© Birkhäuser Verlag, Basel, 2006 Inflamm. res. 55, Supplement 1 (2006) S67–S68 1023-3830/06/01S67-02 DOI 10.1007/s00011-005-0046-9
Inflammation Research
Histamine in selected beer samples C. Wackes, M. Herwald, H. Borck, E. Diel, L. Page, B. Horr, L. Rohn and F. Diel Institut für Umwelt und Gesundheit (IUG) and University of Applied Sciences, FB:Oe, Biochemistry, Marquardstrasse 35, 36039 Fulda, Germany, Fax: ++49 661 9640399, e–mail:
[email protected] Published Online First 20 March 2006
Introduction Recently we have shown that histamine produces only poor sensory characteristics and the average threshold value of “taste” is difficult to determine in normal tap water [1]. However, histamine is an important mediator in inflammation and influences the exocrine and paracrine processes in gastric mucosal cell regulation [2]. The aim of the present study was to assess whether histamine can be measured in commercially available beer and in how far it can be tasted in the test samples. Materials and methods Ten test samples were purchased from supermarkets and beer on draught from typical German bars. Type of beer and alcohol concentration were documented from their labels. Sugar was estimated refractometrically. Immediately after opening the bottles, freshly drawn beer was tasted using standardized sensitivity assessment methods by 10 instructed assessors – trained according to German DIN 10961/ISO 6658 [1]. Triangle test was performed according to ISO 4120 as a double blind method [3]. The reference test samples were diluted in normal tap water (0–5 µM Histamine–HCl; Janssen, Belgium). Specific sensory statistical software FIZZ was used [1]. Histamine and other biogenic amines including tyramine, cadaverine and putrescine were measured fluorimetrically after HPLC (Fig. 1) and thin–layer–chromatography in the beer samples (data not shown here) [4].
Results and discussion Ten instructed assessors indicated a mean sensory threshold for reference histamine 2.5 ±1.5 µM (mean ±SD, n = 10) in normal tap water. This is in good accordance with recently published results [1]. However, a threshold value for histamine in beer could not be evaluated using the Spearman´s correlation test. Furthermore, the triangle test as a double blind identification method resulted in only one of the test persons recognizing the histamine reference correctly. SenCorrespondence to: F. Diel
sitivity criteria such as pharyngeal irritation, tingling tongue, swelling of mucosal tissues in the mouth were defined to be characteristic for the “specific taste” of the biogenic amines. No positive correlation was found between histamine/ biogenic amine concentration in beer and the sensory sensation (R = 0.6; ranking of intensity of biogenic amines in 10 beer samples – data not shown here). The mean histamine concentration was 0.49 mg/l (range 0.44–0.55 mg/l, excluding Guinness beer) in the bottled and 10.3 mg/l (range 0.5–35.2 mg/l) in the drawn beer, respectively. Although “Schwarzer Hahn” showed a particularly soft taste, the histamine level dramatically increased to an extremely high level (35.2 mg/l) 2 h after drawing (Tab. 1). This could be due to bacteria contamination producing histamine [5]. In the drawn beer samples “Schwarzer Hahn” (Fig. 1A) and “Pils hell” (Fig. 1B) the histamine peak could clearly be demonstrated. In the 15 year old “Guinness” beer sample in Fig. 1C, the histamine peak is suppressed and the other biogenic amines seem to dominate the HPLC profile. Sugar and alcohol concentrations in these beers are relatively similar and will contribute to the taste characteristics (Table 1). It can be concluded that only the beer on draught revealed high histamine values (up to 35 mg/l) and only after standing. The sensory characteristics did not correlate to the measured histamine concentrations, using the standardized sensitivity assessment. Acknowledgements. Beer samples were received from Wiesenmühle brewery (Fulda). The project was supported by funds of the AVE e.V.
References [1] Rohn L, Page L, Borck H and Diel F. Can histamine be tasted in wine? Inflamm Res 2005; 54: 66–7. [2] Diel F and Szabo S. Dose–dependent effects of linear and cyclic somatostatin in ethanol–induced gastric erosions: the role of mast cells and increased vascular permeability in the rat. Regulatory Peptides 1986; 13: 235–43. [3] ISO 4120:2440: Sensory Analysis – Methodology – Triangle Test.
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Inflamm. res. Supplement 1 (2006)
type of beer
bottled on draught beer
His* (HPLC) (mg/l)
Sugar** (%)
alcohol*** (%)
Schwarzbier (K.)
+
0.41
5.2
4.8
Kristallweizen
+
0.45
5.2
5.3
Ur–Weisse dunkel
+
0.48
5.5
5.3
Hefeweizen naturtrüb
+
0.56
5.75
5.3
Guinness
+
0.1–1.1
5.9
5.0
Antoniusweizen
+
0.54
5.75
5.3
Wiesenmühlenbier
+
0.68
5.75
4.9
Pils (hell)
+
4.83
6.0
4.9
0.6
5.3
4.9
Schwarzer Hahn
+
Schwarzer Hahn
+
1.1
6.0
4.9
Schwarzer Hahn
+
35.16
5.4
4.9
remarks
Table 1. Histamine and some of the other ingredients in the tested beer samples
15 years old
2 h after drawing
* Histamine was measured fluorimetrically after HPLC separation as described in Fig. 1, average values (n = 2, maximum linear variation ±15 %, except Guinness beer n = 5, ± 46 %; references in perchloric acid 0.5 M: standard correlation 0.9995, detection limit 0.8 ng/ml) ** Sugar was measured semiquantitatively using the refractometer (KRÜSS Optronic, Hamburg) *** as indicated by the brewery
Beer 1 Beer 2 Beer 3 (old)
µ His
µ
A
µ
His
B
[4] Diel F, Horr B, Borck H, Savtchenko H, Mitsche T, Diel E. Pyrethroids and piperonyl–butoxide affect human T–lymphocytes in vitro. Tox Letters 1999; 107: 65–74.
His
Fig. 1. Ion–exchange HPLC profile of three selected beer samples: A “Schwarzer Hahn”: drawn beer sample, 2 h after drawing; B “Pils hell”: freshly drawn; C “Guinness”: bottled (15 years old), freshly opened. Perchloric acid samples were separated in oxalate/tartrate elutant (0.75/0.5 g/l) 1 ml/min; cationic exchange column (LICHROSIL® IC CA, Merck Darmstadt), post column OPT– derivatisation, fluorescence (340/455 nm). The histamine peak is indicated by the arrow. Calculation and statistical evaluation: Jasco® BORWIN program version 1.5 (Groß–Umstadt).
C [5] Chen ECH and Van Gheluwe G. Analysis of histamine in beer. J Am Soc Brew Chem 1979; 37: 0091.
