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Fachbereich fir Bauingenieurwesen, Labor fir Abfallwirtschaft, Siedlungswasserwirtschaft, Umweltchemie (LASU) , Fachhochschule
Mtinster, D-48149 Mtinster, Germany
Pharmazeutisches lnstitut Poppelsdorf, Rheinische Friedrich- Wilhelms- Universittit Bonn, Kreuzbergweg 26, D-531 15 Bonn, Germany
Departamento de Microbiologia, Facultad de Farmacia, Universidad de Granada, 18071 Granada, Spain
dDepartamento de Ingenieria Quimica, Facultad de Ciencias, Universidad de Granada, 18071 Granada, Spain
G. Knupp et al.
278
CCQH
-i-)
HO-j=&
H~cooH
Brenzcatechol
4-Hydroxyphenylacetic
acid
3.4-Dihydroxyphenylacetic
acid
R=H
R = OH
R=H
R=OH
R = OCHs
Cumaric acid
Caffeic acid
Ferulic acid
OH
H
HO
COOH
R
RI =H
Rj=OH
[ RI = OH
R2 = H
Tyrosol
R2=H
R2 = OH
Hydroxytyrosol
3,4-Dihydroxyphenylglycol ]
Fig. 1. Phenolic
compounds
of alpechin
Oily residue:
phenolic
R=H
R=OH
4-Hydroxybenzoic
acid
Protocatechuic acid
described
in the literature.
Water residue
t
Derivatization:
MSTFA
85C I5 min.
IrnD
k
Compound
GCiMS
Fig. 2. Flowchart
of the analytical
method.
transforms
both
hydroxylic
and
carboxylic
functional groups into the corresponding TMSethers and -esters. It has been proven with the help
of reference substances that no intermediate
products due to non-quantitative
reactions are
produced using this reagent. The derivatized residue
is then separated by capillary-CC
and the
corresponding mass spectra are recorded.
When analysing alpechin in this way, a first idea
of the compound identity can be obtained by
comparing
times
in
the
retention
gas
chromatogram with those of reference substances.
Then, the identities of the phenolic compounds
are confirmed by comparing the mass spectra with
those of authentic reference substances. This
procedure is necessary because there are some
cases described in the literature where retention
times of compounds
from alpechin correlate
perfectly with those of reference substances
although they are not identical (Lopez Aparicio et
1977). By using
combined
GC/MS,
al.,
misinterpretations can be avoided.
With this method, extracts of untreated alpechin
from Spain were analysed. The Spanish alpechin
sample was obtained from the Granada area. In
addition, a sample of Italian alpechin from
Borgomaro in Liguria was analysed. Both samples
were kept frozen at -30C
until further
some
examination.
Table
1 gives
more
information about these two samples. It can be
seen that the Spanish alpechin has a chemical
oxygen demand, a biochemical oxygen demand
all of which are
and dry matter content
approximately double that of the Italian alpechin.
So far, 12 phenolic compounds in alpechin from
both origins have been identified using the GC/
MS method.
Fig. 3 shows two typical gasof ethyl acetate extracts of
chromatograms
untreated alpechin, prepared in the same manner.
Peak no. 1 in the chromatograms of Italian and
Spanish alpechin was identified as the TMSTable 1. Spanish
Sample-Important
COD (gl-I)
BODs (gl-)d
Dry matter (gl-)
pH (24C)
Alpechin
Sample
Parameters
vs
Italian
Alpechin
Spanish alpechin
Italian alpechin
49.0
4.2
35.1
4.9
80.4
11.5
73.0
5.2
279
G. Knupp
280
et al.
100
1
100
1:45
Spanish sample
300
5:15
200
3:30
400
7:oo
500
6:45
600
10:30
500
6~45
6dO
10:30
700
12:15
600 Scans
14:00 Time [min]
Italian sample
8
\
I .
100
1:45
200
3:30
300
515
400
7:oo
Fig. 3. Chromatograms
3
1, 15
600 Scans
14:OO Time [min]
of alpechin extracts.
281
Compound
substance
at peak # 3
179
179
73
262
267
1 I
IL L
SO
lb0
260
150
50
300
250
100
150
Fig. 4. EI/MS
of silylated
Ir
532
I300
lmouritv
48
24 h
HO~jCDOH
372
600
10:30
600
14:00
72
Scan
Time [mini
10:30
I.;.bosca
400
7:oo
200
3:30
10:30
14:o0
Time [min]
139 h
ho
i7:oo
250
tyrosol.
330
0~
3:30
200
400
7:oo
,I
m/z
m/z
.L
14:00
Fig. 5. Biodegradation
400
7:oo
200
3:30
Time [min]
of tyrosol
over
139h.
600
10:30
scan
14:00
Time [min
282
G. Knupp et al.
REFERENCES
Balice, V. and Cera, 0. (1984) Acidic phenolic fraction of the
olive vegetation water determined by a GC method.
Grasas y Aceites, 35, 178-180.
Bianchi, G. and Pozzi, N. (1994) 3,4-Dihydroxyphenylglycol,
major
C&z
phenolic
in
Olea
europaea
fruits.