The UV and IR spectra were recorded on Hitachi-UV-3200 and JASCO 320-A spectrometers. The 1H-, 13C NMR and 2D-NMR spectra were recorded on a Bruker AMX-500 spectrometer with tetramethylsilane (TMS) as internal standard. Chemical shifts (δ) are given in ppm relative to tetramethylsilane internal standard. Jeol JMS-700 High Resolution Mass Spectrophotometer was utilized for the mass determination. Electron impact mode of ionization was used, keeping ionization energy of 70ev. Resolution was set up to 10 k direct probe was used with temperature ramp setting, initial temperature 50 °C rise with rate of 32 °C per minute and final temperature set up to 350 °C. Thin layer chromatography (TLC) was carried out on precoated silica gel F254 plates (E. Merck, Darmstadt, Germany); detection was performed at 254 nm and by spraying with p-anisaldehyde/H2SO4 reagent. All chemicals were purchased from Sigma Chemical Company (St. Louis, MO, USA).
Uv 3200
Manufactured by Jasco
The UV-3200 is a compact and reliable UV-Visible spectrophotometer designed for laboratory use. It features a wavelength range of 190 to 1100 nanometers and can perform various spectroscopic measurements, including absorbance, transmittance, and concentration analysis. The instrument is equipped with a high-resolution display and intuitive user interface, making it easy to operate and interpret results.
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2 protocols using uv 3200
1
Spectroscopic Characterization of Organic Compounds
2
Isolation and Characterization of a New Compound
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A Diaion HP-20 column was used to fractionate the butanol extract with an H2O-MeOH gradient solvent system. The fraction eluted with a 1:1 H2O-MeOH solvent mixture was then subjected to polyamide chromatography with a CHCl3-MeOH gradient solvent system, starting with 100% CHCl3 with 10% step increments in the methanol content, providing eleven fractions (P1-P11) obtained from the various extractions. The fractions with the same TLC profile (P5-P9) were combined (10 g) and fractionated using the Sephadex LH-20 column using a gradient elution with an H2O-MeOH solvent system to afford four fractions. The final purification was performed on a reverse phase recycling HPLC with H2O-MeOH (1:1) and an L-80 column providing a new compound 1 (102 mg) and two unresolved fractions A and B. These fractions were separately resolved using an H-80 column with H2O-MeOH (1:1) solvent resulting in the isolation of four known compounds 2-5 (Choudhary et al., 2008) . The structure elucidation was performed using UV (Hitachi UV-3200), IR (JASCO 302-A), NMR (Bruker AMX-500 with tetramethylsilane as internal standard), and HRMS (ESI-QTOF, Applied Biosystems).
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