Bioquant

All results are expressed as dry weight (dw) and the moisture content was determined using the thermo balance (Sartorius MA 40, Goettingen, Germany) at 120 °C. All analytical determinations were made in triplicate.
Protein contents of BSG and of BSG-enriched pasta samples were measured by micro-Kjeldhal nitrogen analysis according to ICC 105/2 method [34 ]. Total dietary fiber (TDF) content was determined using an enzymatic kit for fiber determination (Bioquant, Merck, Darmstadt, Germany) according to the Official Method 991.42 [35 ]. Ash content was determined according to approved method AACC 08-01.01 [36 ]. β-glucan content was evaluated by the Megazyme (Bray, Ireland) Mixed-Linkage Beta-Glucan kit [37 ].
Total antioxidant capacity (TAC) was determined by the “direct method,” according to Martini et al. [38 (link)].

All results are expressed as dry weight (dw), and the moisture content was determined using the thermo balance (Sartorius MA 40, Goettingen, Germany) at 120 °C. All analytical determinations were made in triplicate on cooked (Section 2.5) and freeze-dried pasta.
Protein content of pasta samples was measured by micro-Kjeldhal nitrogen analysis (ICC 105/2 method) [30 ], using as the conversion factor N × 5.7. Resistant starch (RS) content was determined according to the Official Method 2002.02 [31 ], using Resistant Starch Assay Kit (Megazyme, Bray, Ireland). Total dietary fibre (TDF) content was measured using an enzymatic-gravimetric kit for fibre determination (Bioquant, Merck, Darmstadt, Germany) according to the Official Method 991.43 [32 ]. Ash content was determined by the AACC 08-01.01 method [33 ]. Enzymatic method (AACC International Method No. 32.32) [34 ] was used for the determination of fructooligosaccharides (FOS). Total antioxidant capacity (TAC) was ascertained according to [35 (link)].

Ovary tissues were obtained from Sprague-Dawley (SD) rats and homogenized in PBS. This homogenized solution was centrifuged for 10 min at 1200 rpm. The protein level in the homogenized tissue was quantified with Bioquant (Merck, Darmstadt, Germany). The homogenized tissue was treated with 125 mM tert-butyl hydroperoxide (TBH) with or without a test sample and eventually reacted with thiobarbituric acid (TBA) to form the pink adducts of malondialdehyde (MDA). The optical density of the sample solution was measured at 530 nm with a μQuant spectrophotometer (BioTek). All rats used in this experiment were cared for according to ethical regulations on animal research of our university (permit no.: LAC-2017-0295).