nutrient presentation
TRANSCRIPT
STAKEHOLDER PANEL ON INFANT FORMULA & ADULT NUTRITIONALS (SPIFAN)& ADULT NUTRITIONALS (SPIFAN)
MINERALS
Eric Poitevin – Nestlé Research Center (Switzerland)[email protected]
August 24th , 2013Chicago, IL ‐ USA
Background and HistoryInorganicelements
CAS number Atomic Mass (Number)
Classification (Group)
Year of discovery
1st IP (eV)
Calcium (Ca) 7440‐70‐2 40.078 (20) Alkali earth metal (2) 1808 6.113
Magnesium (Mg) 7439‐95‐4 24.305 (12) Alkali earth metal (2) 1755 7.646
Potassium (K) 7440‐09‐7 39.098 (19) Alkali metal (1) 1807 4.34
Sodium (Na) 7440 23 5 22 990 (11) Alkali metal (1) 1807 5 139Sodium (Na) 7440‐23‐5 22.990 (11) Alkali metal (1) 1807 5.139
Phosphorus (P) 7723‐14‐0 30.974 (15) Nonmetal (15) 1669 10.487
Copper (Cu) 7440‐50‐8 63.546 (29) Transition metal (11) 9000 BC 7.726
Manganese (Mn) 7439‐96‐5 54.938 (25) Transition metal (7) 1774 7.432
Zinc (Zn) 7440‐66‐6 65.390 (30) Transition metal (12) 1746 9.394
Iron (Fe) 7439‐89‐6 55.845 (26) Transition metal (8) 3500 BC 7.90
Background and History1800’ Di f j ti l i l• 1800’s Discovery of major essential minerals
• 1867 Liebig‘s first marketed “ideal IF”• 1915 1st SMA IF with minerals added• 1929 Spectrometer with nebulizer and spray chamber• 1929 1st soya IF with minerals added• 1937 1st commercial grating spectrograph (ARL)1937 1 commercial grating spectrograph (ARL)• 1941 1st FDA label declaration (Ca, P, Fe) • 1959 Iron fortification in IF (microcytic anemia)
1960’ H i d IF ( l l t l d C P ti )• 1960’s Humanized IF (renal solute load, Ca:P ratio)• 1974 1st commercial ICP-AES spectrometer• 1981 Codex standard for IF (all minerals regulated)( g )• 2000’s Axial and dual view ICP-AES spectrometers• 2010’s rc/cc-ICP-MS and MIP-AES spectrometers
Background and HistoryMilk Product Water
(%)Fat(%)
Protein(%)
Carbohydrate(%)
Ash/Minerals(%)
Whole liquid 87 3.7 3.3 4.8 0.7
Whole liquid (human) 87.5 3.8 0.9 7.0 0.2
Skimmed liquid 90 <0.1 3.4 4.9 0.75
Whole powder 2‐4 25‐28 25‐27 37‐38 6‐7
Skimmed powder 3‐5 0.7‐1.3 35‐37 49‐52 7.5‐8
Whey powder 3‐6 0 8‐1 5 9‐13 65‐73 7 5‐12Whey powder 3 6 0.8 1.5 9 13 65 73 7.5 12
Infant formula 2‐3 26‐39 10‐18 40‐60 8
Reference:Dairy Powders and Concentrated products A Y Tamine 2009 Wiley‐Blackwell ISBN 978‐1‐4051‐5764‐3Dairy Powders and Concentrated products. A.Y. Tamine.2009.Wiley‐Blackwell. ISBN 978‐1‐4051‐5764‐3.J.H. Kastle. Am J Physiol.,22 ,1908.
Background and HistoryElement Whole milk
(mg/100g)Skimmed milk(mg/100g)
Whey protein powder(mg/100g)
Casein powder(mg/100g)
Ca 114‐123 117‐123 350‐480 1000‐1500
P 103 103 330‐500 800
Na 41‐46 44‐49 280‐460 1200‐1300
K 152‐155 150‐164 650‐1190 1650
Mg 7‐13 70‐150 5‐7 30‐52
Element Cow milk(mg/L)
Human milk(mg/L)(mg/L) (mg/L)
Fe 0.5 0.4
Zn 4 3
C 0 9 0 4Cu 0.9 0.4
Mn 0.03 0.01
Background and HistoryElement Sources Benefits Risks
Ca Dairy products,Legumes, green
Bones and teeth formationNerve stimulation
Stunted growthOsteoporosis, osteomalaciag g
vegetables Cell permeabilityEnzyme activation
pTetanyRickets
P All foods Bones and teeth formationEnergy metabolism enzyme formation
Stunted growthRicketsEnergy metabolism, enzyme formation Rickets
Na All foods Osmotic pressure, Acid‐base balanceCell permeability, nerve stimulation and muscle contraction
Nausea, vomiting, muscularweakness, heart failure
K All foods Same as Na. Protein synthesis and glycogen formation
Rare: nausea, vomiting, cramps
Mg Green vegetables
Structural (bones and teeth) Nerve stimulation
Renal or alcoholism diseaseTremors leading to seizurevegetables
Nuts, grains, milk seafood
Nerve stimulationMuscle contractionEnzyme activation
Tremors leading to seizure
Background and HistoryElement Sources Benefits Risks
Fe Organ meats, fish eggs dairy
Hemoglobin and myoglobin formationEssential component of many enzymes
Anemia, decrease in oxygentransport and cellularfish, eggs, dairy
productsEssential component of many enzymes transport and cellular
immunity, muscle weakness
Zn Oysters, fish, meat, milk, nuts,
Constituent of many enzymesCarbon dioxide transport
Retard growth and sexualdevelopment, poor
grains, legumes Vitamin A utilization immunity and skin lesions, dwarfism, taste sensitivity
Cu Organ meats, oyters, nuts,
Necessary for Fe utilization and hemoglobin formation, constituent of
Anemia, neutropenia, leucopenia, skeletaloyters, nuts,
fruits, driedlegumes
hemoglobin formation, constituent of cytochrome oxidase, involved in boneand elastic tissue development
leucopenia, skeletaldemineralization
Mn Tea, grains, nuts Cofactor of large number of enzymes, anti oxidant involved in normal brain
Reduced growth, abnormalreproduction and boneanti‐oxidant, involved in normal brain
function, reproduction and bonestructure
reproduction and bonedevelopment
Background and History
• Iron deficiency (Anemia) • Zinc deficiency (Retard growth)
Minerals Chemistry• Ca, P: Ca phosphates, Ca caseinate, Ca citrate, ionic form (50% casein bound)
N K f i i f (N i d i h Chl id ) i i f i• Na, K: free ionic forms (Na associated with Chloride) in casein fraction
• Mg: 40% Mg citrate, 16% as free ion and 7% as Mg phosphate
• Fe: 32% Fe citrate, 26% (whey proteins), 24% (caseins) and 14% (fat)
• Zn: 32% bound to casein and the rest bound to colloidal Ca phosphateZn: 32% bound to casein and the rest bound to colloidal Ca phosphate
• Cu: 47% in low Mw fraction, 44% (caseins), 8% (whey proteins) and 2% (fat)
• Mn: 67% bound to caseins, 14% (whey proteins), 18% (low Mw) and 1% (fat)
Regulations
Legislation Infant formulae Follow‐on FormulaeFormulae for special medical
purpose
C d Ali i FAO/WHO (2007) FAO/WHO (1987) C d FAO/WHO (2007)Codex AlimentariusCommission
FAO/WHO (2007)Codex Stan 72‐1981 –Revised Section A
FAO/WHO (1987) Codex Stan 156
FAO/WHO (2007)Codex Stan 72‐1981 –Revised Section B
USA FDA (1985, 2007a,b)Infant Formula Act 21 CFR
Not Applicable FDA (1985, 2007a,b)Infant Formula Act 21 CFRInfant Formula Act 21 CFR
107Infant Formula Act 21 CFR 107
EU EU (2006b) Commission Directive 2006/141/EC
EU (1999) Commission Directive 1999/21/EC
EU (1989) Council Directive 89/398/EC
Australia & New Zealand
Food Standards Australia and New Zealand Standard 2.9.1a Infant Formula Products)
China GB 10765‐2010 (Infant Formula and Follow‐on Formula)China ( )
Recommended Daily Allowances (RDA/AI)Life stage Age Ca (mg/day) P (mg/day) Na (mg/day) K (mg/day) Mg (mg/day)
Infants 0‐6 months 210 (AI) 100 (AI) 120 (AI) 400 (AI) 30 (AI)
Infants 7‐12 months 270 (AI) 275 (AI) 370 (AI) 700 (AI) 75 (AI)
Children 1‐3 years 500 (AI) 460 1000 (AI) 3000 (AI) 80
Children 4‐8 years 800 (AI) 500 1200 (AI) 3800 (AI) 130
Children 9‐13 years 1300 (AI) 1250 1500 (AI) 4500 (AI) 240
Children 14‐18 years 1300 (AI) 1250 1500 (AI) 4700 (AI) 410 (360F)
Adolescents‐adults 19‐30 years31‐50 years
1000 (AI) 700 1500 (AI) 4700 (AI) 420 (320 ‐F)400 (310 ‐F)
Adults 51 ‐70 years 1200 (AI) 700 1300 (AI) 4700 (AI) 420 (320 ‐F)
Adults 70 years+ 1200 (AI) 700 1200 (AI) 4700 (AI) 420 (320 ‐F)
Pregnancy 19‐30 years 1000 (AI) 700 1000 (AI) 4700 (AI) 350Pregnancy 19 30 years31‐50 years
1000 (AI) 700 1000 (AI) 4700 (AI) 350360
Breast feeding 19‐30 years31‐50 years
1000 (AI) 700 1000 (AI) 5100 (AI) 310320
Recommended Daily Allowances (RDA/AI)Life stage Age Fe (mg/day) Mn (mg/day) Zn (mg/day) Cu (µg/day)
Infants 0‐6 months 0.27 (AI) 0.003 (AI) 2 (AI) 200 (AI)
Infants 7‐12 months 11 0.6 (AI) 3 220 (AI)
Children 1‐3 years 7 1.2 (AI) 3 340
Children 4‐8 years 10 1.5 (AI) 5 440
Children 9‐13 years 8 1.9 (1.6‐F) (AI) 8 700
Children 14‐18 years 11 (15‐F) 2.2 (1.6‐F) (AI) 11 (9‐F) 890
Adolescents‐adults 19‐30 years31‐50 years
8 (18‐F) 2.3 (1.8‐F) (AI) 11 (8‐F) 900
Adults 51 ‐70 years 8 2.3 (1.8‐F) (AI) 11 (8‐F) 900
Adults 70 years+ 8 2.3 (1.8‐F) (AI) 11 (8‐F) 900
Pregnancy 19‐30 years 220 2.0 (AI) 11 1000Pregnancy 19 30 years31‐50 years
220220
2.0 (AI) 11 1000
Breast feeding 19‐30 years31‐50 years
290290
2.6 (AI) 12 1300
Tolerable Uptake level (UL)Life stage Age Ca P Na K Mg Fe Mn Zn CuLife stage Age Ca P Na K Mg Fe Mn Zn Cu
Infants 0‐6 months ND* ND ND ND ND 40 ND 4 ND
Infants 7‐12 months ND ND ND ND ND 40 ND 5 ND
Children 1 3 years 2500 3000 1500 ND 65 40 2 7 1Children 1‐3 years 2500 3000 1500 ND 65 40 2 7 1
Children 4‐8 years 2500 3000 1900 ND 110 40 3 12 3
Children 9‐13 years 2500 4000 2200 ND 350 40 6 23 5
Children 14‐18 years 2500 4000 2300 ND 350 45 9 34 8
Adolescents‐adults 19‐70 years 2500 4000 2300 ND 350 45 11 40 10
Adults 70 years+ 2500 3000 2300 ND 350 45 11 40 10y
Pregnancy 14‐18 years19‐50 years
2500 3500 2300 ND 350 45 9 3440
810
Breast feeding 14‐18 years19 50
2500 4000 2300 ND 350 45 11 3440
81019‐50 years 40 10
* ND: Not Determined
Regulations – Codex (Minerals content)Element Product Target population Min Max GUL UnitElement Product Target population Min Max GUL Unit
Calcium Infant formulaFollow‐up
0 – 6 months6 – 12 months
50 90
‐ 140 mg/100 kcal
Phosphorus Infant formulaF ll
0 – 6 months6 12 th
25 60 ‐ 100 mg/100 kcalFollow‐up 6 – 12 months 60 g/
Ca:P ratio Infant formulaFollow‐up
0 – 6 months6 – 12 months
1:1 2:1 ‐ mg/100 kcal
Sodium Infant formulaF ll
0 – 6 months6 12 th
20 20
60 85 ‐ mg/100 kcalFollow‐up 6 – 12 months 20 85 g/
Potassium Infant formulaFollow‐up
0 – 6 months6 – 12 months
60 80
180 ‐ ‐ mg/100 kcal
Magnesium Infant formulaF ll
0 – 6 months6 12 th
5 6 ‐ 15 mg/100 kcalFollow‐up 6 – 12 months 6 g/
Iron Infant formulaFollow‐up
0 – 6 months6 – 12 months
0.45 1 2 ‐ mg/100 kcal
Zinc Infant formulaF ll
0 – 6 months6 12 th
0.50 5
1.5 mg/100 kcalFollow‐up 6 – 12 months 0.5 ‐ g/
CopperManganese
Infant formulaInfant formula
0 – 6 months0 – 6 months
35 1 ‐
120100 µg/100 kcal
Regulations – Entity (Minerals content)Element Product Target population Minimum Maximum UnitElement Product Target population Minimum Maximum Unit
Ca Infant formulaFollow‐up
0 – 6 months6 – 12 months
60 (USA)50 (EU,NZA) 140 (EU) mg/100 kcal
P Infant formulaF ll
0 – 6 months6 12 th
30 (USA)/25‐30*(EU)25 (EU NZA) 30*(EU) 90‐100*(EU)/100(NZA) mg/100 kcalFollow‐up 6 – 12 months 25 (EU,NZA) ‐ 30*(EU) ( )/ ( ) g/
Ca:Pratio
Infant formulaFollow‐up
0 – 6 months6 – 12 months
1.2:1 (NZA) mg/100 kcal
Na Infant formulaF ll
0 – 6 months6 12 th 60 (EU,NZA) mg/100 kcalFollow‐up 6 – 12 months ( , ) g/
K Infant formulaFollow‐up
0 – 6 months6 – 12 months
80 (USA,NZA)60 (EU)
200(USA,NZA)/160(EU)160 (EU)/200 (NZA) mg/100 kcal
Mg Infant formulaF ll
0 – 6 months6 12 th
6 (USA)5 (EU NZA) 15 (EU,NZA) mg/100 kcalFollow‐up 6 – 12 months 5 (EU,NZA) ( , ) g/
Fe Infant formulaFollow‐up
0 – 6 months6 – 12 months
0.15(USA)/0.3‐0.45*(EU)/0.8(NZA)0.6‐0.9*(EU)/0.8(NZA)
3 (USA)/1.3‐1.9*(EU)1.7‐2.5* (EU)/1.5(CHN) mg/100 kcal
Zn Infant formulaF ll
0 – 6 months6 12 th
0.75*(EU)0 75*(EU)
2.4* (EU)/Zn:Cu15:1(NZA)2 4*(EU) /Z C 20 1(NZA) mg/100 kcalFollow‐up 6 – 12 months 0.75*(EU) 2.4*(EU) /Zn:Cu20:1(NZA) g/
CuMn
Infant formulaInfant formula
0 – 6 months0 – 6 months
60 (USA,NZA)/35 (EU)5 (USA), 4.8 (CHN)
100 (EU)/80(NZA)100(NZA) µg/100 kcal
(* Soy protein)
Minerals calculated range for IFElement K
(mg/100g)Ca
(mg/100g)P
(mg/100g)Na
(mg/100g)Mg
(mg/100g)Regulations limits 300‐1000 250‐ 700 75‐500 100‐425 25‐75
Minimum 460 350 300 110 30Minimum 460 350 300 110 30
Maximum 750 640 400 330 55
Concentrate/premixes 1100‐1650 800‐8000 600‐800 500‐1200 75‐4000
Element Fe(mg/100g)
Zn(mg/100g)
Cu(mg/100g)
Mn(mg/100g)
Regulations limits 0.75‐20 2‐9 0.18‐ 0.9 0.005‐0.5
Minimum 1 2 0.2 0.01
Maximum 10 7.1 0.6 0.4
Concentrate/Premixes 20‐10000 50‐9000 5‐4000 0.1‐9000
Minerals ‐ Sample digestion
Digestion techniqueDry sample
size (g)
Digestion time
(hour)
Degree of digestion (way of losses)
Sample throughput (number of
samples per Basic Equipment
Dry Ashing
losses)run)
1-5Muff le furnace
Crucibles 1-10Incomplete
(volatilization) 10-20
Microwave Open vessel
High Pressure Ashing
Wet ashing
< 0.3 High Pressure asher Quartz tubes
1 Very High (retention)
15-30
0.5-2 Microw ave Glass vessel
< 1 Incomplete (volatilization)
5-10
Microwave Closed vessel
< 0.5 Microw ave Teflon vessel
gashing ( )
< 1 High (retention)
20-40
Minerals ‐ AnalysisAnalyte Macro‐and Micronutrients a yteCriterion FAAS XRF ICP‐OES
Typical IDL (µg/L) XRF (in mg/kg)
1‐1000 1‐1000 1‐100
Precision (%) Short term (< 4 h)
0.1‐1 1‐3 0.3‐2
ac o a d c o ut e ts
Dynamic range (orders of magnitude)
1‐2 4‐5 4‐5
Speed for element (excluding sample preparation time)
Fast 5‐30 sec
(nebulization sample uptake: 5‐6 mL/min
Slow 5 ‐ 10 Minutes
Intermediate 1‐2 minutes
(nebulization sample uptake: ~1 mL/minpreparation time) (nebulization sample uptake: 5 6 mL/min (nebulization sample uptake: ~1 mL/minOverall speed
(including multi‐element capability)
Slow limited multielement capability
6 at most
Fast / Non destructive Multielement capability
Fast Multielement capability
Interferences FewHigh matrix depedency match standard needed
Numerous but well characterized Spectral and non spectral Interferences Few match standard needed
Calibration vs ICP‐OES neededLine selection, internal standardization and
ion buffer needed Multi‐element capability
commonly None Up to 6
Yes Yes
Safety Fire hazard, flammable gasRadiation hazards
Precautions for X‐Ray protectionSafe
inert gas ArPrecautions for X Ray protection inert gas Ar
• ICP‐MS with reaction/collision cells (costly, interferences) • MP‐AES (promising technology, only supplier application notes)
Minerals ‐ Official Methods• AOAC 985.35 Codex Type II method: Ca,Mg,Fe,Zn,Cu,Mn,Na,K‐ Flame atomic absorption spectrophotometry after dry ashingValidated: Infant Formula (enteral soy based RTF whey powder)‐ Validated: Infant Formula (enteral, soy‐based , RTF, whey powder)
• AOAC 986.24 Codex Type II method: P‐Spectrophotometry after dry ashing‐ Validated: Infant Formula (enteral, soy‐based , RTF, whey powder)• AOAC 984.27 Codex Type III method: Ca,Cu,Fe,Mg,Mn,P,K,Na,Zn‐ Validated: Infant Formula (RTF, soy‐based, whey powder)( , y , y p )‐ ICP‐AES after acid digestion• ISO 8070:2007 (IDF 119) Codex Type II method: Ca,Mg,Na,KFlame atomic absorption spectrophotometry after acid digestion‐ Flame atomic absorption spectrophotometry after acid digestion
‐ Validated: Skim and whey milk powders, protein concentate, casein
Minerals ‐ Official Methods• Existing Official Methods suffer from:
‐ use of old and non safe digestion systems (ashing, acid digestion with HCLO4)‐ use of old technology (FAAS, Radial ICP‐AES)l k f ifi i ( ilk b d f l i h‐ lack of specificity (non milk‐based formulas with
added salts)lack of sensitivity (high analytical range of‐ lack of sensitivity (high analytical range of
minerals from µg to g/100g)‐ Relative poor multi‐elemental capacity and high p p y gsample throughput
Minerals ‐ Official Methods• New AOAC 2011.14 First Action: Ca,Mg,Fe,Zn,Cu,Mn,Na,K‐ ICP‐AES after microwave digestionValidated: Fortified Food products including IF‐ Validated: Fortified Food products including IF
Minerals ‐ Official Methods• AOAC 2011.14 First Action method: Ca,Mg,Fe,Zn,Cu,Mn,Na,K‐ Validated with 2 IF, NIST Non fat milk powder, chocolate milk powder with complete digestion covering AOAC sectors 5 6 7powder with complete digestion covering AOAC sectors 5, 6,7 ‐ Ring trial with 9 independent Nestlé laboratories‐ Additional data from 2 proficiency‐tests including 4 IF and 4 Adult nutritionals (2012) for final Action
100 % fat ICP-AES analysisMicrowave digestion
2 4
3
1
0 %
pro
tein
0 % carbohydrate
IN 1
OUT
T- connector
Peristaltic pump
To ICP-AES nebuliser
Internal standards
SampleFrom autosampler
From internal standard solution
75 9
86
100 % protein
100 % carbohydrate
0 % fat
IN 2
Dilute to 50 mL, HNO3 10% v/vDF = 1000.5 g + 5 mL HNO3
Minerals ‐ Official Methods• AOAC 2011.14 First Action method : data for Final Action (end 2013)
‐ Data collected from 2 p‐tests on 2012 (29 independant labs)Satisfactory results for 8 IF and adult nutritionals matrices‐ Satisfactory results for 8 IF and adult nutritionals matrices
Test Matrices (4 samples per p‐test) Mineral Number of labs Unit Consensus Value Range
RSDR (%)
Recovery Min‐Max range for labs
(%)Horrat value
Calcium 9 ‐ 21 mg/100g 78 ‐ 603 < 5 84 ‐ 118 0.64 ‐ 1.09
1 Starter Infant Formula, Milk Based
2 Follow‐Up Infant Formula, Whey Enriched
1 Growing Up Milk Powder
Copper 7 ‐ 21 mg/100g 0.14 ‐ 0.44 < 10 86 ‐ 157 0.66 ‐ 0.79
Iron 12 ‐ 22 mg/100g 1.46 ‐ 8.63 < 8 90 ‐ 124 0.57 ‐ 0.80
Magnesium 7 ‐ 21 mg/100g 26.0 ‐ 60.4 < 4 93 ‐ 107 0.51‐ 0.58
M 7 21 /100 0 055 0 313 < 10 91 117 0 37 1 10Growing Up Milk Powder
1 Adult Complete Formula, Hydrolysed Proteins
2 Adult Complete Formula
1 Adult Formula, Blenderized Natural Foods
Manganese 7 ‐ 21 mg/100g 0.055 ‐ 0.313 < 10 91 ‐ 117 0.37 ‐ 1.10
Phosphorus 8 ‐ 21 mg/100g 67 ‐ 425 < 6 71 ‐ 110 0.60 ‐ 1.10
Potassium 8 ‐ 20 mg/100g 124 ‐ 622 < 6 97 ‐ 110 0.66 ‐ 1.19
Sodium 7 ‐ 22 mg/100g 86 ‐ 231 < 5 94 ‐ 108 0.87 ‐ 1.01
Zinc 7 ‐ 20 mg/100g 1.42 ‐ 6.30 < 6 67 ‐ 110 0.54 ‐ 0.64
Fitness for Purpose
Minerals Working Group
Fitness for purpose statement• A multi‐elemental analytical method to determine level of
nine minerals in infant formula and adult nutritionals asd fi d b AOAC SPIFAN i l d idefined by AOAC SPIFAN, in regulated concentration ranges:
Mineral K(mg/100g)
Ca(mg/100g)
P(mg/100g)
Na(mg/100g)
Mg(mg/100g)
R l i li i (IF) 300 1000 250 700 75 500 100 425 25 75Regulations limits (IF) 300‐1000 250‐ 700 75‐500 100‐425 25‐75
Minimum 200 150 25 30 10
Maximum (IF/Adult) 3200 2400 1600 1600 750
Mineral Fe(mg/100g)
Zn(mg/100g)
Cu(mg/100g)
Mn(mg/100g)
Regulations limits (IF) 0.75‐20 2‐9 0.18‐ 0.9 0.005‐0.5
Minimum 0.5 1 0.2 0.005
Maximum (IF/Adult) 85 32 21 0.5
Fitness for purpose statement
• An analytical method to determine level of nine minerals in i f f l d d l i i l i l finfant formula and adult nutritionals using complete safe digestion systems and updated ICP‐AES technology
• An analytical method having the same performances than AOAC 2011.14 validated for IF and adult nutritionals
• The method needs to be capable of dispute resolution
Questions?Questions?