Protein (nutrient)

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This article is about protein as a nutrient. For protein as class of molecule, see protein. For other uses, see bodybuilding supplement.
Amino acids are the building blocks of protein.
Amino acids are necessary nutrients. Present in every cell, they are also precursors to nucleic acids, co-enzymes, hormones, immune response, repair and other molecules essential for life.

Proteins are essential nutrients for the human body.[1] They are one of the building blocks of body tissue, and can also serve as a fuel source. As a fuel, proteins contain 4 kcal per gram, just like carbohydrates and unlike lipids, which contain 9 kcal per gram. The most important aspect and defining characteristic of protein from a nutritional standpoint is its amino acid composition.[2]

Proteins are polymer chains made of amino acids linked together by peptide bonds. During human digestion, proteins are broken down in the stomach to smaller polypeptide chains via hydrochloric acid and protease actions. This is crucial for the synthesis of the essential amino acids that cannot be biosynthesized by the body.[3]

There are nine essential amino acids which humans must obtain from their diet in order to prevent protein-energy malnutrition. They are phenylalanine, valine, threonine, tryptophan, methionine, leucine, isoleucine, lysine, and histidine.[4][2] There are five dispensable amino acids which humans are able to synthesize in the body. These five are alanine, aspartic acid, asparagine, glutamic acid and serine.[2] There are six conditionally essential amino acids whose synthesis can be limited under special pathophysiological conditions, such as prematurity in the infant or individuals in severe catabolic distress.[2] These six are arginine, cysteine, glycine, glutamine, proline and tyrosine.[2]

Humans need the essential amino acids in certain ratios. Some protein sources contain amino acids in a more or less 'complete' sense. This has given rise to various ranking systems for protein sources, as described in the article.

Animal sources of protein include meats, dairy products, fish and eggs. Vegan sources of protein include whole grains, pulses, legumes, soy, and nuts. Vegetarians and vegans get enough essential amino acids by eating a variety of plant proteins.[5] It is commonly believed that athletes should consume a higher-than-normal protein intake to maintain optimal physical performance.[2]

Protein functions in body[edit]

Protein is a nutrient needed by the human body for growth and maintenance.[1] Aside from water, proteins are the most abundant kind of molecules in the body. Protein can be found in all cells of the body and is the major structural component of all cells in the body, especially muscle.[1][6] This also includes body organs, hair and skin. Proteins are also used in membranes, such as glycoproteins. When broken down into amino acids, they are used as precursors to nucleic acid, co-enzymes, hormones, immune response, cellular repair, and other molecules essential for life.[6] Additionally, protein is needed to form blood cells.[1][7]

Protein function in exercise[edit]

Proteins are believed to increase performance in terms of athletics.[7] Amino acids, the building blocks of proteins, are used for building muscle tissue and repairing damaged tissues.[7] Protein is only used as fuel when carbohydrates and lipid resources are low.[7]

Sources[edit]

Animal sources of protein.

Protein can be found in a wide range of food. The best combination of protein sources depends on the region of the world, access, cost, amino acid types and nutrition balance, as well as acquired tastes. Some foods are high in certain amino acids, but their digestibility and the anti-nutritional factors present in these foods make them of limited value in human nutrition. Therefore, one must consider digestibility and secondary nutrition profile such as calories, cholesterol, vitamins and essential mineral density of the protein source.[8] On a worldwide basis, plant protein foods contribute over 60 percent of the per capita supply of protein, on average. In North America, animal-derived foods contribute about 70 percent of protein sources.[citation needed]

Meat, products from milk, eggs and fish are sources of complete protein.[9]

Whole grains and cereals are another source of proteins. However, these tend to be limiting in the amino acid lysine or threonine, which are available in other vegetarian sources and meats. Examples of food staples and cereal sources of protein, each with a concentration greater than 7 percent, are (in no particular order) buckwheat, oats, rye, millet, maize (corn), rice, wheat, bulgar, sorghum, amaranth, and quinoa.

Vegetarian sources of proteins include legumes, nuts, seeds and fruits. Legumes, some of which are called pulses in certain parts of the world, have higher concentrations of amino acids and are more complete sources of protein than whole grains and cereals. Examples of vegetarian foods with protein concentrations greater than 7 percent include soybeans, lentils, kidney beans, white beans, mung beans, chickpeas, cowpeas, lima beans, pigeon peas, lupines, wing beans, almonds, Brazil nuts, cashews, pecans, walnuts, cotton seeds, pumpkin seeds, sesame seeds, and sunflower seeds.[8]

Plant sources of protein.

Food staples that are poor sources of protein include roots and tubers such as yams, cassava and sweet potato. Plantains, another major staple, are also a poor source of essential amino acids. Fruits, while rich in other essential nutrients, are another poor source of amino acids. The protein content in roots, tubers and fruits is between 0 and 2 percent. Food staples with low protein content must be complemented with foods with complete, quality protein content for a healthy life, particularly in children for proper development.[1][10][11]

A good source of protein is often a combination of various foods, because different foods are rich in different amino acids. A good source of dietary protein meets two requirements:[8]

  • The requirement for the nutritionally indispensable amino acids (histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine) under all conditions and for conditionally indispensable amino acids (cystine, tyrosine, taurine, glycine, arginine, glutamine, proline) under specific physiological and pathological conditions
  • The requirement for nonspecific nitrogen for the synthesis of the nutritionally dispensable amino acids (aspartic acid, asparagine, glutamic acid, alanine, serine) and other physiologically important nitrogen-containing compounds such as nucleic acids, creatine, and porphyrins.

Healthy people eating a balanced diet rarely need protein supplements.[5][8] Except for a few amino acids, most are readily available in human diet. The limiting amino acids are lysine, threonine, tryptophan and sulfur-containing amino acids.[10]

The table below presents the most important food groups as protein sources, from a worldwide perspective. It also lists their respective performance as source of the commonly limiting amino acids, in milligrams of limiting amino acid per gram of total protein in the food source.[10] The green highlighted cells represent the protein source with highest density of respective amino acid, while the yellow highlighted cells represent the protein source with lowest density of respective amino acid. The table reiterates the need for a balanced mix of foods to ensure adequate amino acid source.[citation needed]

Food source Lysine Threonine Tryptophan Sulfur containing
amino acids
Legumes 64 38 12 25
Cereals and whole grains 31 32 12 37
Nuts and seeds 45 36 17 46
Fruits 45 29 11 27
Animal 85 44 12 38
Protein milkshakes, made from protein powder (center) and milk (left), are a common bodybuilding supplement.

Protein powders – such as casein, whey, egg, rice and soy – are processed and manufactured sources of protein.[12] These protein powders may provide an additional source of protein for bodybuilders. The type of protein is important in terms of its influence on protein metabolic response and possibly on the muscle's exercise performance. The different physical and/or chemical properties within the various types of protein may affect the rate of protein digestion. As a result, the amino acid availability and the accumulation of tissue protein is altered because of the various protein metabolic responses.[12]

Protein quality[edit]

The most important aspect and defining characteristic of protein from a nutritional standpoint is its amino acid composition.[2] There are multiple systems which rate proteins by their usefulness to an organism based on their relative percentage of amino acids and, in some systems, the digestibility of the protein source. They include biological value, net protein utilization, and PDCAAS (Protein Digestibility Corrected Amino Acids Score). Also see complete protein, nitrogen balance and protein combining. The PDCAAS was developed by the FDA as an improvement over the Protein efficiency ratio (PER) method. The PDCAAS rating is a fairly recent evaluation method; it was adopted by the US Food and Drug Administration (FDA) and the Food and Agricultural Organization of the United Nations/World Health Organization (FAO/WHO) in 1993 as "the preferred 'best'" method to determine protein quality. These organizations have suggested that other methods for evaluating the quality of protein are inferior.[13]

Digestion[edit]

Most proteins are decomposed to single amino acids by digestion in the gastro-intestinal tract.[14]

Digestion typically begins in the stomach when pepsinogen is converted to pepsin by the action of hydrochloric acid, and continued by trypsin and chymotrypsin in the small intestine.[14] Before the absorption in the small intestine, most proteins are already reduced to single amino acid or peptides of several amino acids. Most peptides longer than four amino acids are not absorbed. Absorption into the intestinal absorptive cells is not the end. There, most of the peptides are broken into single amino acids.

Absorption of the amino acids and their derivatives into which dietary protein is degraded is done by the gastrointestinal tract. The absorption rates of individual amino acids are highly dependent on the protein source; for example, the digestibilities of many amino acids in humans, the difference between soy and milk proteins[15] and between individual milk proteins, beta-lactoglobulin and casein.[16] For milk proteins, about 50% of the ingested protein is absorbed between the stomach and the jejunum and 90% is absorbed by the time the digested food reaches the ileum.[17] Biological value (BV) is a measure of the proportion of absorbed protein from a food which becomes incorporated into the proteins of the organism's body.

Newborn[edit]

Newborns of mammals are exceptional in protein digestion and assimilation in that they can absorb intact proteins at the small intestine. This enables passive immunity from milk.

Dietary requirements[edit]

An education campaign launched by the United States Department of Agriculture about 100 years ago, on cottage cheese as a lower-cost protein substitute for meat.

Considerable debate has taken place regarding issues surrounding protein intake requirements.[18][19] The amount of protein required in a person's diet is determined in large part by overall energy intake, the body's need for nitrogen and essential amino acids, body weight and composition, rate of growth in the individual, physical activity level, individual's energy and carbohydrate intake, as well as the presence of illness or injury.[3][12][20] Physical activity and exertion as well as enhanced muscular mass increase the need for protein. Requirements are also greater during childhood for growth and development, during pregnancy or when breast-feeding in order to nourish a baby, or when the body needs to recover from malnutrition or trauma or after an operation.[21]

If not enough energy is taken in through diet, as in the process of starvation, the body will use protein from the muscle mass to meet its energy needs, leading to muscle wasting over time. If the individual does not consume adequate protein in nutrition, then muscle will also waste as more vital cellular processes (e.g. respiration enzymes, blood cells) recycle muscle protein for their own requirements.

According to US & Canadian Dietary Reference Intake guidelines, women aged 19–70 need to consume 46 grams of protein per day, while men aged 19–70 need to consume 56 grams of protein per day to avoid a deficiency.[22] The generally accepted daily protein dietary allowance, measured as intake per kilogram of body weight, is 0.8 g/kg.[18] However, this recommendation is based on structural requirements, but disregards use of protein for energy metabolism.[18] This requirement is for a normal sedentary person.[20]

Several studies have concluded that active people and athletes may require elevated protein intake (compared to 0.8 g/kg) due to increase in muscle mass and sweat losses, as well as need for body repair and energy source.[18][19][20] Suggested amounts vary between 1.6 g/kg and 1.8 g/kg,[19] while a proposed maximum daily protein intake would be approximately 25% of energy requirements i.e. approximately 2 to 2.5 g/kg.[18] However, many questions still remain to be resolved.[19]

Aerobic exercise protein needs[edit]

Endurance athletes differ from strength-building athletes in that endurance athletes do not build muscle mass from training.[citation needed] Research suggests that individuals performing endurance activity require more protein intake than sedentary individuals so that muscles broken down during endurance workouts can be repaired.[23] Although the protein requirement for athletes still remains controversial (for instance see Lamont, Nutrition Research Reviews, pges 142 - 149, 2012), research does show that endurance athletes can benefit from increasing protein intake because the type of exercise endurance athletes participate in still alters the protein metabolism pathway. The overall protein requirement increases because of amino acid oxidation in endurance-trained athletes.[23] Endurance athletes who exercise over a long period (2–5 hours per training session) use protein as a source of 5–10% of their total energy expended. Therefore, a slight increase in protein intake may be beneficial to endurance athletes by replacing the protein lost in energy expenditure and protein lost in repairing muscles. Some scientists suggest that endurance athletes may increase daily protein intake to a maximum of 1.2–1.4 g per kg body weight.[12]

Anaerobic exercise protein needs[edit]

Research also indicates that individuals performing strength-training activity require more protein than sedentary individuals.[3][12][23] Strength-training athletes may increase their daily protein intake to a maximum of 1.4–1.8 g per kg body weight to enhance muscle protein synthesis, or to make up for the loss of amino acid oxidation during exercise.[12][24] Many athletes maintain a high-protein diet as part of their training, and so protein deficiency is less likely among this group than among non-athletes.[24] In fact, some athletes who specialize in anaerobic sports (e.g. weightlifting) assume a very high level of protein intake is necessary, and may over-consume.[7][24] Research indicates that many athletes consume more protein than they need even without the use of protein supplements.[7]

Special populations[edit]

Phenylketonuria[edit]

Individuals with phenylketonuria (PKU) must keep their intake of phenylalanine extremely low to prevent mental retardation and other metabolic complications.

Maple Syrup Urine Disease[edit]

Maple syrup urine disease is associated with genetic anomalies in the metabolism of Branched Chain Amino Acids (BCAAs). They have high blood levels of BCAAs and must severely restrict their intake of BCAAs in order to prevent mental retardation and death.[2]

Excess consumption[edit]

When a high dietary protein intake is consumed, there is an increase in urea excretion, which suggests that amino acid oxidation is increased.[20] High levels of protein intake increase the activity of branched-chain ketoacid dehydrogenase.[20] As a result, oxidation is facilitated, and the amino group of the amino acid is excreted to the liver.[20] This process suggests that excess protein consumption results in protein oxidation and that the protein is excreted.[20] The body is unable to store excess protein.[20][25] Protein is digested into amino acids, which enter the bloodstream. Excess amino acids are converted to other usable molecules by the liver in a process called deamination. Deamination converts nitrogen from the amino acid into ammonia, which is converted by the liver into urea in the urea cycle. Excretion of urea is performed by the kidneys. These organs can normally cope with any extra workload, but, if kidney disease occurs, a decrease in protein will often be prescribed.[26] When there is excess protein intake, amino acids can be converted to glucose or ketones, in addition to being oxidized for fuel.[27] When food protein intake is periodically high or low, the body tries to keep protein levels at an equilibrium by using the "labile protein reserve", which serves as a short-term protein store to be used for emergencies or daily variations in protein intake.[6] However, that reserve is not utilized as longer-term storage for future needs.[6]

Many researchers have also found that excessive intake of protein increases calcium excretion in urine.[6] It has been thought that this occurs to maintain the pH imbalance from the oxidation of sulfur amino acids.[6] Also, it is inconclusive whether bone resorption contributes to bone loss and osteoporosis.[6] However, it is also found that a regular intake of calcium would be able to stabilize this loss.[6]

Another issue arising from over-consumption of protein is a higher risk of kidney stone formation from calcium in the renal circulatory system.[6] It has been found that high animal protein intake in healthy individuals increases the probability of forming kidney stones by 250 percent.[citation needed]

An epidemiological study from 2006 has found no relationship between total protein intake and blood pressure; it did, however, find an inverse relationship between vegetable protein intake and blood pressure.[28]

Testing in foods[edit]

The classic assays for protein concentration in food are the Kjeldahl method and the Dumas method. These tests determine the total nitrogen in a sample. The only major component of most food which contains nitrogen is protein (fat, carbohydrate and dietary fibre do not contain nitrogen). If the amount of nitrogen is multiplied by a factor depending on the kinds of protein expected in the food the total protein can be determined. This value is known as the "crude protein" content. On food labels the protein is given by the nitrogen multiplied by 6.25, because the average nitrogen content of proteins is about 16%. The Kjeldahl test is typically used because it is the method the AOAC International has adopted and is therefore used by many food standards agencies around the world, though the Dumas method is also approved by some standards organizations.[29]

Accidental contamination and intentional adulteration of protein meals with non-protein nitrogen sources that inflate crude protein content measurements have been known to occur in the food industry for decades. To ensure food quality, purchasers of protein meals routinely conduct quality control tests designed to detect the most common non-protein nitrogen contaminants, such as urea and ammonium nitrate.[30]

In at least one segment of the food industry, the dairy industry, some countries (at least the U.S., Australia, France and Hungary), have adopted "true protein" measurement, as opposed to crude protein measurement, as the standard for payment and testing: "True protein is a measure of only the proteins in milk, whereas crude protein is a measure of all sources of nitrogen and includes nonprotein nitrogen, such as urea, which has no food value to humans. ... Current milk-testing equipment measures peptide bonds, a direct measure of true protein."[31] Measuring peptide bonds in grains has also been put into practice in several countries including Canada, the UK, Australia, Russia and Argentina where near-infrared reflectance (NIR) technology, a type of infrared spectroscopy is used.[32] The Food and Agriculture Organization of the United Nations (FAO) recommends that only amino acid analysis be used to determine protein in, inter alia, foods used as the sole source of nourishment, such as infant formula, but also provides: "When data on amino acids analyses are not available, determination of protein based on total N content by Kjeldahl (AOAC, 2000) or similar method ... is considered acceptable."[33]

The limitations of the Kjeldahl method were at the heart of the Chinese protein export contamination in 2007 and the 2008 China milk scandal in which the industrial chemical melamine was added to the milk or glutens to increase the measured "protein".[34][35]

Protein deficiency[edit]

A child in Africa suffering from kwashiorkor – one of the three protein energy malnutrition ailments afflicting over 10 million children in developing countries.[36]

Protein deficiency and malnutrition can lead to variety of ailments including mental retardation and kwashiorkor.[11] Symptoms of kwashiorkor include apathy, diarrhea, inactivity, failure to grow, flaky skin, fatty liver, and edema of the belly and legs. This edema is explained by the action of lipoxygenase on arachidonic acid to form leukotrienes and the normal functioning of proteins in fluid balance and lipoprotein transport.[37]

PEM is fairly common worldwide in both children and adults and accounts for 6 million deaths annually.[2] In the industrialized world, PEM is predominantly seen in hospitals, is associated with disease, or is often found in the elderly.[2]

See also[edit]

References[edit]

  1. ^ a b c d e Hermann, Janice R. "Protein and the Body". Oklahoma Cooperative Extension Service, Division of Agricultural Sciences and Natural Resources • Oklahoma State University: T–3163–1 – T–3163–4. 
  2. ^ a b c d e f g h i j Dietary Reference Intakes: The Essential Guide to Nutrient Requirements, published by the Institute of Medicine's Food and Nutrition Board, currently available online at http://fnic.nal.usda.gov/dietary-guidance/dietary-reference-intakes/dri-reports
  3. ^ a b c Genton, Laurence; Melzer, Katarina; Pichard, Claude (2010). "Energy and macronutrient requirements for physical fitness in exercising subjects". Clinical Nutrition 29 (4): 413–423. doi:10.1016/j.clnu.2010.02.002. PMID 20189694. 
  4. ^ Young VR (1994). "Adult amino acid requirements: the case for a major revision in current recommendations". J. Nutr. 124 (8 Suppl): 1517S–1523S. PMID 8064412. 
  5. ^ a b "Protein in diet". United States National Library of Medicine, National Institutes of Health. 2009. 
  6. ^ a b c d e f g h i Food and Nutrition Board (2005). A Report of the Panel on Macronutrients, Subcommittees on Upper Reference Levels of Nutrients and Interpretation and Uses of Dietary Reference Intakes, and the Standing Committee on the Scientific Evaluation of Dietary Reference Intakes. Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids (Macronutrients). THE NATIONAL ACADEMIES PRESS, Washington, D.C. ISBN 0-309-08537-3. 
  7. ^ a b c d e f Nutrition Working Group of the International Olympic Committee (2003). "Nutrition for Athletes". IOC Consensus Conference on Nutrition for Sport. Lausanne.  Missing or empty |title= (help)
  8. ^ a b c d Vernon Young, Peter Pellett (1994). "Plant proteins in relation to human protein and amino acid nutrition". American Journal of Clinical Nutrition 59: 1203S-l2S. 
  9. ^ Steinke, Waggle et al. (1992). New protein foods in human health: nutrition, prevention and therapy. CRC Press. pp. 91–100. ISBN 978-0-8493-6904-9. 
  10. ^ a b c Amino acid content of foods and biological data on proteins (FAO nutritional studies number 24). Food and Agriculture Organization. 1985. ISBN 92-5-001102-4. 
  11. ^ a b Michael C. Latham (1997). "Human nutrition in the developing world". Food and Agriculture Organization of the United Nations. 
  12. ^ a b c d e f Lemon, PW (June 1995). "Do athletes need more dietary protein and amino acids?". Int J Sport Nutr. 5 Suppl: S39–61. PMID 7550257. 
  13. ^ Boutrif, E., Food Quality and Consumer Protection Group, Food Policy and Nutrition Division, FAO, Rome: "Recent Developments in Protein Quality Evaluation" Food, Nutrition and Agriculture, Issue 2/3, 1991
  14. ^ a b Digestion of Dietary Proteins in the Gastro-Intestinal Tract
  15. ^ Gaudichon C, Bos C, Morens C, Petzke KJ, Mariotti F, Everwand J, Benamouzig R, Dare S, Tome D, Metges CC (2002). "Ileal losses of nitrogen and amino acids in humans and their importance to the assessment of amino acid requirements". Gastroenterology 123 (1): 50–9. doi:10.1053/gast.2002.34233. PMID 12105833. 
  16. ^ Mahe S, Roos N, Benamouzig R, Davin L, Luengo C, Gagnon L, Gausserges N, Rautureau J, Tome D (1996). "Gastrojejunal kinetics and the digestion of [15N]beta-lactoglobulin and casein in humans: the influence of the nature and quantity of the protein". Am J Clin Nutr 63 (4): 546–52. PMID 8599318. 
  17. ^ Mahe S, Marteau P, Huneau JF, Thuillier F, Tome D (1994). "Intestinal nitrogen and electrolyte movements following fermented milk ingestion in man". Br J Nutr 71 (2): 169–80. doi:10.1079/BJN19940124. PMID 8142329. 
  18. ^ a b c d e Bilsborough, Shane; Neil Mann (2006). "A Review of Issues of Dietary Protein Intake in Humans". International Journal of Sport Nutrition and Exercise Metabolism (16): 129–152. Retrieved 6 December 2012. 
  19. ^ a b c d Lemon, Peter (2000). "Beyond the Zone: Protein Needs of Active Individuals". Journal of the American College of Nutrition 19 (5): 513–521. doi:10.1080/07315724.2000.10718974. PMID 11023001. 
  20. ^ a b c d e f g h Tarnopolsky MA, Atkinson SA, MacDougall JD, Chesley A, Phillips S, Schwarcz HP (1992). "Evaluation of protein requirements for trained strength athletes". Journal of Applied Physiology 73 (5): 1986–95. PMID 1474076. 
  21. ^ World Health Organization, Food and Agriculture Organization of the United Nations , United Nations University (2007). "Protein and amino acid requirements in human nutrition" (PDF). WHO Press. Retrieved 8 July 2008. 
  22. ^ "Dietary reference intakes: macronutrients" (PDF). Institute of Medicine. Retrieved 18 May 2008. 
  23. ^ a b c Phillips, Stuart (2006). "Dietary protein for athletes: from requirements to metabolic advantage". Appl. Physiol. Nutr. Metab. 31 (6): 647–654. doi:10.1139/H06-035. PMID 17213878. 
  24. ^ a b c Lemon, Peter (1995). "Do athletes need more dietary protein and amino acids?". International Journal of Sport Nutrition 5: S39–S61. PMID 7550257. 
  25. ^ Ten Have, Gabriella A.M.; Engelen, Marielle P.K.J.; Luiking, Yvette C.; Deutz, Nicolaas E.P. (2007). "Absorption Kinetics of Amino Acids, Peptides, and Intact Proteins". International Journal of Sport Nutrition and Exercise Metabolism 17: S23–S36. 
  26. ^ Born Steve. "Fueling for endurance: ten mistakes endurance athletes make and how you can avoid them". UltraCycling Magazine. 
  27. ^ Smith, Jack L.; Gropper, Sareen Annora Stepnick; Groff, James L. (2009). Advanced nutrition and human metabolism. Belmont, CA: Wadsworth Cengage Learning. ISBN 0-495-11657-2. 
  28. ^ Elliott, Paul; Stamler, Jeremiah; Dyer, Alan R.; Appel, Lawrence; Dennis, Barbara; Kesteloot, Hugo; Ueshima, Hirotsugu; Okayama, Akira; Chan, Queenie; Garside, Daniel B.; Beifan, Zhou (2006). "Association between protein intake and blood pressure: the INTERMAP Study". Archives of Internal Medicine 166 (1): 79–87. doi:10.1001/archinte.166.1.79. PMID 16401814. Retrieved 21 January 2013. 
  29. ^ Dr. D. Julian McClements. "Analysis of Proteins". University of Massachusetts Amherst. Retrieved 27 April 2007. 
  30. ^ Weise, Elizabeth (24 April 2007). "Food tests promise tough task for FDA". USA Today. Retrieved 29 April 2007. 
  31. ^ P.M. VanRaden and R.L. Powell. "Genetic evaluations for true protein". United States Department of Agriculture. Retrieved 27 April 2007. 
  32. ^ Snyder, Alison (August 2007). "Protein Pretense: Cheating the standard protein tests is easy, but industry hesitates on alternatives". Scientific American. Retrieved 9 November 2007. 
  33. ^ "Food energy – methods of analysis and conversion factors". FAO. Retrieved 9 November 2007. 
  34. ^ Stephen Chen (18 September 2008). "Melamine – an industry staple". South China Morning Post. pp. Page A2. 
  35. ^ Moore, J.C.; Devries, Jonathan W.; Lipp, Markus; Griffiths, James C.; Abernethy, Darrell R. (17 August 2010). "Total Protein Methods and Their Potential Utility to Reduce the Risk of Food Protein Adulteration". Comprehensive Reviews in Food Science and Food Safety 9 (4): 330–357. doi:10.1111/j.1541-4337.2010.00114.x. 
  36. ^ "Marasmus and Kwashiorkor". Medscape Reference. May 2009. 
  37. ^ Jeffery Schwartz; Bryant, Carol A.; DeWalt, Kathleen Musante; Anita Courtney (2003). The cultural feast: an introduction to food and society. Belmont, California: Thomson/Wadsworth. pp. 282, 283. ISBN 0-534-52582-2.