Lysozyme Enzymatic Activity in Thick Albumen and Amniotic Fluid

Thick albumen and amniotic fluid samples were thawed (4°C for 12 h). Phosphate buffer was prepared within the Enzymatic Assay of Lysozyme EC 3.2.1.17. (Merck, KGaA, Darmstadt, Germany) (Merck, 2022). The buffer was also used for a Micrococcus lysodeikticus (luteus) bacteria suspension. The absorbance was in the range of 0.6 to 0.7 (λ = 450 nm). From each sample, 1 mL was taken and refilled with buffer to a volume of 10 mL.

For the enzymatic activity of lysozyme analysis, 2.4 mL of the bacterial suspension and 0.1 mL of the thick albumen/amniotic fluid sample were added to the spectrophotometric cuvette and placed in the spectrophotometer (SP830 Plus, Metertech, Taipei, Taiwan). The enzymatic activity of lysozyme (U/ mL) was calculated using the formula: (ΔA450/minTest−ΔA450/minBlank)(df)(0.001)(0.1) (df, dilution factor; 0.001, change in absorbance (ΔA450) as per the Unit Definition; 0.1, volume (mm) of added solution).