EFFECT OF NONI JUICE ON KIDNEY PARAMETERS

BY
ORJI JANE CHIOMA

A PROJECT PROPOSAL SUBMITTED TO THE DEPARTMENT OF

PHARMACEUTICAL TECHNOLOGY, SCHOOL OF INDUSTRIAL AND APPLIED
SCIENCES, FEDERAL POLYTECHNIC NEKEDE OWERRI, IMO STATE.

IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE AWARD OF

HIGHER NATIONAL DIPLOMA (HND) IN PHARMACEUTICAL
TECHNOLOGY.

JUNE, 2022

1
 CHAPTER ONE

INTRODUCTION

1.1. Background of Study

It has been considered that if a drug is effective, it will have side effects. Therefore, herbal

medicines as drugs either have side effects or are ineffective. However, herbal medicines

are generally considered to be safe and effective agents. Therefore, people every year turn

to herbal medicine because they believe plant remedies are free from undesirable side

effects (Philomena, 2011). Medicinal plants represent the most ancient form of medication,

used for thousands of years in traditional medicine in many countries around the world.

The empirical knowledge about their beneficial effects was transmitted over the centuries

within human communities (Khan, 2014). Natural products play a pivotal role as a source

of drug compounds and, currently, a number of modern drugs which are derived from

traditional herbal medicine are used in modern pharmacotherapy (Patwardhan et al., 2008).

According to a report by the WHO (2008), about 80%of the people in developing countries

rely on traditional herbal mixtures to treat different diseases. Most villages in Africa still

depend solely on traditional herbal mixtures as a source of health treatments because of

their beliefs and culturally acceptable indigenous knowledge, accessibility, and

affordability. Many herbal mixtures are indubitably expedient for maintaining good health

or treating diverse diseases (Bandaranayake, 2006; Bodeker et al., 2005). Notwithstanding,

as was stated by Okaiyeto and Oguntibeju (2021) that uncontrolled consumption of herbal

mixtures could lead to liver damage, kidney failure and stomach upsets, diarrhoea, etc.

2
1.2. Statement of Problem

Kidney injury is a significant clinical problem and is considered to be the main cause of

acute renal failure, which can result from shock, partial nephrectomy, or renal

transplantation and can lead to morbidity and mortality (Torres-González et al., 2018).

Multiple pathogenic factors contribute to the eventual death of kidney cells as a result of

I/R, including excessive oxidative stress, actions of proinflammatory cytokines,

recruitment of inflammatory cells, and apoptosis (Torres-González et al., 2018). The use

of herbs in treating diseases has a history of thousands of years. During the past decades,

herbs or their extracts have gained increasing attention worldwide due to their significant

efficacy in treating and preventing some diseases (Prabhakar et al., 2019; Jeong et al.,

2018). Notwithstanding, herbs have been reported to cause kidney injury (Kosanam and

Boyina, 2015), and this, among many causes, is a result of misuse and abuse of herbs

(Corsini and Bortolini, 2013; Sasaki and Yokoi, 2018) which includes dosage and course

of treatment (Devarbhavi et al., 2013; Chen et al., 2013). Since lack of dosage has been a

hindrance to the use of plants as medicine, this research work will help determine the safe

dosage level in the consumption of the herb Noni.

1.3 Justification of the Study

Morinda citrofolia has been associated with its medicinal uses. It continues to be in use,

and as a result of the belief of its medicinal value, its consumption seems to be relatively

high and abused as even many persons tend to consume it in its raw state. It is therefore of

great importance to determine its effect on the largest and one of the most important organs

(the kidney) in human since it is involved in drugs excrretion. The success of this research

3
work will sensitize the masses on the safety and/or toxic level of consumption of the plant,

hence, standardizing its consumption.

1.4 Aim of the Study

This research work is aimed at determining the effect of noni Juice on kidney enzymes

1.5 Objectives of the Study

The objectives of this study will be:

• To carry out plant extraction to obtain crude extract.

• To evaluate the effect of the plant extract on the Kidney enzymes using Urea and

Creatinine urea markers.

4
 CHAPTER TWO

LITERATURE REVIEW

Morinda citrifolia L. belongs to the family Rubiaceae, commonly known as Noni. The

genus Morinda comprises some 80 species which all occur exclusively in tropical climate

zones. It has a long tradition as a medicinal plant in Asia pacific countries. Typical uses

have been reported as a treatment of boils and curs, abscesses, fungal infections,

constipation as well as diarrhoea (Potterat and Hamburger, 2007). It has antimicrobial,

anticancer, antioxidant, anti-inflammatory, analgesic and cardiovascular properties (Chan-

Blanco et al., 2006). Many studies of Morinda citrifolia L. juice and isolated compounds

from the fruit has been published including phenolic, volatile compounds and alkaloids

(Krishnaiah et al., 2012). Of the phenolic compounds, the most important reported are

anthraquinones (damnacanthal, morindone and morindin, etc.) and also aucubin,

asperuloside and scopoletin (Krishnaiah et al., 2012; Thongchai et al., 2019).

It is worthy to note that previous study by Thongchai et al (2019) demonstrated that the

crude extract of M. citrifolia fruit showed an antioxidant activity 0.30 times lower than

vitamin C. Another study by Nacimento et al (2018) in its Results showed that Noni seeds

presented higher percentage of free radical scavenging (FRS) and pulp possesses higher

total phenolic contents (TPC) and antioxidant capacity by DPPH method. Oral

administration of deacetylasperulosidic acid isolated from the Noni fruit has been

demonstrated to reduce lipid peroxidation and enhance superoxide dismutase and catalase

activity (ThyagaRagan et al., 2015). Spectroscopic analysis of Noni fruit revealed several

compounds out of which a neolignan, americanin A, has been found to have potent

antioxidant activity (ThyagaRagan et al., 2015).

5
 CHAPTER THREE

MATERIALS AND METHODS

3.1. Materials

3.1.1. Plant Materials

Fresh Juice of Morinda citrifolia will be brought from the market in Abiaokpor, a suburb

of Abak in the state of Akwa Ibom State, Nigeria. The plant sample will be identified by a

Botanist, Dr. Duru, C.N. of Environmental Biology Federal Polytechnic Nekede.

3.1.2. Animals

Wistar rats (8-12 weeks old) with an average weight of 120.11 g will be used for this study.

These animals will be purchased from a local breeder in Ihiagwa Owerri-West L.G.A of

Imo State. The animals will be kept in well-aerated stainless steel wire cages in the animal

house of the Department of Biochemistry. The rats will be given standard feed for at least

two weeks after purchase to acclimatize them to the laboratory environment before use.

3.1.3 Chemicals and Reagents

3.1.3.1 Chemicals

All chemicals to be used in this study will be of good and analytical grade.

3.1.4 Instruments/Equipment

The equipment used will be obtained from the Departments of Pharmaceutical Technology

Federal Polytechnic Nekede, Owerri. They include:

6
 Centrifuge PAC, Pacific

Glasswares Pyrex

Haematocrit centrifuge Hawksley, Germany

Micropipette Perfect

Refrigerator Thermocool

Spectrophotometer Spectronic 20D, USA

Syringe Lifescan

Water bath Gallenkamp, England

Weighing balance Mettler HAS, USA

3.2. Methods

3.2.1. Experimental design.

The rats will be weighed and mean weight will be obtained. They will be then divided into

5 groups. Group 1 as Normal Control, Group II as Negative control, Group III as a positive

control, Group IV as Test control & Group V as a Test control.

Kidney injury will be induced by ischemia caused by 45 min of occlusion of the renal

pedicle using vascular clamps, after which the clamps will be withdrawn and reperfusion

will be allowed for 15 h. During this period, rats will be allowed access to food and water

ad libitum. Groups III-V will be induced by ischemia for kidney injury.

Group I (Normal control): will be fed with feed and water only

Group II (Negative control): will be induced but was not treated.

Group III (positive control): will be administered standard drug

7
Group IV (Test control): animals will be administered 400mg/kg of Noni fruit juice

orally.

Group V (Test control): animals will be administered 200mg/kg of Noni fruit juice orally.

Animals will be sacrificed 24hours after the last administration. Blood will be taken from

the heart and centrifugated using a centrifuge to separate the hemolytic for analysis.

3.2.2. Kidney Function Test.

Determination of urea concentration

Urea level will be determined using Berthelo’s urease reaction method, as described by

Fawcett (1960).

Principle: Urea in serum is hydrolysed to ammonia in the presence of urease. The

ammonia coupled to sodium nitroprusside in an alkaline medium to form a colour that is

read colorimetrically.

Urea + H2O Urease 2NH3 + CO2

NH3 + hypochlorite + phenol → Indophenol (blue compound)

Reagents: EDTA (R1) (116mmol/l), Sodium nitroprusside (6mmol/l), Urease (1g/l),

Diluted phenol (R2) (120mmol/l), Diluted sodium hypochlorite (R3) (27mmol/l), Sodium

hydroxide (0.4N), Urea standard

8
Determination of Serum Creatinine

Serum creatinine will be determined using direct end point according to Henry (1974) as

described in redox commercial kit.

Principle: the analysis was based on the principle that creatinine reacts with picric acid in

alkaline conditions to form a colored complex, which absorbs at 510nm. The rate of

formation of color (the intensity of colored compound) is proportional to the creatinine

concentration in the sample, in the end point method, the difference in absorbance

measurements after color formation yields a creatinine value corrected for interfering

substances.

Reagents: picric acid (10mM), sodium borate (10mM), sodium hydroxide and surfactant

(240mM), creatinine standard (5.0mg/dl)

3.2.3. Statistical Analysis

The results will be presented as mean ± SD. Total variations present in a set of data will be

estimated by One Way Analysis of Variance (ANOVA) using Duncan post hoc test and p

value less than 0.05 will be considered statistically significant.

9
 CHAPTER FOUR

RESULTS

4.1. EXPECTED OUTCOME

The outcome of the study will be drawn from the result of analysis.

4.2. EXPECTED CONCLUSION

The conclusion of the work will be drawn from the result of the analysis.

BUDGET

S/N ITEMS COST

1. Extraction N 2,000

2. Chemicals and solvents N 10,000

3. Animals N 33,000

4. Cage and feed N 10, 000

5. Kidney enzyme Tests

1. Serum urea N 10,000

2. Creatine urea N 10,000

Grand Total N 75,000

10
 REFERENCES

Bandaranayake, W.M. (2006). Quality Control, Screening, Toxicity, and Regulation of

Herbal Drugs. In Modern Phytomedicine: Turning Medicinal Plants into Drugs;
Wiley: Hoboken, NJ, USA: pp. 25–57.

Bodeker, C., Bodeker, G., Ong, C.K., Grundy, C.K., Burford, G. and Shein, K. (2005).
WHO Global Atlas of Traditional, Complementary and Alternative Medicine;
World Health Organization: Geneva, Switzerland

Chan-Blanco, Y., Vaillant, F., Perez, A.M., Reynes, M., Brillouet, J.M. and Brat, P. (2006).
The Noni fruit (Morinda citrifolia L.): A review of agricultural research, nutritional
and therapeutic properties. J. Food Compos. Anal., 19: 645-654.

Chen, M., Borlak, J. and Tong, W. (2013). High lipophilicity and high daily dose of oral
medications are associated with significant risk for drug-induced liver injury.
Hepatology; 58: 388–396

Corsini, A. and Bortolini, M. (2013). Drug-induced liver injury: The role of drug
metabolism and transport. J. Clin. Pharmacol.; 53: 463–474.

Jeong, S.M., Seo, B.K., Park, Y.C. and Baek, Y.H. (2018). A Review of Complementary
and Alternative Medicine Therapies on Muscular Atrophy: A Literature Review of
In Vivo/In Vitro Studies. Evid. Based Complement. Altern. Med. Ecam.; 2018:
8654719

Khan, H. (2014). Medicinal plants in light of history: Recognized therapeutic modality. J.

Evid. Based Integr. Med.; 19: 216–219.

Krishnaiah, D., Nithyanandam, R. and Sarbatly, R. (2012). Phytochemical Constituents

and Activities of Morinda citrifolia L. In: Phytochemicals: A Global Perspective of
their Role in Nutrition and Health, Rao, V. (Ed.). Chapter 6, InTech Publ., Rijeka,
Croatia, ISBN: 978-953-51-0296-0, pp: 127-150.

Nascimento, L.C.S., Rodrigues, N. da R., Alves, M.P.C., Sabaa Srur, A.U.O., Barbosa
Junior, J.L. and Barbosa, M.I.M.J.(2018). Chemical characterization, nutritional
aspects and antioxidant capacity of noni (Morinda citrifolia L) produced in
northeastern Brazil. International Food Research Journal 25(2): 870-875.

Okaiyeto, K. and Ogutibeju, O.O. (2021). African Herbal Medicines: Adverse Effects and
Cytotoxic Potentials with Different Therapeutic Applications. International
Journal of Environmental Research and Public Health; 18: 1-20.

11
Pakkirisamy, M., Daniel, N., and Mani, J. (2021) Antioxidant and Anti-inflammatory
Activity of Nilavembu kudineer choornam : In vitro Evaluation. Med Aromat
Plants 10: p060

Patwardhan, B., Vaidya, A., Chorghade, M. and Joshi, S. (2008). Reverse pharmacology
and systems approaches for drug discovery and development. Curr. Bioact. Compd.
4: 201–212.

Philomena G. (2011). Concerns regarding the safety and toxicity of medicinal plants - An
overview. J Appl Pharmaceut Sci; 01(6): 40-44.

Potterat, O. and Hamburger, M. (2007). Morinda citrifolia (Noni) fruit-phytochemistry,

pharmacology, safety. Planta Med., 73: 191-199.

Prabhakar, A., Kaiser, J.M., Novitch, M.B., Cornett, E.M., Urman, R.D. and Kaye, A.D.
(2019). The Role of Complementary and Alternative Medicine Treatments in
Fibromyalgia: A Comprehensive Review. Curr. Rheumatol. Rep.; 21: 14.

Sasaki, E. and Yokoi, T. (2018). Role of cytochrome P450-mediated metabolism and

involvement of reactive metabolite formations on antiepileptic drug-induced liver
injuries. J. Toxicol. Sci.; 43: 75–87

Thongchai, T., Parisa, S. and Phutdhawong, W.S. (2019). Major chemical composition of
fruit extracts of Morinda citrifolia L. and their antibacterial, antioxidant and
cytotoxicity properties. J. Applied Sci., 19: 366-375.

ThyagaRagan, S., Rethinam, P. and Pratap, U.P. (2015). Pharmacological properties and
clinical applications of Morinda citrofolia. Intl. J. Noni. Res; 10(1&2):1-18.

Torres-González, L., Cienfuegos-Pecina, E., Perales-Quintana, M. M., Alarcon-Galvan,

G., Muñoz-Espinosa, L. E., Pérez-Rodríguez, E., & Cordero-Pérez, P.
(2018). Nephroprotective Effect of Sonchus oleraceus Extract against Kidney
Injury Induced by Ischemia-Reperfusion in Wistar Rats. Oxidative Medicine and
Cellular Longevity, 1–7. doi:10.1155/2018/9572803

World Health Organization (2008). Traditional Medicines. Retrieved from

http://www.who.int/mediacentre/factsheets/fs134/en/ (accessed on 8 October
2020).

12