Determination of glucose and fructose by enzymatic means.
Suitable for beer, malt drinks, low-alcohol beer, beer-based mixed drinks, non-alcoholic soft drinks, NAB, fruit juices, soft drinks and other beverages.
Glucose and fructose are phosphorylated by the enzyme hexokinase (HK) and adenosine 5'-triphosphate (ATP) to glucose 6-phosphate (G-6-P) and fructose 6-phosphate (F-6-P):
\(\text{Glucose + ATP} \space ^{\underrightarrow{\text{HK}}} \space \text{G-6-P + ADP}\)
\(\text{Fructose + ATP} \space ^{\underrightarrow{\text{HK}}} \space \text{F-6-P + ADP}\)
In the presence of the enzyme glucose-6-phosphate dehydrogenase (G6P-DH), G-6-P is oxidized from nicotinamide adenine dinucleotide phosphate (NADP+) to gluconate-6-phosphate. Reduced nicotinamide adenine dinucleotide phosphate (NADP + H+) is formed:
\(\text{G-6-P + NADP}^+ \space ^{\underrightarrow{\text{G6P-DH}}} \space \text{Gluconate-6-phosphate + NADPH + H}^+\)
The amount of NADP + H+ formed during the reaction is equivalent to the amount of glucose. NADPH + H+ is a measurand and is determined based on its absorbance at 340 nm.
After the reaction is complete, F-6-P is converted to G-6-P by phosphoglucose isomerase (PGI):
\(\text{F-6-P} \space ^{\underrightarrow{\text{PGI}}} \space \text{G-6-P}\)
The amount of NADPH + H+ formed during the reaction is equivalent to the amount of fructose. NADPH + H+ is the parameter being measured and is determined on the basis of its absorption at 340 nm.
G-6-P reacts in turn with NADP+ to form gluconate-6-phosphate and NADP + H+. The additional amount of NADP + H+ formed is equivalent to the amount of fructose and is determined photometrically based on its absorption at 340 nm.
Note:
Alternatively, NAD+/NAD + H+ can be used instead of NADP+/NADPH + H+:
\(\text{G-6-P + NAD}^+ \space ^{\underrightarrow{\text{G6P-DH}}} \space \text{Gluconate-6-Phosphate + NAD + H}^+\)
Determination of the viscosity of beverages
wort, beer, beer-based beverages, NAB, juice, beverages in general
Capillary viscometers measure the kinematic viscosity of Newtonian fluids, unlike falling ball and the rotational viscometers, which directly measure the dynamic viscosity. However, with values for the kinematic viscosity and the density of the test liquid, the dynamic viscosity can be calculated.
The time required for a volumetrically defined amount of fluid (the volume between two marks on the tube of the viscometer) to flow through a capillary of a defined length is measured.
1 square meter per second of a homogenous fluid with a dynamic viscosity of 1 Pa × s and a density 1 kg/m³
The unit for expressing kinematic viscosity (the relationship of viscosity to density = viscoplasticity) is 1 m²/s = 106 mm²/s
The kinematic viscosity is expressed as 1 square meter per second of a homogenous fluid with a dynamic viscosity of 1 Pa × s and a density 1 kg/m³.
Determination of glucose by enzymatic means.
Suitable for wort, beer, malt-based drinks, nutritional beer, beer-based mixed drinks, non-alcoholic soft drinks, NAB, fruit juice, soft drinks, juices, drinks.
Glucose is phosphorylated by the enzyme hexokinase (HK) and adenosine 5'-triphosphate (ATP) to glucose 6-phosphate (G-6-P).
\(\text{Glucose + ATP} \space ^{\underrightarrow{\text{HK}}} \space \text{G-6-P + ADP}\)
In the presence of the enzyme glucose-6-phosphate dehydrogenase (G6P-DH), G-6-P is oxidized by nicotinamide adenine dinucleotide phosphate (NADP+) to gluconate-6-phosphate. Reduced nicotinamide adenine dinucleotide phosphate (NADPH) is formed:
\(\text{G-6-P + NADP}^+ \space ^{\underrightarrow{\text{G6P-DH}}} \space \text{gluconate-6-phosphate + NADPH + H}^+\)
The amount of NADPH+H+ formed during the reaction is equivalent to the amount of glucose. NADPH+H+ is determined based upon its absorbance at 340 nm.
Note:
Alternatively, NAD+/NADH + H+ can be used instead of NADP+/NADPH+ H+:
\(\text{G-6-P + NAD}^+ \space ^{\underrightarrow{\text{G6P-DH}}} \space \text{Gluconate-6-Phosphate + NAD + H}^+\)
Determination of glucose, fructose, sucrose by enzymatic means.
Suitable for wort, beer, malt-based drinks, nutritional beer, beer-based mixed drinks, non-alcoholic soft drinks, NAB, fruit juice, soft drinks, juices, drinks.
The D-glucose content is determined before and after enzymatic hydrolysis of sucrose. D-fructose is measured following D-glucose determination.
D-glucose/D-fructose determination before inversion:
Glucose and fructose are phosphorylated by the enzyme hexokinase (HK) and adenosine-5'-triphosphate (ATP) to glucose-6-phosphate (G-6-P):
\(\text{Glucose + ATP} \space ^{\underrightarrow{\text{HK}}} \space \text{G-6-P + ADP}\)
\(\text{Fructose + ATP} \space ^{\underrightarrow{\text{HK}}} \space \text{F-6-P + ADP}\)
In the presence of the enzyme glucose-6-phosphate dehydrogenase (G6P-DH), G-6-P is oxidized from nicotinamide adenine dinucleotide phosphate (NADP+) to gluconate-6-phosphate. Reduced nicotinamide adenine dinucleotide phosphate (NADP + H+) is formed:
\(\text{G-6-P + NADP}^+ \space ^{\underrightarrow{\text{G6P-DH}}} \space \text{Gluconate-6-phosphate + NADP + H}^+\)
The amount of NADP + H+ formed during the reaction is equivalent to the amount of glucose. NADP + H+ is measurand and is determined based on its absorbance at 340 nm.
After the reaction is complete, F-6-P is converted to G-6-P by phosphoglucose isomerase (PGI):
\(\text{F-6-P} \space ^{\underrightarrow{\text{PGI}}} \space \text{G-6-P}\)
G-6-P reacts in turn with NADP+ to form gluconate-6-phosphate and NADP + H+. The additional amount of NADP + H+ formed is equivalent to the amount of fructose and is determined photometrically based on its absorbance at 340 nm.
Enzymatic inversion:
Sucrose is hydrolyzed to glucose and fructose by the enzyme β-fructosidase (invertase) at pH 4.6:
\(\text{Saccharose + H}{_2}\text{O} \space ^{\underrightarrow{\text{β-Fructosidase}}} \space \text{Glucose + Fructose}\)
The D-glucose determination after inversion (total D-glucose) is carried out as described above.
The sucrose content is calculated from the difference between the glucose concentration before and after enzymatic inversion.
This method describes how to determine whether a lot of grain is contaminated with ergot visually through manual inspection.
Barley intended for the production of malt; therefore, the kernels are to be evaluated on the basis of the characteristics described below.
A representative sample of grain is visually inspected for the presence of ergot sclerotia.
This method describes how to determine the hectoliter weight of barley.
Barley intended for the production of malt is evaluated on the basis of the hectoliter weight.
The hectoliter weight determines how many kilograms 100 liters of barley weighs. For this analysis, the weight of a defined sample volume of barley is determined, and the corresponding hectoliter weight is calculated.
One hectoliter of malting barley generally weighs between 68 and 75 kg, although higher values are not unusual (up to 78 kg).