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Nutraceutical products from banana pseudostem and rhizomeNutraceutical products from banana pseudostem and rhizomeNutraceutical products from banana pseudostem and rhizomeNutraceutical products from banana pseudostem and rhizome

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Chapter 5: Chapter 5: Chapter 5: Chapter 5:

Development of RTS beverage and Development of RTS beverage and Development of RTS beverage and Development of RTS beverage and

dehydrated powder from banana dehydrated powder from banana dehydrated powder from banana dehydrated powder from banana

pseudostem and rhizomepseudostem and rhizomepseudostem and rhizomepseudostem and rhizome


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Introduction

Pseudostem and rhizome of banana plant together contributes 43.48 % of the

biomass. Pseudostem appears to be a rich source of fibre, total carbohydrate and

cellulose. Manimehalai (2005) observed that moisture, protein, fat, minerals, fibre and

carbohydrates content of 93.1, 0.3, 0.03, 1.04, 0.68 and 1.20 g/100 g of pseudostem,

respectively. At present lessthan 2% of pseudostem production is used for human

consumption, the remaining are incinerated and wasted. Similarly, the nutritive value of

rhizome includes moisture, protein, fat, minerals, crude fibre, carbohydrates, calcium,

phosphorus, iron, and energy of 85.1, 0.4, 0.2, 1.4, 1.1, 11.8, 0.025, 0.010, 0.0011 g and

51 kcal/100 g of edible portion respectively (Gopalan et al., 1989). Rhizome is often

cooked and eaten as a vegetable in India, mainly in tribal areas. Use of rhizome as seed

material is highly discouraged due to inexpensive and adequate supply of disease free,

high yielding tissue culture plantlets. In indigenous systems of medicine, pseudostem

juice is a well-known remedy for urinary disorders, stomach troubles like diarrhoea,

dysentery and flatulence and also it helps in treatment for removal of stones in the

kidney, gall bladder, and prostate and is also used as an antidote for snake-bite. Rhizome

extracts were used as a coolant, to treat diabetes, piles, intestinal worms, mental diseases,

acidity, food poisoning, to cure pyorrhea and to heal wounds. Despite rich nutritional and

nutraceutical properties of pseudostem and rhizome, they are wasted and incinerated in

India and elsewhere. Extensive literature survey also supported that no work has been

reported regarding utilization of banana pseudostem and rhizome for food, nutraceutical

or pharmaceutical purpose. Hence, present study is focused on to utilize these vast

quantities of banana plant bio-waste as a source of polyphenols rich RTS beverage and

dehydrated powder, and elucidate their antioxidant properties.


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Materials and methods

Plant material

Banana (Musa AAB var. Nanjanagudu Rasbale) plants (Wealth of India 1962;

Venkatachalam et al., 2006) were identified and harvested from plantations in Mysore

district of Karnataka, India. After harvesting of fruit bunch, pseudostem and rhizome

were separated from plant, transferred to lab and used for preparation of RTS beverage

and dehydrated powders.

Juice extraction and standardization of RTS beverage

The pseudostem and rhizome collected after harvesting of fruit bunch, were

cleaned, washed thoroughly in water, and juices were extracted after passing through the

pulper and strained by using muslin cloth. Control of astringency browning was done by

pretreatment of pseudostem and rhizome juice by 25ppm of L-cystein. Juices were cooled

immediately to 4°C, centrifuged for 10 min at 3000 rpm. RTS beverages with various

strengths of juice (10, 15, 20 and 25%), sugar (10 and 15°brix) and acidity (0.25 and

0.30%) were mixed and filled into pre-sterilized bottles (Ranganna, 2001). Sealed bottles

were pasteurized for 25 minutes in boiling water, cooled and stored at room temperature

for further analysis (Fig.5.1).

Sensory analysis of RTS beverages

Sensory evaluation (colour, appearance, body, taste, flavor and overall quality

with 9 point hedonic scale) for the pseudostem and rhizome RTS beverages was done

with panel consisting of departmental staff and research workers both male and female

aged 22 to 58 years, who had experience in judging fruit and vegetable products and their

sensory quality (Ranganna, 2001).


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Figure 5.1: Banana pseudostem and rhizome RTS beverages. (A)-banana rhizome, (B)-

banana pseudostem, (C)- banana rhizome pieces, (D)- banana pseudostem slices, (E)-

banana rhizome juice,(F)-banana pseudostem juice, (G)-banana rhizome RTS beverages, (H)-banana pseudostem RTS beverages

D

T6 T5 T7 T8 T5 T6 T7 T8

A B

C

E F

G H


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Preparation of Dehydrated products

Different drying methods viz., cabinet drying, vacuum drying and freeze drying

(Table 5.1; fig 5.2 and 5.3.) were employed to obtain powder and were used to study

bioactive components (total phenolic and flavonoid content) and antioxidant activities

using different in vitro assays.

Table 5.1: Drying of banana pseudostem and rhizome using different

methods

Drying methods Drying conditions

Cabinet dryer Drying temperature 40oC

Drying duration 36h

Vacuum dryer

Drying temperature 60oC

Drying pressure 25 mm WC

Drying duration 24h

Freeze dryer

Freezingtemperature - 30oC

Drying temperature - 30oC to + 20o C

Drying duration 16h

Drying pressure 1000 millitorr

Preparation of extracts from pseudostem and rhizome dehydrated powders

The preparation of extracts from pseudostem and rhizome powders using serial

extraction procedure was followed as given in chapter 1 and 2 (materials and methods).

The acetone extract of pseudostem and rhizome exhibited high polyphenolic content and

antioxidant activity as discussed in chapter 1 and 2. Hence, only acetone extract was used

to study the effect of different drying methods on bioactive constituents (total phenolic

and flavonoid content) and antioxidant activities.


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Figure 5.2: Dehydrated powders from banana pseudosyem. A-1, A-2:fresh pseudostem slices, B-1: cabinet dried pseudostem slices, B-2: cabinet dried pseudostem powder, C-1: vacuum dried pseudostem slices, C-2: vacuum dried pseudostem powder, D-1: freeze dried pseudostem slices, D-2: freeze dried pseudostem powder

A-1 A-2

B-1 B-2

C-1 C-2

D-1 D-2


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Figure 5.3: Dehydrated powders from banana rhizome. A-1:cabinet dried rhizome shreds, A-2: cabinet dried rhizome powder, B-1: vacuum dried rhizome shreds, B-2: vacuum dried rhizome powder, C-1: freeze dried rhizome shreds, C-2: freeze dried rhizome powder

A-1 A-2

B-1 B-2

C-1 C-2

A B C D


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Chemical composition of banana pseudostem and rhizome juices, RTS beverages

Total soluble solids (oBrix) and pH of juices were observed using digital

refractometer (% sucrose content) and pH meter respectively. Total acidity (% citric acid)

and tannins were estimated as given by Ranganna (2001). Total suagr, non-reducing and

reducing sugars were determined according to lane and leynons method by using

Fehling’s solution (Ranganna, 2001). The protein and amino acid content were estimated

using Folins-Ciocalteu reagent (Lowry et al., 1951) and ninhydrin (Ranganna, 2001),

respectively.

Bioactive constituents

The juices, RTS beverages and acetone extract of dehydrated powders from

banana pseudostem and rhizome were used to study the total phenolic content (TPC) and

total flavonoid content (TFC) by following folin-ciocalteau colorimetric method

(Kamatou et al., 2010; Prior et al., 2005) and aluminum chloride colorimetric method

(Esmaeili and Sonboli, 2010; Gulcin et al., 2010) respectively. The TPC and TFC of

juices, RTS beverages were expressed as GAE and CE/100 µL concentration,

respectively and in dehydrated powders were expressed as GAE and CE/g of acetone

extract. The detailed methodology is explained in materials and methods section of

chapter 1.

Antioxidant activity

The juices, RTS beverages and acetone extract of dehydrated powders from

banana pseudostem and rhizome were studied for their antioxidant activities using

various methods viz., DPPH radical scavenging activity (DPPH RSA), superoxide radical

scavenging activity (SRSA), metal chelating activity (MCA) and total reducing power


127

(TRP) assay, using catechin, EDTA and ascorbic acid as standards. The antioxidant

activity results of juices, RTS beverages were expressed per 100 µL concentration and

dehydrated powders were expressed as 100 µg/mL of acetone extract as per the

methodology described in chapter 1 and 2 (materials and methods section).

Statistical analysis

Results were expressed as mean ± standard deviation of triplicate analyzes. Data

were analyzed by one-way analysis of variance (ANOVA) and post-hoc mean separations

were performed by Duncan’s Multiple Range Test (DMRT) at p<0.05 using Microsoft

Excel XP (Microsoft Corporation, USA).

Results and discussion

In the present investigation of RTS beverages and dehydrated powders were

prepared at different concentrations from banana pseudostem and rhizome and

characterized for their chemical composition, bioactive contents, antioxidant activity and

sensory quality.

Sensory evaluation

Among the various RTS beverages prepard, only RTS beverage with twenty

percent rhizome (T7) juice and twenty five per cent pseudostem (T8) juice with 15 obrix

TSS and 0.30% acidity content showed higher overall quality score of 8.0±0.26 and

7.1±0.21, respectively (Table 5.2). Therefore, only those concentrations of juices were

subjected to further analysis.


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Table 5.2: Sensory evaluation of banana pseudotsem and rhizome RTS beverages

Treatment

No.

Juice

content

(%)

Pseudostem juice Sensory acore

TSS

(oBrix)

Acidity

(%) Overall quality

1 10 10.16 ± 0.26 0.252 ± 0.01 4.0 ± 0.11a

2 15 10.32 ± 0.15 0.250 ± 0.03 4.3 ± 0.34b

3 20 10.23 ± 0.19 0.252 ± 0.02 5.4 ± 0.29c

4 25 10.17 ± 0.21 0.255 ± 0.03 6.0 ± 0.33d

5 10 15.04 ± 0.28 0.300 ± 0.02 5.5 ± 0.20a

6 15 15.09 ± 0.39 0.306 ± 0.02 5.9 ± 0.25b

7 20 15.14 ± 0.65 0.303 ± 0.04 6.6 ± 0.12c

8 25 15.29 ± 0.19 0.302 ± 0.06 7.1 ± 0.21d

Rhizome juice

1 10 10.66 ± 0.09 0.257 ± 0.05 4.5 ± 0.20a

2 15 10.22 ± 0.12 0.252 ± 0.02 5.5 ± 0.25b

3 20 10.24 ± 0.14 0.255 ± 0.04 6.0 ± 0.16c

4 25 10.07 ± 0.18 0.253 ± 0.02 6.0 ± 0.13c

5 10 15.40 ± 0.22 0.310 ± 0.02 6.5 ± 0.25a

6 15 15.10 ± 0.44 0.311 ± 0.09 7.0 ± 0.21b

7 20 15.34 ± 0.53 0.306 ± 0.05 8.0 ± 0.26c

8 25 15.59 ± 0.33 0.309 ± 0.08 7.2 ± 0.20b

Mean values in a column with different superscripts differ significantly at p<0.05

Chemical composition of banana pseudostem and rhizome juices and RTS

beverages

The chemical composition of banana pseudotsem juice, rhizome juice and its RTS

beverages were presented in able 5.3. Pseudotsem and rhizome juice showed low TSS

(2.9±0.39 and 2.54±0.25obrix) and acidity (0.0069±0.01 and 0.07±0.02%) respectively.

The pH of the pseudostem (6.3±0.15) and rhizome (6.7±0.22) juice were near to neutral

pH, which were suitable for preparing the beverages. This value (pH) is of importance as

measure of the active acidity which influence the flavor or palatability of a product

(Ranganna, 2001). Total sugar, reducing sugar, non-reducing sugar, protein, amino acid

and tannin content (TAE-tannic acid equivalents) of rhizome juice (18.07±0.32,


129

14.12±0.42, 3.95±0.12, 127.99±0.52, 12.34±0.42 and 3.12±0.20 mg/100 mL respectively)

and its RTS (T7) beverage (14.56±0.24, 5.17± 0.20, 9.39±0.55, 25.21±0.67, 3.63±1.01

and 1.08±0.67 mg/100 mL respectively) was higher than pseudostem juice (8.3±0.47,

6.2±0.26, 2.1±0.20, 60±0.45, 8.3±0.26 and 1.95±0.15 mg/100 mL respectively) and its

RTS (T8) beverage (13.61±0.56, 4.87±0.51, 8.74±0.67, 16.61±2.67, 2.15±0.84,

0.72±0.51 mg/100 mL respectively).

Table 5.3: Chemical composition of banana pseudotsem juice, rhizome juice and RTS

beverage

Parameters Pseudostem

juice

Rhizome

juice

T8

(Pseudostem

RTS beverage)

T7

(Rhizome

RTS beverage)

Juice yield (%) 65±2.2b 25±0.15a - -

TSS (˚Brix) 2.9±0.39b 2.54±0.25a 15.29±0.19c 15.34±0.53c

Acidity (%) 0.0069±0.01a 0.07±0.02b 0.30±0.06c 0.30±0.05c

pH 6.3±0.15b 6.7±0.22b 2.94±0.02a 2.90±0.06a

Total sugars♣ 8.3±0.47a 18.07±0.32d 13.61±0.56b 14.56±0.24c

Reducing sugar♣ 6.2±0.26c 14.12±0.42d 4.87±0.51a 5.17± 0.20b

Non reducing sugar♣ 2.1±0.20a 3.95±0.12b 8.74±0.67c 9.39±0.55d

Protein♣ 60±0.45c 127.99±0.52d 16.61±2.67a 25.21±0.67b

Amino acids♣ 8.3±0.26c 12.34±0.42d 2.15±0.84a 3.63±1.01b

Tannins♣ 1.95±0.15c 3.12±0.20d 0.72±0.51a 1.08±0.67b

Mean values in a row with different superscripts differ significantly at p<0.05; ♣mg/100mL


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Bioactive components of banana pseudostem juice, rhizome juice, RTS beverages

and dehydrated powders

Table 5.4: Bioactive components of banana pseudotsem juice, rhizome juice and its

RTS beverage

Dehydrated powder TPC

(mgGAE/100 µL)

TFC

(mgCE/100 µL)

Pseudostem juice 105.21±3.11c 40.17±1.36c

Rhizome juice 312.34±10.24d 74.30±3.32d

T8 52.14±2.04a 11.54±1.50a

(Pseudostem RTS beverage)

T7 58.59±1.06b 21.13±1.27b

(Rhizome RTS beverage)

Mean values in a row with different superscripts differ significantly at p<0.05. TPC-total phenolic content; TFC-total flavonoids

content.

The amount of TPC and TFC were higher in rhizome juice (312.34±10.24

mgGAE and 74.30±3.32 mgCE/100 µL, respectively). Comparatively pseudostem juice

showed lower TPC and TFC (105.21±3.11 mgGAE and 40.17±1.36 mgCE/100 µL,

respectively) (Table 5.4). The polyphenolic content present in plant can vary significantly

due to different factors, such as plant genetics and cultivar, soil composition and growing

conditions, maturity state, and post harvest conditions, and others (Jaffery et al., 2003). In

RTS beverage also, the TPC and TFC inT7 from rhizome juice (58.59±1.06 mgGAE and

21.13±1.27 mgCE/100 µL of juice, respectively) were higher than the RTS from

pseudostem (T8) juice (52.14±2.04 mgGAE and 11.54±1.50 mgCE/100 µL of juice,

respectively). The high polyphenolic content (TPC and TFC) present in these juices

might be responsible for high sesory overall quality score. Polyphenolic antioxidants are

often added to foods for stabilization, prevent off-flavour formation, minimise rancidity,

retard the formation of toxic oxidation products, maintain nutritional quality, and increase

shelf life (Espin et al., 2007).


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Table 5.5: Bioactive components of acetone extract of banana

pseudostem and rhizome dehydrated powder

Dehydrated

powder

TPC

(mgGAE/g of extract)

TFC

(mgCE/g of extract

Pseudostem powder

Cabinet dried 275.01±17.77a 68.84±2.9a

Vacuum dried 279.12±12.62a 72.36±3.5a

Freeze dried 296.98±14.04a 76.47±3.1a

Rhizome powder

Cabinet dried 589.94±18.13a 347.36±20.87a

Vacuum dried 591.92±20.05a 352.15±15.88a

Freeze dried 612.75±22.27a 364.65±17.41a

Mean values in a coloumn with different superscripts differ non significantly at p<0.05. TPC-total

phenolic content; TFC-total flavonoids content

Effect of drying methods on bioactive compounds of banana pseudostem and

rhizome powders was presented in table 5.5. Freeze dried powders from both rhizome

and pseudostem were showed high TPC (612.75±22.27 and 296.98±14.04 mgGAE/g of

acetone extract, respectively) and TFC (364.65±17.41 and 76.47±3.1 mgCE/g of acetone

extract, respectively), followed by vacuum dried powders (591.92±20.05, 279.12±12.62

mgGAE, and 352.15±15.88, 72.36±3.5 mgCE/g of acetone extract, respectively) and

cabinet dried powders 589.94±18.13 and 275.01±17.77 mgGAE, and 347.36±20.87 and

68.84±2.9 mgCE/g of acetone extract, respectively). However, no significant (p>0.05)

difference were observed between the cabinet dried, vacuum dried and freeze dried

powders on phenolic and flavonoid contents.


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Antioxidant activities of banana pseudostem juice, rhizome juice, RTS beverages,

and dehydrated powders

In this study, rhizome juice displayed highest activity in all the antioxidant assays

tested viz., DPPH radical scavenging activity (80.24±2.72%), superoxide radical

scavenging activity (45.29±1.98%), metal chelation activity (42.56±1.86%) and total

reducing power (0.826±0.016 OD at 700 nm) at the concentration of 100 µL, when

compared to pseudostem juice (70.11±3.63%, 34.78±1.66%, 32.17±0.69% and

0.302±0.006 OD at 700 nm, respectively). The DPPH radical scavenging activity of

rhizome juice is comparable to the activity of standard catechin (81.06±1.16%) at 100

µg/mL. Whereas, in RTS beverages, T7 (from rhizome) showed high antioxidant in all

the models tested viz., DPPH radical scavenging activity (51.37±2.16%), superoxide

radical scavenging activity (31.44±1.73%), metal chelation activity (28.40±0.40%) and

Table 5.6: Antioxidant activity of banana pseudotsem juice, rhizome juice and its

RTS beverage

Parameters DPPH RSA

(%)

SRSA

(%)

MCA

(%) TRP

♣♣♣♣

Pseudostem juice 70.11±3.63c 34.78±1.66c 32.17±0.69c 0.302±0.006c

Rhizome juice 80.24±2.72d 45.29±1.98d 42.56±1.86d 0.826±0.016d

T8

36.22±2.14a 24.67±0.91a 23.85±0.23a 0.232±0.009a (Pseudostem RTS

beverage)

T7 51.37±2.16b 31.44±1.73b 28.40±0.40b 0.271±0.005b

(Rhizome RTS beverage)

Standards

Catechin 81.06±1.16 82.74±1.12 NT NT

EDTA NT NT 95.11±1.08 NT

Ascorbic acid NT NT NT 0.794±0.27

Mean values in a row with different superscripts differ significantly at p<0.05; DPPH RSA-1,1-diphenyl-2-picrylhydrazyl

radical scavenging activity, SRSA-superoxide radical scavenging activity, MCA-metal chelating activity and TRP-total reducing

power assay;♣OD at 700 nm; NT-not tested.


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total reducing power (0.271±0.005 OD at 700 nm) at the concentration of 100 µL,

followed by T8 (36.22±2.14%, 24.67±0.91%, 23.85±0.23% and 0.232±0.009 OD at 700

nm respectively). There was a significant difference (p<0.05) between the antioxidant

activity of the juices and RTS beverages tested (Table 5.6). Higher polyphenolic content

in rhizome juice and its RTS beverage (T7) contributed strongest antioxidant activity,

when compare to pseudostem juice and its RTS beverage (T8).

Table 5.7: Antioxidant activityof acetone extract of banana pseudostem and

rhizome dehydrated powder and standards

Dehydrated

powder

DPPH RSA

(%)

SRSA

(%)

MCA

(%) TRP

♣

Pseudostem powder

Cabinet dried 62.69±2.23a 51.49±1.69a 42.45±2.14a 1.267±0.021a

Vacuum dried 65.54±1.98a 53.58±1.07a 54.08±2.02a 1.277±0.020a

Freeze dried 68.57±2.10a 60.81±1.58a 53.84±1.78a 1.724±0.024a

Rhizome powder

Cabinet dried 80.43±1.67a 76.19±1.49a 62.67±1.67a 2.463±0.037a

Vacuum dried 82.29±2.18a 79.24±1.66a 64.31±1.94a 2.458±0.036a

Freeze dried 89.47±2.29a 83.72±2.33a 69.91±1.13a 2.897±0.034a

Standards

Catechin 81.06±1.16 82.74±1.12 NT NT

EDTA NT NT 95.11±1.08 NT

Ascorbic acid NT NT NT 0.794±0.27

Mean values in a colomn with different superscripts differ non significantly at p>0.05; DPPH RSA-1,1-diphenyl-2-

picrylhydrazyl radical scavenging activity, SRSA-superoxide radical scavenging activity, MCA-metal chelating

activity and TRP-total reducing power assay; ♣sample concentration to get 0.5 of absorbance at 700 nm.; NT-not

tested.

Effect of drying methods on antioxidant activity of banana pseudostem and

rhizome powders were presented in table 5.7 Freeze dried powders from both rhizome

and pseudostem showed high antioxidant activity in all the four methods tested viz.,

DPPH radical scavenging activity (89.47±2.29 and 68.57±2.10%, respectively),


134

superoxide radical scavenging activity (83.72±2.33 and 60.81±1.58%, respectively),

metal chelation activity (69.91±1.13 and 53.84±1.78%, respectively) and total reducing

power (2.897±0.034 and 1.724±0.024 OD at 700 nm) at the concentration of 100 µg/mL

of acetone extract, followed by vacuum dried powders (82.29±2.18 and 65.54±1.98%,

79.24±1.66 and 53.58±1.07%, 64.31±1.94 and 54.08±2.02%, and 2.458±0.036 and

1.277±0.020 OD at 700 nm, respectively) and cabinet dried powders (80.43±1.67 and

62.69±2.23%, 76.19±1.49 and 51.49±1.69%, 62.67±1.67 and 42.45±2.14%, and

2.463±0.037 and 1.267±0.021 OD at 700 nm, respectively). However, the difference

between the AOA of the powders from cabinet dried, vacuum dried and freeze dried were

found to be non-significant (p>0.05).

Antioxidant activities of banana pseudostem and rhizome juices, RTS beverages

and dehydrated powders could be related to the nature of polyphenolic compounds

present and their electron transfer or hydrogen donating ability. The results suggest that

the polyphenolic compounds enriched in the pseudostem and rhizome juices, RTS

beverages and dehydrated powders displayed scavenging effect on DPPH and superoxide

anion radical, pro-oxidant ability to reduce Fe3+ to its more active Fe2+ could help prevent

or ameliorate oxidative damage and also capable of complexing with and stabilizing

transition metal ions, rendering them unable to participate in metal catalyzed reaction

(Bourgou et al., 2008). Phenolics and flavonoids are well known for their ability to

donate electron (Karaman et al., 2010).

Conclusion

For the first time nutritional and nutraceutical components of juices, RTS

beverages and dehydrated powders from banana plant bio-waste viz. pseudostem and

rhizome var. Nanjanagudu Rasbale were enumerated. Highly acceptable RTS beverages

were standardized and also dehydrated powders were developed using different drying

methods. High concentration of bioactive polyphenols (phenolics and flavonoids) in

juices, RTS beverages and dehydrated powders of psudostem and rhizome were


135

concurrent to its high antioxidant activities were demonstrated. Thus, juices, RTS

beverages and dehydrated powders from banana plant bio-wastes, with high polyphenolic

content were characterized as good antioxidants by their multiple antioxidant potential

and can be effectively used to produce new age functional beverages, and also rich source

of nutraceutical supplement for food industries. The products developed also provide new

vista for better utilization of banana plant bio-waste.

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