TECHNICAL NOTE

Dumas – A well-established method

for N/protein analysis

Introduction
The determination of the total protein content is an essential tool for quality
control and protein declaration according to international labeling laws in EAS DUMAS METHOD
the food & feed industry and research facilities. Protein content can directly
correspond to product properties and classifications e.g. dough properties, rapid N exceed®
foam formation, the taste of beer, or the differentiation between starch rapid MAX N exceed
and gluten-free starch. In all application areas highly precise, matrix-
independent protein analyses are required.
In principle, protein quantification is achieved via a series of specific and
non-specific physical and chemical reactions followed by suitable detection.
Full method automation as well as accordance to industry standards is
generally desirable in current laboratory operations, which has led to two
different widely accepted primary methods for the determination of total
protein content: the wet chemical method according to Kjeldahl and the
high-temperature combustion method according to Dumas. As a secondary
analysis method near infrared spectroscopy (NIRS) can be found, however,
this method requires a primary method for calibration purposes.
For more than 100 years the Kjeldahl principle was the most commonly used
method and described in the majority of standards for the determination of
total protein content of food products. As a wet chemical analysis it is time
consuming, labor-intensive and requires hazardous and toxic chemicals.
All this is nowadays unwanted with regard to laboratory safety but also for
economic reasons. This explains the trend in recent years that the Kjeldahl
principle is more and more displaced by the Dumas principle.
Combustion versus wet chemistry
The Dumas principle relies on quantitative conversion of
the sample into well-defined gaseous species at 950°C in
an oxygen enriched environment. During the combustion
phase, all nitrogen in the sample is converted to nitrogen
oxides, which are reduced to nitrogen gas and quantified
with a thermal conductivity detector. All other combustion
gases, including excess oxygen, are trapped or absorbed
prior to the quantification. The analysis time can be as
fast as 4 minutes. Figure 1 shows the functional principle
of Elementar’s next generation N/protein analyzer rapid
N exceed, which uses the EAS REGAINER® technology for a
significantly reduced cost per analysis.
Sample Carousel
Blank-Free Ball Valve
Carrier Gas: Carbon Dioxide
REGAINER
Combustion Tube
Furnace
EAS REDUCTOR
EAS
Combustion
Post
3-Fold Drying
Combustion
Figure 1. Functional principle of the rapid N exceed utilizing
the EAS REGAINER technology.
In the first step of the Kjeldahl principle the sample is
digested using concentrated sulfuric acid and a catalyst
at 420°C for 90 minutes. During this process the nitrogen
in the sample is converted to ammonium sulfate. In a
second step sodium hydroxide is added to the ammonium
solution. The released ammonia is distilled off and trapped
in a boric acid solution. The ammonium borate complex
is finally titrated with sulfuric acid or hydrochloric acid.
These two steps together require approximately 5 minutes.
With the initial digestion step including the necessary
heating and cooling time, a complete analysis will need at
least 100 minutes.
Table 1. Selected factors for different sample matrices.
SAMPLE FACTOR
eggs, meat, corn 6.25
milk 6.38
barley, millets, oats, rye 5.83
rice 5.95
wheat 5.70
Comparability of methods
Although the principles of the Kjeldahl and Dumas
methods are different, both determine the total nitrogen
content of the sample. For food samples it is known that
TCD nitrogen and protein content have a linear relationship.
By using a suitable factor, the nitrogen concentration can
therefore be converted to protein content. The factor varies
Detection depending on the relative amount of different proteins
and their amino acid composition (see Table 1).
Both methods do not differentiate between protein and
non-protein nitrogen. In most cases, the results obtained
by the Dumas method are slightly higher compared to the
Kjeldahl method. This is due to a better nitrogen recovery
of the Dumas method compared to Kjeldahl.
There are numerous scientific studies published comparing
the Kjeldahl and Dumas methods for a wide range of
different sample types. Taking into account that the
calculated protein content in both methods is only based
on approximations, the careful selection of the calculation
factor is of utmost importance. A conversion between
Kjeldahl and Dumas values using appropriate coefficients
is possible and suggested by Simonne et al. [1] for selected
food groups. However, in many cases the difference
between both methods is not statistically significant as
was shown e.g. by Željko et al. [2]
Throughput and amount of work
For Dumas analyses, sample preparation is simple and
fast. Samples are weighed into tin cups, wrapped and
placed in the autosampler of the rapid N exceed. Or, in
case of the rapid MAX N exceed, the samples are weighted
into reusable stainless steel crucibles and placed in the
autosampler. In both cases the sample preparation requires
less than a minute before the first measurement can be
started. With up to 90 sample positions in case of the rapid
MAX N exceed, unattended runs of more than 7 hours are
possible. Hence, up to 200 samples can be measured in a
routine daily operation.
In contrast, the sample preparation for Kjeldahl analyses
not only requires weighing of the sample, but also the
addition of an exact amount of sulfuric acid solution
and catalyst tablets. As already mentioned, the complete
analysis will take at least 100 minutes. It is possible to
increase the daily throughput of a Kjeldahl analyzer by
batch processing. This approach has an upper limit of
20 samples per batch, because at this number the time
necessary to perform the distillation/titration step for the
whole batch equals the time for the digestion step. This
sets the upper sample limit at 100 per day for a Kjeldahl
analyzer (see Figure 1). Although both steps are automated,
a manual sample transfer between the two modules is
inevitable. Therefore, the unattended operation time is
only 100 minutes.

Work time Unattended operation time

Dumas
190 min.
rapid N exceed
190 samples
100 min.
rapid MAX N exceed
200 samples
Kjeldahl Manual Operation (Sample Preparation)
150 min.
High-end system, Combustion & Analysis
batches of 20 Digestion
100 samples Distillation & Titration
8:00 9:00 10:00 11:00 12:00 13:00 14:00 15:00 16:00 17:00

Figure 2. Comparison of a typically daily operation with Dumas and Kjeldahl analyzers.

Cost per analysis

Traditionally, Dumas analyzers use heated metals such as It has to be noted that the labor costs are not yet included
copper or tungsten to bind excess oxygen and to reduce in the calculations. Using the Dumas method, twice as
formed nitrogen oxides to N2. The reduction metal is much samples can be analyzed per day with just 2/3 of the
oxidized to inactive metal oxide typically after 200 - 300 working time required compared to Kjeldahl.
samples when considering whole gas analysis. Therefore,
the reduction metal together with the carrier gas are the Table 2. Comparison of reagent usage for 2000 samples.
main price drivers. Elementar’s proprietary EAS REGAINER
RAPID N EXCEED KJELDAHL
technology increases the reduction metal lifetime
significantly and lowers maintenance by a factor of up to
400 g non-toxic EAS REGAINER reagent 40 l concentrated H2SO4
five. Hence, the cost per analysis based on recommended
end user prices amounts to approximately 0.25€, including 260 g copper oxide 120 l NaOH solution
all required reagents and gases in case of the rapid N
exceed and 0.49€ for the rapid MAX N exceed. 15 g catalyst 4000 catalyst tablets
The main price drivers in Kjeldahl analyses are the costs
for chemicals and their disposal and manpower, resulting 90 g drying agent, 65 g corundum 40 l HCl solution, 0.2 N

in an overall cost per analysis of approximately 6.00€.

3.5% of a standard laboratory oxygen
The incurred chemical waste from a typical Kjeldahl 120 l H3BO3 buffer solution
gas cylinder (grade 4.5)
methodology for 2000 samples amounts to 560 liter
compared to a negligible amount of only 430 g solids 1/3 of a standard laboratory carbon
240 l ultrapure water
dioxide gas cylinder (grade 4.5)
for the rapid N exceed, as presented in Table 2. The EAS
REGAINER reagent is exhausted during the analysis and 2000 tin foils or nitrogen free paper
2000 sheets of weighing paper
doesn’t form any solid waste. wrappings
 Standards using the Dumas method
The displacement of the Kjeldahl method by the Dumas method also explains the increasing number of valid standards
world-wide which codify the determination of total protein content according to the Dumas principle.

Table 3. List of the most important international and national standards.

STANDARD DESCRIPTION
Milk and milk products - determination of nitrogen content -
DIN EN ISO 14891:2002-07
routine method using combustion according to the Dumas principle
Food products - determination of the total nitrogen content by combustion according to the Dumas principle and calculation of the crude protein content
DIN EN ISO 16634-1:2009-07
- part 1: oilseeds and animal feeding stuffs
Cereals, pulses, milled cereal products, oilseeds and animal feeding stuffs - determination of the total nitrogen content by combustion
DIN EN ISO 16634-2
according to the Dumas principle and calculation of the crude protein content

ICC STANDARD No. 167 Determination of crude protein in grain and grain products for food and feed by the Dumas combustion principle

AACC METHOD 46.30 Crude protein – combustion method (animal feeds, cereals and oil seeds)

AOAC 990.03-2002 Crude protein in animal feed

AOAC 992.15-1992 (1996) Crude protein in meat and meat products

AOAC 992.23-1992(1998) Crude protein in cereal grains and oilseeds

AOAC 993.13-1996 Nitrogen (total) in fertilizers

AOAC 997.09-2008 Nitrogen in beer, wort, and brewing grains

AOCS BA 4E-93 (11) Generic combustion method for determination of crude protein.

AOCS BA 4F-00 (11) Combustion method for determination of crude protein in soybean meal

ASBC COMBUSTION METHODS Nitrogen by combustion method - beer, wort, malt, barley, brewing grains, adjunct materials and cereals

ICC (International Association of Cereal Science and Technology); ISO (International Organization for Standardization); AACC
(American Association of Cereal Chemists); AOAC (Association of Official Analytical Chemists); AOCS (American Oil Chemists
Society); ASBC (American Society of Brewing Chemists).

Summary
It was shown that the Dumas method has clear advantages
over Kjeldahl regarding laboratory
throughput, labor time, amount of chemical waste and
thus cost-per-analysis. Furthermore, the high level of
automation also results in less risk of errors.
With Elementar’s next generation N/Protein analyzers,
the cost-per-analysis was drastically reduced compared
to “classical” Dumas analyzers. The investment into
safety, sample a rapid N exceed is now economically justified even for
laboratories with only 10 samples per day. It furthermore
gives secureness and confident in possible future growths
of samples. The low initial investment cost for entry-level
Kjeldahl analyzers are nowadays not anymore the key
selling point.

[1] Simonne, A.H., Simonne, E.H., Eitenmiller, R.R., Mills, H.A., Cresman III, C.P. 1997. Could the Dumas method replace the
Kjeldahl digestion for nitrogen and crude protein determinations in foods? Journal of the Science of Food and Agriculture,
73:39-45.
[2] Željko, A.M., Sandra M.J., Nadežda, B.P., Željko, N.Ć., Milica, M.Ž., Comparison of the Kjeldahl method, Dumas method
and NIR method for total nitrogen determination in meat and meat products Journal of Agroalimentary Processes and
Technologies 2015, 21(4), 365-370.

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