Glucose (G8270), lactose (17814), hydrogen peroxide (H1009), Carrez I solution (potassium hexacyanoferrate (II) trihydrate (60279)), Carrez II solution (zinc sulfate heptahydrate (31665)), magnesium chloride (M8266), triethylamine (TEA) hydrochloride (T1502), sodium citrate monobasic (71498), citric acid (27488), sodium phosphate monobasic (71496), sodium phosphate dibasic dihydrate (71645), potassium phosphate monobasic (795488), potassium phosphate dibasic (795496), iodonitrotetrazolium chloride (INT) (I8377), flavin adenine dinucleotide (F6625), Triton X‐100 (T9284), and imidazole (I202) were purchased from Sigma‐Aldrich, Arklow, Ireland. Vivinal GOS (Product # 598227; Lot # 704075) was kindly provided by Friesland Campina, Amersfoort, The Netherlands. Allolactose (O‐LAC6), 4‐nitrophenyl‐β‐d ‐galactopyranoside (O‐PNPBGAL), Glucose oxidase/catalase mixture (E‐GOXCA), hexokinase/Glucose‐6‐phosphate dehydrogenase mixture (E‐HKGDH), 6‐phosphogluconate dehydrogenase (E‐PGDHEC), diaphorase (E‐DIAEC), EcLacZ (E‐ECBGAL), A. oryzae (E‐BGLAN) and MZ104 β‐galactosidase enzyme mixtures were obtained from Megazyme, Bray, Ireland. Note that following the current study, all components required to perform the low‐lactose (LOLAC) assay were made available as an assay kit from Megazyme (K‐LOLAC).
K lolac
Manufactured by Megazyme
K‐LOLAC is a colorimetric assay kit designed for the quantitative determination of L‐lactic acid (L‐lactate) in various samples. The kit employs an enzymatic reaction that converts L‐lactate to pyruvate, which is then coupled to the reduction of a tetrazolium dye, resulting in the formation of a colored compound that can be measured spectrophotometrically.
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Lab products found in correlation
Triton x 100, by Merck Group (1 mentions)
Potassium phosphate dibasic, by Merck Group (1 mentions)
Carrez solution 1, by Merck Group (1 mentions)
Hydrogen peroxide, by Merck Group (1 mentions)
Imidazole, by Merck Group (1 mentions)
Carrez 2 solution, by Merck Group (1 mentions)
Flavin adenine dinucleotide, by Merck Group (1 mentions)
Sodium phosphate dibasic dihydrate, by Merck Group (1 mentions)
Sodium citrate monobasic, by Merck Group (1 mentions)
Sodium phosphate monobasic, by Merck Group (1 mentions)
Potassium phosphate monobasic, by Merck Group (1 mentions)
Uv 1900 spectrophotometer, by Shimadzu (1 mentions)
2 protocols using k lolac
1
Comprehensive Enzymatic Lactose Analysis
2
Quantitative analysis of lactose and glucose
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The concentration of free D-glucose, as well as the D-glucose component of lactose was determined by glucose spectrophotometric method using the lactose test kit (k-LOLAC, Megazyme). The method includes pre-treatment steps to clarify and deproteinate samples and also to remove the high levels of free D-glucose in the samples.
The determinations were performed according to the method for the measurement of lactose in low-lactose and lactose-free products under Standard Method Performance Requirement (SMPRVR) 2018.009 [17] (link). The absorbance reading of the samples was performed at wavelength at 340 nm using UV Vis Shimadzu UV-1900 spectrophotometer. The analyses were performed in triplicate.
The amount of D-glucose was determined based on the relationship:
The amount of lactose was determined based on the relationship:
where: F-dilution factor; The initial lactose concentration and the concentration of D-glucose released were used to calculate the hydrolysis efficiency.
The determinations were performed according to the method for the measurement of lactose in low-lactose and lactose-free products under Standard Method Performance Requirement (SMPRVR) 2018.009 [17] (link). The absorbance reading of the samples was performed at wavelength at 340 nm using UV Vis Shimadzu UV-1900 spectrophotometer. The analyses were performed in triplicate.
The amount of D-glucose was determined based on the relationship:
The amount of lactose was determined based on the relationship:
where: F-dilution factor; The initial lactose concentration and the concentration of D-glucose released were used to calculate the hydrolysis efficiency.
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