# Nutritional, Antioxidant, Sensory, Energetic, and Electrical Properties of Enriched Pasta
## 2 Materials and Methods  
The carrot roots (_Daucus carrota_ L.) were collected in autumn (September 2020), nettle leaves (_Urtica dioica_ L.) in spring (May 2020), and elderberry fruits (_Sambucus nigra_ L.) were collected in autumn (September 2020), in the village of LieSt'any, situated in Central Slovakia at an altitude of 333 m above sea level. The plant parts were botanically identified at the Institute of Food Sciences of the Faculty of Biotechnology and Food Sciences. All parts (fruits, leaves, and roots) were crushed to powder using a mortar. Pasta samples were prepared using an extruder (Gina, Italy) for the production of the extruded pasta. Here, 500 g of wholemal wheat flour was mixed with carrot, nettle leaves, and elderberry powder (3%), one egg (60 g), salt (7 g), and tap water in a rotary shaft mixer at 25 \({}^{\circ}\)C for 20 min, to obtain a dough with 30% moisture content. A screw (length 30 cm; diameter 5.5 cm) that ended with a bronze die (hole diameter, 1.70 mm) was used to extrude the dough into an elbow shape. The screw speed was 50 min\({}^{-1}\). The extrusion pressure was approximately 3.4 bar (0.34 MPa), and the temperature of the pasta after the extrusion was from 27 \({}^{\circ}\)C to 28 \({}^{\circ}\)C. Each type of pasta was prepared separately. Extruded pasta was dried in a drying chamber (8 h at 40 \({}^{\circ}\)C) to a final moisture content of approximately 12.5%. Before analyzing, pasta was homogenized to a particle size of 0.5 mm. The total protein, dry matter, and ash content were determined using the AACC standard method 08-01 [20]. Protein content was detected using the semi-micro Kjeldahl method, and factor 5.7 was used for converting of proteins. The total fat content was determined using a Fat Extractor Ancom XT15 (Ancom, Fairport, NY, USA) according to the method recommended by the producer. Ion exchange chromatography, with a strong cation exchanger and a sodium-citrate elution buffer system, followed by post-column derivatization with ninhydrin and spectrophotometric detection, was used for measuring amino acid composition, according to method described by manufacturer of the amino acid analyzer (Ingos, Prague, Czech Republic). The amino acid standard solution was used for the calibration of the amino acid analyzer. Amino acid tryptophan was not evaluated because this compound was destroyed during acid hydrolysis, and glutamine and asparagine changed to glutamic acid and aspartic acid; thus, they were determined in this form. Antioxidant activity of the samples was evaluated using the DPPH method according to [PERSON] et al. [21] with slight modifications. The spectrophotometric method using the Folin-Ciocalteu reagent was used for total polyphenol content detection according to [PERSON] and [PERSON] [22]; the spectrophotometric method was also used for the total phenolic acid content, according to Farmakopea Polska [23], using Arnova reagent. Total chlorophyll content was determined spectrophotometrically according to the Lichtenthalerand Wellbum method [24]. The absorbance of the reaction mixture was determined using the spectrophotometer Jenway (6405 UV/Vis, England) at 649 nm and 664.5 nm. Anthocyanin content was measured according to the method of [PERSON] and [PERSON] [25] with modifications [26]. For pH 1.0, a sample (0.4 mL) was diluted with 0.025 M of potassium chloride (3.6 mL). For pH 4.5, a sample was diluted (0.4 mL) with 0.4 M of sodium acetate. The absorbance of sample was measured at 520 nm and 700 nm against a blank reagent (distilled water). The concentration of total anthocyanins was expressed as cyanidin-3-glucoside (Cy-3-glc) equivalent. The total carotenoid content was determined from the molar absorption coefficient of \(\beta\)-carotene according to the STN method [27]. The taste panel of 20 evaluators (average consumers) was used for the evaluation of the sensory properties of samples.