IN-VIVO EVALUATION OF ACUTE TOXICITY OF TRIGONELLA …

IN-VIVO EVALUATION OF ACUTE TOXICITY OF

TRIGONELLA FOENUM-GRAECAUM SEEDS AQUEOUS

EXTRACT ON RENAL FUNCTION AND HISTOLOGY OF

THE KIDNEY USING FEMALE RAT MODEL

BY

TAHA ABDUL QAYYUM AL-SINDI

A dissertation submitted in fulfilment of the requirement for the

degree of Master of Medical Sciences

Kulliyyah of Medicine

International Islamic University Malaysia

SEPTEMBER 2017

ii

ABSTRACT

Introduction; Herbal medicine is largely used for prevention and treatment of several

ailments. Trigonella foenum-graecum is one of the popular medicinal plants which is

widely used as a treatment herb as well as condiment. Countless studies have been

done on its beneficial health effects, while there are few studies on its adverse effect.

Aim: This study is aimed to determine the acute oral toxicity effect of high doses of

aqueous seeds extract of Trigonella foenum-graecum (TSA) following an acute oral

toxicity guideline of the Organization for Economic Cooperation and Development.

Methodology: Twenty-four female Sprague Dawley rats were purchased and

randomly divided into 4 groups (6 rats each), control group was received distilled

water, whereas the 3 tested groups received single high dosage of 3 g/kg, 6 g/kg or 9

g/kg body weight of fenugreek seed aqueous extract. Blood withdrawn after 24 hr.,

day 7 and day 14 of giving the required dose. The rats were sacrificed after 14 days,

required organs were obtained. Results: No mortality witnessed among the treated

groups, rats' behavior remain normal, no clinical signs observed. A mild increase in

breathing were sighted for hours in group that received 9g/kg. No significant changes

were detected in the renal functions. Biochemical analysis of blood serum showed

normal levels of creatinine, urea and total protein. Electrolytes such as sodium,

calcium and potassium were assessed and showed no toxicity. Histopathological

examination revealed mild and moderate hemorrhage and inflammation in all tested

groups. Conclusion: Our results suggest that the oral administration of high doses of

TSA extract shows safety of renal functions. However, adverse effects were remarked

on the kidney architecture which were; inflammation, haemorrhage and glomerular

degeneration.

Keywords: Trigonella foenum graecum, Fenugreek, Toxicity, Safety, Traditional

Medicine, Herbs, Extract, acute oral toxicity, toxic effect.

iii

البحث ملخص

في معالجة الأمراض أو تفاديها. الحلبة -حتى الآنوما زال -م الزمن ه منذ قدلاتعمبدأ اس الأعشابطب مقدمة:فوائده العلاجية؛ إحدى هذه الأعشاب الطبية المعروفة باستعمالاته العلاجية والغذائية. أبحاث عديدة أُجريت لاكتشاف

معرفة الأضرار إلىهدف البحث الحالي ي الهدف: المحتملة.النبتة أضرار هناك قصور في أبحاث السمية أو لكن يظل طريقة البحث:الحلبة المائي. بذور لة التي قد يسببها نبات الحلبة؛ وذلك باستخدام جرعات عالية من مستخلص المحتم

إلى أربع مجموعات، بحيث -أيام من وصولها 5وبعد -فأر في التجربة حيث تم تقسيمها عشوائياً 24تم استخدام (. أعُطيت مجموعات التجربة جرعة ختبار الجرعاتلامجموعات 3و ضابطةفئران )مجموعة 6كل مجموعة على تحتوي

كجم. تم سحب /جرام9كجم أو /جرام6كجم، /جرام3لحلبة على التوالي: بذور اواحدة فقط من المستخلص المائي لتم ذبح -بةمدة التجر -يوم 14بعد ساعة، اليوم السابع وكذلك يوم الرابع العشر من التجربة. 24عينات الدم بعد

تغيّر أي لم يحدث و ، التجربةات في فئران مجموعات وفيلم تكن هناك النتائج:للفحص النسيجي. الفئران وأخذ الكلية كجم /جرام9ملاحظة ارتفاع في معدل التنفس في المجموعة التي أعُطيت جرعة عدا ي للفئران،في النمط السلوكواضح

ات في معدلات الكرياتين من المستخلص. تحليل مصل الدم لم يظُهر اليوريا. المعادن مثل الصوديوم والبوتاسيوم و تغيّرخفيف إلى والتهاب . الفحص المجهري لنسيج الكلية أظهر تجمع دموي وم لم يلُحظ أي تغيّر في مستوياتها أيضاً والكالسي

بشكل عام، فإن :الخلاصة .المجموعة الضابطة/متوسط في أنسجة الكلية في مجموعات التجربة مقارنة بالكونترولفي الفحص طفيفة الجرعات الثلاثة لم تظهر سمية في التحليل الدموي. مع ذلك، فإن أنسجة الكلية قد أظهرت تغييّات

النسيج.

.ادةالحسمية اختبار الأعشاب، مستخلص نباتي، ة: الحلبة، سمية، الطب البديل، الكلمات المفتاحي

iv

APPROVAL PAGE

I certify that I have supervised and read this study and that in my opinion, it conforms

to acceptable standards of scholarly presentation and is fully adequate, in scope and

quality, as a thesis for the degree of Master of Medical Sciences.

…………………………………..

Ahmed Kaid Naji Allow

Supervisor

…………………………………..

Hamoud Hussein Al-Faqeh

Co-Supervisor

…………………………………..

Wael Mohamed Yousef

Co-Supervisor

I certify that I have read this study and that in my opinion it conforms to acceptable

standards of scholarly presentation and is fully adequate, in scope and quality, as a

thesis for the degree of Master of Medical Science.

…………………………………..

Roslina Abdul Rahim

Internal Examiner

This thesis was submitted to the Department of Basic Medical Sciences and is

accepted as a fulfilment of the requirement for the degree of Master of Medical

Sciences.

…………………………………..

Zunariah Buyong

Head, Department of Basic

Medical Sciences

This thesis was submitted to the Kulliyyah of Medicine and is accepted as a fulfilment

of the requirement for the degree of Master of Medical Sciences.

…………………………………..

Azmi Md Nor

Dean, Kulliyyah of Medicine

v

DECLARATION

I hereby declare that this thesis is the result of my own investigation, except where

otherwise stated. I also declare that it has not been previously or concurrently

submitted as a whole for any other degrees at IIUM or other institutions.

Taha Abdul Qayyum Al-Sindi

Signature…………………. Date ………………….

vi

INTERNATIONAL ISLAMIC UNIVERSITY MALAYSIA

DECLARATION OF COPYRIGHT AND AFFIRMATION OF

FAIR USE OF UNPUBLISHED RESEARCH

IN-VIVO EVALUATION OF ACUTE TOXICITY OF

TRIGONELLA FOENUM-GRAECAUM SEED AQUEOUS

EXTRACT ON RENAL FUNCTION AND HISTOLOGY OF THE

KIDNEY USING FEMALE RAT MODEl

I declare that the copyright holder of this thesis/dissertation are jointly owned by the

student and IIUM.

Copyright © 2017 Taha Abdul Qayyum Al-Sindi and International Islamic University Malaysia. All

rights reserved.

No part of this unpublished research may be reproduced, stored in a retrieval

system, or transmitted, in any form or by any means, electronic, mechanical,

photocopying, recording or otherwise without prior written permission of the

copyright holder except as provided below.

1. Any material contained in or derived from this unpublished research may

be used by others in their writing with due acknowledgement.

2. IIUM or its library will have the right to make and transmit copies (print

or electronic) for institutional and academic purposes.

3. The IIUM library will have the right to make, store in a retrieval system

and supply copies of this unpublished research if requested by other

universities and research libraries.

By signing this form, I acknowledged that I have read and understand the IIUM

Intellectual Property Right and Commercialization policy.

Affirmed by Taha Abdul Qayyum Al-Sindi

……..……..………… ……………………..

Signature Date

vii

ACKNOWLEDGEMENT

Alhamdulillah for his countless blessing, all praises to Allah alone سبحانه وتعالى, who

guides me through my life, he brought me up to this level. Thank you Allah for your

never-ending grace, mercy, and provision.

My deepest indebtedness gratitude to my beloved parents, my father Dr. Abdul

Qayyum Al-Sindi and my mother Manzoorah, who encouraged me for further studies,

supported me emotionally and financially and kept praying for me. I dedicate this

research to them for their extraordinary support.

I am grateful to my supervisor Assistant Prof. Dr. Ahmed Kaid Allow, whom I

really have had the pleasure to work under his supervision in one of his projects. He is

a friend, advisor and has provided me extensive personal and professional guidance

about both scientific research and life in general. My highest appreciation goes to him.

My heartfelt gratitude also goes to my co-supervisors Assistant Prof. Dr. Hamoud

Hussein Al-Faqeh and Assistant Prof. Dr. Wael Muhamad for their valuable advices.

I’d like also to thank my colleagues for their delightful collaboration,

Mohamed Yehdih, Dr. Asma Saad, Dr. Taher Elshami, Dr. Mahmoud Syed,

Muhammed Saleh, Naeem Mubarak, Majid Abdul Ghaffar, Khaled Benchoula,

Hamzeh AlKhatib, Dr. Abdulrahman Mohammed Abdualkader, Dr. Yousif Ali,

Hosam Tamimi, Dr. Hidayah Ismawi, Dr. Ali Alqodah, Dr. Hamad Alfarisi, Hassan

Sheikh and Dr. Ekhlas Gazem. You supported me greatly and were always willing to

help me. Acknowledgements extends to the lab technicians at physiology and

biochemical labs.

Finally, yet importantly, my sincere thankfulness to my brothers and sisters,

other family members, and friends for their prayers. My endless du'a for the Ummah,

may Allah raise the standings of Islam and the Muslim Ummah.

viii

TABLE OF CONTENTS

Abstract………………………………………………..…………...…….……………ii

Arabic Abstract ……………………………………….……………………………..iii

Approval Page ……………...…………………………………………………...……iv Declaration……………………………………………….……………………………v Copyright Page……………………………………….………………………………vi

Acknowledgement ………………………………………………...…………………vii

Table of Contents ………………………………...…………………………………viii

List of Tables …...……………………………………………………………………..x

List of Figures …………………………………………………...…..………………xii

List of Abbreviations ……………………………………………………………….xiv

List of Symobls ………………………………………………………………………xv

CHAPTER ONE: INTRODUCTION ...................................................................... ..1 1.1 Background ....................................................................................................... ..1

1.2 Research Gap .................................................................................................... ..2 1.3 Research Questions ........................................................................................... ..2

1.4 General Objective ............................................................................................. ..2 1.5 Specific Objectives ........................................................................................... ..2

1.6 Hypothesis ......................................................................................................... ..3

CHAPTER TWO: LITERATURE REVIEW ......................................................... ..4 2.1 Fenugreek: Botanical aspects ............................................................................ ..4 2.2 Nutritional composition of fenugreek seeds ..................................................... ..6

2.3 Fenugreek therapeutic properties ...................................................................... ..8 2.4 Herbal toxicity .................................................................................................. 10

2.4.1 Herbs toxicity on kidney ........................................................................... 13 2.5 Safety evaluation of Fenugreek Consumption .................................................. 14 2.6 Acute Oral Toxicity (AOT) .............................................................................. 17 2.7 Markers for renal function tests (RFT) ............................................................. 17

CHAPTER THREE: METHODOLOGY…………………………………...…….18

3.1 Aqueous extract preparation of Trigonella foenum-graecum (TSA) …...….....18

3.2 Animal study and experimental procedure …………………..………………..23

3.2.1 Route of administration …………………………………………….……23

3.2.2 Biochemichal and histological procedure ……………………….……….25

3.3 Sample size ……………………………………………………...…………….27

3.4 Dosage selection ………………………………………..………….………….28

3.5 Statistical Analysis…………………………………………..…………….…..28

CHAPTER FOUR: RESULTS AND FINDINGS…………………………...…….29

4.1 Mortilaty rate and General Behavioural Signs .................................................. 29

4.2 Effect on Rats' Body weight .............................................................................. 32

4.3 Effects of HDs of TSA extract on organ's weight index ................................... 34

ix

4.4 Effects of HDs of TSA extract on blood glucose .............................................. 35 4.5 Renal function parameters ................................................................................. 36 4.5.1 Effects of HDs of TSA extract on serum creatinine level .......................... 36

4.5.2 Effects of HDs of TSA extract on serum urea level ................................... 38 4.5.3 Effects of HDs of TSA extract on serum total protein level ....................... 39 4.5.4 Effects of HDs of TSA extract on electrolytes ........................................... 42

4.5.4.1 Effects of HDs of TSA extract on serum sodium level ..................... 42 4.5.4.2 Effects of HDs of TSA extract on serum calcium level .................... 43

4.5.4.3 Effects of HDs of TSA extract on serum potassium level ................. 45 4.5.4.4 Effects of HDs of TSA extract on serum chloride level .................... 44 4.6 Microscopic Observation ................................................................................... 46

CHAPTER FIVE: DISCUSSION ............................................................................. 53 5.1 Mortilaty rate and general behavioural signs .................................................... 53

5.1.1 Determination of LD50…….……………………………………………. 52

5.1.2 Behavioral pattern………………………………………………………..52

5.2 Effect of TAS on body weight .......................................................................... 54

5.3 Effects of HDs of TSA extract on organ's weight index ................................... 54 5.4 Effects of HDs of TSA extract on blood glucose ............................................. 54

5.5 Renal function parameters ................................................................................ 54 5.6 The toxicity effect of HDs of TSA extract on renal histology architecture ...... 55

CHAPTER SIX: CONCLUSION, RECOMMENDATION FOR FURTHER

STUDIES ................................................................................................................... 57 6.1 Conclusion......................................................................................................... 57 6.2 Future studies .................................................................................................... 57

6.3 Study limitations ............................................................................................... 58

REFERENCES………………………………………………………………………58

APPENDIX 1: HERBARIUM VOUCHER SPECIMEN INFORMATION ........ 63 APPENDIX 2: ETHICAL APPROVAL LETTER FROM IACUC-IIUM .......... 67

APPENDIX 3: POSTER FOR IRS 2016 ................................................................. 68

x

LIST OF TABLES Table 2.1 The classification of fenugreek plant 4

Table 2.2 The nutrient composition of fresh fenugreek seeds 7

Table 2.3 A list of fenugreek medicinal species 8

Table 2.4 Summary of some of the fenugreek's effects 9

Table 2.5 Summary of LD50 studies of a fenugreek herb 15

Table 2.6 Summary of adverse effects of fenugreek different extractions 16

Table 3.1 Demonstrates the systematic order of chemical agents and the

time needed for tissue processing

27

Table 3.2 The H & E staining scheme 28

Table 4.1 Mortality rate of acute toxicity dose of TSA extract

administered orally to rats

30

Table 4.2 General Signs and behaviors noticed in control and treated

groups

31

Table 4.3 Effect of TSA extract on rats' body weight 32

Table 4.4 Effect of TSA extract on organ/body weight ratio 34

Table 4.5 Effect of TSA extract on plasma glucose 35

Table 4.6 Effect of TSA extract on creatinine level 36

Table 4.7 Effect of TSA extract on urea level 38

Table 4.8 Effect of TSA extract on total protein level 39

xi

Table 4.9 Effect of TSA extract on sodium level 41

Table 4.10 Effect of TSA extract on calcium level 42

Table 4.11 Effect of TSA extract on potassium level 43

Table 4.12 Effect of TSA extract on chloride 45

xii

LIST OF FIGURES Figure 2.1 Leaves and flowers of fenugreek 5

Figure 2.2 A diagram of possible ways of herbal toxicity occurrence 12

Figure 3.1 Fenugreek seeds after cleaning from foreign materials 20

Figure 3.2 Different colures of Fenugreek seeds. 20

Figure 3.3 Foreign materials found between the seeds 21

Figure 3.4 Process of grinding the seeds 21

Figure 3.5 Process of stirring the solution on a hot plate for 24 hours 22

Figure 3.6 Process of centrifuging the aqueous extract 22

Figure 3.7 Storing the samples at -80°C 23

Figure 3.8 Samples in a powder status after freeze drying for 6 days 23

Figure 3.9 Shows the study groups 25

Figure 3.10 Administration of the doses 25

Figure 3.11 Sonicator machine to prepare the TSA doses 26

Figure 4.1 Effect of TSA extract on body weight 33

Figure 4.2 Effect of TSA extract on plasma glucose 35

Figure 4.3 Effect of TSA extract on serum creatinine level 37

Figure 4.4 Effect of TSA extract on serum urea level 38

Figure 4.5 Effect of TSA extract on serum total protein level 40

Figure 4.6 Effect of TSA extract on serum sodium level 42

Figure 4.7 Effect of TSA extract on serum calcium level 43

Figure 4.8 Effect of TSA extract on serum potassium level 44

xiii

Figure 4.9 Effect of TSA extract on serum chloride level 45

Figure 4.10 General view of healthy kidney tissue of the control group (×100) 46

Figure 4.11 General view of healthy kidney tissue of the control group (×400) 47

Figure 4.12 View of kidney tissue of group A receiving dose 3g/kg of TSA

extract (×100)

47


extract (×100)

48


extract (×400) 48

Figure 4.15 View of Glomerulus of group A received dose 3g/kg of TSA

extract (×400).

49

Figure 4.16 General view of kidney tissue of group B received dose 6g/kg of

TSA extract (×40)

49

Figure 4.17 General view of kidney tissue of group B received dose 6g/kg of

TSA extract (×40)

50

Figure 4.18 18 View of glomerulus of group B received dose 6g/kg of TSA

extract (×400)

50

Figure 4.19 General view of kidney tissue of the group C received dose 9g/kg

of TSA extract (×40)

51


of TSA extract (×40)

51


of TSA extract (×100) 52

xiv

LIST OF ABBREVIATIONS

AOT Acute Oral Toxicity

H & E Hematoxylin and eosin

HDs High doses

ICRACU Integrated Center for Research Animal Care and Use

IIUM International Islamic University Malaysia

g/kg Gram per kilogram

KOM Kulliyyah of Medicine

KOP Kulliyyah of Pharmacy

LD50 Median lethal dose

µg Microgram

No. Numbers

RAE Retinol Activity Equivalents

RFT Renal Function Test

rpm Round per minute

SD Standard deviation

TSA Trigonella foenum-graecum seed aqueous

TM Traditional Medicine

xv

LIST OF SYMBOLS

% Percentage

ºC Degree Celsius

_ Minus

= Equal to

< Less than

> More than

± Plus, minus

µ Micro

1

CHAPTER ONE

INTRODUCTION

1.1 BACKGROUND

Traditional medicine (TM) defined as a total of knowledge, skills and practices

gathered for the maintenance of health or the prevention of illness. TM leans on

beliefs, tradition practices and experiences (World Health Organization (WHO),

2002) which may led to unexpected harmful effects to subjects.

The usage of herbs which have therapeutic properties are known since

antiquity. Approximately 25% of drugs prescribed worldwide are derived from

herbs. As utilization of alternative herbs is expanding to treat numerous diseases,

some herbs might cause adverse effects or potential toxicity. (Wachtel-Galor &

Benzie, 2011).

In Africa, for example, up to 80% of the population uses traditional medicine,

48% in Australia, 70% in Canada and 42% in USA (World Health Organization

(WHO), 2002). As well as, 90% of Germans at some point in their life had used a

natural remedy (Hasan et al., 2009). In Malaysia, 88.9% used TM for health

problems and 87.3% for health maintenance, while whole medical system was the

least used, around 1.9% (Siti et al., 2009). The usage of TM is expanding due to the

absence of awareness of local health practices associated with financial and cultural

factors. One of the herbs used widely in treatment is Trigonella foenum-graecum,

commonly known as Fenugreek.

Fenugreek is a worldwide herb used as a culinary herb and in traditional

medicine. Many studies have discovered its curative properties, while not enough

2

toxicity studies are available. This study aims to feed female Sprague Dawley rats

with high doses (HDs) of Trigonella foenum-graecum seeds aqueous (TSA) extract,

and evaluate the acute toxic effects on the renal functions and its histological

changes.

1.2 Research Gap:

Numerous studies were carried out to find the therapeutic effect of fenugreek seeds,

but less information is available about pernicious and the toxicity effect of the plant.

This study is looking for the possible acute toxic effects of high doses of Trigonella

foenum-graecum seeds aqueous (TSA) extract.

1.3 RESEARCH QUESTIONS:

1- What are the physio-chemical changes in the kidney’s functions after

oral administration of high doses of fenugreek seed aqueous extract?

2- What are the evident histopathological changes in the kidney caused by

admission high doses of fenugreek seed aqueous extract?

1.4 GENERAL OBJECTIVE:

The objective of this study is to evaluate the acute toxic effects of high doses of

fenugreek seed aqueous extract on the kidney in female Sprague Dawley rats.

1.5 SPECIFIC OBJECTIVES:

1- To examine the physio-chemical changes of renal functions after

administration of high doses of fenugreek seed aqueous extract.

2- To elucidate the histological changes in the kidney caused by admission

high doses of fenugreek seed aqueous extract.

3- To assess the levels of blood glucose after administration of fenugreek

3

seed aqueous extract.

4- To monitor the changes in the body weight during the experiment

period.

1.6 HYPOTHESIS:

1- Admission high doses of fenugreek seed aqueous extract may lead to

changes in renal function parameters.

2- Admission high doses of fenugreek seed aqueous extract causes

histopathological changes in the kidneys.

3- No changes would be observing in body weight or glucose level after

admission a dose of fenugreek seed aqueous extract.

4

CHAPTER TWO

LITERATURE REVIEW

2.1 FENUGREEK: BOTANICAL ASPECTS

Fenugreek (Trigonella foenum-graecum) is an annual crop known as a condiment

herb and one of the oldest medical plants recognized. Fenugreek classified under

Fabaceae family (Please refer to Table 2.1 for the complete taxonomical position of

fenugreek). Trigonella, a Latin word means ‘‘little triangle” describing the shape of

its flowers (Ahmad et al., 2016; Flammang et al., 2004). The specie foenum-graecum

means “Greek hay”, denotes its utilization as animal food. Carbonized Fenugreek

seeds were discovered in India, used in commerce long time ago 2000 -1700 B.C.

(Acharya et al., 2006). The leaves are utilized as vegetables in the diet, while the

seeds as seasoning (Srinivasan 2006).

Table 2.1: The classification of fenugreek plant

Kingdom Plantae

Division Magnoliophyta

Class Magnoliopsida

Order Fabales

Family Fabaceae / Leguminoseae

Genus Trigonella

Species foenum-graecum Linn.

(Adopted from Snehlata & Dande, 2012)

5

Fenugreek species' number has been debated. Upwards of 260 Trigonella

species may exist (Acharya et al., 2006; Snehlata & Dande, 2012). About 128 species

alone of fenugreek were documented by Vasil'chenko, while 97 species recorded by

Fazli, and 70 species listed by Hector, Hutchinson and, Rouk and Mangesha

(Mehrafarin et al., 2011).

Morphologically, erect plant, massive roots finger-like, cylindrical pinkish

stem to a height of 30–60 cm, toothed leaves. White to yellowish flowers with 5

petals (Please refer to figure 2.1). The flowers are self-pollinated. Pods are light

brown in colour, sickle shaped, contain 10–20 greenish-brown seeds. Mature seeds

colour is golden yellow. The harvest-time of the fenugreek seeds is after 30–35 from

flowering. The Seeds are 5 mm long, soft, hard, and colored yellow to brownish

(Acharya et al., 2008; Ahmad et al., 2016; Srinivasan, 2006b). The seeds are bitter,

but roasting them decreases its bitterness (Ahmad et al., 2016).

(a)

(b)

Figure 2.1 The fenugreek leaves (a) and flowers (b).

(Adopted from gernot-katzers-spice pages.com, 2012)

6

2.2 NUTRITIONAL COMPOSITION OF FEUNGREEK SEEDS

Different parts of fenugreek plant, leaf and seed, have been used widely for

therapeutic use (Acharya et al., 2008). Fenugreek is commonly known as

nutraceutical plant, due to its medicinal value chemical constituents: steroidal

sapogenins, isoleucine and galactomannans. It had been suggested that these three

chemicals work synergistically to produce health impacts (Acharya et al., 2008).

Fenugreek seeds being used for over 2500 years (Srinivasan, 2006). The

seeds contain considerable amount of fiber, protein, phosphorus and many other

functional components. They are also rich source of vitamins, vitamin A, B1, B2, C

(Ahmad et al., 2016). The composition of fresh fenugreek seeds is given in Table 2.2.

7

Table 2.2: The nutrient composition of fresh fenugreek seeds

Component value/

100g Component

value/

100g Component

value/

100g Component

value/

100g

Protein 30 g Chlorine (Cl) 165 mg Potassium (K) 530 mg Triacylglycerol 4.330 g

Fat 7.5 g Manganese (Mn) 1.5 g Cupper (Cu) 33 mg Diacylglycerols 0.280 g

Fiber 50 g Zinc (Zn) 7.0 mg Sulfur (S) 16 mg Monoacylglycerols 0.180 g

Sapogenins 2 g Chromium (Cr) 0.1 mg Folic acid 84 µg Phosphatidylcholine 0.110 g

Trigonelline 380 mg Choline 50 mg Vitamin B1 0.41 mg Phosphatidylethanolamine 0.036 g

Calcium (Ca) 160 mg Vitamin C 43 mg Vitamin B2 0.36 mg Phosphatidylinositol 0.009 g

Magnesium

(Mg) 160 mg β-Carotene 96 µg Vitamin B6 0.6 mg Free fatty acids

0.160 g

Phosphorus (P) 370 g Thiamine 340 µg Niacin 6.0 mg

Iron (Fe) 14 mg Riboflavin 290 µg Vitamin A,

RAE

3.0 µg-

RAE

Sodium (Na) 19 mg Nicotinic acid 1.1 mg Vitamin A 60-100

IU

(Adopted from Srinivasan 2006; Ahmad et al. 2016; Chatterjee et al. 2010)

7

8

2.3 FENUGREEK THERAPEUTIC PROPERTIES

Fenugreek is recorded longtime ago as a therapeutic herb both in traditional Chinese

and Ayurvedic medicines (Acharya et al., 2006). Chinese herbalists used fenugreek

for kidney diseases and also for some male reproductive tract problems (Snehlata &

Dande, 2012). There are 13 species of fenugreek which contain medical properties

listed in Table 2.3.

Whole seed is utilized traditionally to increase lactation (Srinivasan 2006). A

study proved that fenugreek has antioxidant potential to protect vital organs against

diabetes induced oxidative stress damage (Tripathi & Chandra, 2010). See Table 2.4

for some other beneficial effects of fenugreek.

Table 2.3: A list of fenugreek medicinal species:

1 T. foenum-graecum 2 T. balansae

3 T. caerulea 4 T. calliceras

5 T. corniculata 6 T. cretica

7 T. lilacina 8 T. maritima

9 T. occulta 10 T. polycerata

11 T. radiata 12 T. spicata

13 T. spinos

(Adopted from Mehrafarin et al., 2011)

9

Table 2.4: Summary of some of the fenugreek’s effects:

No Fenugreek effect Plant’s part

usage

Some

References

1 Analgesic Effects seeds (Vyas et al.,

2008)

2 Ulcer Protective Effect seeds (Suja et al., 2002)

3

Management of Diabetes:

A. Hypoglycemic Effect seeds (Khosla et al.,

1995)

B. Anti-hyperglycemic Effect leaves (Abdel-Barry et

al., 1997)

4 Immuno-stimulatory Effect Total plant

extract

(Bin-Hafeez et

al., 2003)

5 Serum Cholesterol Lowering

Effect leaves

(Chaturvedi &

Pant, 1987)

6 Lipid-Lowering effect leaves (Annida et al.,

2005)

7 Hypocholesterolemic effect seeds (Rao et al., 1996)

8 Antioxidant Effects seeds (Ravikumar &

Anuradha, 1999)

9 Anti-Inflammatory effects seeds (Vyas et al.,

2008)

10 Increase Lactation

seeds (Hassan et al.,

2012)

11 Anti-Fertility Effect seeds

(Petropoulos,

2004); (Hilles et

al., 2016)

However, these listed effects are fenugreek's effect alone. There are series

studies on fenugreek effects in combination with other compounds. For instance, a

study suggested that fenugreek seeds extract in combination with honey accelerate

IN-VIVO EVALUATION OF ACUTE TOXICITY OF TRIGONELLA …

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