couraging results. The most recent model for ergot alkaloid production during
batch fermentation of Claviceps purpurea based on microbial life as the main
characteristic for microbial development during fermentation process was
proposed [98]. The aging process of the microorganism is represented by life
function, defined in microbial life space which is a measure of space in which
the observer follows the development of a biosystem through physiological and
morphological changes of a microorganism. As a consequence of such an approach, the relativistic theory is recognized. Growth and alkaloid synthesis data
from an industrial fermentation were tested to validate the developed model.
Metabolic flux analysis has not yet been applied to this system. It has been
suggested that an extension of the principles of metabolic control theory would
make it possible to identify rational optimal strategies for improvement of ergot
alkaloid formation [4].
It has also been suggested that the redox state of the cellular cytoplasm is
critical for the activity of coordinated enzymic events and thus for the elaboration of ergot alkaloids.
5
Analytical Methods
A very detailed review of the HPLC methods has been carried out [7]. The
author has described the stationary phase, the mobile phase, flow rate, and
detector system used by researchers since 1973. We would like to describe the
other analytical methods such as the capillary electrophoresis, flow injection
analysis and two-dimensional fluorescence spectroscopy which have found applications in ergot alkaloid research.
Using capillary zone electrophoresis (CZE), the resolution of ergot alkaloid
enantiomers and epimers was obtained [99]. Complete separation of racemic
mixtures in their enantiomers was obtained by using g-cyclodextrin as a chiral
additive in the background electrolyte. An easy and sensitive high performance
capillary zone electrophoresis (HPCZE) method for the determination of
ergovaline in the endophyte-infected fescue seed was reported [100]. With this
method, detection and quantification of ergovaline at low micrograms per
kilogram of the seeds were possible. The simultaneous assay of caffeine and
ergotamine in the pharmaceutical dosage tablet formulations by capillary
electrophoresis was reported [101]. The qualitative and quantitative determination of ergonovine, ergonovinine, ergocorninine, ergocornine, ergokryptine, ergosine, ergocristine, ergocristinine, and ergotamine by using
capillary electrophoresis (CE) was developed [102]. Using a laser-induced
fluorescence detection, the limit of detection of these alkaloids can be improved
30-fold compared to UV detection.
A micellar electrokinetic capillary chromatographic (MECC) method to
separate 17 dihydroergotoxines, aci-alkaloids, and oxidation products has been
described [103]. The authors used novel cationic dimeric (Gemini) surfactants
such as 1,3-bis(dodecyl-N,N-dimethyl ammonium)-2-propanol and 1,3-bis(tetradecyl-N,N-dimethyl ammonium)-2-propanol for the separation in less than
8 min.
16
J. Mukherjee · M. Menge
batch fermentation of Claviceps purpurea based on microbial life as the main
characteristic for microbial development during fermentation process was
proposed [98]. The aging process of the microorganism is represented by life
function, defined in microbial life space which is a measure of space in which
the observer follows the development of a biosystem through physiological and
morphological changes of a microorganism. As a consequence of such an approach, the relativistic theory is recognized. Growth and alkaloid synthesis data
from an industrial fermentation were tested to validate the developed model.
Metabolic flux analysis has not yet been applied to this system. It has been
suggested that an extension of the principles of metabolic control theory would
make it possible to identify rational optimal strategies for improvement of ergot
alkaloid formation [4].
It has also been suggested that the redox state of the cellular cytoplasm is
critical for the activity of coordinated enzymic events and thus for the elaboration of ergot alkaloids.
5
Analytical Methods
A very detailed review of the HPLC methods has been carried out [7]. The
author has described the stationary phase, the mobile phase, flow rate, and
detector system used by researchers since 1973. We would like to describe the
other analytical methods such as the capillary electrophoresis, flow injection
analysis and two-dimensional fluorescence spectroscopy which have found applications in ergot alkaloid research.
Using capillary zone electrophoresis (CZE), the resolution of ergot alkaloid
enantiomers and epimers was obtained [99]. Complete separation of racemic
mixtures in their enantiomers was obtained by using g-cyclodextrin as a chiral
additive in the background electrolyte. An easy and sensitive high performance
capillary zone electrophoresis (HPCZE) method for the determination of
ergovaline in the endophyte-infected fescue seed was reported [100]. With this
method, detection and quantification of ergovaline at low micrograms per
kilogram of the seeds were possible. The simultaneous assay of caffeine and
ergotamine in the pharmaceutical dosage tablet formulations by capillary
electrophoresis was reported [101]. The qualitative and quantitative determination of ergonovine, ergonovinine, ergocorninine, ergocornine, ergokryptine, ergosine, ergocristine, ergocristinine, and ergotamine by using
capillary electrophoresis (CE) was developed [102]. Using a laser-induced
fluorescence detection, the limit of detection of these alkaloids can be improved
30-fold compared to UV detection.
A micellar electrokinetic capillary chromatographic (MECC) method to
separate 17 dihydroergotoxines, aci-alkaloids, and oxidation products has been
described [103]. The authors used novel cationic dimeric (Gemini) surfactants
such as 1,3-bis(dodecyl-N,N-dimethyl ammonium)-2-propanol and 1,3-bis(tetradecyl-N,N-dimethyl ammonium)-2-propanol for the separation in less than
8 min.
16
J. Mukherjee · M. Menge