Unipoint lc software

The instrumentation setup for our natural product peak library generation has been recently published.^{21 (link)} The HPLC stage utilized two Waters 510 pumps and a Waters 717plus autosampler, both controlled with Empower 2 software. Separation was performed on a 250 × 10 mm 5 μm Luna C₁₈ column (Phenomenex). Spectra from three detectors were acquired during peak library fractionation: Waters 996 photo diode array, SEDEX 55 ELSD, and Mariner 5054 ESI-TOF-MS. The mobile phase parameters are CH₃CN (A) and H₂O (B) with a flow rate of 2 mL/min and the following elution conditions: 30 min gradient from 10:90, 10 min isocratic 100:0, 1 min gradient from 100:0 to 10:90, 9 min isocratic 10:90. The injection amount was 15 mg/150 μL. Sample collection was performed using a Gilson 215 liquid handler controlled with Gilson Unipoint LC software. Samples were collected into BD Biosciences 96-deep-well plates, with a working volume of 2 mL (part number: 353966). Fractions were collected every minute. After the LC-MS-UV-ELSD library is collected, a duplicate archive plate is generated for analytical reference using a 12-channel pipet, creating an exact copy and counter balance for centrifugal drying. Plates were dried and concentrated using a Savant AES2010 SpeedVac.

LC-MS based purification was performed using a Waters purification system running Empower 2 software that utilized the following: (a) 717 Autosampler, (b) 510 HPLC pumps, (c) 996 PDA detector. The elution was split between a (1) Sedex model 55 evaporative light scattering detector (ELSD), (2) an Applied Biosystems Mariner electrospray ionization time-of-flight (ESI-TOF) mass spectrometer, and (3) a sample collection tube. Sample collection was performed using a Gilson 215 liquid handler controlled with Gilson Unipoint LC software as reported in detail previously.^{46 (link)} NMR experiments were run on a Varian Unity 500 spectrometer (500 and 125 MHz for ¹H and ¹³C respectively). High accuracy mass spectrometry measurements were obtained using the Applied Biosystems Mariner ESI-TOF mass spectrometer.