Flavins Analysis Service

Flavin is the common name for a group of organic compounds based on

pteridine, formed by the tricyclic heterocycle isoalloxazine. The riboflavin
(RF), vitamin B2, is an essential precursor of flavin adenine dinucleotide
(FAD) and flavin mononucleotide (FMN) co-enzymes. Another compound of
the flavin family is lumiflavin. The flavins have in common the isoalloxazine
ring system where the redox process occurs. The nature of the group
attached to the N-10, is the flavin members differentiation and can influence
the flavin adsorption orientation on the electrode surface, leaving the active
site more or less available. The redox process of the flavins
is thermodynamically reversible, irrespective of whether one or two electrons
per flavin molecule are being transferred, which means that their redox
states (quinone state, semiquinone state, hydroquinone state) must be
considered.
 Figure 1. Chemical structure and nomenclature of the flavins in oxidized state (a)

and redox process of part of flavin structure (b).

Flavins are useful in physiologic systems in that they are stronger oxidizing
agents than NAD+, thus fitting in further along the electron transport chain.
They participate in one or two electron processes (and thus reactions with
free radicals or metal ions), and in reduced form react directly with O (as in
hydroxylation reactions). Riboflavin is thus an enzyme cofactor, or
coenzyme, fundamental to many areas of metabolism, and is intimately
involved in processes by which the oxidation of glucose and fatty acids are
utilized for adenosine triphosphate (ATP) formation, and thus the support of
metabolic processes. Riboflavin coenzyme formation (and thus trapping
within cells), is initiated through phosphorylation by a flavokinase, which is
positively regulated by the most active form of thyroid hormone.
Flavins that are bound to proteins are resistant to degradation. However,
unbound forms are subject to catabolism. Both FAD and FMN are
catabolized by intracellular enzymes in ways directly analogous to the
breakdown of these forms in foods during their absorption across the
intestinal mucosal cell. Thus, FAD is converted to FMN by FAD-
pyrophosphatase (releasing AMP), and FMN is degraded to free riboflavin by
FMN-phosphatases. Both FAD and FMN are split to yield free riboflavin by
alkaline phosphatase.
The degradation of riboflavin per se involves initially its hydroxylation at
the 7α- and 8α-positions of the isoalloxazine ring by hepatic microsomal
cytochrome P-450-dependent processes. It is thought that catabolism
proceeds by the oxidation and then removal of the methyl groups. The liver,
in at least some species, has the ability to form riboflavin α-glycosides. As a
result of this metabolism, human blood plasma contains FAD and FMN as
the major riboflavin metabolites, as well as small amounts of 7α-
hydroxyriboflavin. Side chain oxidation has been observed in bacterial
systems, but not in higher animals.

Figure 2. Riboflavin metabolism and cellular processing pathways.

 The chemistry of flavins has been studied extensively. Riboflavin is a stable
compound with a variety of interesting properties. FMN and FAD are much
more important in cells, but are less stable in solution because of
the presence of phosphate ester and anhydride bonds. Riboflavin
is responsible for the important biological chemistry of all flavins.
One reason that flavoproteins have been so extensively analyzed is that
they can be studied by examining the properties of their cofactor that is
nearly always involved in the core function of each protein. Another reason is
the variety of chemical properties that have been exploited for
important biological functions of considerable diversity.
Flavins work as co-factors and form an integral part of the redox active sites
of many different enzymes involved in dehydrogenation reactions, dioxygen
activation, and electron transfer reactions. The understanding of the versatile
chemistry of flavins and the mechanisms of action of flavoprotein enzymes is
of extreme importance and has been focus of many investigations. Creative
Proteomics provides reliable, rapid and cost-effective flavins targeted flavins
metabolomics services.
Platform
 HPLC
Summary
 Identification and quantification of flavins.
Report
 A detailed technical report will be provided at the end of the whole
project, including the experiment procedure, instrument parameters.
 Analytes are reported as uM or ug/mg (tissue), and CV's are generally<10%.
 The name of the analytes, abbreviation, formula, molecular weight and
CAS# would also be included in the report.