The Effect of Chives (Allium schoenoprasum L.) Leaves Ethyl

Acetate and Hexane Fractions on Human Calcium Kidney Stones

Solubility by In Vitro Method

Siti Morin Sinaga

, Sudarmi

and Iksen

Department of Pharmaceutical Chemistry, Universitas Sumatera Utara, Jl. Tridharma Kampus USU ,Medan, Indonesia

Keywords: Chives, Kidney Stones, Calcium, Fraction, Solubility.

Abstract: The aim of this research was to find out whether the ethyl acetate and hexane fractions had an effect to

dissolve the level of calcium in human kidney stones.The solubility activity of chives leaves ethyl acetate

and hexane fraction were carried out by using the in vitro method which was to test the solubility of the

calcium components of human kidney stones in various concentrations (1%, 2.5% and 5%). The fractions

were added by human kidney stones and incubated on 37

C for four hours. The ammount of calcium

solubility was measured by using atomic absorption spectrophotometer at 422.7 nm. The results showed that

ethyl acetate fraction had a dissolving effect of calcium on the human kidney stone which are respectively

18.98 µg/mL, 32.84 µg/mL and 26.22 µg/mL better than hexane fraction which are respectively 2.02 µg/mL,

2.45 µg/mL and 3.03 µg/mL In these results, the concentration of 2.5% of ethyl acetate fraction showed the

highest dissolving level which is 91.99%. Based on the research, it can be concluded that ethyl acetate

fraction of chives leaves is a new potential for herbal treatment of kidney stones.

1 INTRODUCTION

Kidney stones are one of the most common diseases

in the kidneys. Generally, kidney stones occur

because the body lacks fluids resulting in blockages

in the channels from the kidneys to the bladder. The

stones in the kidney are mainly formed from

chemicals that are usually found in urine such as

calcium, uric acid, phosphate, cystine and others

chemicals compound. Different types of treatments

are applied for kidney diseases such as medicine,

changing of life style, treatment by surgery etc.

(AUA, 2014; Mugni, 2013).

According to Al-Snafi (2013) and Iksen (2015),

chives are believed to be a multifunctional medicine

for various diseases. Chives contain various

phytochemical compounds including alkaloids,

flavonoids, glycosides, steroids, tannins and various

minerals such as potassium, calcium, magnesium

and sodium. Previous research showed that infuse

water of chives has a calcium oxalate solubility [5].

However, until now there is no scientific proof of

human calcium kidney stone of ethyl acetate and

hexane fractions of chives leaves. The purpose of

this study is to study the effect of chives leaves ethyl

acetate and hexane fractions on human calcium

kidney stones solubility.

2 MATERIALS AND METHODS

2.1 Materials

Distilled water, human calcium kidney stones, fresh

chives leaves, nitric acid 65% (Merck), ethanol

solvent (Merck), ethyl acetate solvent (Merck),

hexane solvent (Merck) and standard solution of

calcium 1000 ppm.

2.2 Plant Material Preparation

Fresh chives leaves was collected from local area of

Simalungun distric (North Sumatera, Indonesia) and

authenticated by Herbarium Medanese (MEDA)

Universitas Sumatera Utara. Voucher specimen was

collected and deposited in Pharmacognosy

Laboratory, Faculty of Pharmacy, Universitas

Sumatera Utara.

914

Sinaga, S., Sudarmi, . and Iksen, .

The Effect of Chives (Allium schoenoprasum L.) Leaves Ethyl Acetate and Hexane Fractions on Human Calcium Kidney Stones Solubility by In Vitro Method.

DOI: 10.5220/0010101109140916

In Proceedings of the International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches (ICOSTEERR 2018) - Research in Industry 4.0, pages

914-916

ISBN: 978-989-758-449-7

2.3 Preparation of Fraction

Extraction was done by maceration method using

ethanol solvent. 200 g of powdered chives leaves

were marcerated in 1 L ethanol solvent 24 hours,

then filtered, do it continuously until the filtrate

obtained is clear and colorless. 10 g concentrated

ethanolic extract, then fractioned using ethyl acetate

and hexane solvents to get the ethyl acetate and

hexane fractions (Ditjen POM, 1995; Depkes RI

1995).

2.4 Experimental Design

In this research, samples were divided into 6 groups,

which were namely EA1 (1% ethyl acetate

fraction), EA2 (2.5% ethyl acetate fraction), EA3

(5% ethyl acetate fraction), H1 (1% hexane

fraction), H2 (2.5% hexane fraction), and H3 (5%

hexane fraction). All of these groups were added by

human calcium kidney stone and incubated at 37

for four hours. Calcium level was measured before

and after incubation by using atomic absorption

spectrophotometry method at 422.7 nm wavelenght.

2.5 Calcium Calibration Curve

Calcium calibration curve was prepared by using 6

different concentration which were 0 ppm, 0.2 ppm,

0.4 ppm, 0.6 ppm, 0.8 ppm and 1,0 ppm. All of these

concentrations will be measured by using atomic

absorption spectrophotometry method at 422.7 nm

wavelenght.

2.6 Calcium Determination in Sample

Before measurement of calcium level, the organic

compounds should be destructed by using nitric acid

65% which heated on a hot plate until the sample

solutin become transpicious. After that, the sample

solution measured using air-acetylene flame at

422.7 nm wavelenght.

3 RESULTS AND DISCUSSIONS

3.1 Calcium Calibration Curve

Calcium calibration curve is obtained by measuring

the absorbance of the standard solution at a

wavelength of 422.7 nm for calcium. Calibration

curve of calcium standard solution is as shown in the

figure below (Fig. 1). From the calculation results

obtained the correlation coefficient (r) of 0.9997 and

the regression line equation Y = 0.101443X +

0.000795. These results indicate that there is a linear

relationship between concentration and absorbance,

which is indicated by the value of the correlation

coefficient that is close to 1. The value of the

correlation coefficient obtained meets the

predetermined requirements, namely the value of the

correlation coefficient of not less than 0.995.

Figure 1: Calibration curve of calcium

3.2 The Effect of Chives Leaves Ethyl

Acetate and Hexane Fractions on

Solubility of Human Calcium

Kidney Stones

The level of of human calcium kidney stone

solubility is presented in Table 1. Based on the

results, it can be seen that ethyl acetate fraction

within dose 2.5% gave the highest activity in

dissolving the kidney stone campared to other

fraction. This can happen due to the potassium

which could happen because potassium is able to

push the calcium bond in kidney stone and can be

remove through urine (Putra et al., 2018; Iksen,

2015).

Other possibility is because of the phytochemical

compound especially flavonoid compounds which

are the main compound in ethyl acetate fraction. The

mechanism by flavonoid maybe caused by its

improving the dissolving effect by making a

complex bond with the calcium from kidney stone.

Calcium complex bond with flavonoid will be free

and can be remove through urine (Haro et al., 2017;

Sinaga et al., 2018).

4 CONCLUSIONS

Chives leaves ethyl acetate fraction solution with

2.5% concentration gives the highest activity in

dissolving calcium kidney stone. This can happen

due to the potassium and flavonoids compound from

chives leaves. It is hoped that this study could be an

alternative for the treatment of kidney stones

disease.

The Effect of Chives (Allium schoenoprasum L.) Leaves Ethyl Acetate and Hexane Fractions on Human Calcium Kidney Stones Solubility

by In Vitro Method

915

Table 1: The Effect of Chives Leaves Ethyl Acetate and Hexane Fractions on Solubility of Human Calcium Kidney Stones

No Treatments Calcium Level (μg/mL) Solubility

Before Incubation After Incubation (μg/mL) (%)

1 EA1 (1%) 1.81 20.79 18.98 91.30

2 EA2 (2.5%) 2.85 35.70 32.84 91.99

3 EA3 ( 5%) 3.74 29.97 26.22 87.49

4 H1 (1%) 11.78 13.80 2.02 14.63

5 H2 (2.5%) 13.71 16.17 2.45 15.15

6 H3 (5%) 20.88 23.91 3.03 12.67

ACKNOWLEDGEMENTS

The authors acknowledge the financial support by

Research Institutions, University of Sumatera Utara,

in accordance with the Contract Research

TALENTA (No: 2590/UN5.1.R/PPM/2018)

Universitas Sumatera Utara, Fiscal Year 2018.

Authors also thanks to Kevin and Marselina

Purnama Sari for their help for this research project.

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