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Qualitative phytochemical and physiochemical analysis of Cassia

fistula L. Fruit

Article  in  Medicinal Plants - International Journal of Phytomedicines and Related Industries · September 2014
DOI: 10.5958/0975-6892.2014.00482.1

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 138 Agrawal et al.
Research Article

Qualitative phytochemical and physiochemical analysis of

Cassia fistula L. fruit
K. Agrawal1, Apurva Joshi3, Shivani Ghildiyal 1, M.K. Gautam 2, Mayank Gangwar 2, R.K. Goel2
and V.K. Joshi 1
1Department of Dravyaguna, Faculty of Ayurveda, Institute of Medical Sciences,

Banaras Hindu University, Varanasi-221 005, India

2Department of Pharmacology, Faculty of Modern Medicine, Institute of Medical Sciences,

Banaras Hindu University, Varanasi-221 005, India

3Department of Pharmaceutics, Indian Institute of Technology, Banaras Hindu University,

Varanasi-221 005, India

ABSTRACT

The fruit pulp of Cassia fistula Linn. is known to have purgative properties. A specific process, i.e., keeping well
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ripe fruits buried within sand for seven days and then drying in sun prior to its medicinal use, has been mentioned
in Charaka Samhita. To explore the effect of above classical processing, a comparative physiochemical and
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phytochemical evaluation of classically processed (PD) and non-processed (NPD) fruit pulp of Cassia fistula has
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been done together with HPTLC. Results of the study showed presence of alkaloid, glycosides, steroids, triterpenes,
phenolic compounds, tannins, proteins, amino acids and carbohydrats in both types of samples. The extractive
values of PD and NPD were 18.25 % w/v and 15.65% w/v, respectively. The difference in physiochemical analysis
of PD and NPD was also recorded and the HPTLC of PD sample showed an extra peak around Rf 0.76 in comparison
to NPD. These results signify appreciable changes in classically processed sample of Cassia fistula fruit pulp and
provide scientific evidence for the classical processing. [Medicinal Plants 2014; 6(2) : 138-142]

Keywords : Cassia fistula, physiochemical analysis, phytochemical analysis, HPTLC, fruit pulp

INTRODUCTION long stick-like fruit pod with a pungent odour. Cassia

fistula fruit has been advocated as a mild laxative for
Cassia fistula L. is a flowering plant of family children and pregnant women. It is purgative due to
Caesalpiniaceae and native of India, Mauritius, South the wax aloin and has been reported to treat many
Africa, Mexico, Brazil, China, Nepal, West Indies other intestinal disorders like ulcers (Biswas et al.,
and East Africa (Chopra, 1956; Kirtikar and Basu, 1973; Kirtikar and Basu, 1975; Satyavati and Sharma,
1975; Chatterjee and Pakrashi, 1992). It is an 1989). The fruit has also been reported to be a rich
ornamental plant due to its beautiful bunches of yellow source of potassium, calcium, iron and manganese
flowers. The tree is a medium sized deciduous, 6-9 (Barthakur et al., 1995). The purgative action of fruit
meters tall with straight trunk and spreading branches. pulp of C. fistula is due to presence of anthraquinone
The plant has got golden yellow coloured flowers and (Agrawal et al., 2012). Pharmacological screening of C.
fistula plant was reported to have analgesic, antipyretic
(Patel et al., 1965), sedative (Mazumdar et al., 1998),
Corresponding author : V.K. Joshi anti-hypercholesterolemic (El-Saadany et al., 1991),
e-mail : vkjoshivns@sify.net anti-inflammatory (Suwal, 1993), hypoglycemic
(Esposito et al., 1991), antipyretic (Kashiwada et al.,
Received : January 3, 2014; Accepted : March 24, 2014 1990), anti-rheumatic (Bodding, 1983), anti-tumor
doi: 10.5958/j.0975-6892.5.2.009
(Gupta et al., 2000), hepatoprotective (Bhakta et al.,

Medicinal Plants, 6(2) June 2014

 Qualitative phytochemical and physiochemical analysis of Cassia fistula L. fruit 139

1999), antioxidant (Luximon-Ramma et al., 2002), anti- guidelines on quality control method for medicinal
fungal and anti-bacterial (Dhar and Qasba, 1984) plants (Government of India, 1996; WHO, 1992). The
properties. Extensive studies have been carried out extracts obtained by cold maceration were subjected to
during the past few decades on isolation and preliminary phytochemical screening for the presence
characterization of chemical constituents of various of various phytoconstituents like alkaloids, flavonoids,
parts of C. fistula. Rhein, glucose, sucrose and fructose triterpinoids, carbohydrate, protein, resins, saponins,
were isolated from the fruit pulp (Lal and Gupta, 1972) steroid, tannin, starch and glycosides by standard
whereas f istulic acid, kaempferol and a methods (Gautam and Goel, 2012).
leucopelargonidin tetramer having free glycol unit were
Fluorescence analysis
isolated from the pods and flowers respectively (Agrawal
et al., 1972). C. fistula pod has been reported to have The ground powder was dissolved in the specif ic
very low level of toxicity and no pathogenic effects on reagent, mixed vigorously and filtered through a filter
liver, kidney and testis of rat (Akanmu et al., 2004). paper. Finally, it was visualized under long wave UV
The fruit pulp of C. fistula has been well known for (365 nm) as well as in daylight. The color of the
its mild purgative action as explained in Charaka fluorescence was obtained and compared with the
Samhita (1000 BC to 4th Centuary), the most important reference standard colour (http://en.wikipedia.org/wiki/
information regarding its use has been described in Color_index).
Kalpa sthana. It has been advised that full-grown, ripe
fruits possessing good qualities should be collected Extractive value
and kept buried within sand for a week. Thereafter
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fruits should be taken out, dried in sun and then their Four hundred mg of classically processed (PD) pulp
pulp is stored for use (Agnivesha, 2007). World Health and non processed (NPD) pulp samples were weighed
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Organization has emphasized the need to ensure the separately, in two glass-stopper conical flasks.
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quality of medicinal plant products by using modern Macerated with 100 ml of distilled water for 6 hours by
control techniques and applying suitable standards shaking frequently and then allowed to stand for 18
(Ghildiyal et al., 2012). hours. Filtered rapidly taking care not to lose any
The present study has been undertaken to find out solvent and 25 ml of the filtrate was transferred to tare
the qualitative phytochemical analysis and flat-bottomed dishes and evaporated to dryness on a
physiochemical changes in both classically processed water bath. Dried at 105°C for 6 hours, cooled in
(PD) and non processed (NPD) fruit pulp of C. fistula desiccator for 30 minutes and weighed without delay.
to validate the fact behind the classical processing. The extractable materials were calculated in mg/g of
air-dried material.

MATERIALS AND METHODS High performance thin layer chromatography

(HPTLC)
Plant material HPTLC Analysis was performed on high performance
silica gel 60 F 254 TLC plates. The plates were pre-
The fruits of Cassia fistula L. were collected from the
cleaned by development to the top with methanol and
campus of Banaras Hindu University during the month
dried in fume-hood before use. Sample solutions were
of June 2011. A voucher specimen (CF.12/2011) was
applied to the plate by means of automated spray on
deposited in the Department of Dravaguna, IMS, BHU.
band applicator equipped with 100 ml syringe and
The matured fruits of the plant were divided into two
operated with band length 10 mm, application rate 10
groups. Group-I was kept buried within sand for a week
s ml–1, table speed 10 mm s -1, and distance from bottom
(PD) and group-II was not kept within the sand (NPD).
10 mm. The volume applied for each analysis was 10
Following the stipulated period both the groups were
ml of the sample. Distance between tracks is 10 mm.
dried in sun light, fruit pulp was taken out and were
The plates were developed 8 cm beyond the origin
kept separately in air tight glass jars for further study.
with solution of Toluene: Ethyl acetate: Acetic acid
Physiochemical and phytochemical analysis 50:40:10 solvent system as mobile phase in vapor-
equilibrated twin-trough chamber. After development
The values of physiochemical parameters, viz., moisture the mobile phase was evaporated from the plate by
content, total ash, foreign matter, acid insoluble ash drying in a fume-hood for 10 min. Then, the CAT-3
and sulphated ash, were determined according to WHO software controlling the CAMAG -TLC scanner was

Medicinal Plants, 6(2) June 2014

 140 Agrawal et al.

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Fig. 1. One extra peak was found to be present around Rf 0.76 in Sample 1 (Pink). A significant change in Rf were obtained.
Compounds present in Sample 1 were found to have more Rf than same Compounds present in Sample 2.

used to scan the developed zones under Long and provides information in respect of identif ication,
Short UV and under white light after derivatization. standardization of herbal drug of C. fistula of Ayurvedic
compendia. Correct identification and quality assurance
RESULTS AND DISCUSSION of the starting materials is an essential prerequisite to
ensure reproducible quality of herbal medicine which
The values of physiochemical parameters (Table 1) were will contribute to its safety and efficacy (Ghildiyal et
different in both the samples whereas the phytochemical al., 2012). The physicochemical evaluation of the drug
assessment (Table 2) indicated an identical situation. is an important parameter in detecting adulteration or
In fluorescence analysis study (Table 3) specific colour
appeared with specific reagents. The extractive values Table 1. Physiochemical analysis
of PD and NPD fruit pulp of Cassia fistula were 18.25
% w/v and 15.65% w/v respectively. HPTLC analysis Parameter NPD PD
showed one extra peak present around Rf 0.76 in PD Total ash 9.425% 12.5%
Sample (Fig. 1).
Acid insoluble ash 4.44% 4.32%
Herbal drug which are used in various traditional
Water soluble ash 3.3% 4%
medicine needs detailed investigation with
ethanopharmacological approach. The present study % Extractive value 18.25% w/v 15.65% w/v

Medicinal Plants, 6(2) June 2014

 Qualitative phytochemical and physiochemical analysis of Cassia fistula L. fruit 141

Table 2. Phytochemical analysis of non processed (NPD) commonly known for their anti-oxidant, anti-
and processed (PD) samples of fruit pulp of inflammatory and antimicrobial activities, while
Cassia fistula triterpenoids have been reported to show
Phytoconstituents NPD PD immunomodulatory property (Gautam et al., 2012).
Extractive values are useful for the determination of
Alkaloids + + exhausted or adulterated drugs and also help in
Glycosides + + estimation of specific constituents soluble in particular
Flavonoids - - solvent. The obtained extractive value of classically
Steroidal/triterpenes + + processed (PD) and non processed (NPD) plant material
Phenolic and tannins + + is 18.25%w/v, 15.65% w/v respectively. The
Proteins + + fluorescence analysis is adequately sensitive and
Amino Acids + +
enables the precise and accurate determination over a
satisfactory concentration range without several time-
Sugars + +
consuming dilution steps prior to analysis of
+ = Present; – = Absent pharmaceutical samples (Pimenta et al., 2006). The
fluorescence colour is specific for each compound. A
non-fluorescent compound may fluorence if mixed with
improper handling of drugs. The ash value of the drug impurities that are fluorescent. The colour of the powder
gives an idea of the inorganic composition and other of Cassia fistula was observed both under ordinary
light and UV light, which gave specific colour change
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impurities present in the plant species. Preliminary

phytochemical screening reveals the chemical nature present in Table 3. In HPTLC one extra peak was found
to be present around Rf 0.76 in processed sample (Pink).
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of the extract and be a sign for the further

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pharmacological activity. C. fistula fruit pulp revealed compounds present in Sample 1 (PD) were found to
the presence of alkaloid, glycosides, steroids, have more Rf value than same Compounds present in
triterpenes, phenolic compounds, tannins, proteins, Sample 2 (NPD).
amino acids and carbohydrate. Tannins have been The above findings provide scientific evidence for
reported to possess antioxidant, wound healing and the classical processing of Cassia fistula fruit pulp for
antimicrobial activities. Phenolic compounds are desirable medicinal effect.

Table 3. Fluorescence powder drug analysis of Cassia fistula L.

S.No. Powder + Reagent Fluorescence in daylight Fluorescence under UV (365nm)

1. Powder as such Brown No Fluorescence (NF)

2. Powder + 1N NaOH in methanol Golden red Dark sea green
3. Powder + 1N NaOH in water Peru Medium Sea Green
4. Powder + 1N HCl in methanol Dark golden red Light green
5. Powder + 1N HCl in water Lemon chiffon Medium aquamarine
6. Powder + 1N HNO 3 in methanol Tan Light green
7. Powder + 1N HNO 3 in water Sea shell NF
8. Powder + Iodine (5%) Crimson NF
9. Powder + FeCl 3 (5%) Peru Dark red
10. Powder + KOH (50%) Dark orange Lime green
11. Powder + Ammonia (25%) Golden Olive green
12. Powder + Picric acid (saturated) Yellow Golden
13. Powder + Acetic acid Yellow Yellow green

Medicinal Plants, 6(2) June 2014

 142 Agrawal et al.

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