By

Dr.G.SANDHYARANI
M.PHARM,Ph.D.PDF
 Toshow the anti urolithiaitic
activity of moringa leaves
 Kingdom: Plantae
 Sub kingdom: Tracheobionta
 Super Division: Spermatophyta
 Division: Magnoliophyta
 Class: Magnoliopsida
 Moringa oleifera Lam (syn. M. ptreygosperma Gaertn.) is a tropical plant belonging to
family Moringaceae, native of India which was introduced in Brazil around 1950.
 Moringaceae is a single genus family with 13 known species. Among these oleifera is
most widely used and utilized species (Sengupta and Gupta, 1970; Morton, 1991).
 The tree originated from Agra and Qudh in the northern eastern region of India, south
of Himalayas (Mugal et al., 1999). It is cultivated throughout the plains, especially in
hedges and in house yards, thrives best under the tropical insular climate, and is
plentiful near the sandy beds of rivers and streams (Qaiser, 1973).
 It can grow well in the humid tropics or hot dry lands, can survive destitute soils, and
is little affected by drought (Morton, 1991). Moringa grows best at altitudes up to 600
m but it will grow at altitudes of 1000 m. It tolerates a wide range ofrainfall with
minimum annual rainfall requirements estimated at 250 mm and maximum.
 Moringa leaves have been reported to be a rich source of β-carotene,
protein, vitamin C, calcium and potassium and act as a good source of
natural antioxidants
 Thus enhance the shelf-life of fat containing foods due to the presence
of various types of antioxidant compounds such as ascorbic acid,
flavonoids, phenolics and carotenoids.
 The Moringa plant is perennial, evergreen tree that grows up to 20 ft tall
or ranges from 5 to 10 m (Morton, 1991), with a straight trunk and a
corky, whitish bark.
 Fruit is long and round with green color when it is young and brown
when mature.

 Moringa prefers neutral to slightly acidic soils and grows best in well-
drained loam to clay-loam. It tolerates clay soils but does not grow well if
waterlogged
 CALCIUM CHLORIDE
 SODIUM OXALATE
 DLIUTE HYDROCHLORIC ACID(DIL.HCL)
 PHOSPHATE BUFFER
 ETHANOL
 PHENAZINE METHOSULFATE(PMS)
 TRIS
 NICOTINAMIDE ADENINE DINUCLEOTIDE(NADH)
 We choose the classical model for the study of oxalate crystalization because of its simple and
satisfactory reproducibility.
 This model includes the study of crystallization without inhibitor and with it, in order to assess
the inhibiting capacity of any chemical species used.

 Solution of calciumchloride and sodium oxalate were prepared at the final con-centrations of 5
mmol/L and 7.5 mmol/L respectively in a buffer containing Tris 0.05 mol/L and NaCl 0.15
mol/L at pH 6.5. 950 mL of calcium chloride solution mixed with 100 mL of herb extracts at the
different concentrations (100 mg/mle1000 mg/ml).

 Crystallization was started by adding 950 mL of sodium oxalate solution. The temperature was
maintained at 37 CC. The OD of the solution was monitored at 620 nm.

 The rate of nucleation was estimated by comparing the in-duction time in the presence of the
extract with that of control.28,29.The growth of crystals was expected due to the following
reaction:
CaCl2þNa2C2O4/CaC2O4 2NaCl
 AGGREGATION ASSAY:
 The method used was similar to that described by Atmani and Khan.29 with
some minor modifications. ‘Seed’ CaOxmonohydrate (COM) crystals were
prepared by mixing cal-cium chloride and sodium oxalate at 50 mmol/L.

 Both so-lutions were equilibrated to 60 C in a water bath for 1 h andthen

cooled to 37 C overnight.
 The crystals were harvested by centrifugation and then evaporated at 37 C.
CaOX crys-tals were used at a final concentration of 0.8 mg/ml, buff-ered with
Tris 0.05 mol/L and NaCl 0.15 mol/L at pH 6.5
 Experiments were conducted at 37 C in the absence or presence of the plant
extract after stopping the stirring.
 The percentage aggregation inhibition rate (Ir) was then calcu-lated by
comparing the turbidity in the presence of the extract with that obtained in the
control using following formula
lr=(1 -Turbidity sample/Turbidity control)*100
 •Nucleation
Effect of different conc of leaf extract in
Nucleation assay
2.5

assay
Turbidity= (2.3*absorbence)/L 2
y = -0.3186x + 2.6186
R² = 0.9845

Conc of Abs Turbidity

Turbidity
1.5
Herb extract
turbidity
100ug/ml 0.989 2.2747 1 Linear ()

300ug/ml 0.852 1.9596 Linear (turbidity)

500ug/ml 0.741 1.7043 0.5

700ug/ml 0.621 1.4283

0
1000ug/ml 0.412 0.9476 0 1 2 3 4 5 6

Concentration ug/ml
 Concentratio Effect of different conc of leaf extract
Sample n of herb by Aggregation assay
80
name extract Abs Turbidity
70
control 0ug/ml 0.945 2.1735 y = 13.28x + 21.905
60 R² = 0.9401

% Inhibition
50

Conc of Herb Abs Turbidity IR 40

extract 30
Linear (Series2)

100ug/ml 0.651 1.4973 31.11111 20

500ug/ml 0.432 0.9936 54.28571 10

700ug/ml 0.356 0.8188 62.32804 0

0 1 2 3 4 5
1000ug/ml 0.258 0.5934 72.69841
Concentration ug/ml
 Based on the results of the present study, it could be concluded that all
the extracts tested were found to show stone inhibitory property in all
stages of crystal formation in vitro.
 Aqueous extract of Aerva lanata was found to have maximum
inhibitory potential when compared to all the weed extracts selected.
 Diuretic effect and antilithiatic property were also confirmed by the in
vivo studies. Histopathological studies supported the results.
 The phytochemical analysis revealed that the aqueous extract is a rich
source of antioxidants and secondary metabolites.
 The study confirmed that the bio- synergistic action of the
phytochemicals confer antilithiatic and antioxidant property to the
extract.
 Thus,the aqueous extract of Aerva lanata can be recommended as an
antilithiatic agent to treat kidney stones.
 Utilization of weeds in productive ways, so that people may
benefit from an aspect that has been largely ignored.
 ‘Utilization’ has been recognized as an effective means of
weed management
 Use of weeds for providing effective, non harmful cure that
caters to
people of all groups
 NRK 52E cells can be used as alternate for the in vivo
analyses to study the mechanism of action of the crystals in
the cell
 For use as formulation in home remedies
 Prasad KVSRG, Sujatha D, Bharti K. Herbal drugs in
urolithiasis: a review. Pharmacog Rev. 2007;1(1):175e178.
 Grover PK, Kim DS, Ryall RL. The effect of seed crystals of

hydroxyapatite and brushite on the crystallization of calcium

oxalate in
undiluted human urine in vitro: implications for urinary stone
pathogenesis. Mol Med. 2002;8(4). 2000e2009.