Vacuum pump v 700

Only one sediment sample could be collected in the WWTP effluent channel (W.C., Figure 1), since the bottom at Cl2 (Figure 1) and Dp consisted of small pebbles. To evaluate the degree of pollution of this sediment sample we measured the concentrations of heavy metals (chromium, copper, nickel, lead, cadmium, zinc, cobalt, and mercury), arsenic, molybdenum, mineral-oil hydrocarbons, PAHs, and PCBs.
The methods used for determining heavy metal concentrations (Cr, Cu, Ni, Pb, Cd, Zn, Co, and Hg), arsenic, mineral oil hydrocarbons, and PAHs have been previously described in detail [20 (link)]. The concentration of Mo was determined in the same way as the heavy metals. For PCB detection, the oven-dried samples (5 g) were transferred to a Soxhlet apparatus and extracted with 100 mL of n-hexane. The extracts were evaporated on a rotary evaporator (Rotavapor R-210 with Heating Bath B-491, Vacuum Pump V-700, and Vacuum Controller V-850, Büchi) and concentrated in a stream of nitrogen gas. They were then transferred into a centrifuge tube and cleaned up with concentrated sulphuric acid (min. 96%). If the samples were highly contaminated repeated acid clean-up was employed. PCBs were quantified by a gas chromatography with a ⁶³Ni Electron capture detector (GC-17A-ECD, Shimadzu) according to EPA 8082 [21 ] standard method.

The fruit shells of A. spinosa were dried and then finely grounded to powder (IKA WERE, M20, Germany). A quantity (500 g) of the powdered shell was extracted for two weeks in the dark with 5000 ml of ethanol (70%) at room temperature (25 ± 2°C). The mixture was filtered and the extract was concentrated to dryness under vacuum using a rotary evaporator (BUCHI Switzerland, Rotavapor R-210, Vacuum Pump V-700) at 40°C. The residue (3.5% w/w) was stored at −20°C until use.

Dry leaves from M–ML common and control crops were used for obtaining dry extracts. Based on our previous study [22 (link)], the solvent used for the extraction was 50% ethanol for all plants. Exactly 25 g of plant material were used from every crop and were subjected to two consecutive reflux extraction processes: the first extraction used 1.5 L solvent for 30 min, while the second used 750 mL solvent for 30 min. The two extract solutions were mixed and concentrated in a rotary evaporator (Buchi, Vacuum Pump V-700) and then subjected to a lyophilization process (Christ Alpha 1–2/B Braun, BiotechInt, New Delhi, India). The dry extracts were conserved in a glass vacuum desiccator [50 (link)]. The samples were marked as follows: MM E (Mentha extract from control crop), MF E (Mentha extract from common crop), MLM E (Melissa extract from control crop) and MLF E (Melissa extract from common crop). Each stage of the research includes a presentation of additional tools and experimental setups used in this investigation.

The ethanolic extract was transferred to a balloon and the solvent was evaporated at 40 °C (Water Bath, Büchi, Flawil, Switzerland) under vacuum (Vacuum Pump V-700, Büchi, Flawil, Switzerland). Lastly, the residue was frozen at −80 °C and subsequently lyophilized (Christ Beta 2-8 LD) for a period of 24 h (−43 °C/0.09 mbar), obtaining the ethanolic dry extract of VA ssp. abietis (VA_DE).

Extract condensation was performed under vacuum and heat-assisted evaporation, in temperatures below 35 °C, using a Büchi Rotavapor R-210 apparatus, equipped with a Büchi vacuum pump V-700, Vacuum controller V-850 (all obtained from Büchi, Flawil, St Gallen, Switzerland), and Julabo F12 (Seelbach, Germany) cooling unit.
The estimation of Total Phenolic Content (TPC) and Total Tannin Content (TTC) was implemented using an Infinite^® 200 PRO microplate reader (Tecan Group Ltd., San Jose, CA, USA) and the estimation of Total Carotenoid Content (TCC) was performed using an x-ma 100 spectrophotometer (Human Corporation, Seoul, Republic of Korea).
An Accela Ultra High-Performance Liquid Chromatography system equipped with an autosampler and coupled with a TSQ Quantum Access triple-quadrupole mass spectrometer (Thermo Fisher Scientific, Inc., Waltham, MA, USA) was used for the determination of the phenolic and carotenoid compounds qualitative and quantitative content.