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 9alpha-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.
Lindholm
Johan
J
Center for Surface Biotechnology, Uppsala University, BMC Box 577, SE-751 23 Uppsala, Sweden.
Westerlund
Douglas
D
Karlsson
Karl-Erik
KE
Caldwell
Karin
K
Fornstedt
Torgny
T
D001709:Biotechnology; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005285:Fermentation; D006908:Hydroxyprogesterones / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods*; D019906:Nuclear Magnetic Resonance, Biomolecular; D013056:Spectrophotometry, Ultraviolet
D006908:Hydroxyprogesterones