Professional Documents
Culture Documents
ISSN: 1317-2255
Revistamulticiencias@gmail.com
University of the Western Cape
Venezuela
Camacho, Maria; Torres, Gabriel; left, Peter; Benítez, Betty; Allara, Maria; Céspedes, Euclimar
Determination of biogenic amines in beers that are expended in Maracaibo city-
Venezuela
Multiciencias, vol. 7, No. 3, September to December, 2007, pp. 276-287
University of the Western Cape
Fixed point, Venezuela
Summar
y
Parts per million (ppm) [15], while drinks al-coholicas has A standard of 40 ppm of amines, dissolved in perchloric
suggested a limit of 2 ppm [9]. acid 0.4 M. The derivatization of the standards was made in
The consumption of beer in Venezuela is similar to the the same way as samples.
high consumption of wine in Europe, some authors indicate The chromatography equipment specifications
that it is important to control and monitor the with-had of We used a team of High Performance Liquid
these amines in these drinks, because they can au-increase ®
Chromatography (HPLC) Shimadzu , constituted by a
the problems of public health [4]. The aim of this research val-uvula automatic injection, a controller system SCL-6B,
was to determine the concentration of amines in different two pumps LC-6A, a reverse phase column RP-18
types and brands of beers that are produced and sold in ®
LICHROSPHER Merck , of 12.5 cm in length by 4.5 mm
Venezuela. internal diameter placed at room temperature environment,
a UV-VIS detector SPD-10A, 254 nm and 0.08 AUFS. For
Materials and Methods the analysis of the data was used the soft-ware
chromatographic Class VP.
Sampling The mobile phase to was composed of master-Acetate nio
Samples were selected from two types of beers em- 0.1 M, and acetonitrile (50:50) and the mobile phase B only by
botelladas: Light and Pilsen, obtained from three plants acetonitrile. Using a linear elution program with gra-tooth,
process-sadoras of the country, called plant, plant B and where the mobile phase was in a percentage of 80% in the first
plant C, acquired in supermarkets, liquor and abastos ubi- 19 min, 19 to 22 min in a 20 per cent and 22 to 32 min at 80%,
tered in the city of Maracaibo, Venezuela. The beers were with a flow rate of 0.7 mL/min. [6].
collected from different batches of production between the Quantification of Amines
months of October 2002 to March 2003. For the study gave To determine the concentration of amines
a total of 36 samples at a rate of 6 per plant and type of In the samples, comparing the retention time of each
beer, and triplicate were analyzed to determine the Sample and reference standard for the area,
concentration of amines. By performing the calculation according to the following
formula:
Determination of the amines CM= AM /Astd x Cstd x Fd
Removal of the amines Where:
It was carried out following the methodology of Moret et CM = concentration of the sample, expressed in ppm Am
al = Area of the sample
[17]. A volume of 10 mL of beer was degassed by Standard = concentration of the CSTD, expressed in ppm
ultrasonication, adding 10 mL of a mixture of n-butanol and Astd = Area of the standard
chloroform 1:1 (v/v). The same was placed 4
Fd = dilution factor (5 x10 )
®
In a mixer Lab-Line for 30 minutes, and subsequently
transferred to a separating funnel. To 1mL of the organic Statistical Analysis
phase is added 2 drops of HCl 1M, aforan-dose to 10 ml
with acetonitrile. The data obtained were analyzed using the statistical
Formation of Fluorescent Derivatives package SAS-pa. Analysis of variance (ANOVA) on the
Of the sample previously removed is measured 1mL, the basis of a completely randomized experimental design, with
Which were added 200 L of a solution of NaOH 2N and 300 the aim of determining differences between treatments
L of a buffer of NaHCO 3 saturated. Followed-mind were applied, with a significance of 0.05 [19].
added 2 mL of a solution of chloride dansilo in acetone Statistical Model:
(1mg/mL) and the mixture was carried out at 40°C for 45 And ijk= μ + Pi + Tj + (PT)ij + ijk
minutes. where
Subsequently removed the chloride dansilo resi-dual Yijk = response variable (amines)
adding 100 L of pure ammonia. The 30 minutes-cough this Tj = treatment effect (type of beer)
mixture was adjusted to a volume of 5 mL with acetonitrile Ijk = experimental error
and centrifuged for 5 minutes at 2.500 rpm P= Breweries
®
A centrifuge Dynac Model brand Clay Adams . This
derivative is injected 20L in the HPLC.
MULTICIENCIAS VOL. 7, No. 3, 2007 (276 - 281) / CORE FIXED POINT - The University of the Western
Cape 279
Mg/L in the beer type Light and 0.59 mg/L in the Pilsner Table 3. Concentration* media of histamine and triptami-na
beers, in contrast to the results reported by Zee et al [20] by effect of types of beer.
who found in European and Canadian beers a range of 4,50-
7.27 mg/L in the same types of beers. Type Histamine Tryptamine
In both types of beers (Light and Pilsen) The concentration Light 19.01 18.82
of histamine found exceeds 6 mg/L. This concentration can be Pilsen 19.40 22.92
a predisposing factor for the appearance of headaches, Means followed by different letters in the same column, they are mean-
hypotension, and other toxic effects if ingested in quantities of Significantly different (P< 0.05).
more than 10mg/L [15]. * Average values expressed in ppm.
The Table 4, shows the average and significance for the
va-variables histamine, and tryptamine by effect of plant
and type. The site for the types Light and Pilsen submitted Table 4. Concentration* media of histamine and tryptamine
the lowest concentration of histamine (4.62 and 10.91 ppm), by effect of plants and different types of beer.
respectively not detected significant differences among
them, but, if with regard to the plants B and C, which Plant Type Histamine Tryptamine
presented the highest values of histamine. The plants A and To Light 4.62
to
23.13
to
(P<0.05) between the Ligth and Pil-sen (28.55 and 19.81 Pilsen 27.48
c
28.61
to
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