A culture medium is a source of nutrients such as amino acids, glucose, vitamins, inorganic salts, and serum as a source of hormones and growth factors. Different kinds of media are used depending on the type of cell to be cultured. Currently, Chinese Hamster Ovary (CHO) cell culture media have gained a lot of attention for their application in bioprocessing and pharmaceutical research and development. They are highly used for the production of antibodies, cell therapy drugs and other recombinant proteins. However, in Korea, there is a lack of standard analysis methods for objective verification of ingredients in a culture medium. Thus, it is necessary to establish a method to analyze the components of the existing commercial media to aid the companies in the quantitative analysis of culture media components.
This study focuses on the identification and quantification of different kinds of vitamins in a culture medium consisting of various ingredients. The quantitative analysis of various vitamins in culture media using the HPLC-MS system was optimized. Analysis was performed using an Agilent 1200 series HPLC with an Agilent 6460 triple quadrupole (QQQ) mass spectrometer. LC separation was conducted with 0.1% formic acid in distilled water and 0.1% formic acid in acetonitrile as mobile phase. MS detection was conducted on multi-reaction monitoring (MRM) mode acquisition for analysis confirmation of triple Q MS equipped with an Agilent jet stream ion spray, and data were obtained in both positive and negative ionization modes.
Therefore, the development of analytical methods for various components of the medium can contribute to the domestic production of import-dependent CHO cell culture media.