Using Traceability System to Assess Some Heavy

Republic of Iraq Ministry of Higher Education and Scientific Research Al-Mustansiriyah University College of Science

Using Traceability System to Assess Some Heavy Metals, Nutrients and Microbial Factors In Infants Milk Formula By ICP-MS Technology A Thesis Submitted to the Council of College of Science Al-Mustansiriyah University in Partial Fulfillment of the Requirements for the Degree of Master of Science in Biology/ Botany By

Shiamaa Mohammed Abdel Reda B.Sc. Biology 1998-1999

Under Supervisor

Assist Professor Dr.Reyam Naji Ajmi 2016 A.D

1437 H

‫جوهىريت العراق‬ ‫وزارة التعلين العالي والبحث العلوي‬ ‫الجاهعت الوستنصريت‬ ‫كليت العلىم‬

‫أستخدام نظام التتبع لتقٌٌم بعض المعادن الثقٌلة‬ ‫والمغذٌات والعوامل المٌكروبٌة فً حلٌب الرضع‬ ‫بواسطة تقنٌة الحث بالبالزما‬ ‫رسالة‬ ‫مقدمة إلى كلٌة العلوم‪/‬الجامعة المستنصرٌة‬ ‫كجزء من متطلبات نٌل درجة الماجستٌر فً علوم الحٌاة‪ /‬نبات‬ ‫من قبل الطالبة‬

‫شٌماء محمد عبد الرضا االسدي‬ ‫بكالوريوس عموم حياة‪ /‬الجامعة المستنصرية‬

‫بأشراف‬ ‫أ‪.‬م‪ .‬د‪ .‬ريام ناجي عجمي‬ ‫هجري ‪3417‬‬

‫مٌالدي‪1036-‬‬

‫بسم هللا الرحمن الرحيم‬ ‫ون ِ‬ ‫ِ‬ ‫ِ‬ ‫َّ‬ ‫ِ‬ ‫َّ‬ ‫َّ‬ ‫يها‬ ‫ف‬ ‫ق‬ ‫ت‬ ‫م‬ ‫ل‬ ‫ا‬ ‫د‬ ‫ع‬ ‫و‬ ‫ي‬ ‫ت‬ ‫ل‬ ‫ا‬ ‫ة‬ ‫ن‬ ‫﴿مثَ ُل ا ْل َج‬ ‫ْ‬ ‫ُ‬ ‫َ‬ ‫ُ َ َ‬ ‫ُ‬ ‫َ‬ ‫أَ ْنهار ِم ْن م ٍ‬ ‫اء َغ ْي ِر َآ ِس ٍن َوأَ ْن َه ٌار ِم ْن‬ ‫ٌَ‬ ‫َ‬ ‫ط ْع ُم ُه َوأَ ْن َه ٌار ِم ْن َخ ْم ٍر‬ ‫لَ َب ٍن لَ ْم َيتَ َغ َّي ْر َ‬ ‫َّ‬ ‫ِ‬ ‫ِ‬ ‫ٍ‬ ‫ِ‬ ‫َّ‬ ‫ِ‬ ‫س ٍل‬ ‫ع‬ ‫ن‬ ‫م‬ ‫ار‬ ‫ه‬ ‫ن‬ ‫َ‬ ‫أ‬ ‫و‬ ‫ين‬ ‫ب‬ ‫ار‬ ‫لش‬ ‫ل‬ ‫ة‬ ‫ذ‬ ‫لَ‬ ‫ْ‬ ‫َ َ ٌَ ْ َ َ‬ ‫صفًّى﴾‬ ‫ُم َ‬ ‫صدق هللا العظيم‬ ‫} سورة محمد اآلية‪{51 :‬‬

Dedication

To my mother And My father ᾽s Soul

Shiamee

Acknowledgements In the first place, I would like to express my deep appreciation and gratitude to my supervisor, Dr. Reyam Naji for her scientific advice, assistance and encouragement during my study and introducing me to the Quality Control in field environmental contamination. I would like to express my deep thanks especially to Dean of Faculty of Science and Head of the Department of Biology and to all staff members in Science College at Al- Mustansiryah University to creating opportunity to study. My great appreciation to Director-General of UNESCO, Madam Irina Bokova for providing me partial funding to accomplish this thesis and publication two researches in international journals with high-impact Factor. Special thanks to General Company for Foodstuff Trading Department of Quality Control / Commerce, Director Mr .Ali Hamid , Mr ,Ali Ghazi , Mr .Hussein Ali and other . Also the Laboratories staff in The Ministry of Science and Technology.

Finally, I would like to express my thanks to my parent and brothers Maitham and his wife Duha for their encouragement.

Shiamee

List of Content Title English Abstract List of Contents List of Tables List of Figures List of Pictures List of Abbreviations Chapter One/ Introduction 1.1 Introduction

Page I IV VI VI VII VII 1

Chapter Two/ Literature Review 2. Literature Review 2.1 Background to Milk 2.2.1 Infant Formula Feeding 2.1.2 Elements in Infant Formula Milk 2.1.2.1 Zinc Zn 2.1.2.2 Lead Pb 2.1.2.3 Cadmium Cd 2.1.2.4 Copper Cu 2.1.2.5 Iron Fe 2.1.2.6 Arsenic As 2.1.3. Principle of Inductively Coupled Plasma Mass Spectrometry (ICPMS) 2.1.4. MilkoScan™ FT 120 System 2.1.5. Microbial Quality of Infant Formula Milk Powder 2.2. Background, Defining Traceability System 2.2.1. Relationship between Food Contamination and Traceability System 2.3 Improve Safety and Quality and Environmental 2.3.1 Link between Traceability and QA & QC 2.3.2 Industry Concerns, Health Risk in Safety Quality of Food 2.4. Traceability Technologies Applied in Milk 2.5. Milk and Traceability and Elements Chapter Three/ Materials & Methods

6 6 7 8 9 10 11 12 13 14 15 16 18 19 21 23 24 25 27 29

3.1. The Materials & Methods

30

3.1.1. Instruments 3.1.2. Chemical Materials and Media 3.3. Sample Collection 3.3.1. Preparing Experimental and Analyses 3.3.2. Sample Preparation analysis Milk by Topwave analytic Jena type 3.3.3. Determination of Elements by ICP-MS

30 30 31 31 33 35

IV

3.3.3.1. Statistical Method 3.3.4. Determination of Fat, Protein, Lactose and TS, SNF By using MilkoScan FT120 3.3.4.1. Calibration of MilkoScanTM FT 120 3.3.4.2. Standards and approvals 3.3.4.3. Statistical Method 3.3.5.Microbial Quality and Prepare Culture Media 3.3.5.1. Preparation of Milk Samples 3.3.5.2. Enumeration of Total Viable Bacteria 3.3.5.3. Enumeration of Total Coliform Bacteria 3.3.5.4. Salmonella 3.3.5.5. Yeasts and Molds General Counting Method 3.3.5.6. Statistical Method 3.4. Traceability System Chapter Four / Results and Discussion

36 37

4-1: Elements 4.1.1. Zn (Zinc) 4.1.2. Pb ( Lead) 4.1.3. Cd ( Cadmium ) 4.1.4. Cu ( Copper) 4.1.5. Fe ( Iron )

45 45 49 52 55 58

4.1.6. As ( Arsenic ) 4.2. Nutrients: (Fat, Protein, Lactose, Total solid and Solid Nonfat) 4.3. Microbial Quality of infant formula milk powder 4.4. Traceability System 4.4.1. Test Summary Report Template 4.4.2. Reference Environment Checklist 4.4.3. Application Quality Assurance Traceability Chapter Five/ Conclusions and Recommendations 5.1 Conclusions 5.2 Recommendations References Appendix Arabic Abstract

61 64 69 70 70 71 71

V

37 38 38 39 39 39 40 40 41 41 41

73 73 74

List of Tables No. 3-1

Title The Instruments that Used in this study and Industrial Companies with

Page 30

Origin 3-2

3-4 3-5 4-1 4-2 4-3

Chemical Materials and Media used in this Study with Industrial Companies with Origin Parameter Operating for the Determination of ( Zn, Pb, Cd, Cu, Fe, As ) elements in Infants formula milk Sample Traceability Matrix Privacy Information of Lifecycle Milk (Internal Traceability ) The Principal Components Analysis of Zn Concentration in Milk FAO/WHO. WHO, Geneva, 2004 The Principal Components Analysis of Pb Concentration in Milk

42 43 Appendex 47 Appendex

4-4

The Principal Components Analysis of Cd Concentration in Milk

Appendex

4-5 4-6 4-7 4-8

The Principal Components Analysis of Cu Concentration in Milk The Principal Components Analysis of Fe Concentration in Milk The Principal Components Analysis of As Concentration in Milk Results of the Analysis with the MilkoScan FT 120 of Concentration

Appendex Appendex Appendex Appendex

3-3

30 33

Milk study 4-9

The Relationship for all the Nutrients Under Study.

Appendex

4-10 Microbial Quality (cfu /g) in Different Infant Formula Samples

Appendex

Compared to IQS Standards. 4-11 Traceability System Application of Milk Types Under this Study

Appendex

4-12 Checklist Milk Types Under this Study

Appendex

4-13 Quality Assurance Traceability of Milk under this study

71

List of Figures No.

Title

Page

4-1 4-2

Zn Concentration in Three Time Analyses Samples Pb Concentration in Three Time Analyses Samples

49 50

4-3 4-4

Cd Concentration in Three Time Analyses Samples Cu Concentration in Three Time Analyses Samples

52 55

VI

4-5 4-6 4-7

Fe Concentration in Three Time Analyses Samples As Concentration in Three Time Analyses Samples MilkoScan FT 120 Correlation of Fats , Protien and Lactose in the Milk Study MilkoScan FT 120 Correlation of Total solid and Solid nonfat in the Milk Study. The Relationship for all the Nutrients Under Study Diagram of Data entry into the Program as an Evidence Base for Infant Milk more than 6 Months

4-8 4-9 4-10

59 61 65 66 68 72

List of Pictures Title

Page

2-1

ICP-MS ‘ELAN 6000’ PE Sciex Protocol (J. Sucharová etal 2006 )

16

2-2

MilkoScan™ FT 120 Protocol ( References )

18

2-3

Relationship between Traceability System and Quality and Safety

26

2-4

Developments Traceability System

28

3-1

Topwave Analytic Jena AG (Ministry of Commerce / General Company for Food stuff Trading / Quality Control Division)

35

List of Abbreviations No

Title

QA,QC

Quality Assurance, Quality Control

ICP MS

Inductively Coupled Plasma Mass Spectrometry

FDA

Food and Drug Administration

AAP

American Academy of Pediatrics

VII

FAO

Food Agriculture Organization

WHO

World Health Organization

MCT

Medium-Chain Triglyceride

PTWI

Provisional Tolerable Weekly Intake

RDA

Recommended Diet Intake

CFUM

Colony Forming Unit per Mill

IDFS

International Dairy Foods Association

FSMA

Food Safety Modernization Act

FSA

Food standards agency

GMO

Genetically Modified Organisms

HACCP ADA

Hazard Analysis Critical Control Point American Dietetic Association

CRM

Certificate Reference Material

RF

Radio Frequency

Pe

Prediction error

IQS

Iraqi Quality Standards

VRBLA

Violet Red Bile Lactose Agar

SDA

Sabouraud Dextrose Agar

TBGB

Tetrathionate Brilliant Green Broth

XLDA

Xylose Lysine Desoxycholate Agar

IQ

Intelligence Quotient

VIII

It

was

reached

internal

traceability

system

by

identifying

contaminants in infant formula milk powder item before the body is exposed and this reduces the cost of diseases related post-marketing to give accurate information on the impact of pollutants and health risks, especially caused by the accumulation of elements using the induction plasma ICP-MS technique which modern technology is a high-resolution sensitivity , selectivity and analysis of multi-component and set up new traceability model of those elements in the food component of primary (milk) . Compared with the World Health Organization (WHO), Food Agriculture Organization (FAO) and the IQS (Iraqi Quality Standards) as a monitoring product quality control (milk) to consumers as a dietary example of the first class in the food chain. It was selected five types in infant formula milk powder (over 6 months) circulating in the local markets and pharmacies, which are used as substitutes for breast milk during the first day of opening the box to the expiration date .These studies included traceability concentration of elements (Zn, Pb, Cd, Cu, Fe and As) in infant formula milk powder of three stages from opening date of box to expiration date. Results showed that the concentrations of lead growing successively and this constitutes a risk to the consumer compared with the standard limitations (0.002ppm ) [(-0.0407 ± 0.0103) , (-0.00206 ± 0.0129), (0.0292 ± 0.0129)] ppm Respectively Pb Test1, Pb Test 2, Pb Test 3. The rest of the elements (Zn, Cd, Cu, Fe and As) not constitute high levels, It was within the allowable standard limits. Nutrients

included

traceability

concentration

of

(Fat,

Protein,

Lactose, Total solid and Solid Non-fat) in infant formula milk powder of two stages from opening date of box to expiration date. Results showed I

that the average composition of Milk types were (29.27% fat, 21.90% protein, 45.66% Lactose 2.39 Total solid and 0.78% Solid Non-fat) . There was a medium positive correlation between the ratio of fat to protein in milk (r = 0.91-1.61)(0.77- 0.92) between Total Solid and Solid non-fat and medium negative correlation and the ratio of fat and protein to Lactose ( r=0.92 )in milk . It was identified using standard analytical methods; a MilkoScan ™ FT 120 using Standard methods due Is a fast and costeffective and are used widely in Dairy laboratories all over the world, which should make it universally accessible environment for scientists to study in Imported milk (non-local). This result indicated that all milk powder

companies

understudied

are

following

the

legal

and

quality

powder

due

to

standard of World and Iraqi composition powder milk. Microbial

Quality

of

infant

formula

milk

the

importance of infant feeding it was examining the microbiological quality of infant milk powder. A total of five types of milk during the first day of opening the box powders such as Total Coliform count (TCC),Total Salmonella count ( TSC), Total plate Count (TPC) and Yeasts and Molds Count (YMC). Results were obtained respectively; Total

plate count

(