Biological value

Last updated

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. It captures how readily the digested protein can be used in protein synthesis in the cells of the organism. Proteins are the major source of nitrogen in food. BV assumes protein is the only source of nitrogen and measures the amount of nitrogen ingested in relation to the amount which is subsequently excreted. The remainder must have been incorporated into the proteins of the organisms body. A ratio of nitrogen incorporated into the body over nitrogen absorbed gives a measure of protein "usability" – the BV.

Contents

Unlike some measures of protein usability, biological value does not take into account how readily the protein can be digested and absorbed (largely by the small intestine). This is reflected in the experimental methods used to determine BV.

BV uses two similar scales:

  1. The true percentage utilization (usually shown with a percent symbol).
  2. The percentage utilization relative to a readily utilizable protein source, often egg (usually shown as unitless).

The two values will be similar but not identical.

The BV of a food varies greatly, and depends on a wide variety of factors. In particular the BV value of a food varies depending on its preparation and the recent diet of the organism. This makes reliable determination of BV difficult and of limited use — fasting prior to testing is universally required in order to ascertain reliable figures.

BV is commonly used in nutrition science in many mammalian organisms, and is a relevant measure in humans. [1] It is a popular guideline in bodybuilding in protein choice. [2] [3]

Determination of BV

For accurate determination of BV: [4]

  1. the test organism must only consume the protein or mixture of proteins of interest (the test diet).
  2. the test diet must contain no non-protein sources of nitrogen.
  3. the test diet must be of suitable content and quantity to avoid use of the protein primarily as an energy source.

These conditions mean the tests are typically carried out over the course of over one week with strict diet control. Fasting prior to testing helps produce consistency between subjects (it removes recent diet as a variable).

There are two scales on which BV is measured; percentage utilization and relative utilization. By convention percentage BV has a percent sign (%) suffix and relative BV has no unit.

Percentage utilization

Biological value is determined based on this formula. [4] [5]

BV = ( Nr / Na ) * 100

Where:

Na = nitrogen absorbed in proteins on the test diet
Nr = nitrogen incorporated into the body on the test diet

However direct measurement of Nr is essentially impossible. It will typically be measured indirectly from nitrogen excretion in urine. [6] Faecal excretion of nitrogen must also be taken into account - this part of the ingested protein is not absorbed by the body and so not included in the calculation of BV. An estimate is used of the amount of the urinary and faecal nitrogen excretion not coming from ingested nitrogen. This may be done by substituting a protein-free diet and observing nitrogen excretion in urine or faeces, but the accuracy of this method of estimation of the amount of nitrogen excretion not coming from ingested nitrogen on a protein-containing diet has been questioned.

BV = ( ( Ni - Ne(f) - Ne(u) ) / (Ni - Ne(f)) ) * 100

Where:

Ni = nitrogen intake in proteins on the test diet
Ne(f) = (nitrogen excreted in faeces whilst on the test diet) - (nitrogen excreted in faeces not from ingested nitrogen)
Ne(u) = (nitrogen excreted in urine whilst on the test diet) - (nitrogen excreted in urine not from ingested nitrogen)

Note:

Nr = Ni - Ne(f) - Ne(u)
Na = Ni - Ne(f)

This can take any value from 0 to 100, though reported BV could be out of this range if the estimates of nitrogen excretion from non-ingested sources are inaccurate, such as could happen if the endogenous secretion changes with protein intake. A BV of 100% indicates complete utilization of a dietary protein, i.e. 100% of the protein ingested and absorbed is incorporated into proteins into the body. The value of 100% is an absolute maximum, no more than 100% of the protein ingested can be utilized (in the equation above Ne(u) and Ne(f) cannot go negative, setting 100% as the maximum BV).

Relative utilization

Due to experimental limitations BV is often measured relative to an easily utilizable protein. Normally egg protein is assumed to be the most readily utilizable protein and given a BV of 100. For example:

Two tests of BV are carried out on the same person; one with the test protein source and one with a reference protein (egg protein).

relative BV = ( BV(test) / BV(egg) ) * 100

Where:

BV(test) = percentage BV of the test diet for that individual
BV(egg) = percentage BV of the reference (egg) diet for that individual

This is not restricted to values of less than 100. The percentage BV of egg protein is only 93.7% which allows other proteins with true percentage BV between 93.7% and 100% to take a relative BV of over 100. For example, whey protein takes a relative BV of 104, while its percentage BV is under 100%.

The principal advantage of measuring BV relative to another protein diet is accuracy; it helps account for some of the metabolic variability between individuals. In a simplistic sense the egg diet is testing the maximum efficiency the individual can take up protein, the BV is then provided as a percentage taking this as the maximum.

Conversion

Providing it is known which protein measurements were made relative to it is simple to convert from relative BV to percentage BV:

BV(relative) = ( BV(percentage) / BV(reference) ) * 100
BV(percentage) = ( BV(relative) / 100 ) * BV(reference)

Where:

BV(relative) = relative BV of the test protein
BV(reference) = percentage BV of reference protein (typically egg: 93.7%).
BV(percentage) = percentage BV of the test protein

While this conversion is simple it is not strictly valid due to the differences between the experimental methods. It is, however, suitable for use as a guideline.

Factors that affect BV

The determination of BV is carefully designed to accurately measure some aspects of protein usage whilst eliminating variation from other aspects. When using the test (or considering BV values) care must be taken to ensure the variable of interest is quantified by BV. Factors which affect BV can be grouped into properties of the protein source and properties of the species or individual consuming the protein.

Properties of the protein source

Three major properties of a protein source affect its BV:

Amino acid composition is the principal effect. All proteins are made up of combinations of the 21 biological amino acids. Some of these can be synthesised or converted in the body, whereas others cannot and must be ingested in the diet. These are known as essential amino acids (EAAs), of which there are 9 in humans. The number of EAAs varies according to species (see below).

EAAs missing from the diet prevent the synthesis of proteins that require them. If a protein source is missing critical EAAs, then its biological value will be low as the missing EAAs form a bottleneck in protein synthesis. For example, if a hypothetical muscle protein requires phenylalanine (an essential amino acid), then this must be provided in the diet for the muscle protein to be produced. If the current protein source in the diet has no phenylalanine in it the muscle protein cannot be produced, giving a low usability and BV of the protein source.

In a related way if amino acids are missing from the protein source which are particularly slow or energy consuming to synthesise this can result in a low BV.

Methods of food preparation also affect the availability of amino acids in a food source. Some of food preparation may damage or destroy some EAAs, reducing the BV of the protein source.

Many vitamins and minerals are vital for the correct function of cells in the test organism. If critical minerals or vitamins are missing from the protein source this can result in a massively lowered BV. Many BV tests artificially add vitamins and minerals (for example in yeast extract) to prevent this.

Properties of the test species or individual

Under test conditions

Variations in BV under test conditions are dominated by the metabolism of the individuals or species being tested. In particular differences in the essential amino acids (EAAs) species to species has a significant effect, although even minor variations in amino acid metabolism individual to individual have a large effect.

The fine dependence on the individual's metabolism makes measurement of BV a vital tool in diagnosing some metabolic diseases.

In everyday life

The principal effect on BV in everyday life is the organism's current diet, although many other factors such as age, health, weight, sex, etc. all have an effect. In short any condition which can affect the organism's metabolism will vary the BV of a protein source.

In particular, whilst on a high protein diet the BV of all foods consumed is reduced — the limiting rate at which the amino acids may be incorporated into the body is not the availability of amino acids but the rate of protein synthesis possible in cells. This is a major point of criticism of BV as a test; the test diet is artificially protein rich and may have unusual effects.

Factors with no effect

BV is designed to ignore variation in digestibility of a food — which in turn largely depends on the food preparation. For example, compare raw soy beans and extracted soy bean protein. The raw soy beans, with tough cell walls protecting the protein, have a far lower digestibility than the purified, unprotected, soy bean protein extract. As a foodstuff far more protein can be absorbed from the extract than the raw beans, however the BV will be the same.

The exclusion of digestibility is a point of misunderstanding and leads to misrepresentation of the meaning of a high or low BV

Advantages and disadvantages

BV provides a good measure of the usability of proteins in a diet and also plays a valuable role in detection of some metabolic diseases. BV is, however, a scientific variable determined under very strict and unnatural conditions. It is not a test designed to evaluate the usability of proteins whilst an organism is in everyday life — indeed the BV of a diet will vary greatly depending on age, weight, health, sex, recent diet, current metabolism, etc. of the organism. In addition BV of the same food varies significantly species to species. Given these limitations BV is still relevant to everyday diet to some extent. No matter the individual or their conditions a protein source with high BV, such as egg, will always be more easily used than a protein source with low BV.

In comparison to other methods known

There are many other major methods of determining how readily used a protein is, including:

These all hold specific advantages and disadvantages over BV, [7] although in the past BV has been held in high regard. [8] [9]

In animals

The Biological Value method is also used for analysis in animals such as cattle, poultry, and various laboratory animals such as rats. It was used by the poultry industry to determine which mixtures of feed were utilized most efficiently by developing chicken. Although the process remains the same, the biological values of particular proteins in humans differs from their biological values in animals due to physiological variations. [10]

Typical values

Common foodstuffs and their values: (Note: this scale uses 100 as 100% of the nitrogen incorporated.)

Common foodstuffs and their values: [18] (Note: These values use "whole egg" as a value of 100, so foodstuffs that provide even more nitrogen than whole eggs, can have a value of more than 100. 100, does not mean that 100% of the nitrogen in the food is incorporated into the body, and not excreted, as in other charts.)

By combining different foods it is possible to maximize the score, because the different components favor each other:

Criticism

Since the method measures only the amount that is retained in the body critics have pointed out what they perceive as a weakness of the biological value methodology. [21] Critics have pointed to research that indicates that because whey protein isolate is digested so quickly it may in fact enter the bloodstream and be converted into carbohydrates through a process called gluconeogenesis much more rapidly than was previously thought possible, so while amino acid concentrations increased with whey it was discovered that oxidation rates also increased and a steady-state metabolism, a process where there is no change in overall protein balance, is created. [22] They claim that when the human body consumes whey protein it is absorbed so rapidly that most of it is sent to the liver for oxidation. Hence they believe the reason so much is retained is that it is used for energy production, not protein synthesis. This would bring into question whether the method defines which proteins are more biologically utilizable.

A further critique published in the Journal of Sports Science and Medicine states that the BV of a protein does not take into consideration several key factors that influence the digestion and interaction of protein with other foods before absorption, and that it only measures a protein's maximal potential quality and not its estimate at requirement levels. [23] Also, the study by Poullain et al., which is often cited to demonstrate the superiority of whey protein hydrolysate by marketers, measured nitrogen balance in rats after three days of starvation, which corresponds to a longer period in humans. [24] The study found that whey protein hydrolysate led to better nitrogen retention and growth than the other proteins studied. However the study's flaw is in the BV method used, as starvation affects how well the body will store incoming protein (as does a very high caloric intake), leading to falsely elevated BV measures. [25]

So, the BV of a protein is related to the amount of protein given. BV is measured at levels below the maintenance level. This means that as protein intake goes up, the BV of that protein goes down. For example, milk protein shows a BV near 100 at intakes of 0.2 g/kg. As protein intake increases to roughly maintenance levels, 0.5 g/kg, BV drops to around 70. [25] Pellet et al., concluded that "biological measures of protein quality conducted at suboptimal levels in either experimental animals or human subjects may overestimate protein value at maintenance levels." As a result, while BV may be important for rating proteins where intake is below requirements, it has little bearing on individuals with protein intakes far above requirements.

This flaw is supported by the FAO/WHO/UNU, who state that BV and NPU are measured when the protein content of the diet is clearly below that of requirement, deliberately done to maximize existing differences in quality as inadequate energy intake lowers the efficiency of protein utilization and in most N balance studies, calorie adequacy is ensured. And because no population derives all of its protein exclusively from a single food, the determination of BV of a single protein is of limited use for application to human protein requirements. [26]

Another limitation of the use of Biological Value as a measure of protein quality is that proteins which are completely devoid of one essential amino acid (EAA) can still have a BV of up to 40. This is because of the ability of organisms to conserve and recycle EAAs as an adaptation of inadequate intake of the amino acid. [27]

Lastly, the use of rats for the determination of protein quality is not ideal. Rats differ from humans in requirements of essential amino acids. This has led to a general criticism that experiments on rats lead to an over-estimation of the BV of high-quality proteins to man because human requirements of essential amino acids are much lower than those for rats (as rats grow at a much faster rate than humans). Also, because of their fur, rats are assumed to have relatively high requirements of sulphur-containing amino acids (methionine and cysteine).

As a result, the analytical method that is universally recognized by the Food and Agriculture Organization (FAO), World Health Organization (WHO), the U.S. Food and Drug Administration (FDA), the United States Department of Agriculture (USDA), United Nations University (UNU) and the United States National Academy of Sciences when judging the quality of protein in the human is not PER or BV but the Protein Digestibility Corrected Amino Acid Score (PDCAAS), as it is viewed as accurately measuring the correct relative nutritional value of animal and vegetable sources of protein in the diet. [28] [29]

See also

Related Research Articles

Isoleucine Chemical compound

Isoleucine is an α-amino acid that is used in the biosynthesis of proteins. It contains an α-amino group, an α-carboxylic acid group, and a hydrocarbon side chain with a branch. It is classified as a non-polar, uncharged, branched-chain, aliphatic amino acid. It is essential in humans, meaning the body cannot synthesize it, and must be ingested in our diet. Isoleucine is synthesized from pyruvate employing leucine biosynthesis enzymes in other organisms such as bacteria. It is encoded by the codons AUU, AUC, and AUA.

A nutrient is a substance used by an organism to survive, grow, and reproduce. The requirement for dietary nutrient intake applies to animals, plants, fungi, and protists. Nutrients can be incorporated into cells for metabolic purposes or excreted by cells to create non-cellular structures, such as hair, scales, feathers, or exoskeletons. Some nutrients can be metabolically converted to smaller molecules in the process of releasing energy, such as for carbohydrates, lipids, proteins, and fermentation products, leading to end-products of water and carbon dioxide. All organisms require water. Essential nutrients for animals are the energy sources, some of the amino acids that are combined to create proteins, a subset of fatty acids, vitamins and certain minerals. Plants require more diverse minerals absorbed through roots, plus carbon dioxide and oxygen absorbed through leaves. Fungi live on dead or living organic matter and meet nutrient needs from their host.

An essential amino acid, or indispensable amino acid, is an amino acid that cannot be synthesized from scratch by the organism fast enough to supply its demand, and must therefore come from the diet. Of the 21 amino acids common to all life forms, the nine amino acids humans cannot synthesize are phenylalanine, valine, threonine, tryptophan, methionine, leucine, isoleucine, lysine, and histidine.

The net protein utilization, or NPU, is the ratio of amino acid mass converted to proteins to the mass of amino acids supplied. This figure is somewhat affected by the salvage of essential amino acids within the body, but is profoundly affected by the level of limiting amino acids within a foodstuff.

Vegetarian nutrition Nutritional and human health aspects of vegetarian diets

Vegetarian nutrition is the set of health-related challenges and advantages of vegetarian diets.

Whey protein Protein supplement

Whey protein is a mixture of proteins isolated from whey, the liquid material created as a by-product of cheese production. The proteins consist of α-lactalbumin, β-lactoglobulin, serum albumin and immunoglobulins. Whey protein is commonly marketed as a protein supplement, and various health claims have been attributed to it. A review published in 2010 in the European Food Safety Authority Journal concluded that the provided literature did not adequately support the proposed claims. For muscle growth, whey protein has been shown to be slightly better compared to other types of protein, such as casein or soy.

Protein digestibility-corrected amino acid score (PDCAAS) is a method of evaluating the quality of a protein based on both the amino acid requirements of humans and their ability to digest it.

Bodybuilding supplements are dietary supplements commonly used by those involved in bodybuilding, weightlifting, mixed martial arts, and athletics for the purpose of facilitating an increase in lean body mass. Bodybuilding supplements may contain ingredients that are advertised to increase a person's muscle, body weight, athletic performance, and decrease a person's percent body fat for desired muscle definition. Among the most widely used are high protein drinks, pre-workout blends, branched-chain amino acids (BCAA), glutamine, arginine, essential fatty acids, creatine, HMB, whey protein, ZMA, and weight loss products. Supplements are sold either as single ingredient preparations or in the form of "stacks" – proprietary blends of various supplements marketed as offering synergistic advantages.

Nitrogen balance is a measure of nitrogen input minus nitrogen output.

A complete protein or whole protein is a food source of protein that contains an adequate proportion of each of the nine essential amino acids necessary in the human diet.

Protein (nutrient) Nutrient for the human body

Proteins are essential nutrients for the human body. They are one of the building blocks of body tissue and can also serve as a fuel source. As a fuel, proteins provide as much energy density as carbohydrates: 4 kcal per gram; in contrast, lipids provide 9 kcal per gram. The most important aspect and defining characteristic of protein from a nutritional standpoint is its amino acid composition.

Protein combining or protein complementing is a dietary theory for protein nutrition that purports to optimize the biological value of protein intake. According to the theory, vegetarian and vegan diets may provide an insufficient amount of some essential amino acids, making protein combining with multiple foods necessary to obtain a complete protein food. The terms complete and incomplete are outdated in relation to plant protein. The position of the Academy of Nutrition and Dietetics is that protein from a variety of plant foods eaten during the course of a day supplies enough of all essential amino acids when caloric requirements are met.

Soy protein A protein that is isolated from soybean

Soy protein is a protein that is isolated from soybean. It is made from soybean meal that has been dehulled and defatted. Dehulled and defatted soybeans are processed into three kinds of high protein commercial products: soy flour, concentrates, and isolates. Soy protein isolate has been used since 1959 in foods for its functional properties.

Lactalbumin, also known as "whey protein", is the albumin contained in milk and obtained from whey. Lactalbumin is found in the milk of many mammals. There are alpha and beta lactalbumins; both are contained in milk.

Thomas Burr Osborne (chemist)

Thomas Burr Osborne was a biochemist who, with Lafayette Mendel, independently discovered Vitamin A, though Elmer McCollum and Marguerite Davis were ultimately given credit, as they had submitted their paper first by three weeks. He is known for his work isolating and characterizing seed proteins, and for determining protein nutritional requirements. His career was spent at the Connecticut Agricultural Experiment Station.

Cecile Hoover Edwards African American nutritional researcher

Cecile Hoover Edwards was an American nutritional researcher whose career focused on improving the nutrition and well-being of disadvantaged people. Her scientific focus was on finding low-cost foods with an optimal amino acid composition, with a special interest in methionine metabolism. She was also a university administrator, serving as dean of several schools within Howard University between 1974 and 1990.

International Year of Pulses

2016 was declared as the International Year of Pulses by the sixty eighth session of the United Nations General Assembly on December 20, 2013. The Food and Agriculture Organization (FAO) of the United Nations has been nominated to declare a year for pulses, more commonly known as legumes.

Protein quality is the digestibility and quantity of essential amino acids for providing the proteins in correct ratios for human consumption. There are various methods that rank the quality of different types of protein, some of which are outdated and no longer in use, or not considered as useful as they once were thought to be. The Protein Digestibility Corrected Amino Acid Score (PDCAAS), which was recommended by the Food and Agriculture Organization of the United Nations (FAO), became the industry standard in 1993. FAO has recently recommended the newer Digestible Indispensable Amino Acid Score (DIAAS) to supersede PDCAAS. The dairy industry is in favor of this, because while PDCAAS truncates all protein types that exceed the essential amino acid (EAA) requirements to 1.0, DIAAS allows a higher than 1.0 ranking: while for example both soy protein isolate and whey isolate are ranked 1.0 according to PDCAAS, in the DIAAS system, whey has a higher score than soy.

Vegetarian and vegan dog diet Adequate meat-free or animal-free nutrition

As in the human practice of veganism, vegan dog foods are those formulated with the exclusion of ingredients that contain or were processed with any part of an animal, or any animal byproduct. Vegan dog food may incorporate the use of fruits, vegetables, cereals, legumes including soya, nuts, vegetable oils, as well as any other non-animal based foods. The omnivorous domestic canine has evolved to metabolize carbohydrates and thrive on a diet lower in protein. A vegan diet is nutritionally adequate for dogs if properly formulated and balanced. Dogs can also thrive on a vegetarian diet.

Essential amino acids(EAAs) are amino acids that are necessary to build proteins in an organism. The source of complete EAAs are both animal and plant-based food.

References

  1. Thomas, K. Über die biologische Wertigkeit der stickstoff-substanzen in 1909 verschiedenen Nahrungsmitteln. Arch. Physiol., 219.
  2. Optimum Sports Nutrition: Your Competitive Edge, A Complete Nutritional Guide For Optimizing Athletic Performance; Chapter 12. by Dr. Michael Colgan
  3. The Great Animal Versus Vegetable Protein Debate What Is The Best Protein For Muscle Growth?
  4. 1 2 Mitchell, H.H. (1923). "A Method of Determining the Biological Value of Protein". Journal of Biol. Chem. 58 (3): 873.
  5. Chick H., Roscoe, M.H. (1930). "The biological values of proteins: A method for measuring the nitrogenous exchange of rats for the purpose of determining the biological value of proteins". Biochem J. 24 (6): 1780-2.
  6. Fixsen, M.A.B. "The biological value of purified caseinogen and the influence of vitamin B2 upon biological values, determined by the balance sheet method". Biochem J. 1930; 24(6): 1794–1804.
  7. S.G. Srikantia (August 1981). "The Use Of Biological Value Of A Protein In Evaluating Its Quality For Human Requirements". Joint FAO/WHO/UNU Expert Consultation on Energy and Protein Requirements Rome, 5 to 17 October 1981. Food and Agriculture Organization of the United Nations.
  8. Mitchell, H.H. A method for determining the biological value of protein. 1924 J. Biol. Chem., 58, 873. http://www.jbc.org/cgi/reprint/58/3/873.pdf
  9. Mitchell, H.H. and G.G. Carman. The biological value of the nitrogen of mixtures 1926 of patent white flour and animal foods. J. Biol. Chem., 68, 183.
  10. Recent developments in protein quality evaluation by Dr E. Boutrif.
  11. Hoffman, Jay R.; Falvo, Michael J. (2004). "Protein – Which is Best" (PDF). Journal of Sports Science and Medicine. 3 (3): 118–30. PMC   3905294 . PMID   24482589.
  12. 1 2 3 4 5 6 7 [Soybeans: Chemistry and Technology (copyright 1972) (b) Synder HE, Kwon TW. Soybean Utilization. Van Nostrand Reinhold Company, New York, 1987]
  13. 1 2 3 "ERRP | Expired Registration Recovery Policy". Archived from the original on 2016-03-04. Retrieved 2007-09-10.
  14. Eggum BO, Kreft I, Javornik B (1980). "Chemical-Composition and Protein-Quality of Buckwheat (Fagopyrum esculentum Moench)". Qualitas Plantarum Plant Foods for Human Nutrition. 30 (3–4): 175–9. doi:10.1007/BF01094020.
  15. 1 2 Jolliet, P. "Enteral nutrition in intensive care patients: a practical approach." Intensive Care Medicine (1998).
  16. Ruales J, Nair BM. "Nutritional quality of the protein in quinoa (Chenopodium quinoa, Willd) seeds." Plant Foods Hum Nutr. 1992 Jan;42(1):1-11.
  17. 1 2 3 "Microsoft PowerPoint - The Nutritious Egg" (PDF). Archived from the original (PDF) on 2007-08-07. Retrieved 2007-09-10.
  18. "Protein, Which Is Best" (PDF). JSSM . Retrieved 2007-10-31.
  19. Protein Quality-Report of Joint FAO’/WHO Expert Consultation, Food and Agriculture Organisation, Rome, FAO Food and Nutrition Paper 51, 1991.
  20. 1 2 3 4 "Biologische Wertigkeit Tabelle der besten Eiweiss Quellen -" (in German). 2012-06-01. Retrieved 2018-04-03.
  21. Joint FAO/WHO/UNU Expert Consultation on Energy and Protein Requirements, The use of biological value of protein in evaluatiing its quality for human requirements, S.G. Srikantia, University of Mysore.
  22. Testosterone Nation, The Protein Roundtable, August 24, 2000 Archived March 12, 2007, at the Wayback Machine .
  23. Journal of Sports Science and Medicine (2004) 3, 118-130
  24. Poullain, MG et al. Effect of whey proteins, their oligopeptide hydrosylates and free amino acid mixtures on growth and nitrogen retention in fed and starved rats. J Parenteral and Enteral Nutrition (1989) 13: 382-386
  25. 1 2 Pellett, PL and Young, VR. Nutritional evaluation of protein foods. United Nations University, 1980.
  26. The Use Of Biological Value Of A Protein In Evaluating Its Quality For Human Requirements.
  27. Said, A.K. and Hegsted, D.M., J. Nutr., 99, 474, 1969
  28. FAO/WHO (1991) Protein Quality Evaluation Report of Joint FAO/WHO Expert Consultation, Food and Agriculture Organization of the United Nations, FAO Food and Nutrition Paper No. 51, Rome.
  29. Schaafsma, G. (2000) 'The protein digestibility-corrected amino acid score. Journal of Nutrition 130, 1865S-1867S