Professional Documents
Culture Documents
H. Steinhart* / G. B i e r n o t h
University of Hamburg, Institute of Biochemistry and Food Chemistry, Grindelallee 117, 20146 Hamburg, Germany
Key Words
Gas chromatography
Column liquid chromatography
Foodstuffs quality
Trace analysis
Summary
Quality of food is the utmost goal of all food producers.
It is determined by chemical, physical, microbiological,
and even psychological aspects. The chemical and
microbiological quality aspects are especially responsible for health and welfare of the consumer. Quality of
food, therefore, at all times has to be ensured and be enhanced as far as possible. From the chemical point of
view, foods contain a great number of major and minor
components. That determine the nutritional value of a
food product. Furthermore, taste and flavour are the
result of many food minor components. Foods also may
contain unwanted compounds like residues and contaminants. Which represent undesirable aspects of food
quality.
With chromatography as a powerful analytical tool in
food chemistry, the quality of food products cannot only
be characterized but also be enhanced. Examples of
enhancing quality of food by applying chromatography
to cheese, margarine, meat, and other products is reported.
Introduction
Food Quality
For all food producers the quality of their products is an
important goal. Especially with foods, quality is what the
consumer asks for and by selling quality the producers
can keep and gain market shares.
Chromatography as VersatileMethodforFood
Analysis
By chromatographic methods, nearly all volatile and
soluble compounds in foods can be identified and quantified. This makes chromatography a powerful tool in
food analysis. In some areas like flavour research and
control, chromatographic methods are essential, because chromatographic analysis is the method of choice
for volatile compounds. But due to its versatility,
specificity, sensitivity, and simplicity, chromatography is
applied to all areas of food analysis. Nowadays its high
separation potential combined with sensitive detection
techniques made chromatography the most widely used
analytical method in food analysis. It can be used especially for trace analysis with very low detection limits.
Original
0009-5893/97
11-07
$ 03.00/0
11
Therefore, by applying chromatographic methods within food analysis, not only control of food quality became
possible but also its enhancement. For these reasons
chromatographic methods are widely applied in food research and development.
.o
pregnenolone
progesterone
OH
testosterone
Figure 1
Chemical structures of some steroidal hormones.
Chromatographia Vol.45,1997
Original
Flavour Compounds
Flavour compounds are especially suited for chromatographic analysis due to their volatility. Numerous food
have been identified by gas chromatography.
In the meantime, more tricky flavour problems can be
solved by chromatography, e.g. the question whether a
flavour component is natural or synthetic. Special forms
of chromatography may be necessary to solve such a
problem, making use of differences in the molecule like
chirality or the isotope ratio of specific atoms in the
compound [5].
Chromatographia Vol.45,1997
13
Chemical formula
methylamine
ethylamine
propylamine
butylamine
ethanolamine
2-phenylethylamine
4-arninobutyrica c i d
putrescine
cadaverine
dopamine
histamine
tryptamine
serotonine
tyramine
CH3-NH2
CH3-CH2-NH2
CH3-CH2-CH2-NH2
CH3-CH2-CH2-CH2-NH2
HO-CH2-CH2-NH2
C6H5-CH2-CH2-NH2
NHE-CH2-CH2-CH2-COOH
NH2-(CH2)4-NH2
NH2-(CHE)5-NH2
3,4-(OH)2-C6H5-CH2-CH2-NH2
4-imidazolyl-CH2-CH2-NH2
3-indolyl-CH2-CHE-NH2
3-(5-OH-indolyl)-CH2-CH2-NH2
3-HO-C6H5-CH2-CH2-NH2
l-glycine
l-alanine
t~-aminobutyricacid
norvatine
1-serine
1-phenylalanine
1-glutamic acid
l-omithine
l-lysine
l-dopa
l-histidine
l-tryptophan
5-hydroxy-l-tryptophan
l-tyrosine
The flavour of the wheat-fed carp contained more octanal, methional, and (E,Z)-2,6-nonadienal, whereas the
carp living on plankton had more (Z)-4-heptenal, 1octen-3-on, and 2-acetyl-2-thiazolin. This is a result of
differences in the composition of fatty and amino acids
in the two types of carp. Fatty and amino acids act as
flavour precursors because they are partially metabolized to flavour active c o m p o u n d s . - It was thus established by chromatographic analysis, that the quality of
carp flavour can be influenced and improved by the nature of the feed.
Original
symptoms after eating foods high in amine concentration. To avoid this, foods with low concentrations of
biogenic amines are aimed at.
A study was carried out on the biogenic amine concentration in Emmental cheeses, together with an
examination of the influence of cultures and bacteria
strains on the production of amines, and finally the effect of processing parameters.
These investigations required a lot of reliable analyses
of biogenic amines in cheese. A work-up procedure was
developed comprising an extraction step, which
separated the amines from the protein and fat, followed
by a solid phase extraction step, to separate interfering
substances. Interfering amino acids were separated by
elution on cationic exchange columns. Prior to the final
HPLC analysis amines were derivatized with phthalic
acid dialdehyde and removed by gradient elution
[12].The separated amines were detected fluorometrically. The detection limit of amines was found to be 525 pmol/ml.
As result of the measurements on Emmental cheese
samples, a variation in the content of the main biogenic
amines histamine, tyramine, putrescine, and cadaverine
from 30 up to 1800 mg/kg were found. Samples from the
inner part of cheese generally showed a lower amine
content than samples from outer part. The bacteriological status of the raw milk used to make the cheese was
identified as the main factor in the difference in amine
content. A positive correlation was found between
enterobacteria content of the milk and biogenic amines
content of the cheese. Furthermore an effect of different
types of enterobacteria on amine formation was found.
Although Emmental cheese produced in summer contained larger amounts of amines on average compared
with cheeses produced in winter time, these differences
were not marked [13].
The analytical data showed that both the ability of the
bacteria strains to form and to metabolize the biogenic
amines are responsible for the total content of amines.
This means that the amine content of the cheeses is the
sum of amine formation and decomposition. Bacteria
strains with high formation potential for specific
biogenic amines were identified and others were identified which had a high decomposition potential. Furthermore, in mixtures of bacteria strains, as usually applied in Emmental production, synergism with respect
to increased or decreased amine formation was measured. The selection of the cultures used was found to
have a strong influence and could be chosen to reduce
biogenic amine content in the cheese.
Process parameters were also investigated and it was
found that traditional methods like working in copper
rather than steel vessels and rind ripening in place of foil
ripping proved favourable to low amine content. Ripening time and temperature were also found to have a distinct influence on amine formation [14].
Original
Chromatographia Vol.45,1997
15
NH2
N,,~ -~COOH
NH2
tryptophan
1,1"-ethylidenebistryptophan(EBT)
3-phenylaminoalanine
(PAA)
Figure 2
Chemical structures of tryptophan, 1,1-ethylidenebistryptophan (EBT) and 3-phenylaminoalanine (PAA).
[~~NH2
Oxlndolylalanlne
oxidationof
pyrro~
COOH
(OIA)
OH
COOH
cOOH
hydroxylation
Tryptophan
oxidative
ofindole
ofindole
(TRP)
Hydroxytryptophan
(OH-TRP)
cleavage~
~_~ L ~ 2
O NH2
N..Fonnylkynurenlne
~COOH
Kynurenlne
(Kyn)
Figure 3
Irradiation products of tryptophan [22]
Furthermore, in the context of biotechnological production, the analytical investigations were of great importance in indicating that the EMS-contributing contaminants are formed during purification and not during
biosynthesis with gene-modified microorganisms.
16
Chromatographia Vol.45,1997
Original
Conclusions
Examples have been given where chromatographic
methods have been successfully applied to the control
and enhancement of food quality. There is no doubt that
advanced chromatographic methods will also be used
for this end in the future.
In modern quality control, analysis is carried out more
and more during production at the different production
steps. The Green light for continuation of production is
only given when various analytical parameters of the intermediates are shown to be within specification. For
this purpose chromatographic methods have to be
adopted to the demands of on-line control. Because
there is not much time for analytical work during production, rapid methods are needed. Further work on
method development seems to be necessary to achieve
this goal.
Time can especially be saved by shortening or even
omitting the work-up procedure before chromatographic analysis. This can, for instance, be achieved by
intelligent coupling of chromatographic separation and
detection methods. There are detection methods available which detect substances to be analysed very selectively so that otherwise interfering substances do not
disturb or at least to a lesser extent. In this case these
substances need not to be separated before chromatographic analysis, i. e. work-up can be shortened or
omitted.
Original
References
[1] M. Hartwig, S. Hartmann, H. Steinhart, Z. Lebensm. Unters.
Forsch. 201, 533 (1995).
[2] S.M. Grundy, N. Engl. J. Med. 323, 2307 (1990).
[3] A. Pfalzgraf, M. Timm, H. Steinhart, Z. Ern~ihrungswiss.33,
24 (1994).
[4] A. Pfalzgraf, H. Steinhart, Dtsch. Lebensm. Rundsch. 91, 113
(1995).
[5] A. Mosandl, Lebensmittelchemie 49, 130 (1995).
[6] D. Lehmann, B. Maas, A. Mosandl, Z. Lebensm. Unters.
Forsch. 201, 55 (1995).
[7] D. Bartschat, D. Lehmann, A. Dietrich, A. Mosandl, R. Kaiser,
Phytochem. Anal. 6, 130 (1995).
[8] B. Weber, B. Maas, A. Mosandl, J. Agric. Food Chem., in
press.
[9] R. Braunsdorfer, U. Hener, S. Stein, A. Mosandl, Z. Lebensm.
Unters. Forsch. 197, 385 (1993).
[10] S. Schlater, H. Steinhart, Lebensmittelchemie, in press.
[11] D.M. Beutling (ed.), "Biogene Amine in der Ern~hrung",
Springer-Verlag Berlin Heidelberg New York, 1996.
[12] K.D. Petridis, H. Steinhart, Z. Lebensm. Unters. Forsch. 201,
256 (1995).
[13] K.D. Petridis, H. Steinhart, Dtsch. Lebensm. Rundsch. 92,
114 (1996).
[14] K.D. Petridis, H. Steinhart, ibid., 92, 142 (1996).
[15] E. A. Belongia, C. W. Hedberg, G. J. Gleich, K. E. White,
A. N. Mayeno, D. A. Loegering, S. L. Dunnette, P. L. Pirie,
K. L. MacDonald, M. T. Osterholm, N. Engl. J. Med. 323,
357 (1990).
[16] A.N.Mayeno, G.J. Gleich,TrendsBiotechnol.138,346(1994).
[17] E . M . Kilbourne, Epidemiol. Rev. 14, 16 (1992).
[18] R. H. Hill, S. P. Caudill, R. M. Philen, S. L. Bailey, W. D.
Flanders, W. J. DriskeU, M. L. Kamb, L. L. Needham, E. J.
Sampson, Arch. Environ. Contam. Toxicol. 25, 134 (1993).
[19] E Sato, Y. Hagiwara, Y. Kawase, Arch. Toxicol.69, 444 (1995).
[20] T.J. Simat, K. D. Eulitz, H. Steinhart, GIT Fachzeitschrift
17