advanced journal of chemistry-section b · 2021. 2. 21. · complexometry titration conductometry...

* Corresponding author: Akram Hassan E-mail: [email protected] Tel number: +92554845248

© 2020 by SPC (Sami Publishing Company)

Advanced Journal of Chemistry-Section B Natural Products and Medical Chemistry

Journal homepage: http://www.ajchem-b.com/

Original Research Article

The Electrochemical Study of the Effects of Herbal Tea on Calcium Oxalate Renal Stones

Akram Hassan*, Hamza Ayaan

Department of Chemistry, Aga Khan University, Pakistan

INTRODUCTION

G R A P H I C A L A B S T R A C T

A R T I C L E I N F O

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Objective: Present study designed to evaluate the effectiveness of an extract obtained from Cleome dolichostyla on calcium oxalate crystallization in vitro.

Materials and Methods: In this laboratory research, after taking the aqueous extract of 5 grams of Cleome dolichostyla by 100 ml of distilled water at gentle heating, the formation and dissolution of calcium oxalate in the absence and presence of extract was investigated by complexometric and conductometric methods.

Results: The effect of the extracts on the prevention of sediment formation was studied and the results of the conductometric titration curve method for leaves of Cleome dolichostyla showed a significant difference (175.0 µmol) between the concentration of calcuim in the absence (450.0 µmol) and presence of the extract (625.0 µmol) and for seeds, the difference was (38/0 µmol) in the absence (450/0 µmol) and presence (488/0 µmol) of the extract. In the study of the effect of extract on the stone dissolution, the results of the complexometry method showed that the amount of calcium measured by the dissolution of calcium oxalate crystals for the leaves of Cleome dolichostyla in the presence of extract, was 2.5 µmol higher than that in the absence of extract and for seeds was 6.0 µmol higher.

Conclusion: According to this study, Cleome dolichostyla aqueous extract dissolves calcium oxalate kidney stone, but cannot prevent the formation of them.

Article history: Submitted: 2020-04-29 Revised: 2020-05-24 Accepted: 2020-06-13 Published: 2020-06-22 Manuscript ID: AJCB-2004-1030 DOI: 10.22034/ajcb.2020.109313

K E Y W O R D S Cleome dolichostyla Calcium oxalate Complexometry titration Conductometry Kidney stone

,

G R A P H I C A L A B S T R A C T

A B S T R A C T

Advanced Journal of Chemistry, Section B, 2020, 2(3), 120-127

mailto:[email protected]

http://www.ajchem-b.com/


Akram Hassan, Hamza Ayaan/Ad. J. Chem. B, 2020, 2(3), 120-127

121 | P a g e

Urinary stone formation affects 10-12% of the

population in industrialized countries. Their

prevalence has increased in recent years, while

the age of onset has decreased. Diet and

environment play an important role in kidney

stone diseases, possibly by changing the

composition of the urine [1-3].

Renal stones have different types, including

calcium oxalate, calcium phosphate, ureic acid,

magnesium ammonium phosphate (Struvite) and

cysteine stones, that about 75-80% of urinary

stones are composed of calcium oxalate and

calcium phosphate salts [4-10]. Of course, other

unusual kidney stones such as xanthine,

dihydroxy adenine, silicate and matrix have also

been reported [10-16].

However, for the treatment of patients with

kidney stones, various therapies such as the use of

chemical drugs or surgeries have been used, but,

so far, there is no effective and safe drug

treatment that results in complete treatment or

prevention of stone formation [17-23]. Today,

because of the side effects and harmful effects of

chemical drugs, the use of herbal products has

attracted the attention of researchers [24-31].

In recent researches, the effects of some

medicinal herbs such as barberry, Nigella Sativa,

Juniperus excelsa and Allium jesdanum have been

investigated in clinical or laboratory form on

kidney stone. This study was conducted to

investigate the effect of Cleome dolichostyla

extract on the formation and dissolution of

calcium oxalate kidney ston. Cleome dolichostyla is

an annual herb, which grows wild in barren lands

and belongs to the family Cleomaceae. This plant

contains some mineral compounds, including Ca,

Mg, Fe, Zn, Cu and Mn and fatty acids such as

linoleic acid, palmitic acid, stearic acid and

linolenic acid [31-37].

MATERIAL AND METHODS

In this research, at first, Cleome dolichostyla

plant was collected from the county of Lahore

province, and then washed some of plant with

double distilled water and dried in laboratory

temperature and away from sunlight for 4 days,

then weighed the amount of 5.0 g of leaf and seed

of the plant and mixed with 75.0 ml of double

distilled water at 65 ° C temperature and placed

them toward the steam of boiling water for 120

minutes. The resulting blends were filtered with

Whatman filter paper, and extended the solution

volume under the filter to 100 ml with the help of

double distilled water. At the end, we put the

resulting aqueous extracts at a temperature of 4.0

° C for conducting relevant experiments. The

materials used in the titration are: Sodium oxalate

(Na2C2O4), Dihydrate calcium chloride

(CaCl2.2H2O), Ammonia) NH3(, Hydrochloric acid

(HCl), Ethylenediaminetetraacetic acid (EDTA),

Calcium oxalate (CaC2O4), Eriochrome Black T,

Double-distilled water [21-23].

Meanwhile, ammonia buffer solution with pH

=10.0 was prepared from adding the hydrochloric

acid to ammonia solution and adjusting by pH

meter. Standard calcium solutions with specific

densities were prepared by dissolving the

appropriate amounts of calcium chloride in

double distilled water (Fig. 1).

RESULTS AND DISCUSSION

The result of the end points (Equivalence)

obtained from the complexometry titration of the

solutions that were mentioned above with the

EDTA standard solution are presented in Table

1 and 2. The consumption volume of EDTA for

the solution of the aqueous extract of Cleome

dolichostyla leaves and seeds, saturated with

calcium oxalate, in comparison with their total

volume of consumption in the titration of the two

blank and extract solutions lacking calcium

oxalate shows a significant increase (p <0.05).



122 | P a g e

Fig. 1. The process of extracting and complexometric titration of the Cleome dolichostyla

Fig. 2. The process of complexometric titration of the blank

Table 1. Results of complexometric titration of Cleome dolichostyla leaves with EDTA(0.005M)

Titrand Blank

Extract without

CaC2O4

Extract with

CaC2O4

Volume of titrant

(EDTA) 0.3 9.3 10.1

Table 2. Results of complexometric titration of Cleome dolichostyla seeds with EDTA(0.005M)

Titrand

Blank Extract Extract with

Without CaC2O4 CaC2O4

Volume of titrant

(EDTA) 0.3 8.5

10.0



123 | P a g e

Fig. 3. Conductometric titration of CaCl2(0.025M) solution.

Fig. 4. Conductometric titration of CaCl2(0.025M) solution in absence and presence

of leaf extract of Cleome in absence and presence of seed extract of Cleome

dolichostyla with Na2C2O4 (0.025M) dolichostyla with Na2C2O4 (0.025M)

Comparison of the volume of consumption

of sodium oxalate solution related to the points

of equivalence showed a significant difference

(p <0.05).

CONCLUSION

In this research, with the aim of achieving a

herbal medicine with the least side effects, we

study the effect of aqueous extract of leaf and

seed of Cleome dolichostyla plant on the

formation and dissolution of calcium oxalate

kidney stones. This is important because in

pharmacy science, a comprehensive chemical

medicine which can prevent the formation of

kidney stones and treating them with the least

effects is not available. Since most kidney

stones are generally calcium oxalate, this

compound has little solubility in water (Ksp =

1.7 × 10-9). Therefore, the presence of calcium

ions (Ca2 +) and oxalate (C2O42-), even at

relatively low density in the kidney's aquatic

environment, can lead to calcium oxalate

formation or in other words, kidney stones

[27-30].

Meanwhile, if we can use some herbal

extracts to incorporate compounds into the



Page | 124

kidney environment which those compounds

in the interaction with renal salts prevent the

formation or dissolution of the sediment, we

have taken an effective step in the treatment

and prevention of kidney stone disease.

Therefore, in this research, the effect of Cleome

dolichostyla on the formation and dissolution

of calcium oxalate stones has been studied. In

this regard, two methods of complexometry

and conductometric titration have been used.

In the complexometry method, the density

of Ca2+ existed in the solution in the presence of

Eriochrome Black T detector was determined

by gradual increase of EDTA ligand in an

ammonia buffer environment with pH=10

based on the following reaction:

Ca2+ + EDTA4-↔[Ca(EDTA)]2-

In the test related to the leaf and seed of the

plant, in the presence of the extract, this change

is from light brown to slime green. The end

point of titration for non-extract blank

solutions is obtained by changing the color of

the solutions from violet to blue [31-32].

The results of the complexometry

presented in Table 1 and 2 show that the

volume of EDTA used in the titration of the

aqueous extract of leaf and seed of calcium

oxalate (10.1 and 10 ml, respectively), which is

higher for both extracts than the total volume

of EDTA for the blank (0.3 ml) and the non-

calcium oxalate extracts (9.3 and 8.5 ml,

respectively), Therefore, for calcium density in

solutions, the following relation is true:

µmolCa2+(leaf extract containing CaC2O4)>

[µolCa2+(blank) +µmolCa2+(leaf extract without

CaC2O4 )] 10.1>0.3 +9.3

µmolCa2+(seed extract containing CaC2O4)>

[µolCa2+(blank) +µmolCa2+(seed extract without

CaC2O4 )] 10.0>0.3 +8.5

If the extract in the solution increases the

solubility of calcium oxalate salt, we expect

that the calcium ion density calculated in

titration with EDTA is higher than the blank

solution. Possibly, the presence of some

interactions between the active ingredients of

the extract with oxalate and calcium ions

increases the dissolution of calcium oxalate

salts. Therefore, it can be concluded that the

aqueous extract of leaf and seed of both cases

increased the dissolution of calcium oxalate in

the solution. Therefore, in patients with

calcium oxalate kidney stones, these extracts

can be recommended to dissolve kidney

stones. Another important issue for kidney

stone disease is the problem of recurrence and

getting afflicted to kidney stones in people who

has improved. Therefore, in this research, we

used the conductometric titration method to

study the effect of aqueous extract of leaf and

seed in the process of preventing the formation

of calcium oxalate sedimentation. In this

titration, with the gradual increase of the

titrant to the titrating solution, the electrical

conductivity is measured by the conductivity

meter, which has a direct relation with the

density and type of ions presented in the

solution.

In this research, the effect of the presence

of extracts in the process of the formation of

calcium oxalate sedimentation in calcium ion

solutions was investigated in titration with

sodium oxalate solution. Since the effect of

titration for each calcium ion of Ca2+two

Sodium ion Na+ is introduced into solution,

therefore the electrical conductivity of the

solutions is gradually increased with gradual

increase of sodium oxalate solution until the

sediment gradually and with gentle slope is

completed. After the equivalence point and

with the continuation of the titration, the

sedimentation process is completed and,

therefore, without the removal of ion from the

solution, only the density of both sodium and

oxalate ions increases in such a way that it

compensates for the dilution of the solution

due to increased volume, So the electrical



Page | 125

conductivity increases with a steeper slope.

Fig. 3 and 4 show the changing process of the

electrical conductivity of the reading (blank,

the extract of leaf and seed Cleome

dolichostyla) containing calcium chloride as

compared to the titrated sodium oxalate. The

study of these titrations shows that the

consumption volume of titrated sodium

oxalate solution for the samples containing

both extracts is higher than the blank sample.

In fact, the higher the amount of calcium

available for the solution to react with oxalate

ions, the higher the titrant intake to reach the

equivalence point which indicates the possible

existence of Ca2+ions in the leaf and seed

extracts. Although sodium oxalate does not

have the ability to break the bond between

calcium ions and the compounds of the extract,

but the presence of acidic compounds in the

extract causes the coordinating of calcium ions.

In the complexometric titration, firstly 5.0 ml of

aqueous extract of leaf and seed of Cleome

dolichostyla was mixed with 20 ml of double

distilled water and 0.01 g of solid calcium

oxalate. Both mixtures were mixed at room

temperature for 90 minutes by magnetic stirrer

and were passed through a glass filter.

Subsequently, 3.0 ml of ammonia buffer was

added to each of the following solutions and

titrated in the presence of Eriochrome Black T

reagent with EDTA 0/005 M. The volume end

points of these titrations were compared with

consumption volumes of EDTA in the titration of

blank solutions (saturated solution of calcium

oxalate in water) and solutions without calcium

oxalate [16-19]. In conductometric titration by

using JENWEY conductivity meter (Model

4510), first separately, 25.0 ml of calcium

chloride solution of 0.025 M containing aqueous

extract of leaf and seed was prepared and

stirred at room temperature for 90 minutes and

filtered with a glass filter. Then conductometric

titration of under the filter solution and the

blank solution (a solution of 0.025 M calcium

chloride in water) with sodium oxalate 0.025 M

was performed.

ACKNOWLEDGMENTS

We want to appreciate the department of

herbal medicine at the University of Lahore for

their great supports.

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HOW TO CITE THIS ARTICLE

Akram Hassan, Hamza Ayaan, The Electrochemical Study of the Effects of Herbal Tea on Calcium Oxalate Renal Stones, Ad. J. Chem. B, 2 (2020) 120-127 DOI: 10.22034/ajcb.2020.109313 URL: http://www.ajchem-b.com/article_109313.html



advanced journal of chemistry-section b · 2021. 2. 21. · complexometry titration conductometry...

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