EFFICACY TRIAL OF INSTANT BEVERAGE MIXES DEVELOPED USING NON DAIRY FOOD INGREDIENTS IN VITRO AND IN IN VIVO STUDIES
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Date
2021
Authors
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Publisher
AAU, Jorhat
Abstract
The present study entitled ―Efficacy trial of Instant Beverage Mixes
developed using non dairy food ingredients: in vitro and in vivo studies‖ was
undertaken to formulate Instant Beverage Mixes namely Cereal Based Instant Beverage
Mix (CBIBM), Pulse Based Instant Beverage Mix (PBIBM), Vegetable Based Instant
Beverage Mix (VBIBM) and Fruit Based Instant Beverage Mix (FBIBM). The product
was developed using non dairy raw ingredients i.e. black rice (Oryza sativa L.), malted
lentil (Lens culinaris), sweet potato (Ipomoea batatas) and mulberry (Morus nigra)
through RSM approach using D optimal mixture design to optimize the formulations.
The objectives of the present study were (i) Development of instant beverage mix using
non dairy food ingredients (ii) Estimation of physicochemical characteristics,
antioxidant and in vitro thrombolytic activity of the developed Instant Beverage Mixes
(iii) Determination of in vivo efficacy in terms of hypoglycaemic and hepatoprotective
property of the developed product using standard animal model protocols. The
nutritional characteristics of the developed mixes were determined using standard
procedures. The free radical scavenging activity was measured by DPPH method as
outlined by Vani et al. (1997). Total phenol, flavonoids, alkaloids and carotenoids were
determined following the methods outlined by Sharma et al. (2008), Aiyegroro and
Okoh (2010), Mallick and Singh (1980), Anaya (1999), respectively. The water holding
capacity of the Instant Beverage Mixes was determined by the method outlined by
Sosulski et al. (1976), bulk and tapped density by the method outlined by Szulc and
Lenart (2016), cohesiveness by the method outlined by Hausner (1967), flow property
by the method outlined by Ribeiro (2020), Dispersibility by the method outlined by
Shittu and Lawal (2007), Wettability by the method outlined by NIRO (1978),
Hygroscopicity by the method outlined by Vivek et al. (2020), and colour was
determined by the method outlined by Lamberts et al. (2006). Glycemic index was done
according to the method outlined by Wolever and Jenkins (1987). The protein Efficacy
Ratio was determined with the method outlined by Buamah et al. (1975). The
thrombolytic property was determined by the method outlined by Hussain et al. (2014).
For the hypoglycemic trial, supplementation was carried for a period of 21 days and
blood glucose was determined using Coral Glucose estimating kit. For the
Hypolipidemic and hypocholesterolemic effects of the developed Instant Beverage
Mixes, supplementation was done for 28 days and estimation was done with the use of
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standard commercial kits coral: HDL-D-160 ml, LDL-D-160 ml, Cholesterol 250 ml,
Triglycerides 250 ml. Hepatoprotective effects of the developed Instant Beverage Mixes
were assessed by using standard commercial kits coral: SGOT-160 ml, SGPT-160 ml.
The shelf life was evaluated in terms of peroxide value (AOAC, 1975) and free fatty
acid (AOAC, 1970) of the Instant Beverage Mixes over a time period of 60 days.
Through Response Surface Methodology (D optimal Design) four formulations were
optimized namely CBIBM (40:20:20:20), PBIBM (55:10:16:19), VBIBM
(45:20:15:20), and FBIBM (35:25:15:25). These formulations were subjected to in vitro
and in vivo efficacy trial. Results of proximate analysis of the raw ingredients revealed
that, black rice, malted lentil, sweet potato and mulberry flour were rich in crude fibre,
protein, crude fat, total mineral and total carbohydrate content. They were also rich in
phytonutrients like total alkaloid, flavonoid, phenolic compound and carotenoid. The
moisture content of the four formulations was within the permissible limit of >11 as per
FSSAI recommendations. PBIBM had significantly higher (p>0.05) crude protein
(15.40±0.24 g/100g), crude fibre (11.74±0.04g/100g) and total mineral (1.53±0.12
g/100g) content compared to other formulations. The carbohydrate content (80.51±0.45
g/100g) and energy value (358.55±3.81kcal) was significantly higher (p>0.05) in
VBIBM. The total mineral content of PBIBM was significantly higher (p>0.01) in
comparison to other formulations. The iron, zinc, calcium, phosphorus, and magnesium
content were 7.80±0.05mg/100g, 3.30±0.21, 56.12±0.02 mg/100g, 180.53±0.26
mg/100g and 249.33±2.08, respectively. FBIBM had a significantly higher (p>0.05)
total alkaloid content of 7.37±0.30 HE mg/100g and phenolic content of 795.45±1.22
GAE mg/100ml.CBIBM had the highest flavonoid content of 309.14±1.35 QE mg/100g
and VBIBM had the highest carotenoid content of 3.07±0.35 mg/100g. Free radical
scavenging property of CBIBM, PBIBM, VBIBM and FBIBM was 63%, 77%, 51%
and 56%, respectively. The highest water holding capacity of 76.00 ±1.00%, bulk
density 0.62±0.01g/ml, and tapped density value of 0.81±0.01g/ml was observed in
PBIBM followed by CBIBM, FBIBM and VBIBM. Better flow property of
12.50±0.50% and poor cohesiveness of 1.14±0.04 was observed in VBIBM. The
highest dispersibility value of 80.38±1.58% was observed in VBIBM. Highest
hygroscopicity value of 70.46±1.42%, and quick wet ability of 46.91±1.41 sec. was
observed in PBIBM. The Glycemic Index of CBIBM, PBIBM, VBIBM and FBIBM
were 37.70, 35.10, 41.72 and 40.39, respectively. Supplementation studies revealed that,
all the subgroups supplemented with test diets namely CBIBM, PBIBM, VBIBM,
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FBIBM showed a significant decrease (p>0.05) in blood glucose level. However,
highest significant decrease (p>0.05) was observed in D3 fed with 70% of PBIBM
(from 296.00±29.30 mg/dl to 194.83±26.55 mg/dl). All four formulations namely
CBIBM, PBIBM, VBIBM and FBIBM had the ability to lower the triglyceride,
cholesterol and LDL level. However, group D3 supplemented with 70% of PBIBM
significant (p<0.05) reduced the triglyceride from 5.16 mg/dl to 17.37 mg/dl,
cholesterol from 8.28 mg/dl to 15.22 mg/dl and LDL level from 2.25 mg/dl to 10.16
mg/dl compared to the other groups. Similarly, it was observed that D3 fed with 70% of
PBIBM had significant (p<0.05) HDL maintaining the property from 5.23 mg/dl to
17.42 mg/dl as per the recommended reference range by ICMR (2017) all the four
formulations also had the ability to lower the ALT, AST enzyme level, however, group
D3 supplemented with 70% of PBIBM significant (p<0.05) reduced the ALT from 1.24
U/L to 3.31 U/Land AST level from 1.41 IU/L to 4.85 IU/L. The Protein Efficiency
Ratio (PER) value of CBIBM, PBIBM VBIBM and FBIBM was 2.0, 2.2, 1.77 and 1.48,
respectively. All of the formulations significantly (p<0.05) linearly increased the body
weight of weaning rats and PBIBM showed a higher impact on body weight. The
thrombolytic activity of FBIBM, CBIBM, PBIBM and VBIBM were 44.89±5.12%,
40.46±3.99%, 32.43±2.88%, 28.84±2.89% when compared with control and standard
drug. After 60 days of storage, the FFA and Peroxide value of the formulations were
within the permissible limit of peroxide value of 4g/100g of sample as per Codex
Alimentarious, 2005 and FFA content of >10 millimoles per kg fat as per FSAAI 2011
guidelines. The findings of this present study provide ample evidences and validate that
the four Instant Beverage Mix (CBIBM, PBIBM, VBIBM and FBIBM) developed from
non dairy foods are not only rich in terms of nutritional quality but significantly exerted
hypoglycemic, hypolipidemic, hepatoprotective effects. It also substantiates that
selection of the right foods for the formulation of ready to eat foods can manage and
control the blood glucose and lipid profile, thereby exerting additional health benefits in
management and prevention of non communicable diseases.