DETERMINATION OF ETHANOL IN BEER BY GAS CHROMATOGRAPHY (DIRECT INJECTION)—COLLABORATIVE TRIAL

A method employing gas chromatography for the determination of ethanol in beer has been collaboratively tested by the Analysis Committee of the Institute of Brewing. It was judged that precision values were independent of concentration over the range 0.93 to 6.05% V/V ethanol. Repeatability (r₉₅) and reproducibility (R₉₅) values of 0.061 and 0.136 respectively, were obtained over this range. At a mean level of 9.17% V/V, the r₉₅ and R₉₅ values were 0.154 and 0.284 respectively. This was probably due to dilution errors as the sample had to be diluted to bring it within the linear range of the method. A comparison of the precision values given by the gas chromatographic method, with those obtained in 1991/1992 by 8 laboratories in a major brewing company using 12 sample pairs, for the IOB Recommended Distillation Method, revealed that there is no significant difference between the precision data for the two methods.     

QUANTITATIVE DETERMINATION OF BITTER SUBSTANCES IN BEER,BY THIN LAYER CHROMATOGRAPHY

A method is described which allows the separation of hop bitter components and derivatives present in beer by combined thin-layer chromatography on a mixture of silica gel and cellulose. This method can be used to evaluate the purity of beer bitterness and to recognize the beer hopping.     

DETERMINATION OF LOWER BOILING POINT VOLATILE COMPOUNDS IN BEER BY HEADSPACE GAS CHROMATOGRAPHY — COLLABORATIVE TRIAL

The Analysis Committee has collaboratively tested local routine headspace gas chromatographic methods for the determination of the lower boiling point volatile compounds in beer. The repeatability values (r₉₅) were dependent upon mean concentration (m) for acetaldehyde and alcohols but not for esters, whilst reproducibility values (R₉₅) were dependent upon concentration in all cases. The range of values of m and the estimates of r₉₅ and R₉₅ (mg/litre) for each compound were, respectively: acetaldehyde (5–8, 0.21m, 0.60m); propanol (9–23, 0.25m, 2.5+0.57m); isobutanol (5–22, 0.56+0.085m, 1.7+0.15m); methylbutanols (45–105, 0.14m, 0.22m); ethyl acetate (10–54, 3.1, 2.1+0.29m); isoamyl acetate (0.8–4.7, 0.36, 0.20+0.58m); and ethyl hexanoate (0.13–0.36, 0.073, 0.10+0.91m). No advantage was gained by diluting beer samples containing 9% V/V ethanol to 4% ethanol (used for the calibration mixtures) prior to analysis, but use of a standard method of sample preparation decreased most of the R₉₅ values. No recommendation is made in this interim report.     

THE DETERMINATION OF ALCOHOL IN BEER USING THE SCABA AUTOMATED METHOD (COLLABORATIVE TRIAL)

The Institute of Brewing Analysis Committee has organised two collaborative trials for the analysis of alcohol using the ServoChem Automatic Beer Analyser (SCABA). The mean repeatability (r₉₅) values for the two trials were 0.051% and 0.034% V/V respectively. The value for the second trial could however be judged to be dependent on concentration (m) at 0.008m% V/V. The Reproducibility (R₉₅) values of both trials were judged to be dependent on concentration at 0.027m% V/V and 0.025m% V/V respectively. The combined data for the two trials gave r₉₅ = 0.044% V/V and R₉₅ = 0.026m% V/V.     

DETERMINATION OF NITRATE IN BEER BY ION CHROMATOGRAPHY

¹A method, relying on ion chromatography, for the determination of nitrate in beer, has been collaboratively tested by members of the European Brewery Convention (EBC) and the Brewery Convention of Japan (BCOJ). Precision values were judged to be acceptable. Repeatability (r₉₅) and reproducibility (R₉₅) values were 1.5, 0.96, 5.1, and 9.3, 10.4, 13.5 respectively for corresponding mean levels of 6.5, 26.2 and 52.8 mg/litre. However, r₉₅ and R₉₅ values of 1.5 and 2.3 respectively were obtained for an aqueous solution of nitrate ions at a mean level of 20.7 mg/litre. The determination of nitrate is recommended for use and inclusion in Analytica-EBC, as an additional analyte in the International Method which relies on ion chromatography for estimating chloride, sulphate and phosphate.     

ESTIMATION OF FERMENTABLE CARBOHYDRATES — COLLABORATIVE TRIAL

A procedure relying on high performance liquid chromatography, for the estimation of fermentable carbohydrates in wort, has been tested collaboratively and adopted by the Institute of Brewing Analysis Committee as a Recommended Method. A radially compressed cartridge containing silica modified with tetraethylenepentamine is used with an eluting solvent of acetonitrile/water. The results of a preliminary trial indicated that better precision is achieved using manual measurements of peak height and omitting an internal standard. In the main trial where 5 laboratories analysed 5 sample pairs the repeatability (r₉₅) and reproducibility (R₉₅) values found with their mean concentration levels (m) to which they are applicable were: fructose (0.0256 + 0.0660m) and (0.0711 + 0.0691m) for 0.17–1.01 g/100 ml; glucose 0.090m and 0.139m for 0.80–2.07 g/100 ml; sucrose 0.095 and (0.0746 + 0.1663m) for 0.12–3.88 g/100 ml; maltose 0.219 and 0.917 for 4.17–6.09 g/100 ml; maltotriose 0.122 and 0.236 for 0.97–1.19 g/100 ml. The precision values for the sums of the total fermentable sugars were 0.578 and 1.040 for 6.71–14.24 g/100 ml.     

ESTIMATION OF ALGINATE,CARRAGEENAN AND FURCELLARAN IN BEER

Quantitative and semi-quantitative methods are given for estimating alginate, carrageenan and furcellaran at low concentrations in beer. A number of commercial beers and experimental beers known to have been brewed with either copper finings or auxiliary finings containing these acidic polysaccharides as active ingredients were analysed for alginate, carrageenan and furcellaran residues. The results show that alginate and carrageenan do not occur in finished beers at levels greater than 1 mg/litre. Using a serological technique which is more sensitive for furcellaran, it was found that levels of furcellaran in most finished beers were below 0.5 mg/litre, though a few beers contained this polysaccharide at about 0.5 mg/litre.     

COLLABORATIVE TRIAL ON THE DETERMINATION OF BETA-GLUCANASE IN MALT BY VISCOMETRIC AND DYE-LABELLED METHODS

The results of a collaborative trial to compare a dye-labelled beta-glucan method with that of a viscometric procedure, for estimating the beta-glucanase content of malt, have demonstrated that overall the precisions of both methods are similar. There has been an improvement in the precision of the viscometric method (which is not a Recommended Method) compared with values obtained in previous trials. The Institute of Brewing Analysis Committee judged that the results obtained by the dye-labelled beta-glucan method were not sufficiently precise to permit adoption as a Recommended Method. However, since this procedure has advantages of speed and ease of operation, it is suggested as an alternative method to that relying on viscometry. It is envisaged that a further collaborative trial will be carried out when sufficient experience of the new method has been obtained. The repeatability (r₉₅) and reproducibility (R₉₅) values found were 0.190 m and 0.327 m over the concentration (m,U/kg) range 92–266 U/kg for the dye-labelled method, and 0.173 m and (91.4 + 0.0717 m) over the concentration (m,IRV) range 233–701 IRV units for the viscometric method.     

DETERMINATION OF DIMETHYL SUPLHIDE IN BEER BY GAS CHROMATOGRAPHY — COLLABORATIVE TRIAL

The Analysis Committee has collaboratively tested both routine headspace gas chromatographic methods and a standardised method for the determination of dimethyl sulphide in beer using the flame photometric and flame ionisation detectors available in the participating laboratories. The mean repeatability value (r₉₅) was 7.8 μg/litre for the concentration range 12–65 μg/litre, and the reproducibility value (R₉₅) showed the relationship R₉₅ = 18.1 + 0.452m μg/litre to the concentration (m). No differences in precision were attributed either to the methods or the types of detectors used.     

THE DETERMINATION OF DIMETHYL SULPHIDE IN BEER BY HEADSPACE GAS CHROMATOGRAPHY — A COLLABORATIVE INVESTIGATION OF PRECISION

A method for the determination of dimethyl sulphide in beer by headspace gas chromatography has been collaboratively tested within ten laboratories of one brewing group at 3 levels from 19.8 to 52.4 μg/litre. The repeatability values (r₃₅ were found to be independent of concentration, but a strong linear relationship was found between the mean values (m) and the reproducibility value (R₃₅). The precision data can be summarised as r₃₅ = 3.3 μg/litre, R₃₅ = 3.66 + 0.168 μg/litre .     

COLLABORATIVE DETERMINATION OF THE PH OF BEER AND WORT

A method for the determination of pH in wort and beer has been tested for precision by the Analysis Committee of the Institute of Brewing . Over the pH range 3.94 to 5.04 it was judged that precision values were independent of the pH value of the sample. Values for r₉₅ and R₉₅ were 0.025 and 0.129 respectively .     

VICINAL DIKETONES IN BEER. PART II: INFLUENCE OF SULPHUR DIOXIDE IN THE DETERMINATION OF DIACETYL IN BEER

The accuracy of the determination of diacetyl in beer using o-phenylenediamine is impaired by the presence of sulphur dioxide. This effect may be overcome when the distillate from the beer is collected in strongly acidic solution¹.     

COLLABORATIVE DETERMINATION OF BEER FOAM STABILITY BY RUDIN AND NIBEM

Two methods, the “Rudin” method and the “Nibem” method have been tested for the determination of foam stability of beer by the Analysis Committee of the Institute of Brewing . For the Rudin method, over the range 90 to 102 seconds, it was judged that precision values were independent of the foam stability of the sample. Values for r₉₅ and R₉₅ were 6 and 22 seconds, respectively . For the Nibem method, precision was also independent of the foam stability of the sample over the range 213 to 246 seconds. Values for r₉₅ and R₉₅ were 22 and 70 seconds, respectively . The two methods ranked the foam stability of the three beers tested differently. This is to be expected, bearing in mind the different principles of the two procedures .     

DETERMINATION OF VICINAL DIKETONES IN BEER BY GAS CHROMATOGRAPHY (HEADSPACE TECHNIQUE)—COLLABORATIVE TRIAL

Two methods for the determination of vicinal diketones in beer have been collaboratively tested by the Analysis Committee of the Institute of Brewing and are recommended for use. Both methods employ gas chromatography and are essentially the same, except that one relies on the use of a packed column and the other on a capillary column. For diacetyl it was judged that repeatability (r₉₅) values were independent of concentration over the range 0.04 to 0.19 mg/litre. Over this range, r₉₅ values for diacetyl of 0.028, 0.020 and 0.026 mg/litre were obtained for capillary, packed columns and combined results respectively. Values for reproducibility (R₉₅) were judged to be dependent on the mean level (m). R₉₅ values were 0.032 + 0.68 m, 0.01 + 0.47 m and 0.005 + 0.67 m were obtained for capillary, packed columns and combined results respectively. For both methods the r₉₅ and R₉₅ values for 2,3-pentanedione were judged to be independent of concentration over the range 0.02 to 0.07 mg/litre. For capillary columns, packed columns and combined results respectively, r₉₅ values were 0.009, 0.009, 0.010 mg/litre and R₉₅ values were 0.037, 0.042 and 0.038 mg/litre.     

A CHEAP,DIRECT METHOD FOR THE ESTIMATION OF DISSOLVED AIR IN BEER

A plastic medical syringe is used to collect a beer sample and to apply vacuum to de-gas it. The beer is then replaced with sodium hydroxide solution and alkali-insoluble gas is displaced into a calibrated plastic capillary tube. Dissolved-air contents in the range 2 ml/litre to 50 ml/litre have been estimated with a repeatability adequate for many process investigations.     

DETERMINATION OF DIACETYL IN BEER USING DIFFERENTIAL-PULSE POLAROGRAPHY

A Polarographie method is proposed for the determination of diacetyl and other vicinal diketones in beer. Similar to the EBC official spectrophotometric method, diacetyl is derivatised with o-phenylene-diamine, producing 2,3-dlmethylquinoxaline, a polarographically active compound . The Polarographic method allows discrimination between diacetyl and methylglyoxal and a detection limit ten times lower than the EBC method is obtained. In addition, problems of erratic results for diacetyl may be avoided and data are in good agreement with those obtained by gas chromatography with electron capture detection and headspace sampling . It is suggested that high results obtained with the spectrophotometric method may be attributed to an erroneous procedure used in the spectrophotometric calculations. An alternative procedure is proposed, with which lower values for diacetyl in beer are obtained, similar to those found with other methods. These low diacetyl values are near the detection limit of the spectrophotometric method .     

DETERMINATION OF CARBON DIOXIDE IN BEER

A rapid, simple and inexpensive method is described for the determination of the carbon dioxide content of beers.     

DETERMINATION OF TOTAL CARBOHYDRATE IN BEER

A colorimetric procedure for the determination of total carbohydrate in beer has been evaluated in a collaborative trial. The method is recommended by the Analysis Committee.