The elementary composition (CHNS) of samples was analyzed using a “Vario Macro Cube” (Elementar) with a TCD detector. Total organic carbon (TOC) was analyzed using a “Vario Macro Cube” (Elementar) with an NDIR detector. Organic matter was measured by weight loss on ignition at 550 °C in a muffle furnace for 5 h. The pH was measured in an aqueous extract (1/4, m/v) using a glass electrode with pH–conductometer CPC 501 (Elmetron) according to Polish standard PN–Z–15011–3:2001.
Vario macro cube
Manufactured by Elementar
Sourced in Germany
The Vario MACRO cube is a compact and robust elemental analyzer designed for the determination of carbon, hydrogen, nitrogen, and sulfur in a wide range of solid and liquid samples. It features a unique cube-shaped design and offers high-precision analysis with minimal sample preparation.
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115 protocols using vario macro cube
1
Elemental Composition and Organic Matter Analysis
2
Demineralization of Coal Samples for FTIR Analysis
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In
the FTIR analysis, to remove the potential effects of minerals
on it, the coal samples needed to be acid-washed using HCl and HF
solutions in an atmospheric environment as described in a previous
study.10 Generally, the acid treatment
under such conditions would not cause significant structural changes.11 The procedure was called demineralization. These
demineralized samples were dried for 12 h in a vacuum room at 60 °C
and stored under a nitrogen atmosphere.9 (link)The equipment used for elemental analysis was the Elementar
vario MACRO cube (Elementar Analysensysteme GmbH, Germany). The measured
elements were carbon, hydrogen, nitrogen, and sulfur in the demineralized
samples, and the oxygen element in the samples was obtained by the
subtraction method. According to the GB/T 476-208 criterion, the ultimate
analysis results of these demineralized samples were determined.
the FTIR analysis, to remove the potential effects of minerals
on it, the coal samples needed to be acid-washed using HCl and HF
solutions in an atmospheric environment as described in a previous
study.10 Generally, the acid treatment
under such conditions would not cause significant structural changes.11 The procedure was called demineralization. These
demineralized samples were dried for 12 h in a vacuum room at 60 °C
and stored under a nitrogen atmosphere.9 (link)The equipment used for elemental analysis was the Elementar
vario MACRO cube (Elementar Analysensysteme GmbH, Germany). The measured
elements were carbon, hydrogen, nitrogen, and sulfur in the demineralized
samples, and the oxygen element in the samples was obtained by the
subtraction method. According to the GB/T 476-208 criterion, the ultimate
analysis results of these demineralized samples were determined.
3
Analytical Methods for Forage Constituents
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The protocols of the Association of Official Analytical Collaboration [31 ] were applied to analyze dry matter (DM, method 934.01), ash, organic matter (OM, method 942.05), and ether extract (EE, method 920.39). Total nitrogen (N) (N × 6.25 = Crude Protein, CP) was analyzed by Elementar Vario Macro Cube (Elementar, Hanau, Germany). The neutral detergent fibre (NDFom) contents were analyzed as previously described [30 ] without using amylase, sodium sulphite, and dekalin while acid detergent fibre (ADFom) and lignin (ADLom) were determined as previously reported [32 (link)]. The NDFom and ADFom contents were quantified without ash. Metabolizable energy (ME) was estimated by utilizing the equation of a previous study [33 ].
Total phenols (TP) and total tannins (TT) were examined by the Folin Ciocalteu procedure as explained previously [34 ] with tannic acid (Fisher Scientific, Loughborough, UK) as the standard reference. The procedure of [35 ] was applied for total saponin (TS) measurement by utilizing diosgenin (Molekula Ltd., Gillingham, UK) as a standard. A UV/VIS-spectrophotometer (Libra S12, Biochrom Ltd., Cambridge, UK) was employed in the TP, TT, and TS analyses.
Total phenols (TP) and total tannins (TT) were examined by the Folin Ciocalteu procedure as explained previously [34 ] with tannic acid (Fisher Scientific, Loughborough, UK) as the standard reference. The procedure of [35 ] was applied for total saponin (TS) measurement by utilizing diosgenin (Molekula Ltd., Gillingham, UK) as a standard. A UV/VIS-spectrophotometer (Libra S12, Biochrom Ltd., Cambridge, UK) was employed in the TP, TT, and TS analyses.
4
PCDD/F Analysis in Soil Samples
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PCDD/Fs were analyzed, according to an EPA 1613
B (1994)37 method, by a commercial laboratory
(INDAM, Castel Mella BS, Italy). In brief, samples were extracted
with an accelerated solvent extractor (ASE) (Thermo Scientific Dionex
ASE 350) and analyzed with HRGC/HRMS (Thermo Scientific TRACE GC Ultra
coupled with Thermo Scientific DFS MS). Seventeen congeners and PCDD/F
classes were determined. The 17 congeners measured were those with
Toxicity Equivalency Factor (TEF).38 (link) For
more details on the selected congeners and analytical methods, please
seeSupporting Information S1 . Concentrations
of PCDD/Fs will be reported as individual congener concentrations,
which means not translated in TEQ using TEF values, unless reported.
Organic carbon (OC) in soil samples was determined, according to the
UNI EN 13137 method (2002)39 (after the
removal of carbonates), with an elementar analyzer Elementar vario
MACRO cube [Elementar Analysensysteme GmbH, Langenselbold (Germany)].
B (1994)37 method, by a commercial laboratory
(INDAM, Castel Mella BS, Italy). In brief, samples were extracted
with an accelerated solvent extractor (ASE) (Thermo Scientific Dionex
ASE 350) and analyzed with HRGC/HRMS (Thermo Scientific TRACE GC Ultra
coupled with Thermo Scientific DFS MS). Seventeen congeners and PCDD/F
classes were determined. The 17 congeners measured were those with
Toxicity Equivalency Factor (TEF).38 (link) For
more details on the selected congeners and analytical methods, please
see
of PCDD/Fs will be reported as individual congener concentrations,
which means not translated in TEQ using TEF values, unless reported.
Organic carbon (OC) in soil samples was determined, according to the
UNI EN 13137 method (2002)39 (after the
removal of carbonates), with an elementar analyzer Elementar vario
MACRO cube [Elementar Analysensysteme GmbH, Langenselbold (Germany)].
5
Pretreatment for TOC and TON Analysis
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Sample pretreatment before total organic carbon (TOC) analysis: An appropriate amount of sediment or suspended particle samples were weighed and placed into a centrifuge tube and added with 20 mL HCl (3 mol/L) to react fully for the removal of inorganic carbon. Samples were washed with ultrapure water to neutral, freezedried, ground, and passed through a 100-mesh (0.15 mm) nylon sieve. Pretreated samples were stored in clean polythene bags.
Sample pretreatment before total organic nitrogen (TON) analysis (Zhang et al. 2018 (link)): An appropriate amount of sediment or suspended particle samples were weighed into a centrifuge tube and added with sufficient amounts of 2 mol/L KCl and 0.5 mol/L HCl to react fully for the removal of inorganic nitrogen. Samples were washed with ultrapure water to neutral, ground, freeze-dried, and passed through a 100-mesh (0.15 mm) nylon sieve. Pretreated samples were stored in clean polythene bags.
The contents of TOC and TON were determined using the Elementar (elementar vario MACRO cube, Elementar Analysensysteme GmbH, Germany). C/N was the ratio of TOC to TON.
Sample pretreatment before total organic nitrogen (TON) analysis (Zhang et al. 2018 (link)): An appropriate amount of sediment or suspended particle samples were weighed into a centrifuge tube and added with sufficient amounts of 2 mol/L KCl and 0.5 mol/L HCl to react fully for the removal of inorganic nitrogen. Samples were washed with ultrapure water to neutral, ground, freeze-dried, and passed through a 100-mesh (0.15 mm) nylon sieve. Pretreated samples were stored in clean polythene bags.
The contents of TOC and TON were determined using the Elementar (elementar vario MACRO cube, Elementar Analysensysteme GmbH, Germany). C/N was the ratio of TOC to TON.
6
Degree of Substitution in Aldehyde-Chitosan Schiff Bases
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The degree of substitution (DS) was determined with an element analyzer (Elementar vario MACRO cube, Elementar Corp., Frankfurt, Germany). The content of C and N was recorded, and the DS was calculated with Equation (1) [25 (link)]:
where (C/N)0 and (C/N) are the carbon-to-nitrogen ratio of CS and the fabricated aldehyde-chitosan Schiff bases and n is the number of carbons in aldehyde, which is 8, 9, 10, 7, and 10 for VL, CA, CN, BZ, and CT, respectively. Also, the degree of deacetylation in the original CS was calculated as 99.1% according to the literature [25 (link)].
where (C/N)0 and (C/N) are the carbon-to-nitrogen ratio of CS and the fabricated aldehyde-chitosan Schiff bases and n is the number of carbons in aldehyde, which is 8, 9, 10, 7, and 10 for VL, CA, CN, BZ, and CT, respectively. Also, the degree of deacetylation in the original CS was calculated as 99.1% according to the literature [25 (link)].
7
Comprehensive Characterization of Carbon Materials
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A Fourier−transform infrared spectrometer (FTIR, Germany, BRUKER−Vertex 70) and UV–Vis absorption spectra (UV−Vis, USA, PE−Lambda 750) were used to determine the types of functional groups in lignin molecules. The contents of different elements (C, H, N, S) in the materials were determined by using the element analyzer (Germany, Elementar−vario MACRO cube). The thermal stability of the materials was tested using an SDT Q600 analyzer (TA Instruments Inc., New Castle, DE, USA). X−ray diffraction (XRD; Germany, Brooker−Bruker D8 ADVANCE) was used to characterize the crystal structure of the materials. The surface composition of the samples was determined by X−ray photoelectron spectroscopy (XPS). Raman spectroscopy, using a Raman microscope, was used to characterize the defect degree of the materials. The morphologies of the carbon materials were characterized by high−resolution transmission electron microscopy (HRTEM) (Japan, JEOL−JEM 2100 F). The specific surface area and pore size distribution were determined by a nitrogen adsorption–desorption test (BET, USA, Mack−ASAP2460).
8
Sediment Carbon and Nitrogen Analysis
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The inorganic carbon in the sediment samples were removed by pretreatment with HCl (3 mol/L), and inorganic nitrogen was removed by pretreatment with KCl (2 mol/L) and HCl (0.5 mol/L) (Zhang et al. 2018b ). The pretreated samples were then freeze-dried and sieved using a 100-mesh (0.15 mm) nylon sieve. The TOC and totol organic nitrogen (TON) were analyzed using the Elementar (Elementar vario MACRO cube, Elementar Analysensysteme GmbH, Germany). C/N refers to the TOC to TON ratio.
9
Photosynthesis and Biomass Analysis
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The net photosynthetic rate of leaves was analyzed using a portable photosynthesis measurer (Li-6000, Li-COR Inc., USA) before the end of the experiment. Five measurements were carried out per pot between 10:00 and 12:00 a.m. under the same light intensity of 1000 μmol m -2 s -1 on a sunny day (Fan et al. 2013) .
Plants were sampled to estimate the biomass and plant uptake in the initial and at the end of the experiment. Every single plantʼs height and weight were measured, and the mean height and weight in each pH condition treatment were calculated. All plants were excavated, washed, and dried. Then, the harvested plant biomass was separated into root dry weight and total dry weight.
The root weight ratios (RWRs) were calculated, and relative growth rate (RGR) was measured according to the equation
Then, the whole plants were ground and analyzed for TN with a CHNS/O analyzer (Elementar vario Macro cube, Elementar Analysensysteme GmbH, Hanau, Germany).
Plants were sampled to estimate the biomass and plant uptake in the initial and at the end of the experiment. Every single plantʼs height and weight were measured, and the mean height and weight in each pH condition treatment were calculated. All plants were excavated, washed, and dried. Then, the harvested plant biomass was separated into root dry weight and total dry weight.
The root weight ratios (RWRs) were calculated, and relative growth rate (RGR) was measured according to the equation
Then, the whole plants were ground and analyzed for TN with a CHNS/O analyzer (Elementar vario Macro cube, Elementar Analysensysteme GmbH, Hanau, Germany).
10
In Vitro Digestibility Analysis
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After the completion of a 2-step digestion, 3 digesta replicates were filtered through Nylon mesh (30 μm, Narae Plus, Seoul, Republic of Korea) and the filtrates were dried at 60 °C for 24 h in a drying oven (NB-901M, N-BIOTEK, Bucheon, Republic of Korea). The weights of dried samples before the digestion (a), filter (Nylon mesh) (b), samples with filter after the digestion, filtration, and drying (c) were obtained after drying at 105 °C for 4 h (Method 934.01, AOAC, 2007 ), and used to calculate dry matter digestibility (%) following the equation by Boisen and Fernández (1995) (link).
For the determination of CP digestibility (%), dried filtrate (25 mg) was used in an Elemental Analyzer (vario MACRO cube, Elementar, Hesse, Germany) for the quantification of nitrogen (Biancarosa et al., 2017 (link)). The analyzed nitrogen content was multiplied by 6.25 to obtain the crude protein content (Marco et al., 2002 (link)). Crude protein digestibility (%) was calculated following the equation: where a = CP in the dried filtrate (%); b = DM digestibility; and c = CP in the sample (%).
For the determination of CP digestibility (%), dried filtrate (25 mg) was used in an Elemental Analyzer (vario MACRO cube, Elementar, Hesse, Germany) for the quantification of nitrogen (Biancarosa et al., 2017 (link)). The analyzed nitrogen content was multiplied by 6.25 to obtain the crude protein content (Marco et al., 2002 (link)). Crude protein digestibility (%) was calculated following the equation: where a = CP in the dried filtrate (%); b = DM digestibility; and c = CP in the sample (%).
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