1H-NMR spectra were measured using a JEOL 400 MHz spectrometer at 293 K. The chemical shifts given in ppm are internally referenced to the residual solvent signal. HPLC-MS data were obtained using a Dionex UltiMate 3000 system on a Phenomenex Gemini C18 column (150 x 3.0 mm, 5μm) coupled to a Thermo LCQ Deca XP Max with electrospray ionization. Solvents used for HPLC: 0.05% HCO2H in H2O and 0.05% HCO2H in CH3CN. Electronic absorption spectra were measured using a Varian Cary 50 UV-Vis spectrophotometer. The UiO67-[RuOH2-]@FTO films were digested in 5 mL of conc. HNO3 for ICP-MS analysis.
Ultimate 3000 system
Manufactured by Phenomenex
The Ultimate 3000 system is a high-performance liquid chromatography (HPLC) instrument designed for analytical and preparative applications. It features a modular design with various interchangeable components, including pumps, autosamplers, detectors, and column compartments. The system is capable of delivering precise and accurate solvent flow rates, enabling efficient separation and analysis of a wide range of compounds.
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Lab products found in correlation
Cary 50 uv vis spectrophotometer, by Agilent Technologies (1 mentions)
400 mhz spectrometer, by JEOL (1 mentions)
Lcq deca xp max, by Thermo Fisher Scientific (1 mentions)
Tentagel r ram, by Rapp Polymere (1 mentions)
Api 2000, by Thermo Fisher Scientific (1 mentions)
Kinetex c18 column, by Phenomenex (1 mentions)
4 protocols using ultimate 3000 system
1
Characterization of UiO67-[RuOH2-]@FTO Films
2
HPLC Characterization of SMC and TSMC
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HPLC analysis was performed to check the chromatogram profile of SMC and TSMC prior to being used in the biological assays. The samples were filtered in a syringe-driven filter unit (phobic PTFE 0.20 μm, Millipore), and 30 μl was injected using a chromatogram method, which was developed to facilitate HPLC fractionation of a partially purified sample. A Dionex Ultimate 3000 system and a Phenomenex C-18 (4.6 × 250) Luna Column was used. The samples were injected at a flow of 1 ml/min. The final conditions used for analytic HPLC were: at zero min. – 50% methanol, 50% water; at 45 min. – 95% methanol, 5% water; and at 60 min. – 95% methanol, 5% water. The temperature of the column was 37 °C, and UV detection was carried out at 315 nm.
3
Synthesis and Purification of bMT2 Peptide
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The N-terminal fragment of MT2 (Met1-Ser32) corresponding to the b-domain (bMT2) was synthesized on TentaGel R RAM resin (loading 0.19 mmol g À1 , Rapp Polymere GmbH) using a Liberty 1 microwave-assisted synthesizer (CEM, USA) according to the standard Fmoc strategy described in detail before. 32, 33 The resinattached peptide was cleaved from the resin by 2 h incubation with a mixture of TFA/thioanisole/EDT/anisole (90/5/3/2, v/v/v/v), followed by precipitation in cold (À20 1C) Et 2 O. The crude peptide was purified by HPLC (Dionex Ultimate 3000 system) on a Phenomenex C18 column (Gemini-NX 5 mm, 110 Å) using 0.1% TFA with a 0-45% ACN gradient in 30 min and then lyophilized. The identity of the obtained pure peptide was confirmed by ESI-MS (API 2000 Applied Biosystems, USA). The average mass calculated for the b-domain was 3252.3/3252.8 Da.
4
Analytical HPLC Characterization of Plant Extracts
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Characterization of the plant extract and the isolated fractions was performed by analytical HPLC using a Thermo Scientific Ultimate 3000 system connected to a Kinetex™ C-18 column (5 µm, 100 Å, 150 × 4.6 mm, Phenomenex) and a PDA-100 Photodiode Array Detector. The elution programme was set as follows: 0% B (0 min), 10% B (1 min), 21% B (11 min), 28% B (12 min), 32% B (16 min), 36% B (17 min), 44% B (21 min), 48% B (22 min), 75% B (26 min), 100% B (27 min), 100% B (29 min), 0% B (29.1 min), 0% B (32 min), with a mobile phase consisting of eluent A (trifluoroacetic acid/acetonitrile/water 0.1 : 5 : 94.9 v/v/v) and eluent B (trifluoroacetic acid/acetonitrile 0.1 : 99.9 v/v), using a flow rate of 0.8 ml min -1 and UV detection at 280 nm. The extracts were analyzed at a concentration of 1 mg ml -1 . The absorption maxima of the single compounds were determined by performing a 3D scan using different wavelengths which are described for these components in the literature. 11 For further investigations we chose 280 nm, which was found to be the (or near the) absorption maximum for the majority of compounds of C. cajan.
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