Varian unity inova 500 spectrometer

¹H NMR experiments were carried out on a Varian UNITY INOVA 500 spectrometer (Agilent Technologies, CA, USA) operating at 499.839 MHz for proton. All the spectra were acquired at 300K using a standard 1D-NOESY pulse sequence for water presaturation with a mixing time of 1 ms and a recycle time of 6 sec. Spectra were recorded with a spectral width of 6000Hz, a 90° pulse, and 128 scans. Prior to Fourier transformation the free induction decays (FID) were multiplied by an exponential weighting function equivalent to a line broadening of 0.5 Hz and data were zero-filled to 64K. Chemical shifts were referred to the TSP single resonance at 0.00 ppm. All spectra were phased and baseline corrected using MestReNova software (Version 9.0.0, Mestrelab Research S.L.).

¹H NMR experiments were performed at 300 K with a Varian UNITY INOVA 500 spectrometer (Agilent Technologies, Inc., Santa Clara, CA, USA), operating at a frequency of 499.83 MHz. One-dimensional (1D) ¹H NMR spectra were acquired using a standard pulse sequence (1D NOESY), with pre-saturation during relaxation and mixing time for water suppression. Both the milk and urine spectra were acquired with 128 scans with 64 k data points, a spectral width of 6000 Hz, a recycle time of 3.5 s, and a mixing time of 1 ms. A 0.3 Hz line-broadening factor was applied to each spectrum prior to Fourier transformation. After phased and baseline correction, the chemical shift scale was set by assigning a value of δ = 0.00 ppm to the signal for the internal standard TSP.

Optical rotations were recorded with a JASCO DIP 1000 digital polarimeter (JASCO, Tokyo, Japan). UV spectra were recorded on a JASCO V-530 spectrophotometer (JASCO, Tokyo, Japan), and CD spectra were recorded on a JASCO J-810 spectrometer (JASCO, Tokyo, Japan). IR spectra were obtained on a JASCO FT/IR 300-E spectrometer (JASCO, Tokyo, Japan). ¹H-, ¹³C-, HSQC, and HMBC NMR experiments were recorded using a Varian Unity INOVA 500 spectrometer (Agilent Technologies, Inc., Santa Clara, CA, USA). HRESIMS were determined on Waters Synapt HDMS LC/MS mass spectrometer (Waters Corp., Milford, MA, USA). TLC was carried out on Merck silica gel F₂₅₄-precoated glass and RP-18 F_254S plates (Merck, Darmstadt, Germany). Medium pressure liquid chromatography (MPLC) was performed using Lobar^TM C₁₈ column (10 × 240 mm, 40–63 μm, Merck, Darmstadt, Germany) and silica gel (40–63 μm, Merck). HPLC was performed on a Hewlett-Packard Agilent 1100 Series (Agilent Technologies, Inc., Santa Clara, CA, USA) HPLC System composed of a degasser, a binary mixing pump, a column oven and a DAD detector using Waters SunFire™ (Waters Corp., Milford, MA, USA) (4.6 × 150 mm, 5 μm) and SunFire™ Prep C₁₈ column (10 × 150 mm, 5 μm) with acetonitrile (solvent A) and water containing 0.1% formic acid (solvent B).