Effectiveness of Green Coconut (Cocos nucifera L.) Water against

Heavy Metal Levels in the Blood of Rattus norvegicus

Yuanita Rachmawati

, Ika Mustika

, Esti Tyastirin

, Reni Ida Wati

, Ayudya Fitri Arifa

Humairah Azzahra

, Aimmatul Maghfiroh

, Meilinda Rodhiya Idrus

, Marsono

Mohamad Khafid Abdullah

, Lailatul Amaliyah

, Islamiati Ningrum

Science and Technology Faculty, Universitas Islam Negeri Sunan Ampel of Surabaya

Health and Psychology Faculty, Universitas Islam Negeri Sunan Ampel of Surabaya

Keywords: Cocos nucifera L. water, Heavy Metal (Pb, Hg, Cd,), Rattus norvegicus

Abstract: The environmental pollution can endanger human health if it comes from waste that has high toxicity such as

heavy metal waste. Heavy metals that are often found in our environment are metal lead (Pb), mercury (Hg),

cadmium (Cd), arsenic (As), and chromium (Cr), but Pb, Hg, and Cd are the most common causes of human

health problems. Green coconut (Cocos nucifera L.) water is widely consumed and believed to be an antidote.

Green coconut water itself is one of the local wisdoms of herbal plants. Even though substances contained in

coconut water are unknown to the ordinary people, coconut water is believed to be able to neutralize toxins

in the body. The purpose of this research is to examine the effectiveness of green coconut (C. nucifera L.)

water in reducing level of heavy metals (Pb, Hg, and Cd) in the blood of Rattus norvegicus L. The Pb, Hg,

and Cd groups were treated each 2 mg/mL of 3 mL/week, while the combination group was given Pb-Hg-Cd

each of 1 mg/mL as much as 3 mL/week. Treatment of heavy metals was carried out 3 times in a week for 4

weeks, while coconut water treatment to the Cocos group was given every day. The results showed that there

were significant differences between the average Cd levels (p.value 0.009) and the average Hg levels (p.value

0.007). Cd level in the Cd group was found to be 0.223 ppm, whereas in the Cd-Cocos group, it was 0.096

ppm. The average Hg level in the Hg group was 276.15 ppm while in the Hg-Cocos group was 81.81 ppm. In

the Pb group, there was a difference in the mean of treatment results, even though the difference was not

significant in the Kruskal-Wallis test. This proves that C. nucifera can effectively reduce heavy metal levels,

especially in Cd and Hg, in the blood of Rattus norvegicus.

1. INTRODUCTION

Environmental pollution is a classic problem that

continues to haunt until now. One of the pollutions

that still becomes a concern is due to heavy metals.

Water in Surabaya, in Kalimas sediment, to be exact,

contained Pb (103,219-138,621 ppm) and Hg

(11,984-13,525 ppm), according to a survey

conducted by Ariestya in 2008. Similar research

conducted on sea cucumber in the coastal area of

Kenjeran found metal level of Hg ranged from 0.099

to 0.112 ppm and Pb 9.86 to 15.27 ppm on sea

cucumber (Lestari, 2015). In Surabaya, Kupang

traditional foods contain heavy metals as well; Pb

metal is 1,281 ppm and Cd is 1,254 ppm (Indasah,

2011). The measurement results of heavy metal

concentrations in air ambient in 10 major cities in

Indonesia found that Surabaya was ranked first in Pb

pollution, followed by Tangerang and Jakarta. The

average Pb level in ambient air in Surabaya is 10.43-

2664.2 ng/m

(Mukthar, 2013). In 2017, a research

conducted in the area of Bandung found that one of

the rice fields there contained heavy metal Hg of 92.2

mg / L. This value is above the Hg quality standard,

which is 25.9 mg/L (Komarawidjaja, 2017).

Metal pollution can endanger human health.

Health problems can be caused by heavy metals,

depending on the level of poisoning experienced by

the individual. For example, Pb Poisoning can attack

the central nerve, inhibit enzyme reactions, shorten

the life of red blood cells, increase iron (Fe) content

in blood plasma, damage the Cerebrum, and inhibit

fetal growth (Ratmini, 2009).

Lead entering through breathing process will

also circulate to all tissues and organs of the body.

More than 90% of lead absorbed by the blood is

Rachmawati, Y., Mustika, I., Tyastirin, E., Wati, R., Arifa, A., Azzahra, H., Maghﬁroh, A., Idrus, M., Marsono, ., Abdullah, M., Amaliyah, L. and Ningrum, I.

Effectiveness of Green Coconut (Cocos nucifera L.) Water against Heavy Metal Levels in the Blood of Rattus norvegicus.

DOI: 10.5220/0008907200002481

In Proceedings of the Built Environment, Science and Technology International Conference (BEST ICON 2018), pages 113-118

ISBN: 978-989-758-414-5

113

bound to red blood cells (Rahmawati et al., 2016).

Consumption of lead 0.2-2 mg/day can cause food

poisoning. While an increase of lead level by 10µg/dl

in blood can result in an IQ reduction of 6-9 points

(Anggraini, 2014). Meanwhile, mercury level in the

tissues of 0.1-1 ppm can cause disruption of body

function.

Cocos nucifera L. is an important member of the

family Arecaceae (palm family), usually called

coconut (Aragao, 2002). The plant is originally from

Southeast Asia (Malaysia, Indonesia, and the

Philippines) and the islands between the Indian and

Paciﬁc Oceans (Purseglove, 1972). All parts of the

coconut can be used. C. nucifera L. gives human

many benefits, especially from its water. Compared

to other types of coconut, water of green coconut (C.

nucifera L.) contains more bioactive enzymes such as

tannins. Tannins are anti-toxic substances, so that

green coconut water is often considered capable of

decomposing and removing toxins in the body

(Candra, 2016).

The Javanese believe that water of green

coconut (C. nucifera L.) contains an antidote, without

knowing what substances are in the water. Green

coconut water itself is one of traditional medicines

based on local wisdom and is believed to be able to

neutralize toxins in the body.

The results of giving young coconut water to

painting workers exposed to Pb for 2 weeks have

shown significant reduction of the average Pb level in

the blood, from 35,009 µg/dl to 9,0089 µg/dl (Candra,

2016). Research conducted on workers exposed to

mercury (Hg) in the gold mining area in Selogiri has

showed an increase in the activity of glutathione

peroxidase (GPx) enzymes after the treatments of

coconut water as an antioxidant (Zulaikha et al.,

2015).

The effectiveness of coconut water in reducing

the level of Cadmium (Cd) in the blood has not been

widely studied. This study will measure , the

effectiveness of C. nucifera L. in reducing the level

of heavy metals (Cd, Pb and Hg) in the blood of

Rattus norvegicus or white rats.

2. METHOD

The research was conducted at the Integrated

Laboratory of UIN Sunan Ampel Surabaya from

October to December 2017. The experimental animal

used for this study was Rattus norvegicus L., or white

rats, that were controlled for all treatments.

Experimental research method was conducted as

Complete Randomly Design (CRD) with the

treatment group, namely ‘the control group’, ‘metal

treatment group of Pb, Hg, and Cd’, ‘the treatment

group of green coconut water’, and ‘the combination

of the 3 treatments of heavy metals with green

coconut water’. Each group has 3 repetitions.

Heavy metal treatments were as follows: Lead

(Pb) group was given Pb 2 mg/mL as much as 3

mL/week; Mercury (Hg) group was given 2 mg/mL

of Hg as much as 3 mL/week; Cadmium (Cd) group

was given Cd 2 µg/mL as much as 3 mL/week as

much as 3 ml/week; Combination group was given

Pb+Hg+Cd each of 1 µg/mL, 3 mL/week. Heavy

metal treatment was given 3 times in 1 week for 4

weeks. Coconut water was given to the Cocos group

every day.

The data in this study were primary data,

obtained from direct measurements of total Hg levels

in blood (THg), total Pb in blood (TPb), and total

levels of Cd in blood (TCd) of Rattus norvegicus L.

taken intracardially. Heavy metal levels testing in rat

blood used the Atomic Absorbance

Spectrophotometry (AAS) method.

The measurement results of metal content in the

blood were in the form of data ratios. Comparison

between levels of heavy metals in experimental

animals with the provision of mineral water and

coconut water and the metal content after the

treatment of giving coconut water was then tested

using statistical analysis. The results of the data were

statistically analyzed using Kruskall-Wallis and

Kolmogorov-Smirnov tests, while further testing was

done with Mann-Whitney if there was significance in

the results.

3. RESULT AND DISCUSSION

3.1 Cadmiun Analysis

On table 1 showed measurement results of cadmium

heavy metals in various treatments, including control

group, control group with coconut water (Control-

Cocos), Cd-treatment group, Cd-treatment group

with coconut water (Cd-Cocos), group combination

of Pb + Hg + Cd (Mix) treatment, and combination

treatment group Pb + Hg + Cd with coconut water

(Mix-Cocos).

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Table 1: Result of average Cd concentration

Treatment

Average Cd

Concentration

(ppm)

P-Value

Control 0.003

0.009

Control-Cocos 0.003

Cd 0.223

Cd-Cocos 0.096

Mix 0.069

Mix-Cocos 0.014

unsignificantly different

Based on the calculation of Kruskall-Wallis test,

a significance value of 0.009 (smaller than α) was

obtained. So, it can be concluded that there is a

significant difference in the average Cd level between

each group. Further analysis with Mann-Whitney

showed a significant difference (P-Value 0.04)

between the Cd group and the Cd-Cocos group, and a

significant difference between the groups of Mix and

the mix-Cocos group. It can be seen in the table that

the average Cd level in the treatment group was 0.223

ppm, while in the Cd-Cocos group, the average Cd

lowered to 0.096 ppm. It can be said that the

administration of coconut water treatment can reduce

cadmium level in the blood of rats.

Cadmium accumulated in the body can cause

nephrotoxicity (kidney toxicity), symptoms of

proteinuria, glycosuria, and aminoaciduria

accompanied by a decrease in the renal glomerular

filtration rate. Cadmium causes osteomalacia

symptoms because of the balance between calcium

and phosphate in the kidneys (Muhajir, 2009).

3.2 Mercury Analysis

Table 2: Result of Average Hg Conentration

Treatment

Average Conentration

(ppm)

P-Value

Control 0

0.007

Control-Cocos 0

Hg 276.15

Hg-Cocos 81.81

Mix 65.39

Mix-Cocos 13.11

unsignificantly different

Table 2 has showed measurement results of

mercury metals in various treatments, including

control group, control group with coconut water

(Control-Cocos), Hg-treatment group, Hg-treatment

group with adding of coconut water (Hg-Cocos),

group combination of Pb + Hg + Cd (Mix) treatment,

and combination treatment group Pb + Hg + Cd with

coconut water (Mix-Cocos).

Kolmogorov-Smirnov test showed that the data

were not normally distributed (P-value is smaller than

0.05), so that non-parametric analysis was done by

Kruskall-Wallis test. The test results showed that

there was a significant difference in the average Hg

level between each treatment group. The significant

different groups were the Hg group with the Hg-

Cocos group, the mixed group, and the mix-Cocos

group.

The content of mercury in the body is very

dangerous. Mercury poisoning can interfere

withblood components, namely an increase in the

levels of amino acids levulinie (ALA) in the blood

and urine, increased level of

 protoporphyrin in red

blood cells, reduced number of red blood cells, the

cause of hemopoietic, and increased level of

hematrocytes in the blood (Aryani, 2013).

3.3 Lead Analysis

The Table 3 has showed measurement results of lead

in various treatments, including control group,

control group with adding of coconut water (Control-

Cocos), Pb-treatment group, Pb-treatment group with

adding of coconut water (Pb-Cocos), group

combination of Pb + Hg + Cd (Mix) treatment, and

combination treatment group of Pb + Hg + Cd with

coconut water (Mix-Cocos).

Table 3: Result of Average Pb Conentration

Treatment

Average

Conentration (ppm)

P-Value

Control 0

0.081

Control-Cocos 0

Pb 0.12

Pb-Cocos 0.04

Mix 0.11

Mix-Cocos 0.03

unsignificantly different

Based on the Kolmogorov-Smirnov test, the

results showed that the average data distribution of Pb

levels was not normal, so the analysis was continued

with the Kruskall-Wallis test. Significance value of

0.081 showed that there was no significant difference

in the average Pb level between groups. Although the

statistical analysis showed no significant differences,

there was a decrease in the level of lead concentration

in the blood of rats treated with coconut water. In

Effectiveness of Green Coconut (Cocos nucifera L.) Water against Heavy Metal Levels in the Blood of Rattus norvegicus

115

coconut water, there are many amino acids such as

glutamate acid, arginine, leucine, lysine, proline,

aspartate acid, alanine, histidine, phenylalanine,

serine, cysteine and tyrosine. This allows the

reduction of lead levels in the blood (Anggraini,

2014).

The research of Jesuorsemwen et al. in 2016



determined the ameliorative effect of coconut water

on haematobiochemical changes due to lead

poisoning in wistar albino rats. For six weeks, sixty

rats were assigned to four groups. Lead 0.10g/l and

75 ml coconut (C. nucifera L.) water were given

orally for six weeks with the mean values of red

blood cells, mean corpuscular volume, mean

corpuscular haemoglobin, mean corpuscular

haemoglobin concentration, red blood cell

distribution width and platelets

, white blood cells,

lymphocytes, total cholesterol, triglyceride, high

density lipoprotein-cholesterol, low density

lipoprotein-cholesterol, low density lipoprotein-

cholesterol/ high density lipoprotein-cholesterol, and

total cholesterol/high density lipoprotein-cholesterol

indicate that coconut water could ameliorate effects

of lead toxicity.

The Javanese believe green coconut water (C.

nucifera L.) is an antidote to poison without knowing

what it is inside coconut water. This belief is passed

down through generations, so that when symptoms of

poisoning occur, parents often recommend

consuming green coconut water. Green coconut water

itself is one of traditional medicines based on the

local wisdom which are believed to be able to

neutralize toxins in the body. Throughout history,

humans have used medicinal plants therapeutically.

Minerals, plants, and animals traditionally have been

the main sources of drugs. The constituents of C.

nucifera have some biological effects, such as

antihelminthic, anti-inﬂammatory, antinociceptive,

antioxidant, antifungal, antimicrobial, and antitumor

activities (Lima et al., 2015).

Compared to other types of coconut, green

coconut water contains more bioactive enzyme called

tannins. Tannins are anti-toxic substances so that

green coconut water can decompose and remove

toxins in the body. Coconut water, aside from being a

fresh drink, also contains various minerals, vitamins,

and sugars as well as essential amino acids, so that it

can be categorized as a highly nutritious soft drink

and can cure various diseases.

Micronutrients in coconut water, such as

inorganic ions and vitamins, play vital roles in

helping the antioxidant defense system of the human

body (Evans & Halliwell, 2001). Some evidence

points toward an antioxidant action of coconut water.

From the research done by Loki & Rajamohan in

2003, administering coconut water (6 mL/100 g of

body weight) to female rats intoxicated with carbon

tetrachloride recovered the action of antioxidant

enzymes (superoxide dismutase and catalase levels)

and decreased lipid peroxidation. Coconut water is

also rich in L-arginine (30 mg/dL), which

signiﬁcantly reduces the generation of free radicals

(Boger et al., 1995) and has antioxidant activity (Salil

& Rajamohan, 2001). It is rich with ascorbic acid (15

mg/100 mL) as well, which decreases lipid

peroxidation in rats (Das et al., 2001). Furthermore,

many parts of C. nucifera plants have proven to

contain phenolic compounds and ﬂavonoids that

support antioxidant activity.

Coconut water contains antioxidants, such as

vitamin C, vitamin B1, vitamin B6, amino acids like

methionine, L-Arginine, selenium, cytokines, and

minerals. Lead that enters the body will generally

moves freely in the form of Pb (II) ions and

accumulates in body tissues. Accumulation occurs

due to the high affinity between Pb (II) ions and

sulfhydryl groups in amino acids. In coconut water,

there are many amino acids such as glutamate acid,

arginine, leucine, lysine, proline, aspartate acid,

alanine, histidine, phenylalanine, serine, cysteine and

tyrosine . This makes it possible to reduce blood lead

levels.

Basically, almost all heavy metals, such as Hg

and Cd, have high affinity for sulfhydryl (-SH) groups

in amino acids. The decrease in these groups causes a

decrease in the antioxidant system which is

characterized by a decrease in the activity of the

enzyme glutathione peroxidase (GPx). Decreased

glutathione (GSH) and H2O2 buildup will cause

oxidative stress on cells. Giving natural antioxidants

is an alternative to protect cell damage that occurs due

to metal poisoning. One source of natural

antioxidants is water of green coconut (C. nucifera

L.). As mentioned earlier, in coconut water there is an

amino acid in the form of methionine. Methionine

acts as a precursor in the formation of cysteine, which

is the main compound in synthesizing gluthionin in

the body. In addition, in coconut water there is L-

arginine which plays a role in mercury detoxification.

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L-arginine is a source of nitric oxide (NO) which can

inhibit xanthine oxide (XO) and increase SOD

(superoxide dismutase). SOD is needed to reduce

superoxide production in the body. C. nucifera L.

has been recognized as an entity with multiple usages.

Each of its components is biologically active in one

way or other.

"Every disease has a cure" (hadith of Muslim

history). Narrated by Muslims from the hadith Abu

Zubair, Jabir bin Abdillah reported, the Prophet SAW

said: "Each disease has a cure. If a cure is applied to

the disease, then it is relieved by the permission of

Allah SWT. "In Sahih Al-Bukhari and Muslim from

'Atha, from Abu Hurairah, that he said: "While in

Musnad Imam Ahmad mentioned the hadith from

Ziyad bin Ilaqah, from Usman bin Shaikh it was

reported that he told, "One time I was with the

Prophet SAW, then suddenly came a number of

Bedouins. They asked, "Salam Messenger of Allah,

should we not treat sickness?" He replied, "Treat

sickness, for Allah has not created any disease except

He has also created the cure, except for one disease."

They asked, "What is the disease, O Messenger of

Allah?" He replied, "Old age." In other words it is

stated, "Every time Allah sends down disease, Allah

definitely delivers the cure. There are only people

who know it and there are people who do not know it.

The publication of the results of this study is

expected to, at least, help to affirm the belief of the

Javanese community in general that green coconut

water is indeed proven to be able to ward off toxins,

particularly toxins caused by the accumulation of

heavy metals in the blood. Allah SWT has created a

Coconut Plant with its water which can be used as a

medicine.

4. CONCLUSION

The results have showed that there were significant

differences between the average Cd levels in (p-value

0.009) and the average Hg levels (P-value 0.007). Cd

level in the Cd group was found to be 0.223 ppm,

whereas in the Cd-Cocos group, it was 0.96 ppm. The

average Hg level in the Hg group was 276.15 ppm

while in the Hg-Cocos group was 81.81 ppm. In the

Pb group, there was a difference in the mean of

treatment results, even though it was not significant

in the Kruskal-Wallis test. This proves that C.

nucifera L. can effectively reduce the level of heavy

metal, especially Cd and Hg, in the blood of Rattus

norvegicus.

ACKNOWLEDGEMENT

We would like to thank The Ministry of Religion

Affair of The Republic of Indonesia for the research

funding provided in 2017.

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