Uv vis spectrophotometer uv mini 1240

Scolices obtained from hydatids and hydatid cysts (the cyst originating from the lung after surgical removal) were washed five times in normal saline (0.9% sodium chloride). Soluble scolex antigen (SSA) was prepared after the disruption of whole scolices by frequent freezing and thawing. Scolices were further ultrasonically disrupted in a microcentrifuge ultrasonic disintegrator (Soniprep 150, MSE, Heathfield, UK).
The suspension was centrifuged at 12 000 rpm and the supernatant was collected. The deposit was suspended in phosphate-buffered saline (PBS, pH 7.2) and ultrasonically disintegrated. The supernatant was collected and added to the earlier supernatant. The combined supernatant was used as SSA antigen.²³ The protein concentration of the antigen was estimated by the Biuret method using a spectrophotometer (UV-Vis Spectrophotometer UV/mini 1240, Shimadzu, Kyoto, Japan).^{38 (link)}

The free radical scavenging activity method (DPPH: 2,2-diphenyl-1-picrylhydrazyl) was used to assess the antioxidant activity of the oil extracts [21 (link)]. In the reaction tubes, 375 μL of ethanol and 125 μL of DPPH solution were added to 500 μL of the crude extract and dilutions. For each concentration, a blank was prepared by mixing 500 μL of sample and 500 μL of ethanol without adding the DPPH solution. A control tube was also prepared by mixing 500 μL of the solvent (without sample), 375 μL of ethanol, and 125 μL of the DPPH solution. After shaking, tubes were placed in the dark at 25 °C for 1 h. Then, the absorbance values were measured at 517 nm by a spectrophotometer (Shimadzu, UV-VIS Spectrophotometer-UV mini 1240), and the radical scavenging activity was determined using the following formula: $$\mathrm{S}\mathrm{c}\mathrm{a}\mathrm{v}\mathrm{e}\mathrm{n}\mathrm{g}\mathrm{i}\mathrm{n}\mathrm{g} \mathrm{a}\mathrm{c}\mathrm{t}\mathrm{i}\mathrm{v}\mathrm{i}\mathrm{t}\mathrm{y} \%=(\mathrm{A} \mathrm{C}\mathrm{o}\mathrm{n}\mathrm{t}\mathrm{r}\mathrm{o}\mathrm{l}+\mathrm{A} \mathrm{s}\mathrm{a}\mathrm{m}\mathrm{p}\mathrm{l}\mathrm{e}-\mathrm{A} \mathrm{b}\mathrm{l}\mathrm{a}\mathrm{n}\mathrm{k})/\mathrm{A} \mathrm{c}\mathrm{o}\mathrm{n}\mathrm{t}\mathrm{r}\mathrm{o}\mathrm{l}\times 100$$
where A control, A sample, and A blank were the absorbance at 517 nm of the control, sample, and blank tubes, respectively.

Human CA isoforms (hCAI and hCAII) were purified by the Sepharose - 4B - L - tyrosine - sulfanilamide affinity segregation method as reported [34, 35]. Bradford technique was used to measure protein concentrations at 595 nm [36]. Inhibitory effects of the compounds were investigated by measuring the esterase activity according to Verpoorte et al. [37] as described in previous [38–40]. The hCA activity was determined by measuring the conversion of thep-nitrophenyl acetate substrate to
p-nitro phenolate at 348 nm by the spectrophotometer (UV - VIS Spectrophotometer, UVmini-1240, Shimadzu Corporation, Kyoto, Japan) [41]. Acetazolamide (AZA) was used as a control drug. Lineweaver–Burk plot was used to calculate inhibition constants (Ki) of the compounds [33]. All chemicals were purchased from Sigma-Aldrich Chemie GmbH.