Asian J.

Dairy & Food Res, 35(1) 2016: 81-84 AGRICULTURAL RESEARCH COMMUNICATION CENTRE
Print ISSN:0971-4456 / Online ISSN:0976-0563 www.arccjournals.com

Study on effect of custard apple leaf extract on physico-chemical

properties of aonla juice
Mundhe Sushilkumar S.*, Sawate Arvind R., Patil Bhanudas M., Kshirsagar Rajesh B. and S.P. Kulkarni
Department of Food Engineering,
College of Food Technology, VNMKV, Parbhani - 431 401, India.
Received: 30-04-2015 Accepted: 09-01-2016 DOI: 10.18805/ajdfr.v35i1.9257
ABSTRACT
The leaf extract was prepared by shade drying of leaves for 4-5 days, the dried leaves were pulverized and boiled in water
bath with distilled water to get the liquid extract. Further the liquid extract was centrifuged and then supernatant was spray
dried to get powdered extract. Sweetened aonla juice formulation was made with different proportion of custard apple leaf
extract varying from 0 to 15 g in increasing order from control (0) to A1 (5g), A2 (10g) and A3 (15g) respectively. The
prepared juices were analyzed for physico-chemical composition and sensory evaluation. The retention of quercetin
[(a flavonol, is a flavonoid, in other words, a plant pigment with a molecular structure like flavone. It is found in fruits,
vegetables, leaves and grains. It can be used as an ingredient in supplements, beverages, or foods having nutraceutical
value.)] in aonla juice sample A1, A2, A3 was 15.20, 32.80 and 49.75 mg per 1000 ml. Sample A2 (10g custard apple leaf
extract) scored highest score for overall acceptability was 8.8.
Key words: Aonla juice, Custard apple leaf extract, Quercetin, Sensory evaluation.

INTRODUCTION of diabetes, with continued therapy ensuring up to 80%

Custard apple (Annona squamosa) (family:
Annonaceae) and Tribulusterrestris (family: Zygophyllaceae)
are two common plants of India and well described for their
traditional medicinal values. Anti-diabetic potential of both
the plant has been investigated by different researchers.
Methanol extract of aerial parts of Tribulusterrestris possess
anti-diabetic effect against streptozotocin induced diabetes
(El-Tantawy et al., 2007) and anti-diabetic potential of
saponins isolated from the plant also investigated in alloxan
induced diabetic mice ( Li et al.,2002). Different researcher
also investigated the anti-diabetic potential of ethanolic and
aqueous extract of Annona squamosa leaf against different
type 1 and type 2 diabetic model (Gupta et al.,2005, Kaleem
et al., 2008). Annona squamosa L. (Annonaceae), commonly
known as custard apple is a native of West Indies and is
cultivated throughout India, mainly for its edible fruit. The
plant is attributed with medicinal properties, (Shirwaikar et
al., 2004).
The young leaves of annona squamosa are used
extensively for its anti-diabetic activity by tribal men in and
around the villages of Aligarh district in the state of Uttar
Pradesh, India (Atique et al., 1985) and also by the people
of Chota Nagpur district in the state of Bihar, India (Topno,
1997).
In Aligarh district villagers take a mixture of 4–5
newly emerged young leaves along with five grains of black
pepper (Piper nigrum) early in the morning for the treatment
*Corresponding author’s e-mail: sushil.mundhe@gmail.com.
positive results. The treatment is particularly common and
popular in the Lodha community where the plant is
considered to be a holy fruit. The formula is being
successfully used by some Unani and Allopathic physicians
and is in existence till date. Black pepper is reported to be
commonly used as a bioavailability enhancer (Atal et al.,
1981) and piperine isolated from pepper fruits has been
patented as a bioavailability enhancer (Majeed et al., 1998).
The aqueous leaf extract of annona squamosa has also been
reported to ameliorate hyperthyroidism which is often
considered as a causative factor of diabetes mellitus (Sunanda
and Anand, 2003).
Quercetin may also induce insulin secretion by
activation of L-type calcium channels in the pancreatic -
cells (Bardy et al., 2013).Quercetin is shown effective against
numerous conditions. According to the University of
Michigan Health System, studies prove Quercetin’s ability
to fight and help treat prostatitis, allergies and type 1 and
type 2 diabetes. (University of Michigan Health System,
2014). At present, an increased consumer interest in diet and
health is being observed. The food producers tend to produce
safer food choices to meet consumers’ demand. A healthy
food is a food believed to be highly beneficial to health which
generally contains lots of fruits and vegetables and is low in
fat, salt and sugar that may contribute to high risk of health
problems regarding to diabetes, obesity, high blood glucose/
cholesterol, teeth decay and cancer (Knecht 1990).
82 ASIAN JOURNAL OF DAIRY AND FOOD RESEARCH
MATERIALS AND METHODS
Custard apple Leaves, distilled water, ethyl alcohol,
spray dryer, pulverizer, Whatman No.1 filter paper, fresh
aonla, mesh, blender Centrifuge, Spectrophotometer.
Preparation of custard apple (annona squamosa) leaf
extract: The aqueous extract was prepared by cold
maceration of 50 g of the shade-dried leaf powder in 1000
ml of distilled water and allowed to stand overnight and
boiled for 5-10 hours till the volume was reduced to half of
its original volume. The solution was then cooled, filtered,
concentrated and spray dried in spray drier at temp- 130 0c
(yield 7 g) and the powder stored in a refrigerator at 2-8°C
for subsequent experiments. Preparation of custard apple
(annona squamosa) leaf extract is given in figure 1. (Kaleem
et al., 2006)
Selection of custard apple (annona squamosa) leaves
Leaves of custard apple plants were shade dried (For 5-6 days)
Pulverized and passed through sieve 40 mesh
Cold Maceration
(Allowed to stand overnight)
Boiled till the volume was reduced to half of its original volume
(For 5-10 hours)
Solution was then cooled and filtered
(By using Whatman No.1 filter paper)
Centrifugation of filtrate was carried out
(at 10000 rpm for 15 min)
Discarded sediment to get supernatant
Leaf extract powder preparation
(By using spray drier)
FIG 1: Flow sheet for preparation of custard apple (annona squamosa) leaf
extract (Kaleem et al., 2006)
Formulation of custard apple leaf extract fortified
sweetened aonla juice: Fresh, fully ripe, sound aonla were
used for extraction of juice (Sasi Kumar et al, 2013). Each
fruit was cleaned, thoroughly washed, blanched and blended
in a laboratory blender to a pulp and the juice extracted was
blended with custard apple leaf extract and stored
refrigerated temperature. Further formulation process is
given in figure 2.
TABLE 1: Formulation of custard apple leaf extract fortified SweetenedAonla juice
Aonla juice Quantity of Juice Aspartame
Control 750 ml
A1 750 ml
A2 750 ml
A3 750 ml
A1 - 5 g custard apple leaf extract incorporated sample
A2 - 10 g custard apple leaf extract incorporated sample
A3 - 15 g custard apple leaf extract incorporated sample
Aonla Fruits
Washing
Lye Treatment (2% NaOH at 100°C for 5 minutes)
Removal seed and Separation of fruit segments
Addition of water and Passing through mixer
Fruit pulp passing through strainer
Addition of custard apple leaf extract 250 mg/kg body weight
(Pornrutet al, 2008)
Addition of low calorie sweetener
Aonla juice
FIG 2: Flow sheet for the preparation of sweetened anola juice (Srivastava
and Sanjeev Kumar, 2006)
Determination of quercetin in prepared Aonla juice: It
was done by taking absorbance (AU) readings in du- plicate
at 362 nm using a double beam spectrophotometer and
compared with standard sample of quercetin solution.
Sensory evaluation of custard apple leaf extract fortified
Sweetened Aonla juice: Custard apple leaf extract fortified
Sweetened Aonla juice was organoleptically evaluated by panel
members by using nine point Hedonic Scale.
RESULTS AND DISCUSSION
Formulation of custard apple leaf extract fortified
Sweetened aonla juice: Recipe for preparation of artificially
sweetened aonla juice, the formulation was made with
different proportion of custard apple leaf extract varying from
0 to 15 g in increasing order from control (0) to A1 (5g), A2
(10g) and A3 (15g) respectively.The recipe of aonla juice
was standardized by considering the quantity of ingredients
like aonla juice, aspartame, and citric acid as per the process
adopted by (Srivastava and Sanjeev Kumar, 2006). The
artificial sugar like aspartame was used as it is 200 times
sweeter than sucrose and the amount (4g) is used as
proportion to the sweetness ratio with sugar. (NICUS, 2013).
The proportion of custard apple leaf extract used in the aonla
juice was decided by considering the permissible intake of
250 mg / kg body weight as suggested by (Pornrut et al.
2009). The data with respect to formulation of custard apple
leaf extract fortified aonla juices were presented in Table 1.
Retention of quercetin in prepared aonla juice: In aonla
juice custard apple leaf extract was fortified at the level of
Citric acid WaterIn liter Leaf extract
4 g 11 g 4.2 —
4 g 11 g 4.2 5g
4 g 11 g 4.2 10 g
4 g 11 g 4.2 15 g
 Volume 35 Issue 1 (2016)
0 g to 15 g. In sample A1 (1g) of custard apple leaf extract was
fortified and the retention of quercetin in sample A1 was 15.20
mg. and in sample A2 (2g) of custard apple leaf extract was
fortified and the retention of quercetin in sample A2 was 32.80
mg. whereas in sample A3 (3g) of custard apple leaf extract
was fortified and the retention of quercetin in sample A3 was
49.75mg.In aonla juice the retention of quercetin was slightly
reduced may be due to the accumulation of quercetin at the
bottom during storage of aonla juice. Data with respect to
retention of quercetin in aonla juice was presented in Table 2.
TABLE 2: Retention of quercetin in prepared aonla juice
Retention mg / 1000 ml of juice
Control —
A1 15.20
A2 32.80
A3 49.75
Effect of addition of different proportions of custard
apple leaf extract on chemical properties of Aonla juice:
The aonla juice samples contain TSS in control sample
(7 0Bx), sample A 1 (7.1 0Bx), sample A 2 (7.2 0Bx), A 3
(7.30Bx).Acidity of control sample was (0.38 per cent)
similarly sample A1 (0.37 per cent), A2 (0.37 per cent), A3
(0.34 per cent). The control sample of aonla juice contains
(pH 3.8) similarly sample A1, A2 and A3 (pH 3.9, 4.0 and
4.1) respectively. The pH of control sample is statistically
significant over other samples whereas highest pH was
recorded by sample A3.
The reducing sugar content of sweetened aonla juice
were 4.7 percent whereas reducing sugar content of control
83
sample (2.95 per cent), A1 (2.97 per cent), A2 (2.98) and A3
(2.99 per cent) respectively. The reducing sugar content of
all the samples was found to be less as sugar was replaced
by artificial sweetener.
The non-reducing sugar content of sweetened aonla
juice was also less. The non-reducing sugar content of control
sample (2.71 per cent), A1 (2.73 per cent), A2 (2.72) and A3
(2.71 per cent) respectively. The non-reducing sugar content
of all the sample was found to be less whereas lowest non-
reducing sugar content score was recorded by control sample
and A3.Total sugar content of sweetened aonla juice were
control (5.66), A1, A2 and A3 (5.70 per cent) respectively.
The ash content of sweetened aonla juice control
(0.03 percent), A1 (0.047per cent), A2 (0.064 per cent) and
A3 (0.81 per cent). Similar result for chemical properties are
reported by Pareek et al, (2012). The data with respect to
effect of addition of custard apple leaf extract on chemical
properties of aonla juice were presented in Table 3.
The prepared sweetened aonla juice containing
different proportions of custard apple leaf extract were
organoleptically evaluated with regards to colour and
appearance, flavor, taste and mouth feel and overall
acceptability. The scores rated by the judges are summarized
in Table 4.
CONCLUSION
It can be finally concluded that organoleptically
acceptable aonla juice having higher nutraceutical value
containing flavonoid Quercetin (16.72 mg/g) can be prepared.
Sample A2 (10g custard apple leaf extract) scored highest score
for overall acceptability was 8.8 on 10 point hedonic scale.

TABLE 3: Effect of addition of custard apple leaf extract on chemical properties of aonla juice
Sample TSS Acidity pH Reducing Sugar Non-reducing sugar TotalSugar Ash
(0Bx) (%) (%) (%) (%) (%)
Control 7.0 0.38 3.8 2.95 2.71 5.66 0.030
A1 7.1 0.37 3.9 2.97 2.73 5.70 0.047
A2 7.2 0.36 4.0 2.98 2.72 5.70 0.064
A3 7.3 0.34 4.1 2.99 2.71 5.70 0.081
Mean 10.65 0.45 3.98 3.31 3.076 6.39 0.063
SE 0.064 0.012 0.025 0.008 0.004 0.01 0.010
CD@ 5 % 0.20 0.039 0.077 0.026 0.014 0.03 0.035
A1 - 5 g custard apple leaf extract incorporated sample
A2 - 10 g custard apple leaf extract incorporated sample
A3 - 15 g custard apple leaf extract incorporated sample

TABLE 4: Organoleptic evaluation of sweetened aonla juice

Sample Color and appearance Flavor Taste and mouth feel Overall Acceptability
C 8.0 8.0 8.0 8.0
A1 8.3 8.5 8.2 8.1
A2 9.0 8.5 8.4 8.8
A3 7.5 7.0 7.0 7.1
Mean 8.3 8.1 8.1 8.1
S.E ± 0.08 0.09 0.10 0.07
C.D. at 5% 0.28 0.29 0.32 0.23
84 ASIAN JOURNAL OF DAIRY AND FOOD RESEARCH
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