Development, validation and application of an HPLC method for reduced vitamin C qualification in HBOCs solution

Abstract

A rapid and sensitive high-performance liquid chromatography (HPLC) method for vitamin C (Vc) determination was developed in hemoglobin-based oxygen carriers (HBOCs) solution. After polymerized hemoglobin precipitation, Vc could be tested out within 5 min. The standard curve was linear in Vc quantity of 0.00–3.00 μg. Vc recovery was 103.56–108.68%, and relative standard deviation values of the repeatability and precision were below 1.00%. The limits of detection and limits of quantification were 0.002 ppm and 0.01 ppm, respectively. Furthermore, the application of established HPLC methods in methemoglobin reduction by Vc under different pH indicated that higher pH values could accelerate the reduction. This study suggested the potential application of established HPLC method in HBOCs research and development.

Keywords:

• Hemoglobin-based oxygen carriers
• high-performance liquid chromatography
• methemoglobin
• reduction
• vitamin C

Declaration of interest

This research was financially supported by the Sichuan International Cooperation and Exchange Research Program (No. 2014HH0065), Sichuan Science and Technology Support Program (No. 2014SZ0123), Scientific Research Program of Sichuan Provincial Health and Family Planning Commission (No. 150256).