FT-IR spectra of the prepared 1–6 year(s) old bamboo lignin samples were recorded on a spectrophotometer (Tensor 27, Bruker Optics, Karlsruhe, Germany) in the range of 1800–800 cm−1 using a KBr disc containing finely ground samples (1%) [21 (link)]. Lignin acetylation was carried out with reference to relevant literature [22 (link)], and the molecular weight of acetylated lignin was determined by GPC. In short, 1 mg acetylated lignin was dissolved in 1 mL of tetrahydrofuran (≥99.0%, HPLC, Sigma, Oliver Township, MI, American), and then filtered through an organic phase filter and measured by GPC (Nexera UHPLC/HPLC System Shimadzu, Kyoto, Japan) [21 (link)]. Experiments were repeated three times to obtain the average values. NMR spectra (13C NMR and 2D-HSQC spectra) of the lignin samples were obtained on a 600 MHz Bruker Avance (Tensor 27, Bruker Optics, Karlsruhe, Germany). The 13C NMR analysis conditions were: 100 mg lignin dissolved in 0.5 mL DMSO-d6, sampling time was 1.35 s, relaxation time was 1.5 s, scanned 3000 times; 2D-HSQC analysis conditions were: 60 mg lignin dissolved in 0.5 mL DMSO-D6, the sampling time was 0.17 s, the relaxation time was 1.5 s, sampled 128 times, and the scanning time was 6 h [23 (link)].
Nexera uhplc hplc system
Manufactured by Shimadzu
Sourced in Japan
The Nexera UHPLC/HPLC System is a high-performance liquid chromatography (HPLC) instrument designed by Shimadzu. It is capable of performing both ultra-high performance liquid chromatography (UHPLC) and conventional HPLC analyses. The system is equipped with advanced features to ensure accurate and efficient separation of complex samples.
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Lab products found in correlation
Tensor 27, by Bruker (1 mentions)
Lc 30ad pump, by Shimadzu (1 mentions)
Inertsustain c18 column, by GL Sciences (1 mentions)
Cto 30a column oven, by Shimadzu (1 mentions)
Sil 30ac autosampler, by Shimadzu (1 mentions)
Lcms 8040 triple quadrupole mass spectrometer, by Shimadzu (1 mentions)
Qtrap 6500 mass spectrometer, by AB Sciex (1 mentions)
4 protocols using nexera uhplc hplc system
1
Bamboo Lignin Characterization by FT-IR, GPC, and NMR
2
UHPLC Analysis of Compounds
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Separations were conducted using a Shimadzu Nexera UHPLC/HPLC system consisting of a DGU-20AR degasser, two LC-30AD pumps, an SIL-30AC autosampler, and a CTO-30A column oven (Shimadzu Scientific Instruments, Kyoto, Japan). A GL Sciences InertSustain C18 column (2.1 mm× 150 mm; 2 μm) was used, and the column temperature was 40°C. The mixture of 10 mmol/l aqueous formic acid solution and methanol was employed as the mobile phase, using the following gradient: 1% methanol for 0.1-10 min, 1%-50% methanol over 10-15 min, 50% methanol over 15-17 min, and 1% methanol over 17-24 min. The flow rate was 0.2 ml/min, and the injection volume was 1 μl.
3
Quantification of NAD+ in Meibomian Glands
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Concentrations of NAD+ were determined by LC–MS/MS, using a Shimadzu Nexera UHPLC/HPLC System (Shimadzu) coupled to a LC–MS-8040 triple quadrupole mass spectrometer (Shimadzu). Meibomian gland cells, selectively isolated into 50% methanol (see above), were sonicated with chloroform (1:1). After centrifugation, supernatants were filtered (0.22 µm) and freeze-dried. The samples were resolved in water before analysis and separated through a COSMOSIL Packed Column 5C18-PAQ (2.0 mm × 150 mm, Nacalai). NAD+ was detected using the multiple reaction monitoring mode and identified by comparison of its LC retention time and MS2 (link) fragmentation pattern (m/z 663.85 > 136.05) with those of the authentic standard. NAD+ levels were quantified based on the peak area compared to a standard curve. The values of NAD+ were normalized to the tissue volume (mm3) captured by laser microdissection.
4
Quantification of Docetaxel in Cells
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Cells (200,000 cells/well) were placed in a 24-well plate and cultured for another week at 100% confluency. Thereafter, 0.125 mg/mL of DTX was added as a stimulant at 48, 72, 96, and 120 h. The cells were washed with double-distilled water three times and collected in a centrifuge tube for ultrasonic crushing for 15 min. The final samples were sent to the Institute of Clinical Pharmacology of Central South University for HPLC-MS/MS detection of DTX residue. Briefly, the standard stock solution of DTX, 0.552 mg/mL, was diluted with methanol/water (1:1, v/v) to prepare a set of DTX calibration solutions with various concentrations and then deproteinized with acetonitrile. After centrifugation at 15,000 rpm for 10 min at 4 °C, 100 μL of the supernatant was used for HPLC-MS/MS analysis to establish the standard curve. Tested samples were also diluted with methanol/water (1:1, v/v) and deproteinized by addition of acetonitrile. Analyses were performed after centrifugation as above. All chromatography experiments were performed at ambient temperature using the Nexera UHPLC/HPLC system (Shimadzu, Japan). The QTRAP 6500 mass spectrometer produced by AB Sciex (Concord, ON, Canada) was used for analyte detection and operated in the positive ion mode using multiple reaction monitoring. The precursor-product ion transitions were monitored at m/z 830.3→549.2 for DTX.
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