By Baico Biotech 丨 Textile Enzyme 丨 BIO-SCOURING ENZYME

In the cellulase activity detection system with sodium carboxymethyl cellulose (CMC-Na) as the substrate, the reducing sugar produced by the catalytic reactions react with DNS reagent in a boiling water bath to form a brownish-red substance, 3-amino-5- nitrosalicylic acid. The absorbance value of the solution is subsequently measured by a spectrophotometer.

However, not all absorbance values are usable. Excessively low or high values will cause distorted test results and reduce the accuracy of cellulase activity assays. According to China light industry standard QB/T 2583-2023, the absorbance value should be controlled within the range of 0.33 to 0.35. Baico Biotech BBT enzymes is a textile enzyme and household detergent enzyme manufacturer in China. BBT Enzyme specifies that the absorbance value shall be maintained around 0.3, and the dilution ratio of the enzyme solution needs to beadjusted once the value deviates from the specified range.

According to Lambert-Beer law, absorbance is directly proportional to the concentration of the absorbing substance in the solution. However, this linear relationship does not hold perfectly across all concentration ranges. A high absorbance value is generally caused by an under dilution of the enzyme solution. An excessively high enzyme solution concentration promotes the production of large amounts of the brownish-red product 3-amino-5-nitrosalicylic acid during the catalytic reaction. A higher concentration of the absorbing substance leads to denser molecules and greater interparticle interaction, deviating from the linear relationship between absorbance and concentration. Direct calculation via the linear standard equation under such conditions will result in a lower value. Furthermore, a high enzyme concentration may indicate that the substrate CMC-Na is nearly depleted, meaning the catalytic reaction is not proceeding kinetically, also leading to inaccurate results.

A low absorbance value is usually caused by an over dilution of the enzyme solution. The influences from instrumental noise, baseline drift, ambient stray light, trace impurities in solution and other factors get amplified. Minor environmental variations can cause considerable fluctuations in test results.

Controlling the absorbance value within a reasonable range can effectively avoid systematic errors and linear failures, ensuring accurate and reliable cellulase activity results. It is an indispensable and critical quality control step in the DNS colorimetric detection method for cellulase activity.

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