content (mg (GAE)/100 g) 220.7 ± 2.5a 170.00 ± 4.52b 89.41 ± 2.11c 49.25 ± 0.17a 58.56 ± 0.25b 68.77 ± 0.34c
Antioxidant potential (% free radical scavenging activity) 89.24 ± 1.3a 82.25 ± 0.95b 64.8 ± 0.65c 27.50 ± 0.25a 28.65 ± 0.24b 36.50 ± 0.32c
Antimicrobial activity (mm ZoI) 34.60 ± 0.80a 41.30 ± 1.0b 18.70 ± 0.85c 16.00 ± 0.43a 16.20 ± 0.22a 20.80 ± 0.35b
Sensory attributes of developed beverages
Appearance 6.65 ± 0.10a 7.95 ± 0.09b 8.00 ± 0.10b
Flavor 6.80 ± 0.08a 7.89 ± 0.07b 7.90 ± 0.08b
Overall acceptability 6.85 ± 0.06a 7.88 ± 0.08b 7.92 ± 0.07b
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*Values are mean of five determinations ± Standard Error

The values followed by the same lower case letter, in the same row for individual parameter (fresh fruits & squash) are not significantly different (Tukey's post hoc test one way test at P < 0.05)

Fig. 2.

Fig. 2

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Effect of aonla juice fortification on sensorial attributes of developed beverages

Comparative Quality attributes of the blended beverages

The physico-chemical and nutritional quality of Aloe vera leaves and fresh fruits (apricot and aonla) was evaluated and has been presented in Table 2. The post hock analysis clearly revealed that blending of apricot and Aloe vera pulp led to an increase in the nutritional and sensorial profile of the developed beverage. The best blend (40% AP + 60% AV) selected from experiment-1 significantly had higher total phenolic (58.56 mg/100 g), ascorbic acid (9.35 mg/100 g) and antioxidant potential (28.65% free radical scavenging activity) compared to the AV squash (49.25 mg/100 g, 3.89 mg/100 g and 27.50% free radical scavenging activity), respectively. Further, the blended beverage had strong antimicrobial activity against Staphylococcus aureus (20.80 mm zone of inhibition). Notably, the fortification of apricot blended AV squash fortified with AJ further improved the nutritional quality of the prepared beverage and the specific effect was observed as higher phenolic content (68.77 mg/100 g), ascorbic acid content (26.83 mg/100 g) and antioxidant potential (36.50% free radical scavenging activity) of the fortified beverage. Blending significantly improved the phytochemical potential of the beverage as the proportionate level of replacement of the contents with AJ might contain higher level of the phytochemicals. Similar results were also observed by Sharma and Thakur (2017). The blending has also exerted a slightly positive but non-significant effect on the sensory attributes of the beverages. The post hoc analysis revealed that there was a continuous increase in the sensory scores of the beverages at each stage of blending i.e. the appearance rating increased from the initial scores of 6.65 ± 0.10 (Aloe vera squash) to 8.00 ± 0.09 (apricot blended Aloe vera squash) followed by a non-significant decrease to 7.95 ± 0.80 (aonla juice fortified apricot blended Aloe vera squash). Similarly, the flavor scores increased from 6.80 ± 0.08 to 7.89 ± 0.07 to 7.90 ± 0.08 and the overall acceptability scores from 6.85 ± 0.06 to 7.88 ± 0.08 to 7.92 ± 0.07 in the control (Aloe vera), apricot blended Aloe vera and aonla juice fortified apricot blended Aloe vera squash, respectively (p < 0.05). Many workers have reported blending as a suitable technique for making more palatable and nutritious beverages from different raw materials (Bhardwaj and Pandey, 2011; Ahmed et al., 2016; Mishra and Sangma, 2017; Sharma et al., 2018). Similar trends were also observed by Balaji and Prasad (2014) in kinnow-aonla blended RTS beverages.

Effect of stevioside on quality of Vitamin C enriched AV-AP beverage

The suitability of preparing low-calorie functional squash was also evaluated by replacing sucrose with an equivalent level of sweetness of stevioside and standardization of the product through sensory evaluation as presented in Table 3. The addition of stevioside resulted in significant changes in the sensory attributes of resultant squash. Generally, the scores for color, consistency, flavor and overall acceptability decreased with an increase in the proportion of stevioside used as well as the storage period. The post hoc analysis using Tukey's HSD test revealed that the effect of the storage period on the quality attributes was most significant as the color, consistency, flavor and overall acceptability scores had distinctive, lower but still acceptable values after 3 and 6 months of storage. Further, the addition stevioside in various concentrations resulted in significant changes in the sensory quality of the developed beverage, while the post hoc analysis also revealed that the differences in the reported values for different sensory attributes were non-significant till the addition of stevioside up to 75% level. Further, increasing the concentration of stevioside beyond 75% resulted in a significant decrease in the sensory quality of the squash with a slightly bitter aftertaste which is not desirable. Overall, the honestly significant difference revealed that the replacement of conventional sugar with stevioside up to 75% resulted in comparatively non-significant changes in the sensory quality of the developed beverage during storage. The bitter aftertaste of stevioside at higher concentrations gave a negative impact on sensory attributes and might have led to reduction in the score for overall acceptability (OAA) as has also been reported by Balaswamy et al. (2014). Our findings are also in conformity with the results of Singh and Singh (2014) and Sharma et al. (2018) who have attempted the development of various Aloe vera based beverages.

Table 3.

Effect of storage conditions (21 ± 2 °C, 65 ± 5% RH) and sucrose replacement with stevioside on the physico-chemical and sensorial attributes of the optimized squash containing AV (60) + AP (40) + AJ (7)

Parameter Storage in months Sucrose (100%)* Sucrose (75%) + Stevioside (25%) Sucrose (50%) + Stevioside (50%) Sucrose (25%) + Stevioside (75%) Stevioside (100%)
Sensory attributes recorded on 9-ponit hedonic scale
Color 0 7.85 ± 0.05aA 7.82 ± 0.03aA 7.83 ± 0.04aA 7.80 ± 0.04aA 7.73 ± 0.09aB
3 7.64 ± 0.09bA 7.66 ± 0.05bA 7.64 ± 0.08bA 7.66 ± 0.06b 7.51 ± 0.06bB
6 7.59 ± 0.07bA 7.56 ± 0.10cbA 7.58 ± 0.06bA 7.52 ± 0.03bA 7.45 ± 0.07bB
Consistency 0 8.15 ± 0.09aA 8.08 ± 0.04aA 7.94 ± 0.07aB 7.80 ± 0.08aC 7.73 ± 0.05aC
3 8.00 ± 0.07bA 7.96 ± 0.07bA 7.84 ± 0.04bB 7.75 ± 0.05bC 7.54 ± 0.06bD
6 7.95 ± 0.06bA 7.84 ± 0.05cB 7.79 ± 0.06cbC 7.78 ± 0.07cbC 7.48 ± 0.03cD
Flavor 0 8.10 ± 0.01aA 8.00 ± 0.02aB 7.94 ± 0.03aC 7.90 ± 0.06aDC 7.72 ± 0.07aE
3 7.98 ± 0.04bA 7.92 ± 0.09baB 7.90 ± 0.04baB 7.85 ± 0.03aC 7.68 ± 0.02aD
6 7.75 ± 0.06cA 7.70 ± 0.06bA 7.68 ± 0.05cA 7.64 ± 0.04bA 7.54 ± 0.01bB
Overall acceptability 0 8.04 ± 0.07aA 8.05 ± 0.07aA 8.05 ± 0.05aA 8.06 ± 0.03aA 7.89 ± 0.02aB
3 7.86 ± 0.04bA 7.84 ± 0.04bA 7.90 ± 0.06bA 7.90 ± 0.05bA 7.78 ± 0.03bB
6 7.72 ± 0.09cA 7.70 ± 0.06cA 7.82 ± 0.04cB 7.84 ± 0.07cbB 7.50 ± 0.06cC
Physio-chemical parameters
TSS (%) 0 40.00 ± 0.72aA 35.40 ± 0.45aB 26.50 ± 0.33aC 15.26 ± 0.38aD 5.78 ± 0.11bB
3 40.46 ± 0.61aA 35.58 ± 0.23aB 26.63 ± 0.41aC 15.38 ± 0.43aD 5.84 ± 0.10aB
6 40.58 ± 0.38aA 35.61 ± 0.52aB 26.75 ± 0.17aC 15.52 ± 0.47aD 5.88 ± 0.08aB
Acidity (%) 0 1.31 ± 0.02aA 1.28 ± 0.02aB 1.25 ± 0.05aC 1.25 ± 0.07aC 1.24 ± 0.03aD
3 1.29 ± 0.01aA 1.23 ± 0.03bB 1.22 ± 0.03bC 1.18 ± 0.02bD 1.18 ± 0.01bD
6 1.26 ± 0.01bA 1.21 ± 0.01bB 1.21 ± 0.02bC 1.18 ± 0.04cD 1.17 ± 0.02bE
pH 0 3.83 ± 0.05aA 3.79 ± 0.03aA 3.74 ± 0.02aB 3.73 ± 0.02aB 3.73 ± 0.05aB
3 3.84 ± 0.03aA 3.81 ± 0.01aA 3.75 ± 0.06aBA 3.76 ± 0.01aB 3.75 ± 0.03aB
6 3.83 ± 0.02aA 3.81 ± 0.02aA 3.75 ± 0.05aB 3.76 ± 0.04aB 3.74 ± 0.02aB
Ascorbic acid (mg/100 g) 0 19.22 ± 0.02aA 19.26 ± 0.05aBA 19.25 ± 0.05aB 19.28 ± 0.06aB 19.36 ± 0.03aC
3 16.24 ± 0.03bA 16.27 ± 0.01bA 16.32 ± 0.04bB 16.30 ± 0.02bB 16.42 ± 0.04bC
6 14.31 ± 0.01cA 14.35 ± 0.04cBA 14.34 ± 0.01cB 14.38 ± 0.03cB 14.42 ± 0.05cCB
Reducing sugar (%) 0 13.83 ± 0.08aA 11.11 ± 0.05aB 8.10 ± 0.03aC 4.41 ± 0.04aD 2.77 ± 0.04aE
3 13.76 ± 0.05aA 10.78 ± 0.03bB 7.64 ± 0.03bC 4.12 ± 0.03bD 2.61 ± 0.04bE
6 13.40 ± 0.01bA 10.43 ± 0.04cB 7.32 ± 0.04cC 4.05 ± 0.07cbD 2.54 ± 0.08cbE
Total sugar (%) 0 34.27 ± 0.08aA 26.76 ± 0.12aB 21.23 ± 0.12aC 12.48 ± 0.11aD 4.36 ± 0.05aE
3 34.22 ± 0.09aA 26.470.08bB 20.71 ± 0.06bC 12.03 ± 0.03bD 4.27 ± 0.09baE
6 33.75 ± 0.04bA 26.20 ± 0.10cB 20.42 ± 0.07cC 10.20 ± 0.03cD 4.18 ± 0.05cE
Total phenolic content (mg (GAE)/ 100 g) 0 70.10 ± 0.85aA 70.11 ± 1.12aA 70.15 ± 1.09aA 70.15 ± 1.21aA 70.18 ± 0.82a
3 68.50 ± 1.35bA 68.53 ± 1.02bA 68.57 ± 0.95bA 68.59 ± 1.09bA 68.61 ± 0.90bA
6 63.93 ± 1.02cA 63.97 ± 0.52cA 64.01 ± 1.05cA 64.07 ± 1.00cA 64.09 ± 1.12c
Antioxidant activity (%) 0 38.16 ± 0.09aA 38.18 ± 0.11aA 38.26 ± 0.09BA 38.32 ± 0.05aB 38.34 ± 0.07aB
3 36.15 ± 0.08bA 36.17 ± 0.06bA 36.21 ± 0.01bA 36.23 ± 0.02bB 36.25 ± 0.11bB
6 33.24 ± 0.01cA 33.25 ± 0.03cA 33.28 ± 0.01cA 33.30 ± 0.03cB 33.31 ± 0.06cB

Energy Value

(Kcal/100 g)
0 142.0 ± 1.12Aa 116.19 ± 0.92bA 85.26 ± 1.02cA 56.10 ± 0.98dA 22.36 ± 0.96eA
3 141.50 ± 1.03Aa 115.72 ± 1.02bA 85.01 ± 0.92cA 55.23 ± 0.98dA 21.90 ± 0.94eA
6 140.16 ± 0.96aA 114.20 ± 1.01bA 82.98 ± 0.90cA 54.76 ± 1.02dA 20.30 ± 0.92eA
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*The combination of the sweeteners (sucrose and stevioside) with pre optimized squash, containing 70% Aloe vera + 30% apricot pulp and 7% aonla juice. Measurements were made after standardization of the product after 0, 3 and 6 months of storage at ambient condition

The values followed by the same lower case letter, in the same column and parameter and by the same upper case letter in the same row are not significantly different at 95% confidence level (Tukey's HSD test at P < 0.05).

The data on physico-chemical characteristics of low-calorie aonla fortified Aloe vera-apricot blended functional squash is presented in Table 3. It is inferred from the data in Table 3 that during progression in the storage period from 0 to 6 months there were significant changes in the ascorbic acid, total sugar, phenolic content and antioxidant activity of the beverage which decreased from 19.22 to 14.31 (%), 34.27 to 33.75 (%), 70.10 to 63.93 (mg (GAE)/ 100 g) and 38.16 to 33.24 (%), respectively. Further, the post hoc analysis also revealed that there was not any significant change in the parameters like acidity, pH, ascorbic acid content, total phenolics and the antimicrobial activity of the developed beverage with increase in the replacement level of the sucrose with the stevioside from 0 to 100 per cent. However, the replacement significantly affected the parameters like TSS, total and reducing sugars and the energy values of the beverage which justifies the concept of low-calorie functional beverage development. It was reported that during all the storage periods with increase in the stevioside concentration the values of TSS decreased from around 40 to slightly above 5, reducing sugars from around 13 to slightly higher than 2, total sugars from around 34 to slightly higher than 4 and the energy values from about 140 to around 20 kcal which shows about 8, 6, 8 and 7% reduction in the TSS, reducing sugars, total sugars and the energy values, respectively.

The decreased TSS and sugars might be due to the fact that non-nutritive sweeteners including stevia/ stevioside do not add solids to the finished products and thus reports no increase in the TSS and the sugar values. Similar observations have also been reported earlier by Sharma and Thakur (2017). However, the slight mathematical variation in the proportionate quantity of the TSS and sugars by addition of stevoside as sugar replacer might be due to its binding/degradation during processing and preparation in some of the treatments (Adesh et al., 2012; Singh and Singh, 2014).

HPLC analysis of aloin and stevioside content

The quantitative estimation of aloin content of the prepared beverages was performed by using high pressure liquid chromatography (HPLC). The HPLC analysis revealed that the aloin peaks of pure Aloe vera squash (control) corresponds to retention time (15 min) on the other hand, the peak was not detected in other samples where respective replacement of the AV was made with AP and was further fortified with AJ. Further, it is also evident from the graph that even in the control samples the aloin was present in a very small volume, while the replacement of AV led to its further reduction to almost non-significant amount. The aloin content in AV squash (control) was found to be 1.8 ppm. Whereas, in low-calorie AJ fortified AV –AP squash the aloin content was found to be 0.9 ppm (Fig. 3a, b) The aloin content in the developed beverages were very much under the upper permissible limit of 10 ppm as prescribed for oral ingestion of the Aloe vera based products (Sharma et al., 2018).

Fig. 3.

Fig. 3

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HPLC chromatograms of Aloe vera-apricot squash (a, b) for aloin content and aonla fortified low calorie Aloe-vera-apricot squash (c, d) for standard stevioside and in beverage sweetened with stevioside

Stevioside was used for the replacement of conventional sugar to make the functional beverage suitable health drink for diabetic people. The stevioside (steviol glycoside) contained rebadioside-A (163.5 µg/mL), stevioside (354.1 µg/mL) and rebadioside-F (234.43 µg/ml) (Fig. 3c, d).The stevioside content in the aonla fortified Aloe vera-apricot squash was recorded as 0.76 mg/mL on the preparation day which had no reportable change even after 6 months of storage (21 ± 2 ºC temperature and 65 ± 5% RH) indicating the heat and storage stability of stevioside. Further, it can be inferred from the HPLC chromatograms of low-calorie aonla fortified Aloe vera-apricot squash that the stevioside peak had almost the same retention time as that of standard stevioside indication no change in its chemical form and availability. Similarly, estimation of stevioside in the commercial samples of stevia leaf and powder were also estimated using HPLC technique by Khan et al. (2011) who also have reported the storage stability of the product.

Conclusion

The squash containing 25 per cent fruit part (prepared by mixing 60% AV gel and 40% AP) with a TSS of 40°B was standardized as the best blend with highest level of utilization of Aloe vera as the functional ingredient. Further, for vitamin-C enrichment aonla juice fortification up to 7 per cent was found most acceptable without any significant effect on the sensory quality and acceptability of the developed beverage. For creating a functional beverage for the diabetic people the conventional sugar in the functional squash can be replaced with 75% equi-sweetness level with stevioside without the characteristic bitter aftertaste of stevioside. It is concluded that the developed beverage containing 25 per cent fruit part (AV + AP), 25ºB TSS, 7% AJ and 75% sugar replacement with stevioside had the highest sensorial quality and better physico-chemical properties. Further, the beverage can be recommended as a healthy drink for the general masses and a specialty drink for diabetic people.

Authors' contributions

Rakesh Sharma and Satish Kumar conceptualized the idea of the project and accordingly planned the outline of the work. The resource availability was ensured by Rakesh Sharma. Geney Burang collected the relevant data and performed the research experiments in the laboratory under the supervision of Rakesh Sharma. HPLC analysis for aloin and stevioside was carried out by Y P Sharma. The analysis and drafting of document was done by Rakesh Sharma and Vikas Kumar. The manuscript was edited and accordingly restructured by Rakesh Sharma and Satish Kumar. Before final submission, all authors have contributed significantly in the review process and proofreading of the manuscript.

Funding

This work did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Data availability

The data supplied has been generated in the laboratory and used in the manuscript.

Declarations

Conflicts of interest

There is no potential Conflicts of interest/Competing interests reported among the authors.

Ethical approval

No animal trials were involved in the study and ethical approval was not required.

Consent to participate

I will review at least three manuscripts of my own specialization submitted to JFST.

Consent for publication

The research submitted is the original work carried by the authors and is not under consideration in any other journal.

Footnotes

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