The certification of the mass fractions of vitamins in four reference materials

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Wholemeal flour (CRM 121), milk powder (CRM 421), lyophilized mixed vegetables (CRM 485) and lyophilized pigs liver (CRM 487): B1, B6 and folate in CRM 121; B1, B2, B6, B12 and folate in CRMs 421 and 487, and B1, B6, folate and carotenoid in CRM 485
Agricultural and fisheries research

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ISSN 1018-5593
European Commission
ber information
REFERENCE MATERIALS
"THE CERTIFICATION OF THE MASS FRACTIONS
OF VITAMINS IN FOUR REFERENCE MATERIALS:
WHOLEMEAL FLOUR (CRM 121),
MILK POWDER (CRM 421),
LYOPHILIZED MIXED VEGETABLES (CRM 485)
AND LYOPHILIZED PIGS LIVER (CRM 487)" European Commission
ber information
REFERENCE MATERIALS
'THE CERTIFICATION OF THE MASS FRACTIONS
OF VITAMINS IN FOUR REFERENCE MATERIALS:
WHOLEMEAL FLOUR (CRM 121),
MILK POWDER (CRM 421),
LYOPHILIZED MIXED VEGETABLES (CRM 485)
AND LYOPHILIZED PIGS LIVER (CRM 487)"
B„ B6 and Folate in CRM 121; B„ B2, B,,, B„ and Folate in CRMs 421 and 487,
and Β,, Β,, Folate and Carotenoid in CRM 485
P.M. FINGLAS('), K.J. SCOTT('), CM. WITTHÖFT, H. VAN DEN BERG(2),
and I. DE FROIDMONT­GÖRTZf)
(') Institute of Food Research, Norwich, (UK)
C) TNO Nutrition and Food Research Institute, Zeist (NL)
C) Standards, Measurements and Testing Programme, European Commission, Brussels (B)
Directorate­General
Science, Research and Development
1998 EUR 18320 EN EUROPEAN COMMISSION
LEGAL NOTICE
Neither the European Commission nor any person acting on behalf of the Commission
is responsible for the use which might be made of the following information
A great deal of additional information on the European Union is available on the Internet.
It can be accessed through the Europa server (http://europa.eu.int).
Cataloguing data can be found at the end of this publication.
Luxembourg: Office for Official Publications of the European Communities, 1999
ISBN 92-828-5959-2
© European Communities, 1999
Reproduction is authorised provided the source is acknowledged.
Printed in Belgium EUROPEAN COIVI IVI ISSION
Edith CRESSON, Member of the Commission responsible
for Research, Innovation, Education, Training
and Youth
DG \ 11/(711.3 - Programme Measurements & Testing ABSTRACT
This report describes the work performed to prepare and certify four BCR reference materials
|CRMs 121 (Wholemeal flour), 421 (milk powder), 485 (lyophilized Mixed Vegetables), and
lyophilized Pig's Liver (CRM 487)] for a range of vitamins. A previous report (EUR-18039)
describes the certification of vitamins D, and E in CRM 421. The reference materials may be
used for the quality control of analytical measurements for nutritional labelling, and for the
production of detailed food composition data for research. The certified mass fractions
(expressed in mg/kg dry matter) with the uncertainties of measurement (in parentheses) are listed
as follows:
Vitamin CRM 121 CRM 421 CRM 485 CRM 487
(Wholemeal (Milk Powder) (Mixed (Pig's Liver)
flour) Vegetables)
Certified Value Certified Value Certified Value Certified Value
(Uncertainty) (Uncertainty) (Uncertainty) (Uncertainty)
mg/kg dry matter mg/kg dry matter mg/kg dry mg/kg dry matter
matter
B, (thiamin)1 4.63 (0.39) 6.51 (0.48) 3.07 (0.34) 8.6(1.1)
B2 (riboflavin) ­ 14.5 (0.6) ­ 106.8 (5.6)
B6 (total 4.10(1.02) 6.66 (0.85) 4.8 (0.8) 19.3 (2.9)
pyridoxine)2
B,2 ­ 0.034 (0.005) ­ 1.12(0.09)
Folate (total) 0.50 (0.07) 1.42(0.14) 3.15(0.28) 13.4(1.3)
Trans α-Carotene ­ ­ 10.5 (0.6) ­
Total e ­ ­ 9.8 (0.7) ­
Trans ß-Carotcne ­ ­ 23.7(1.5) ­
Total ß-Carotene ­ ­ 25.6(1.2) ­
Lutein ­ ­ 12.5(0.8)
­
Lutei n+Zeaxanthin 22.3(1.3) • 1
expressed as thiamin chloride hydrochloride,
expressed as pyridoxine hydrochloride. Indicative values (expressed as mg/kg dry matter) are presented for 5-methyltetrahydrofolic
acid (5-MTIIF) in CRM 421 (milk powder) and 485 (mixed vegetables):
Vitamin CRM 421 CRM 485
(Milk powder) (Mixed Vegetables)
Indicative Value (Uncertainty) Indicative Value (Uncertainty)
mg/kg dry matter mg/kg dry matter
0.25 (0.02) 5-MTHF 2.14(0.42)
Additional information values (expressed as mg/kg dry matter) are also presented for 5-MTHF
in CRMs 121 and 487, and carotenoids in CRM 485:
Vitamin CRM 121 CRM 485 CRM 487
(Wholemeal Flour) (Mixed Vegetables) (Pig's Liver)
Information Value (range) Information Value Information Value
(range) (range)
mg/kg dry matter
mg/kg dry matter mg/kg dry matter
5-MTHF 0.04 (O.03-O.O8) - 2.6(1.9-3.8)
trans-Lycopene - 13.8(10-20) -
total-Lycopene - 14.2(10-23) -
Zeaxanthin - 9.7(6-13) -CONTENTS
Page
ABSTRACT 1
CONTENTS 3
ABBREVIATIONS 6
1. INTRODUCTION 9
1.1 Background and Need for CRMs
1.2 Choice of Materials for CRMs
1.3 Dates of Preparation, Homogeneity & Stability Testing, and 9
Certification Measurements
2. PARTICIPANTS 11
2.1 Preparation of Material
2.2 Homogeneity and Stability Studies
2.3 Analyses for Certification, Indicative and Additional 1
Information Results
3. PRELIMINARY INTERCOMPARISONS3
3.1 B-group vitamins 1
3.2 Folate4
3.3 Carotenoids
4. PREPARATION OF THE MATERIALS 16
4.1 Wholemeal Flour (CRM 121 )
4.2 Milk Powder (CRM 421)
4.3 Mixed Vegetables (CRM 485)7
4.3.1 Selection of materials
4.3.2 Preparation of material 18
4.3.3 Analysis
4.3.4 Preparation of main batch9
4.4 Pig's Liver (CRM 487) 20 HOMOGENEITY 21
5.1(¡encrai21
5.2MethodsforHomogeneity Studies22
5.1VitaminsΒ,&niacin in CRM 12122
5.3.1 Pre-packaging homogeneity study 22
5.3.2 Main homogeneitystudy24
5.4 Vitamins A, D3, B2 & B12inCRM42126
5.5 Carotenoids in CRM 48526
5.6 Vitamins A, B„B2& R inCRM48727
6. STABILITY 30
6.1GeneralDesign30
6.2B-vitaminsinCRM 12132
6.3VitaminsA,C,D, & E, and niacin in CRM 421 38
6.4 Carotenoids in CRM 48538
6.5 Vitamins A & Li, B-group, C & folateinCRM48746
6.6 Summary of stability findings and recommendations54
7. ANALYTICAL METHODS 55
7.1 DesignoftheCertificationExercise55
7.2 AnalyticalMethodsforCertification55
7.3 Calibration,PurityofStandards & Traceability 56
7.4 Drymatter56
8. RESULTS 81
8.1TechnicalEvaluation81
8.1.1VitaminB,81
8.1.2nB281
8.1.3VitaminB681
8.1.4 Total folate & 5-MTHF 81
8.1.5 Carotenoids82
9. EVALUATION OF THECERTIFICATIONEXERCISE112
9.1 Statistical Evaluation112
9.2 Certified Values and Uncertainties124
9.3 Indicative Values and s126
9.4 Information Values 126
10. REFERENCES127