The egg yolk was freeze-dried using a lyophilizer (NAI; Shanghai, China). The egg yolk was ground, placed in a bag, which was sealed and stored in a refrigerator at −80°C. Then, 2.5 g of lyophilized yolk sample and 5 mL of deionized water were added to a 50-mL centrifuge tube that was placed in a 50°C ultrasonic machine for 30 min after cooling. The yolk sample in the centrifuge tube was washed 3 times with 30 mL of dichloromethane. The contents of the centrifuge tube were placed in a separatory funnel and filtered 3 times. Chromatographic separation was performed using an Alliance 2695 HPLC instrument (Waters, Milford, MA). The analysis time was 10 min, which was followed by a re-equilibration time of 2 min, for a total run time of 12 min. The flow rate of the mobile phase was 1 mL/min, the injection volume was 10 μL, the column temperature was maintained at 30°C, and ASTA was detected at a wavelength of 474 nm.
Alliance 2695 hplc instrument
Manufactured by Waters Corporation
Sourced in United States
The Alliance 2695 HPLC instrument is a high-performance liquid chromatography system designed for analytical separation and quantification of chemical compounds. It features a modular design, advanced solvent and sample management, and precise temperature control to ensure reliable and reproducible results.
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Lab products found in correlation
2 protocols using alliance 2695 hplc instrument
1
Quantification of Lyophilized Egg Yolk ASTA
2
Size Exclusion Chromatography for Antibody Characterization
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Size exclusion chromatography for the antibody was performed on an Alliance 2695 HPLC instrument (Waters Corporation, Milford, MA, USA) with UV/Vis detection at 280 nm using a Tosoh Biosciences TSK G3000SWXL 7.8 mm ID × 30 cm L column (Tosoh Bioscience GmbH, Griesheim, Germany) at 20-25 • C. The mobile phase of 0.1 M potassium phosphate, pH 6.8 ± 0.1 was maintained at a flow rate of 0.5 mL/min. Samples were diluted to 10 mg/mL and maintained at 5 • C prior to injection of 20 µL. Peak areas were integrated with respect to the baseline with Empower 3 software (Waters Corporation, Milford, MA, USA), and high molecular weight species (HMWs), low molecular species (LMWs), and monomer content were reported as percent of the total peak area.
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