Rowth price with respect towards the culture time was accomplished plus the maximal particular development rate may be obtained from the curve. Lipid productivity was calculated making use of the Orexin A Protocol following equation: Lipid productivity (P, mg/L/day) = (2 – 1)/(t2 – t1) (1)exactly where 1 and 2 were the lipid production (mg/L) on days t1 and t2 , respectively. The lipid productivity was calculated for every 3-day interval as well as the maximal lipid productivity will likely be obtained after all calculations. 2.five. Fatty Acids Analysis The fatty acid content in the microalgae was estimated as the volume of fatty acid methyl esters (FAMEs), which was obtained applying the direct esterification process. The obtained crude lipid was saponified with KOH-CH3 OH then FAMEs were ready by methylene esterification with boron trifluoride in methanol and extracted with nhexane . The qualitative and quantitative analyses on the lipid sample have been carried out utilizing a GC-2030AF gas chromatograph (Shimadzu, Tokyo, Japan) equipped having a DB-Wax capillary column (30 cm, 0.32 mm ID, 0.five film thickness) in addition to a flame ionization detector (FID). The temperature system was as follows: initially kept 120 C for three min; then enhanced to 190 C at 10 C/min, exactly where the temperature was maintained for 1 min; then elevated to 230 C at 2 C/min and held there for 15 min using a split ratio of ten:1. Nitrogen was utilized as a carrier gas at a flow price of three mL/min. Fatty acids were identified by comparing the peak and retention time in the reference common 21-component fatty acid methyl ester mix (NU-CHEK-PREP, INC). The relative and absolute contents of fatty acids were quantified by the location normalization approach using methyl heptadecanoate purchased from Sigma-Aldrich (Merck Co., Kenilworth, NJ, USA) as an internal normal. two.six. Oxidative Strain Assessment and Determination of Antioxidant Enzyme Activities The algal cultures have been collected by centrifugation at ten,000 rpm for 10 min at four C, then the pellet was resuspended in two mL of 0.05 M sodium phosphate buffer option (PBS, pH 7.8) and immediately disrupted by sonication (JY92-IIDN, Ningbo Scientz Biotechnology CO., LTD, Ningbo, China) for five min with a 3 s pause following every 3 s pulse in an ice bath. The cell homogenate was centrifuged at 4000 rpm for ten min at 4 C as well as the supernatant was recovered for the assessment of enzyme activities, the malondialdehyde (MDA) level, and also the hydrogen peroxide (H2 O2) content. The soluble protein content material was determined working with the approach of Bradford . The H2 O2 content was measured by a commercial assay kit (Parsaclisib Inhibitor Nanjing Jiancheng Biology Engineering Institute, Nanjing, China) following the manufacturer’s directions as well as the outcome was expressed as mmol H2 O2 /mg protein. The lipid peroxidation (LPO) was determined by measuring the MDA (a productNanomaterials 2021, 11,5 ofof lipid peroxidation) content, which was measured according to the thiobarbituric acid (TBA) technique . The MDA content was expressed as nmol MDA/mg protein. Catalase (CAT) activity was estimated by recording the decrease inside the absorbance of H2 O2 at 240 nm applying a UV is spectrophotometer (UV2150, Unico (Shanghai) Instruments Co., LTD., Shanghai, China). One unit of CAT activity (U) was defined because the quantity of enzyme expected to decompose 1 ol of H2 O2 per mg of soluble protein per minute . The enzyme activities have been calculated per mg of protein. The activity of superoxide dismutase (SOD) was measured based on its ability to inhibit the photochemical re.