1260 infinity 2 quaternary system

To identify allantoin in yam extract, HPLC was conducted using an Agilent 1260 infinity II quaternary system equipped with a G7129A vial sampler and a WR G7115 Adiode array detector (Agilent, Waldbronn, Germany) and a ZORBAXNH2 column (4.6 × 150 mm, 5-micron). Chromatographic separation was performed using a gradient solvent system consisting of acetonitrile (HPLC grade, Merck, Darmstadt, Germany) (B) and water (HPLC grade, Merck) (A). The gradient program was as follows: 0 min, 25% B; 5 min, 17% B; 10 min, 17% B. The injection volume was 10 μL and the column eluent was monitored at UV 200 nm while chromatography was performed at 30 °C with a flowrate of 1.0 mL/min. The HPLC pattern of allantoin in yam water extract has been reported in the literature [9 (link),26 (link)].

100 μCi (10 μL) of ⁶⁴Cu (Mallinckrodt Institute of Radiology, Washington University School of Medicine, St. Louis, Missouri, USA) was mixed with 10 μg (10 μL) of YbT in 0.1 M ammonium acetate (pH 7) to bring the reaction volume to 100 μL. The mixture was vortexed for 10–15 seconds before incubating in a thermomixer with 800 rpm agitation at 37°C for 1 hour. Labeling efficiency and radiochemical purity were determined using radio-HPLC (Agilent 1260 Infinity II Quaternary System with a Flow-RAM radio-HPLC detector) on a C18 column (Agilent Poroshell 120 EC-C18 column, 3 × 50 mm, 4 μm). The mobile phase consisted of water (A) and acetonitrile (B) with a gradient of 5%–95 % B over 15 min using a flow rate of 1 mL/min. The retention time of ⁶⁴Cu-YbT was 6.35 minutes, and a radiochemical yield of greater than 95% was achieved and used without further purification.