Making a Claim: Factors impacting Protein Quality and a New Way for Measuring David W. Plank May 4, 2016 Sr. Technical Manager 1
Overview • Demand for high-protein foods • Need for alternative protein sources • Protein quality regulations around the world • Requirements for protein claims in the U.S. • PDCAAS protein quality test • New animal safe PDCAAS test • Future development in protein quality measurement 2
Protein Demand Continues through 2016 • Protein continues to be a hot trend in the food industry — Snacks, meals, beverages and more — Focus has shifted to protein from plant sources — FAO has declared 2016 the international year of the pulses o Low-cost source of protein and dietary fiber The 68th UN General Assembly declared 2016 the International Year of Pulses Source – MINTEL 2013 Source-FAO 2016 Source – Dairy Foods 2015 3
Animal protein may not be sustainable for supporting further growth of protein foods LAND Usage WATER Usage CO 2 EMISSIONS Source: www.waterfootprint.org Source: http://www.huffingtonpost.com/ 4 Source: breakingnews.ewg.org
Current Common Protein Sources (pulses) Lower Quality Higher Quality 5
Some Potential Future Protein Sources * * * * WSV = waste stream valorization 6
Alternative Protein Sources • How can I compare these sources? • How will they impact the final product? • What concerns should I have? 7
Five regulatory authorities of the world for protein quality Canada European Union (Health Canada) Codex Alimentarius United States (FDA-USDA-FTC) Codex Alimentarius Codex Alimentarius Australia – New Zealand (FSANZ) Source - Lewis (2012) Br J Nutr. 108, S212-S221 8
Protein quality testing around the world Alternatives for Different Regulatory Regions 1. amino acid composition or reference amino acid profile; 2. protein digestibility-corrected amino acid score (PDCAAS); 3. protein efficiency ratio (PER); 4. protein rating; 5. reference protein without amino acid profile or method of determination. Test generally required for protein content declaration • United States – PDCAAS • Canada – PER • EU – PDCAAS depending on food • FSANZ – PDCAAS depending on food • CODEX – PDCAAS depending on food; Moving to DIAAS Source - Lewis (2012) Br J Nutr. 108, S212-S221 9
U.S .S. . Regulations for Protein Claims • Food product must contain a minimum of 10% of the daily value of quality protein on both a per serving and per RACC (Reference Amount Customarily Consumed) basis • The amount of quality protein must be determined by the PDCAAS method which includes both amino acid and digestibility measures • The total protein claimed may be based on the total crude protein (Dumas combustion or Kjeldahl x 6.25) ( 21CFR101.9(7) ) 10
Protein Claims - Overview Threshold Requirement: >10% DV of Protein (high quality) per RACC and per serving as determined by PDCAAS Examples: • Nutrient Content — Good Source of Protein, With Protein, Made With Xg of Protein, Contains Protein, Source of Protein • Statements of Fact — Xg of protein (apart from NF panel) • Structure Function — Protein helps build/maintain/repair muscles • Food Combination Claims — Xg of protein when made with/eaten with milk, yogurt, chicken, etc. o Requires full context: A serving of product X when eaten with Y provides Xg of protein
Elements of f PDCAAS Protein Quality Measurement • Amino Acid Analysis — Acid hydrolysis for majority of amino acids — Performic acid oxidation for cysteine and methionine o Alternative approaches e.g. reduction/carboxymethylation ― Base hydrolysis for tryptophan • Determination of crude protein (nitrogen x 6.25) • Calculation of limiting amino acid relative to ideal protein source • Determination of protein digestibility PDCAAS Value = Limiting Amino Acid Value x Digestibility 12
Ideal Protein – Contains all amino acids essential for human nutrition Food and Agriculture Organization of the United Nations 13
What molecular weight should be used for Amino Acids from Protein? Free Amino Acids Protein synthesis H 2 O (Water 18 g/mol) Hydrolysis by Amino Acid Analysis splits the amino acids in the protein apart by adding water back to the peptide bond so they can be measured. 14
Guidance of FAO/WHO on Amino Acid Molecular weight to be used for Protein FAO/WHO 2001 Rome Working Group Consultation recommended that protein should be measured as the sum of individual amino acid residues (the molecular weight of each amino acid less the molecular weight of water). 15
Effect of including water in Amino Acid Calculations Minus Water Plus Water Dehydrated Hydrated Amino Acid Ratio to FAO Amino Acid Ratio to FAO Amino Acid Content 2011 Nutrition Content 2011 Nutrition Minus Water Plus Water (g/100g) (g/100g) L-Cysteine + Crude Protein (%) = 2.72 1.254 3.14 1.446 80.4 80.4 L-Methionine* L-Tryptophan* 1.06 1.551 1.06 1.551 Animal Safe Digestibility = 0.88 0.88 L-HydroxyProline 0.00 0.00 L-Aspartic acid 6.27 7.26 First Limiting Amino Acid = L-Lysine L-Lysine L-Threonine* 2.31 0.926 2.72 1.091 L-Serine 2.79 3.37 L-Glutamic Acid 11.68 13.31 Amino Acid Score = 0.667 0.761 L-Proline 2.78 3.30 L-Glycine 2.38 3.14 0.59 0.67 PDCAAS Value = L-Alanine 3.19 4.00 L-Valine* 3.96 1.144 4.67 1.352 L-Isoleucine* 3.08 1.198 3.57 1.388 L-Leucine* 5.52 1.041 6.40 1.207 L-Tyrosine + 7.31 1.748 8.16 1.953 L-Phenylalanine* L-Lysine* 3.06 0.667 3.49 0.761 Potential of over-declaring L-Histidine* 1.62 1.006 1.83 1.138 protein content! L-Arginine 5.54 6.18 16% higher Total Protein by Amino Acid Analysis = 65.27 75.59 result 16
Digestibility Measurement: Rat PDCAAS Method Sacrifice Rats must be Live Rats of appropriate age for studies 17
Drawbacks to rat method for digestibility measurement Tim imin ing o 2-3 month turnaround Cost o $5,000+/sample Method o Use of animals for product testing 18
Additional drawbacks to rat PDCAAS method • Large amount of sample required (1 to 1.5 kg) • Cost of full proximate analysis o Formulation of balanced rat feed • Over-fortification with protein o Processing effects on score unpredictable o Significant over-use of expensive ingredients 19
Development of ASAP-Quality Score Method ( Animal-Safe Accurate Protein Quality Score ) 1 2 3 De Determine Sim imulate Rep eport Amino Acid cid Hu Human Res esult lts Composit Co ition Dig Diges estion • • Digest proteins into amino % Quality Protein • Measure using AOAC acids method 994.12 • Amino Acid Profile • React amino acids with • Digestibility Score Ninhydrin and measure • Correct measurement with Amino Acid profile from Step 1 20 Method od Re Refer erenc nce – US Pat Appl No. 14/599,050: IN VITRO METHOD FOR ESTIMATING IN VIVO PROTEIN DIGESTIBILITY, Plank, DW.
ASAP-Quality score digestion overview 21
Correlation of ASAP-Digestibility Score to Rat Digestibility Score 22
Correlation of ASAP-Quality Score to Final Rat PDCAAS Value 23
Advantages of ASAP-Quality Score • Does not use animals for testing • High correlation to animal test • Significantly reduces testing time o 15-days versus 2 to 3 months • Significantly reduces costs o $1,500 per sample versus $5,000+ per sample • Eliminates ingredient waste o Reduces need for over-fortification by food developers 24
How to improve a PDCAAS result Low Quality Protein High Quality Protein 25
Protein Complementation 2 individually inferior proteins combined in the right proportion to result in delivery of a complete protein . Food Limited Amino Acid (LAA) Complement Beans Methionine Grains, nuts, seeds Grains Lysine, Threonine Legumes Nuts/seeds Lysine Legumes Legumes Tryptophan, Methionine Grains, nuts, seeds Corn Tryptophan, lysine Legumes 26
Combining complementary protein compositions Pea protein concentrate Brown rice protein concentrate Complements LAA= Met & Cys LAA= Lys 60% pea + 40% rice = 1.053 Uncorrected amino acid score % pea % rice 0 20 40 60 80 100 27
Medallion Complementation Database • Contains amino acid analysis data and digestibility data from a wide range of ingredients o Source of data is primarily General Mills o Customer data never added to database unless specific permission granted • Database can be used to model best ingredient matches for highest protein quality • Targeting Q3 2016 for general availability to Medallion customers 28
ASAP-Quality Score Next Steps • Conduct International Collaborative Study o AOAC Validation o Official acceptance as alternative for animal testing • Develop correlation to DIAAS Protein Quality Method o New FAO international standard for protein quality 29
DIAAS Protein Quality Method • D igestible I ndispensable A mino A cid S core (DIAAS) • Based on true ileal digestibility of each amino acid • Measured at end of small intestine • Excludes large intestine fermentation • Preferably determined in humans • Alternatively: • Growing pigs (FAO recommended) • Growing rats 30
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