Open Access Journal Volume: 1.

World Journal of Gastroenterology, Hepatology

and Endoscopy
Evaluation of In Vitro Antiurolithiatic Activity of Manilkara zapota Seeds
*Sanjuna Chiluveri
Prasad Mukkera
Anjali Mittapalli
Sandhya Narayanolla Department of Pharmacognosy, Vishnu Institute of Pharmaceutical Education
Y. Manasa and Research, Narsapur, Medak, Telangana, India
M. Srikanth
J. Himabindhu
Dr. K. Ramanjaneyulu

Article Information
Article Type: Research Article *Corresponding author: Citation: Chiluveri Sanjuna (2019)
Journal Type: Open Access Evaluation of In Vitro Antiurolithiatic
Chiluveri Sanjuna
Activity of Manilkara zapota Seeds. World J
Volume: 1 Issue: 1 Department of Pharmacognosy, Gastroenterol Hepatol Endosc, 1(1);1-3
Manuscript ID: WJGHE-1-108 Vishnu Institute of Pharmaceutical
Education and Research, Narsapur,
Publisher: Science World Publishing Medak
Received Date: 25 April 2019
Accepted Date: 7 May 2019
Published Date: 29 May 2019

Copyright: © 2019, Sanjuna C, et al., This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0
International License, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source
are credited.

ABSTRACT
The present study was undertaken to evaluate the in vitro antiurolithiatic activity of the medicinal plant seeds of Manilkara zapota.
Methanolic extracts showed their maximum efficiencies in the dissolution of calcium oxalate crystals. Our results have clearly indicated that the
methanolic extracts of M. zapota of seeds were quite promising for further studies in this regard. In this study Neeri was used as standard drug.

KEYWORDS: In vitro antiurolithiatic activity, Methanolic extract, Urolithiasis, Manilkara zapota, Neeri

INTRODUCTION
Urolithiasis is commonly referred as stone formation in any part of the urinary tract such as kidneys, ureters, urinary bladder and urethra.
It is one of oldest, most frequent and highly recurrent disease and was initially found in the tombs of Egyptian mummies dating back to 4000
BC. Epidemiological studies revealed that urolithiasis is more common in men than in women and is more prevalent between the ages of 20
to 40 in both sexes. Calcium containing uroliths are known as brushite, whewellite, weddellite, whitlockite and carbonate apatite. Struvite and
newberyite are magnesium containing whereas ammonium acid urate, mono sodium urate monohydrate, uric acid anhydrous, uric acid mono
and dehydrate are commonly existing urate stones. Medicinal plants are considered as a rich source of therapeutic agents due to the belief and
observations regarding their traditional use for the prevention of various ailments. Various research findings and data from different part of the
globe are contributing and helping the scientific community in evaluating and establishing the pharmacological activities of these plants [1].
Urinary stone formation affects 10-12% of the population in industrialised countries. From epidemiological data, Calcium Oxalate (CaOx)
is the most common component of the calculi. The formation of such concretions involves several physicochemical events, e.g. nucleation,
growth and aggregation, but the mechanism(s) of these processes remain incompletely understood. It is widely known that a patient who has
one kidney stone is more likely to develop another. Therefore, the prevention of recurrence (Often up to 60%) is crucial. Unfortunately, despite
considerable progress in medical therapy, there is no satisfactory drug to treat kidney stones. Many patients still undergo surgery to remove the
stones; thus in Morocco, as in many countries, most patients (≈70%) use medicinal plants as an alternative therapy for many diseases, including
lithiasis [2].
Nowadays stone formation is the oldest and serious painful urologic disease with significant prevalence in the population due to change
in lifestyle and dietary factors. Stone formation or lithiasis is characterized by calculi formation. It has two main types such as nephrolithiasis
and Urolithiasis. Calculi formation in urinary bladder, ureter or any part of urinary tract rather than kidney is known as Urolithiasis while
nephrolithiasis is characterized calculi formation in kidney. Generally, calcification for the formation of bone and teeth takesplace in controlled
biological situations. Uncontrolled pathological crystallization occurs when solvent becomes supersaturated formation of precipitates in the
body called as kidney stones [3].
There are many theories that explain the pathogenesis of stone formation, for example, the super saturation theory and the inhibitors
theory. Super saturation occur when there is an abundance of solute in a solution. Although, Urolithiasis is a multifactorial dis-ease, nutrition,

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especially fluid intake, with several underlying disorders of colour end point obtained. The amount of remaining undissolved
metabolism: that is why diet is an important treatment, especially calcium oxalate is subtracted from the total quantity used in the
in the prevention of recurrences. Epidemiological studies reveal that experiment in the beginning to know the total quantity of dissolved
about 80% of all kidney stones are composed of calcium salts (75% calcium oxalate by various solvent extracts [5].
calcium oxalate), while about 5% are pure uric acid. Kidney stones
have previously been linked to higher rates of high blood pressure,
RESULTS AND DISCUSSION
obesity, diabetes and other heart disease risk factors. Researchers
have speculated that the Dietary Approach is to Hypertension (DASH) Drug therapy has developed in response to population health
style diet can, also prevent kidney stones. The main components of care [6] needs. There are many crucial areas in medicine such as liver
the DASH diet includes fruit, vegetables, nutsandle gumes, low fat diseases, arthritis, old age related problems, certain viral infections
dairy, whole grains, and lower intake of salt, sweetened drinks, red and cancer where the conventional medicine is devoid of satisfactory
meat and processed meat. The DASH style diet may reduce stone treatment. These are among the promising areas of research and
risk by increasing urinary citrate and volume. The small associations development of medicines from the vast highly potential plant
between higher DASH score and lower relative super saturation of resources. Plants are also attractive sources for the development of
calcium oxalate and uric acid suggesting that unidentified stone novel and very effective and safe therapeutic agents against kidney
inhibitors in dairy products and/ or plants. procumbens. Herbal medicines are also in great demand in the
developed world for primary health care because of their efficacy,
Sapotaceae is a family of some 35-75 ill-defined genera and
safety and lesser side effects [7]. Unlike allopathic medicines which
about 800 species, most are tropical trees. Manilkara zapota (L.) Van
target is only one aspect of urolithiatic pathophysiology, most of plant
Royen (Synonyms: Manilkara zapotilla, Manilkara achras, Mimusopus
based therapy have been shown to be effective at different stages of
manilkara, Achras zapota, and Achras sapota), growing abroad, was
stone pathophysiology [8]. About 80% of the world populations rely
subjected to chemical study resulted in isolation of flavonoids,
on the use of traditional medicine which is predominantly based
tannins (Mainly from unripe fruits), triterpenes, and saponins
on plant materials [9]. Plant based drug discovery programmes
(Mainly from the seeds). Also, the plant was reported to exhibit
continue to provide an important source of new drug leads [10].
antioxidant, antimicrobial, and analgesic activities [4].
Lithiasis (stone formation) is an important cause for acute and
chronic renal failure, includes both nephrolithiasis (stone formation
MATERIALS AND METHODS in kidney) and urolithiasis (Stone formation in ureter or bladder or
both). Among the various kinds of stones identified, calcium stones
Plant Material occur mainly in Men, while phosphate stones formation is more in
The seeds of Manilkara zapota was collected in the month of march women [11].
2019 from Venkatapur village, Medak dist. of Telangana, India. The This study evaluates the antiurolithiatic activity methanolic
plant was authenticated by M. Mallareddy [M.sc, M.phil in botany] extract of seeds of M. zapota. The percentage i.e. 99% of CaOx
retired lecturer in botany. The seeds were washed with tap water dissolution was observed in methanolic extraxct. Of seds of M. zapota
and dried under shade. Methanolic extract of M. zapota was found to be more effective in
Preparation of Plant Extract dissolution of calcium oxalate than standard drug Neeri. From this
study, it was observed that methanolic extracts of showed their
The seeds were shade dried and powdered. The crude plant extract highest dissolution of calcium oxalate. Methanolic extract was found
was prepared by Soxhlet extraction method. 50 g of powdered to be effective in dissolution of calcium oxalate. This study has
plant material was extracted with 500 ml of ethanol. The process of given primary evidence for M. zapota as the plant which possess
extraction was carried out up to 6 cycles, till the solvent in siphon lithotriptic property. This in vitro study has given lead data and
tube of an extractor became colorless. The two extracts were filtered shown that Aqueous and Ethanolic extracts are quite promising for
separately, and evaporated to dryness using rotary evaporator. further studies in this regard.
Further the dried extracts were maintained in a refrigerator at 4°C
for further antiurolithiatic activity. Table 1: Shows % dissolution of Calcium Oxalate (CaOx) by
Manilkara zapota leaves extracts
Chemicals Used
Neeri, Sodium oxalate, Tris buffer, calcium chloride, Potassium % of dissolution of calcium oxalate
permanganate (KMnO4), Sulphuric acid (H2SO4).
S. No GROUPS Manilkara zapota
Investigation of In Vitro Antiurolithiatic Activity Test by 1. Blank 0
Titrimetry
2. Positive Control 81
The experimental kidney stones of CaOx were prepared in the
laboratory by taking equimolar solution of calcium chloride 3. Methanolic extract 99
dehydrate in distilled water and sodium oxalate in 10 ml of 2 N
H2SO4. Both were allowed to react in sufficient quantity of distilled
water in a beaker, the resulting precipitate was calcium oxalate.
The precipitate was freed from traces of sulphuric acid by ammonia
solution, washed with distilled water and dried at 60°C. The
dissolution percentage of calcium oxalate was evaluated by taking
exactly 1 mg of calcium oxalate and 10 mg of the extract, packed it
together in semi permeable membrane of egg as shown in the model
designed given below. This was allowed to suspend in a conical flask
containing 100 ml of 0.1 M Tris buffer. First group served as blank
containing only 1 mg of calcium oxalate. The second group served as
positive control containing 1 mg of calcium oxalate and along with
the 10 mg standard drugs, i.e. Neeri. The 3rd, groups along with 1
mg of calcium oxalate contain methanolic and aqueous, extracts. The
conical flasks of all groups were kept in an incubator preheated to
37°C for 2 h. Remove the contents of semi permeable membranes
from each group into separate test tubes, add 2 ml of 1Nsulphuricacid Figure 1: In vitro experimental model setup to evaluate
to each test tube and titrated with 0.9494 N KMnO4 till a light pink antiurolithiatic activity

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CONCLUSION
In vitro urolithiasis has been performed on the selected plant M.
zapota by using the standard drug, Neeri. The work was performed
by using in vitro antiurolithiatic model for calculating percentage
dissolution of kidney stone. Methanolic seed extracts of M. zapota
shows highest dissolution than standard drug Neeri. This study has
given primary evidence for M. zapota as the plant which possess
antiurolithiatic property.

ACKNOWLEDGEMENT
We sincerely thankful to our principal Dr. A. Ramesh and staff
members, Director and chairman of our college Vishnu Institute of
Pharmaceutical Education and Research (VIPER) for supporting us.

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Figure 1(c): Egg membrane along with the contents suspended into
the 0.1 M Tris buffer

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