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6,1999 1389
Despite the absence of method performance statistics, ni- sample handling are described for many materials in the Offi-
trogen test results obtained by using Kjeldahl analyses are rou- cial Methods of Analysis of AOAC INTERNATIONAL (29).
tinely used as reference values for estimation of protein in The last issue relates to the expected method performance.
dairy products other than milk. This is an entirely valid ap- As previously noted, the method performance of the Kjeldahl
proach if the properties of the dairy materials under study, as total nitrogen method for milk is well documented. The results
they pertain to Kjeldahl analysis, are understood by both the of Kjeldahl analyses in individual laboratories or in an
researcher and the analyst. Sample origin, homogeneity, sta- interlaboratory study should be evaluated against the perfor-
bility, perishability, handling, and composition are of critical mance documented for the collaborative study of the official
importance, as are the size of the test portion and normality of method (1, 2, 6). A discussion of how to interpret collabora-
the titrant used in the Kjeldahl analysis. tive study statistics is given for those unfamiliar with the ter-
minology (30). As the nitrogen level of the material decreases
below that expected for the total nitrogen in milk, repeatability
distillation, the total volume in the receiving flask should be "hard to digest" amino acid because of its ring structure.
greater than or equal to 200 mL at the conclusion. Amino acid standards should not be predryed before use be-
cause decomposition may occur.
Recovery
Amino acids can be added directly to Kjeldahl tubes
Detecting Nitrogen Loss (weigh a container with the amino acid, pour into tube, weigh
empty container), but this may be difficult to do with tradi-
Nitrogen loss is determined by using ammonium sulfate
tional Kjeldahl flasks because of the relatively narrow necks.
standard (>99% pure). Ammonium sulfate should be predried
An alternative procedure is to weigh the amino acid directly
in an oven (100°C overnight) and cooled to room temperature
in a desiccator before use. Sucrose is added to the digestion onto a piece of ashless filter paper (Whatman No. 42 or equiv-
flask to simulate the process of digestion by providing organic alent), fold the filter paper containing the amino acid, and drop
material that will consume sulfuric acid in an amount similar it down the neck of the flask. If this procedure is used, the
Rounding the amount of nitrogen in the test portion and, second, that
there will be enough sulfuric acid remaining at the end of di-
Weights should be recorded to 4 decimal places and titra- gestion (if the solids content of the test portion is too high,
tion volumes to 2 decimal places. Calculation of percent nitro- there will not be enough sulfuric acid remaining, resulting in
gen should be carried out to at least 4 decimals before conver- low nitrogen recovery). Criteria for proper selection are dis-
sion to protein (N x 6.38). Report final protein values to cussed below.
3 decimals and nitrogen values to 4 decimals.
Titrant Normality
Procedure
If the level of nitrogen in the distillate is low, it is a simple
When all else fails, the procedure, apparatus, and reagents matter to switch titrants from 0. IN HCl to 0.01N HCl. The dis-
being used should be compared with the specifications in
Titration error Use class A buret for titration; check normality of HCI; ch
"
LYNCH & BARBANO: JOURNAL OF AOAC INTERNATIONAL VOL. 82, No. 6,1999 1397
c Consumed H 2 S0 4 =
O _ 0)
11 o 75
O 0
O
2
0
to 0
CO
g H 2 S0 4 consumed by digestion of organic matter:
[(g fat x 18) + (g protein x 9) + (g lactose x 7)]
3 CO
•a 3 CO O) o o
0 O Q. 2 CO 0 Available H 2 S0 4 = added H 2 S0 4 - consumed H 2 S0 4
o to o °> 0
* to
2 c
Q. g fi-
ll If (c) Goals.—At least 20 g H 2 S0 4 calculated as available.
8
J
g> o> (d) Examples.—(1) Whole milk.—3.S% fat, 3.3% pro-
0 .—
'0 g 5 C?
5
3
cT O •== = tein, 5.0% lactose, 5 g test portion:
^)
to
CD O) 0 2 3 -S
c > r ' 0)
T3
4- E £ 0
O CO
Added H 2 S0 4 = 44.0 g (traditional system)
&s ; Q.-0
CO _0
•D D.
CO " D
!
S o =
M i Consumed H 2 S0 4 =
the end of digestion is more complex than described previ- 10-15 g (28). To come up with a figure for "available" acid, a
ously. In addition to the amount of acid required to digest the liberal assumption was made that 5 g is lost to volatihzation.
solids in the test portion, a certain amount is lost to volatiliza- Thus, the amount of available acid by calculation is approxi-
tion. The amount lost to volatilization is a function of time, mately 20 g (15 g left as residual acid and 5 g for volatilization).
temperature, and, particularly in the case of block digestors, The actual amount of residual sulfuric acid remaining after
aspiration rate. Aspiration rate, especially, has a strong influ- digestion can be directly determined if the analyst feels adven-
ence on acid loss and can vary widely among and even within turesome. After digestion, quantitatively transfer the contents
laboratories. A further consideration is the ratio of sulfuric of the Kjeldahl flask or tube to a 500 mL volumetric flask us-
acid to potassium sulfate and the amount of sulfuric acid re- ing distilled water. Cool to 20°C and bring to volume with
quired to convert potassium sulfate to acid sulfate. 20°C distilled water. Mix well. Titrate a 25 mL aliquot with
The calculated minimum amount of acid left at the end of standardized IN NaOH using a phenolphthalein indicator.
digestion (20 g) used previously as a benchmark for determin- Calculate the amount of residual sulfuric acid as follows: