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Lt200 system

Manufactured by HACH

The LT200 system is a laboratory instrument designed for the analysis and measurement of various water quality parameters. It is capable of performing automated tests and providing accurate results. The LT200 system is suitable for use in water treatment facilities, environmental laboratories, and other settings where water quality monitoring is required.

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2 protocols using lt200 system

1

Lignin Quantification and Characterization

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Five mg of CWR were digested with 2.6 mL of 25% acetyl bromide in glacial acetic acid for 2 h at 50 °C using a HACH LT200 system. Samples were cooled on ice and transferred into 10 mL of 2 M NaOH plus 12 mL glacial acetic acid. The reaction tube was rinsed with glacial acetic acid and 1.75 mL of 0.5 M hydroxylammonium chloride was added. Each volume was adjusted to 30 mL with glacial acetic acid, centrifuged (3000 g, 15 min) and lignin content was measured spectrophotometrically (280 nm, extinction coefficient ε = 22.9 g− 1 L cm− 1).
Lignin was characterized following the nitrobenzene oxidation method previously described [69 (link), 78 (link)]. The products after nitrobenzene oxidation of ten mg of CWR were derivatized with Bis(trimethylsilyl)trifluoroacetamide and analysed by gas chromatography coupled to mass spectrometry (GC-MS). A HP-5MS column (30 m × 0.25 mm, 0.25 μm, Agilent) was used on a 7890B-5977A GC-MS system (Agilent). Salicylic acid-D4 was used as internal standard.
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2

Lignin Characterization by Nitrobenzene Oxidation

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Lignin was characterised on 3 or 4 biological replicates using the nitrobenzene oxidation method [64 (link)]. 10 mg of CWR were digested with 2 mL of 2 M NaOH and 30 μL nitrobenzene at 165 °C for one hour (Hach LT200 system). After centrifugation, ca. 1500 μL of supernatant was collected and 10 μl of vanillin-D3 (Sigma-Aldrich) at 10 mg/mL in 1,4-dioxan were added as a surrogate standard. Nitrobenzene was removed by four washing steps with ethyl acetate (1 mL, vortexing/centrifugation cycle). The pH of the solution was adjusted to 2–3 by adding approximately 200 μL of 6 N HCl solution. The oxidation products were recovered by two successive extractions with 1 mL ethyl acetate (vortexing/centrifugation cycle) followed by cleaning with 500 μl of saturated NaCl solution and drying with Na2SO4. The GC-MS analysis was performed after trimethylsilylation, realized by addition of 50 μl of Bis(trimethylsilyl)trifluoroacetamide (BSTFA) to 50 μL of dried extract and derivatization at 60 °C for 30 min. Quantitative analyses were performed using a HP-5MS column (30 m × 0.25 mm, 0.25 μm, Agilent) installed in a 7890B-5977A GC-MS system (Agilent). Injection was done at 250 °C in splitless mode. The oven program started at 40 °C for 5 min, increased to 230 °C at 10 °C/min, then to 320 °C at 40 °C/min and was kept at 320 °C for 10 min. Salicylic acid-D4 was used as internal standard.
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