The extraction of amino acids was performed using sprouts homogenized in 1 mL of 80% (v/v) aqueous ethanol, supplemented with norvaline, to control the loss of amino acids during extraction. Amino acid content was assessed using the Waters Acquity UPLC-tqd system (Waters Corporation, Milford, MA, USA) along with a 2.1 × 50 amide BEH column, as described by AbdElhawad et al. [35 (link)]. Nitrogen levels were assessed using a CN element analyzer (NC-2100; Carlo Erba Instruments, Milan, Italy).
Nc 2100
Manufactured by Carlo Erba
Sourced in Italy
The NC-2100 is a laboratory instrument designed for the analysis of nitrogen and carbon content. It operates on the principle of thermal combustion and is capable of providing accurate measurements of these elemental components in various sample types.
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Lab products found in correlation
4 protocols using nc 2100
1
Amino Acid Extraction and Analysis
2
Elemental Analysis of Plant Samples
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C and N contents were measured with a CN element analyser (NC-2100, Carlo Erba Instruments, Milan, Italy). For Macro-minerals and trace elements, 100 mg DW plant material was digested in a 5∶1 ratio of HNO3/H2O in a microwave oven and determined by mass spectrometry (ICP-MS, Finnigan Element XR, Scientific, Bremen, Germany). A mixture of standards was prepared in 1% nitric acid.
3
Nitrogen Content and Enzyme Activities
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For total nitrogen (N) content, fine ground sprout samples (0.2 g) were digested in H2SO4–H2O at 260 °C, and the nitrogen levels were assessed with a CN element analyzer (NC-2100, Carlo Erba Instruments, Milan, Italy). Glutamine synthetase (GS, EC 6.3.1.2), glutamate synthase (EC 1.4.7.1), and glutamine 2-oxoglutarate aminotransferase (GOGAT, EC 1.4.7.1) enzyme activities were measured by monitoring the reduction of NADH at A340. GDH determining 2-oxoglutarate-dependent NADH oxidation. GS activity was detected by assessing the formation of γ-glutamyl hydroxamate at A340. GOGAT activity was estimated by following the glutamine-dependent NADH oxidation at A340. The activity of the nitrate reductase (NR, EC 1.7.1.1) enzyme was measured by following the nitrite-dependent NADH oxidation (A340) [45 ]. Protein concentrations were determined according to Lowry et al. [46 (link)].
4
Soil Chemical Properties Analysis
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Chemical properties of the soil (pH and electrical conductivity) were measured in diluted aqueous soil extract (1:5 w/v) using a pH meter (AD 3000) and a conductivity meter (Jenway 3305), respectively. Following Zhang et al. [31 (link)]), the phenolic content was estimated spectrophotometrically (Shimadzu UV-Vis 1601 PC, Kyoto, Japan). Twenty grams of soil were stirred vigorously with bi-distilled water followed by filtration and the filtrate was used for estimation of phenolic content. The content of mineral nutrients in soil rhizosphere were determined by shaking the excavated root very gently to separate from the bulk soil; then, the soil attached to the fine roots (2 mm thick layer) was obtained by brushing. About 20 g of soil rhizosphere were extracted in aqueous HNO3 solution (80%) and were detected by using ICP-MS (Finnigan Element XR, Scientific, Beremen, Germany) [32 ]. A total of 20 g of soil was mixed in a flask with 100 mL of distilled water and shaken for 12 h. Then, the soil extract was filtered and used to determine organic acid and phenolic compounds [33 (link)]. The percentage of CaCO3 was estimated according to Brown et al. [34 (link)]. The contents of carbon (C) and nitrogen (N) were measured by CN element analyzer (NC-2100, Carlo Erba Instruments, Milan, Italy).
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