Use of soybean products in animal feeding. Part II – Quality control

use of soybean meal on animal feeding - Part2 - Quality control1


G. Fondevila

G. G. Mateos

J. L. Archs

L. Cámara

The quality of soybean meal (SBM) for animal feeding depends of the balance betweeen heat inactivaction of antinutritional factors, and the Maillard reactions rendering insoluble proteins due to excessive heat treatment. To estimate SBM quality, the feed industry uses three methods: measuring urea activity, KOH protein solubility, and protein dispersion index (PDI). This article analyses the advantages and disadvantages of each test.  



Soybean quality _ bag iconSoybeans contain variable amounts of antinutritional factors (ANF), including trypsin inhibitors (TI). They have negative effects on the activity of the enzymes trypsin and chymotrypsin, interfering with amino acid (AA) digestibility.

The antitrypsin factors are thermolabile, so their content and activity are reduced after heat processing. Therefore, the adequate processing of soybean increases AA digestibility and significantly improves the nutritional value of the soybean meal (SBM).

  • It should be considered that excess heat reduces the presence of IT, but accelerates the Maillard reaction, resulting in reduced protein digestibility, particularly Lysin and Cysteine (Fontaine et al., 2007; González-Vega I 2011).
  • Insufficient heating would leave a great part of the TI without being

To know the real quality of SBM protein, it is not enough to analyze TI levels. It is also important to determine the amount of Lys that remains active at the end of the heating process. SBM with TI values lower than 1-2 mg/g could indicate overheated products if the incidence of Maillard reactions is high.

The existing techniques to determine and estimate both TI and Maillard reactions are complicated and expensive. Therefore, it is fundamental to use correctly and carefully the methods available in the laboratory


Methods to control protein quality in soybean meal used in animal feeding

There are several methods to control the protein quality in SBM. Among them, the most used ones in the animal feed industry are:

  • Urease test
  • KOH protein solubility (KOH)
  • Protein Dispersion Index (PDI)

Urease test

soybean quality animal _ Urease activity test

Soybeans contain variable amounts of urease, a thermolabile enzyme whose destruction by heat treatment has, roughly speaking, a similar dynamic to the inactivation of TI (Balloun, 1980; Waldroup et al., 1985). The urease test measures the increase in pH due to ammonia production from urea in the presence of urease. The pH increases in unprocessed or poorly processed meals. This method indirectly measures the degree of heat processing applied to the SBM and, therefore, the degree of TI destruction.

Characteristics of the method

  • It is simple and of low cost, reason why it is the most used method in the feed
  • The main limitation of the method is that it does not include negative values. A zero value does not allow us to know whether the heat processing has been “perfect”, and destructed all ANF,  or if it has been excessive.

High urease values indicate raw or poorly processed SBM, while values close to zero can indicate well processed or overheated SBM.

KOH protein solubilityKOH protein solubility test

The solubility of the globulins, which are the major storage proteins in beans, is high but it decreases with heat processing. Before the processing, the bean protein is

very soluble, and the KOH value is above 90-95%.

The KOH protein solubility test allows us to measure soybean protein solubility in a 0.2% KOH solution, indirectly determining the degree of protein denaturation due to heat processing. The method is effective to determine the overheating of the bean or meal, however it is less effective in detecting poorly processed beans (Anderson- Haferman et al., 1992).

It should be noted that the handling of the samples during analysis – grinding size and time, stirring, temperature, pH of the solution, time, etc. – may influence the results (Whittle and Araba, 1992). In any case, the KOH index is less affected by the storage time than other methods such as PDI (Protein Dispersion Index) (Serrano et al., 2013).


Protein Dispersion Index (PDI)

PDI measures the solubility in water of the nitrogen present in soybean samples.

TPDI doya in waterhe method allows to determine the optimal level of processing of a bean or SBM. According to Hsu and Satter (1995), Batal et al. (2000) and Dudley-Cash (2001), It is more accurate than the urease test and, sometimes, than the KOH protein solubility test.

As indicated for the KOH protein solubility method, the handling and sample processing can affect the laboratory results. In particular, the duration of storage tends to reduce PDI values much more than its effects on KOH and TI (Sueiro et al., 2009a, b; Serrano et al., 2013) (Table 1). For this reason, PDI values of a SBM individual sample are possibly lower when it is analysed in the country of destination than it would be if analysed in the country of origin.

Table 1. Influence of duration of storage of soybean meal on protein quality indicators (UA = Urease activity (mg N/g); KOH = Protein solubility in KOH (%); PDI = Protein Dispersion Index (%); ATI = Activity of trypsin inhibitor (mg/g DM)) .

Influence of duration of storage of soybean meal on protein quality indicators


Despite the different existing criteria (Lee et al., 2007; from Coca-Sinova et al., 2008), KOH protein solubility, and to a lesser extent PDI, better estimate the quality of SBM processing than urease activity. It is important to consider that SBM from USA shows average KOH and PDI values 4-5 points higher than those for SBM from Brazil or Argentina (García-Rebollar et al., 2016).

Based on the present knowledge, it would probably be more convenient or more adequate to use SBM with urease values close or equal to zero and KOH protein solubility values above 85-88%.These KOH values are higher than the current recommendations put forward by most authors. Table 2 shows the acceptable values of soybean protein quality (KOH protein solubility, PDI, urease activity and ATI), according to van Eys (2012).

Table 2. Values of soybean meal quality indicators most commonly accepted by the animal feed industry (van Eys, 2012)

Table 2_Accepted values



About antinutritional factors

We need to consider….

pancreas enlargmentTrypsin inhibitors (TI)

For TI, the values are presented as activity units (ATI) and not as TI units (TIU).

  • The units are not equivalent, but it is estimated that they are closely correlated (1 x TIU = 1.9 x ATI).
  • In practice, the TI found in beans belong to two different groups, namely Kunitz and Bowman-Birk. These types differ in antinutritional effects and their sensitivities to heat. Therefore, a reduction in their levels (mg/g) does not necessarily indicate a proportional improvement in the inactivation and productive results.
  • There are different effects of TI on the physiology of different animal species.

The trypsin inhibitors cause pancreatic hypertrophy in poultry, since the birds try to compensate for the enzyme loss. This is not observed in pigs.


Stachyose and raffinose


The oligosaccharides (stachyose and raffinose), simple carbohydrates that cannot be digested by the animal but can be fermented in the large intestine, are additional relevant ANF found in SBM.

  • The excess of these oligosaccharides may lead to problems due to the uncontrolled growth of microorganisms in the cecum.
  • Moderate concentrations of oligosaccharides can be fermented to produce extra energy (volatile fatty acids), particularly in adult pigs.
  • Also, the short chain fatty acids resulting from the fermentation of oligosaccharides reduce the pH of the large intestine, what could bring benefits for intestinal health, particularly in the case of butyric acid.
  • In fact, at low concentrations the oligosaccharides are considered as prebiotics instead of ANF, in some circumstances (Grizard and Barthomeuf, 1999; Conway, 2001; Rycroft et al., 2001).


  • The use of soybean meal at high levels in animal diets requires an exhaustive control of its quality.
  • The origin of the beans, the type of processing, and storage time and conditions directly affect the characteristics of SBM.
  • For the proper estimation of SBM nutritional  value, it sis fundamental to correctly use the different test methods.
  • The presence of TI, which has consequences on animal physiology, as well as the presence of other ANF such as the oligosaccharides, should be considered when formulating diets for monogastric animals.
  • Despite its limitations, KOH protein solubility could be the method of choice to control the quality of soybean meal for animal feeding. In fact, the determination of urease activity values is of little practical value in developed countries, which have well established soybean processing industries.

Further reading 

Use of soybean products in animal feeding. Part I – Nutritional value

Use of soybean products in animal feeding. Part III- Origin








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