Use of liquid chromatography–diode-array detection and mass spectrometry for rapid product identification in biotechnological synthesis of a hydroxyprogesterone

In exploratory scale biotechnological process development, the product must be rapidly identified although a reference compound may not always be available. LC–diode-array detection and MS were used for this purpose in a process producing 9α-hydroxyprogesterone from progesterone as substrate. The electrospray ionization mass spectrometer was combined with an ion trap mass spectrometer for the second generation MS. The preliminary identification, which could be carried out within the course of a day, confirmed that the product was a hydroxyprogesterone. The final identification step, which was much more material intensive and hence time consuming, involved a two-step preparative separation to yield quantities necessary for definitive product identification based on 1H- and 13C NMR.