The data analysis and chemometric models were performed using Simca (Version 13.0, Umetrics, Umeå, Sweden) and Matlab (Version R2017a, Mathworks, Natick, MA, USA). Omnic (Version 8.2, Thermo Fisher, Waltham, MA, USA) and UV Probe (Version 2.34, Shimadzu, Tokyo, Japan) were applied to treat the FT-MIR and UV-Vis spectral data.
Uv probe
Manufactured by Shimadzu
Sourced in United States, Japan
The UV Probe is a spectroscopic analysis instrument manufactured by Shimadzu. It is designed to measure the ultraviolet and visible light absorption spectra of liquid or solid samples. The UV Probe provides accurate and reliable data for identifying and quantifying various chemical compounds.
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6 protocols using uv probe
1
Chemometric Analysis of Spectral Data
2
Bone Specimen Transmittance Analysis
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The transmittance of the bone specimen was obtained using a SolidSpec‐3700 integrating sphere (Shimadzu). The bone sample was placed at the entrance of the sphere, and its transmittance at the 500‐900 nm wavelengths were analyzed with a UV Probe (Shimadzu).
3
UV-vis Spectroscopic Analysis of Samples
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The UV-vis spectroscopic analysis was carried out using Shimadzu UV-2450 (SHIMADZU, JAPAN) with UV Probe, Japan. The spectrum was recorded using 1 cm quartz cell that has a slit width of 1.0 nm. The wavelength ranges chosen for recording the spectrum was 190-800 nm. The absorbance spectra (in the range of 0.2–0.9) and absorbance maximum (λmax) were recorded.
4
MALDI-TOF MS Analysis of Oligomers
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MALDI/TOF-MS analysis was performed using a new method established in this study with the technical support by the Tsinghua University (Beijing, China) Analysis Center. Sinapic acid (SA) was chosen as the MALDI matrix for oligomer detection, and α-Cyano-4-hydroxycinnamic acid (CHCA) was chosen as the MALDI matrix for peptide detection. The matrix solution was prepared by dissolving 20 mg 3-HPA and 45 mg dihydrogen ammonium citrate (DHAC) in a 1 mL mixture solution of acetonitrile/water (1:1, v/v). When testing an oligomer, we did not mix SA with the sample, but instead dried the three layers of SA-oligomer-SA on a steel plate to keep the original properties of the oligomer. MALDI-MS analysis was performed on AXIMA Performance (Shimadzu Scientific Instruments) using a standard stainless steel plate with manual pipetting. This instrument was equipped with a 337 nm nitrogen laser. The mass spectrum was acquired in the negative linear mode with an acceleration voltage of 20 kV, and the vacuum pressure in the ion source was (3–5) × 10−6 Torr. Data evaluation was done using the mass spectrometry software, UV Probe (Shimadzu Scientific Instruments).
5
Rubber Reflectance Spectroscopy
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The reflectance of the rubbers was measured with an integrating sphere-installed spectrometer (SHIMADZU CORP., SolidSpec-3700 DUV). Each rubber was cut into a size of (L30 × W15 × H3, mm) for the measurements. The range of the wavelength and its resolution for the measurements were set 400 -700 nm and 0.5 nm, respectively. The reflectance was analyzed with the software installed into the spectrometer (SHIMADZU CORP., UVprobe).
6
Mitochondrial Swelling Assay with Diclofenac and CsA
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Mitochondrial swelling was evaluated using a previously described method (Masubuchi et al., 2002) , with modifications. Mitochondria (0.5 mg/mL) were incubated in a reaction buffer composed of 125 mM sucrose, 65 mM KCl, 5 mM succinate, 10 mM HEPES-KOH, and 2.5 µM rotenone. The absorbance at 540 nm was then monitored for 150 sec or 1,500 sec using a UV Probe (Shimadzu Corporation), after adding 100 µM diclofenac and/or 5 μM cyclosporin A (CsA).
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