SPECIFICATON OF SAMRONG (SCAPHIUM SCHAPHIGERUM) FRUIT GEL
Wichittra Phlicharoenphon1*, Wandee Gritsanapan1, Mansuang Wuthi-udomler2, Pongtip
Sithisarn1 #
Department of Pharmacognosy1, Department of Microbiology2, Faculty of Pharmacy, Mahidol
University, Bangkok, Thailand
*ladynamfon.fon@gmail.com # pongtip.sit@mahidol.ac.th
Abstract
Samrong (Scaphium scaphigerum (G.Don) Guib. & Planch.) is a tree in Sterculiaceae
family. The gel from the fruits of samrong is widely used as foods and drinks for mild
laxative effect and weight control. The main constituent in samrong fruit gel is a dietary fiber
which can swell almost 10 times of its original size. This fiber will not be digested by the
enzymes in human gastrointestinal tract that lead to the satiety of the consumer. This study
was conducted in order to set up the specification of samrong fruit gel including the
appropriate preparation method, the physical and chemical properties such as the appearance,
swell volume, total acidity, viscosity and thin layer chromatographic (TLC) and infrared (IR)
fingerprints. The Microbial limits of samrong fruit gel were also determined. From various
preparation methods, the gel from samrong fruits prepared by soaking the fruits in water until
the gel was fully formed then boiled the gel with water and dried the gel in a hot air oven at
65-70 ºC for 8 hours promoted the gel with the best physical characteristics and yield. The
suitable particle size of samrong fruit gel that promoted high swell volume (158 .05 ± 0.0 0
ml) was 841 µm. Using the titration method, total acidity of the gel was 0.57 ml of 0.03 N
potassium hydroxide (KOH) while the viscosity was 1.18 cps. TLC fingerprint of the
hydrolyzed fraction of samrong fruit gel showed the chromatographic bands corresponded to
some monosaccharides including galactose, arabinose and rhamnose. IR (KBr disc)
fingerprint promoted the peaks at wave number corresponded to the functional groups of
hydroxyl (OH), ketone (C=O), C-O, CH3 and CH. From the microbial limit test, there was no
pathological bacterial including Bacillus cereus, Clostridium spp., Escherichia coli,
Pseudomonas aeruginosa, Salmonella spp., Staphylococcus aureus and Candida albicans in
samrong fruit gel.
Keywords : Samrong, Scaphium scaphigerum, swell volume, total acidity, viscosity, TLC
fingerprint, IR fingerprint, microbial limit, specification
Introduction
Samrong (Scaphium scaphigerum (G.Don) Guib. & Planch) is a tree in Sterculiaceae
family. The gels from the fruits of this plant are traditionally used as laxative and weight
control agents and for the treatments of cough and sore throat and using as expectorant1).
After soaking in water, samrong seed coats become a brown glass jelly which can be used as
a bulk laxative and promote the satiety. This gel contains polysaccharide PP-III, which the
subunits are manosaccharides; galactose, arabinose and rhamnose(1-3). Even though there are
a lot of beverages and dessert preparations, especially healthy drinks for weight control from
samrong fruit gel, however there is no report about the specification and the quality control of
this gel before. Therefore, this experiment was set up in order to develop a specification of
samrong fruit gel including suitable preparation method, physical and chemical properties
including viscosity, swell volume, chromatographic and spectroscopic fingerprints (TLC and
IR) using methods according to USP 36 monograph. The contaminations of pathological
microbial were also investigated.
Methodology
Plant material : Samrong fruits were purchased from Chantaburi province, Thailand in
April, 2013. The specimen was compared with the authentic plant material of Forest
Herbarium, Wildlife and Plant Conservation Department of Thailand. Samrong fruits were
prepared by cleaning and cutting upper and lower parts of the fruits.
Preparation of samrong fruit gel by different methods
Samrong fruits were divided into 2 groups, first group were original air dried fruits
(G1) and the second group were G2 which were dried in a hot air oven (65-70 ºC) for 8 hours
before used. Ten grams of each G1 and G2 are separately prepared as follows;
Method 1 : Samrong fruits G1 were boiled in distilled water at 100˚C for 60 minutes.
The swollen samrong fruit gel was passed through a sieve to separate other contaminants,
then weighed and measured the volume of the gel. The physical properties of samrong fruit
gel were then observed.
Method 2 : Samrong fruits G2 were boiled in distilled water at 100˚C for 60 minutes.
The swollen samrong fruit gel was passed through a sieve to separate other contaminants,
then weighed and measured the gel volume. The physical properties of samrong fruit gel
were then observed.
Method 3 : Samrong fruits G1 were boiled in 0.05% sodium hydroxide solution at
100˚C for 60 minutes. The swollen samrong fruit gel was passed through a sieve to separate
other contaminants then weighed and measured the gel volume. The physical properties of
samrong fruit gel were then observed.
Method 4 : Samrong fruits G2 were boiled in 0.05% sodium hydroxide solution at
100˚C for 60 minutes. The swollen samrong fruit gel was passed through a sieve to separate
other contaminants, then weighed and measured the gel volume. The physical properties of
samrong fruit gel were then observed.
Method 5 : Samrong fruits G1 were soaked in 0.05% sodium hydroxide solution at
room temperature for 60 minutes. The swollen samrong fruit gel was passed through a sieve
to separate other contaminants, then weighed and measured the gel volume. The physical
properties of samrong fruit gel were then observed.
Method 6 : Samrong fruits G2 were soaked in 0.05% sodium hydroxide solution at
room temperature for 60 minutes. The swollen samrong fruit gel was passed through a sieve
to separate other contaminants, then weighed and measured the gel volume. The physical
properties of samrong fruit gel were then observed.
Method 7 : Samrong fruits G1 were soaked in distilled water at room temperature for
60 minutes. The swollen samrong fruit gel was passed through a sieve to separate other
contaminants, then weighed and measured the gel volume. The physical properties of
samrong fruit gel were then observed.
Method 8 : Samrong fruits G2 were soaked in distilled water at room temperature for
60 minutes. The swollen samrong fruit gel was passed through a sieve to separate other
contaminants, then weighed and measured the gel volume. The physical properties of
samrong fruit gel were then observed.
Method 9 : Samrong fruits G1 were soaked in distilled water at room temperature for
15 minutes. Then the contaminations were separated. The swollen samrong fruit gel was
rinsed and boiled in distilled water at 100˚C for 60 minutes. The swollen samrong fruit gel
was left on a sieve until the water was drained. Samrong fruit gel was passed through a fabric
filter then weighed and measured the gel volume. The physical properties of samrong fruit
gel were then observed.
The method that promoted samrong fruit gel with good physical properties such as
yield, physical appearance and odor was used to prepare samrong sample for further studies.
Evaluation of suitable drying method of samrong fruit gel dried powder.
Samrong fruit gel from a suitable gel preparation method was separately dried using
various drying methods including dried in a hot air oven, spray drying, and freeze drying.
Then the yield and swell properties of the dried gel were determined. The suitable method for
drying samrong fruit gel was then selected.
Evaluation for the suitable particle size of dried samrong fruit gel powder.
Samrong fruit gel powder was prepared into 2 particle sizes, used the sieves number
18 and 20, respectively. The obtained samrong fruit powder was evaluated for physical and
swell properties. The suitable particle size of samrong fruit gel powder was then selected for
analyzed the specification as described below.
Evaluation of samrong fruit gel properties for setting up the specification.
Swell volume
Samrong fruit gel powder (0.5 g) was placed in 100 ml-glass-stopper cylinder. The
water was added to a total volume of 100 ml. The cylinder was inverted upside down every 4
hours and stayed for 16 hours and determined for the gel volume. The samrong fruit gel
should be produced at least 80 ml per 1 g of samrong fruit gel powder (4).
Total acidity
The supernatant from samrong fruit gel powder from swell volume test (40 ml) was
placed in an erlenmeyer flask, then phenolphthalein T.S. (1 ml) was added and the solution
was titrated with 0.03 N sodium hydroxide (NaOH) solution and the end point was observed.
The volume of NaOH solution being used to reach the end point should not more than 1.8 ml
(4).
Viscosity
The viscosity of samrong fruit gel and samrong fruit gel powder were determined by
brookfield viscomiter machine (4).
Infrared spectrometric (IR) fingerprint
The infrared spectra of samrong fruit gel powder was identified by IR (KBr disc)
techniques.
Thin layer chromatographic (TLC) fingerprint
Samrong fruit gel was hydrolyzed and submitted for TLC analysis, using 1-propanol :
ethyl acetate : water (4:0.5:0.5 v/v/v) as a solvent system and 10% sulfuric acid in methanol
as a spraying reagent with standard authentic monosaccharides.
Preparation of samrong fruit gel: Samrong gel powder (5 g) was swelled in
water (1000 ml) for 16 hours then the gel was separated. Samrong gel (5 g) was mixed with
95% ethanol (210 ml) and left at 4˚C for 12 hours then filtered. The obtained precipitate was
hydrolyzed by adding 15 ml of 5% sulfuric acid solution. The solution was heated on the
water bath (90˚C) for 2 hours. The solution was cooled and neutralized using barium
carbonate. The mixture was filtered and a supernatant was used for TLC analysis (5).
TLC condition: The hydrolyzed fraction of samrong fruit gel and the
authentic monosaccharides were analyzed by thin layer chromatography using the condition
as follow;
Adsorbent : Silica gel GF60
Solvent system: 1-propanol : ethyl acetate : water (4:0.5:0.5 v/v/v)
Detection: 10% sulfuric acid in methanol. After spraying, the TLC plate was
heat at 100˚C for 10 minutes (5).
Evaluation of microbial limits
Samrong fruit gel powder was evaluated for total aerobic microbial count, specific
bacteria and total yeast and mold count followed the methods according to ASEAN guideline
on stability study of drug product, 2005 (6).
Results
The preparation for the suitable samrong fruit gel powder
As shown in Table 1, samrong gels prepared from various preparation methods
showed different physical characteristics and yields. Samrong gel obtained from method 9
promoted the best physical characteristics and yield (7650.00 % v/w of dried fruits). So this
method was considered as suitable preparation method.
Table.1 Yield and physical characteristics of samrong fruit gel from various preparation
methods
Methods
Weight (g)
Volume (ml)
Physical characteristic
1
6239.0 ± 185.3
6250.0 ± 70.7
Brown glass gel with specific odor
2
4642.5 ± 175.4
4600.0 ± 141.4
Brown glass gel with specific odor
3
1181.0 ± 21.2
1250.0 ± 70.7
Black brown viscous liquid
4
1095.5 ± 57.3
1050.0 ± 70.7
Black brown viscous liquid
5
239.0 ± 18.4
250.0 ± 28.3
Black brown viscous liquid
6
135.0 ± 48.0
140.0 ± 42.4
Black brown viscous liquid
7
1564.8 ± 123.8
1550.0 ± 70.7
Brown glass gel
8
1282.8 ± 42.2
1300.0 ± 0.00
Brown glass gel
9
7626.0 ± 90.5
7650.0 ± 212.1
Swollen brown glass gel with the specific
odor, smooth sensations without the
contaminants
Evaluation of suitable drying method of samrong fruit gel dried powder.
It was found that drying samrong fruit gel using hot air oven promoted the dried light
brown glassy gel with the highest yield (329 mg / 100 g of fresh gel) while the freeze drying
and spray drying methods promoted the dried gels with lower yields (237 and 46 mg / 100 g
of fresh gel, respectively). Therefore, drying with hot air oven was chosen as suitable drying
method.
Evaluation for the suitable particle size of dried samrong fruit gel powder.
The gel prepared from method 9 was dried in a hot air oven at 65-70 ºC for 8 hours
then powdered through the sieve No.20 and 18 to promote the gel powders with the particle
sizes of 841 and 1000 µm, respectively. The gel powder with the particle size of 8 4 1 µm
promoted the gel with higher swell volume (Table 2). Therefore, this gel powder was selected
for further investigations.
Table.2 Swell volume of samrong fruit gel powders with different particles sizes
Powder size of
dried powder
841µm
1000 µm
Weight (g)
Volume (ml)
145.57 ± 5.54
132.36 ± 3.70
158.05 ± 0.00
151.60 ± 0.51
Physical characteristic of
obtained gel
Brown glass gel
Brown glass gel
Evaluation of samrong fruit gel properties for setting up the specification
Total acidity
The average volume of 0.3 N sodium hydroxide solution used for the titration was
0.57 ± 0.06 ml. This total acidity value is less than the maximum criteria according to
psyllium hemicelluloses monograph in USP 36 (1.8 ml).
Viscosity
Samrong fruit gel promoted the viscosity of samrong gel 4.55 x 103 centipoises (cps).
corresponded to the viscosity of xanthan gum in USP 36 monograph which indicated that the
viscosity at 24 °C should not less than 600 cps.
Thin layer chromatography (TLC) fingerprints
Determination of the phytochemical properties of hydrolyzed fraction of samrong
fruit gel was conducted by TLC detected with 10% sulfuric acid in methanol spraying
reagents. As shown in Figure 1 there were the chromatographic bands corresponding to those
of the standard sugars including galactose, arabinose and rhamnose (track 3, 4 and 5
respectively) corresponds to the report of polysaccharides PP-III which contained the subunit
sugars of galactose, arabinose and rhamnose (1-3).
1
2 3 4
5
6
Figure.1 TLC chromatogram of hydrolysis fraction of samrong fruit gel and standard sugars;
1= hydrolysis fraction of samrong fruit gel, 2 = glucose (Rf = 0.40), 3 = galactose (Rf =
0.32), 4= arabinose (Rf = 0.44), 5 = rhamnose (Rf = 0.60), 6 = manose (Rf = 0.46), stationary
phase = silica gel GF254, solvent system = 1-propanol: ethyl acetate: water (4:0.5:0.5),
detector = 10% sulfuric acid in methanol and heat at the temperature of 100˚C for 10 minutes.
Infrared (IR) fingerprint
As shown in Figure 2, IR (KBr disc) fingerprint of samrong fruit gel powder
promoted the peaks at wave number corresponded to some functional groups including
hydroxyl (OH), ketone (C=O), C-O, CH3 and CH suggesting the polysaccharides structures.
The IR data was shown in Table 3.
%T
2
4
3
1
56
7
Wavenumber (cm-1)
Table.3 IR spectra (KBr disc) and functional groups of samrong fruit gel powder
Wavenumber (cm-1)
Functional group
3378.27 (1)
OH stretching
2937.21 (2)
CH stretching
1739.63 (3)
C=O stretching
1379.01 (4)
-CH3 bending
1253.46 (5)
1148.12 (6)
1037.40 (7)
C-O stretching (ester, ether)
C-O stretching (secondary, tertiary alcohols)
C-O stretching (primary alcohol)
Microbial limits test
Using methods according to ASEAN guideline on stability study of drug product,
2005 (5), samrong fruit gel powder promoted negative results to all tested pathological
bacteria including Bacillus cereus, Clostridium spp., E. coli, P. aeruginosa, S. aureus,
Salmonella spp. and C. Albicans as shown in Table 4 (6).
Table.4 Investigation for some pathological bacteria in samrong fruit gel powder
Organisms
Result of Detection
Bacillus cereus
Clostridium spp.
E. coli
P. aeruginosa
S. aureus
Salmonella spp.
C. albicans
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Discussion and Conclusion
The suitable preparation method to prepare samrong fruit gel was soaking samrong
fruits in water until the gel was fully formed, then further boiled with water. Drying the gel in
a hot air oven at 65 -70 º C for 8 hours was the appropriate drying method and the suitable
particle size of samrong fruit gel powder was 841 µm. The physical, chemical and microbial
contamination properties of samrong fruit gel powder was evaluated and specified as
indicated in Table 5.
Table.5 Specification of samrong fruit gel
Specification
Preparation method
Drying method
Particle size of gel powder
Property
Soaked samrong fruit in distilled water at room temperature for 15
minutes. Then the contaminations were separated. The swollen
samrong fruit gel was rinsed and boiled in distilled water at 100 oC
for 60 minutes. The swollen samrong fruit gel was left on a sieve
until the water was drained. Samrong fruit gel was passed through a
fabric filter.
Drying in a hot air oven at 65-70 ºC for 8 hours.
841 µm
Physical characteristic of the gel
Brown glass gel with the specific odor, smooth sensations without
the contaminants
Swell volume of dried gel powder Not less than 80 ml per 1 g of dried gel powder
Total acidity
Not more than 1.8 ml
Viscosity
Not less than 600 cps.
TLC fingerprint*
(conditions as mentioned before)
The hydrolysed fraction of the gel shows specific TLC fingerprint
with the bands corresponded to galactose, arabinose and rhamnose.
IR fingerprint*
(KBr disc)
IR fingerprint of dried gel powder shows the peaks at wave number
corresponded to the functional groups of hydroxyl (OH), ketone
(C=O), C-O, CH3 and CH
The gel powder promotes negative results to the pathological
bacterial including Bacillus cereus, Clostridium spp., E. coli, P.
aeruginosa, S. aureus, Salmonella spp. and C. Albicans
contamination tests.
Microbial limit*
(ASEAN guideline methods)
Acknowledgments
The authors would like to express their gratitudes to Thailand Research Fund (TRF)
for financial support.
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