9-anthryldiazomethane

Biotin does not exhibit UV absorbance. It neither shows fluorescence nor electrochemical activity. Therefore, it needs to be derivatized. 4-Bromomethylmethoxiycoumarin (BMMC) [590], 9-anthryldiazomethane (ADAM) [591], and 1-pyrenyldiazomethane (PDAM) [592] have been used as precolumn reagents to convert biotin to fluorescent absorbing derivatives. Instead, to obtain derivatives that are UV detectable, hydrazines are used, such as 2-nitrophenylhydrazine hydrochloride... 

Anthryldiazomethane (ADAM) has been widely used for HPLC of various biologically important carboxylic acids. 

Nimura, N. and Kinoshita, T., Fluorescent labeling of fatty acids with 9-anthryldiazomethane (ADAM) for high performance liquid chromatography, Anal. Lett., 13, 191, 1980. 

One important question about YTX is if the toxin enters the cellular cytosol. This matter is important to clarify where the toxin induce the effect. In a set of experiments done with YTX labeled with 9-anthryldiazomethane (ADAM), the entry of YTX either in lymphocytes and enterocytes has been studied. The toxin is going inside both cellular models even the kinetic uptake was higher and faster in enterocytes (unpublished results). 

The polymerization of methyl methacrylate initiated by the products of the thermal degradation of 9-anthryldiazomethane can serve as an example of the synthesis of a polymer vrith terminal LM using an initiator-containing luminescent group ... 

The preparation of labeled polyalkylidenes is another example . Polyalkyl-idenes are hydrocarbon polymers of the f CH(R)4n type formed by polymerization or copolymerization of diazoalkanes RCHN2 they are of interest as model polymers Polyalkylidenes of various structures containing LM (Table 1, LMg) can be obtained by copolymerization of diazomethane (or its mixture with other diazoalkanes) and 9-anthryldiazomethane . ... 

When 9-anthryldiazomethane and the polymer have a common solvent, the homogeneous method is the most convenient one, since it ensures the highest yield and leads to the most homogeneous distribution of LM in the polymer (in the case when the contents of carboxylic groups in macromolecules greatly exceeds the required amount of LM) ... 

For water-soluble natural or synthetic polymers, one of the heterophase methods can be used l In this case, a considerable portion of 9-anthryldiazomethane is consumed by side reactions and this loss must be considered when choosing the amount of the reagent. 

These methods of the reaction between 9-anthryldiazomethane and carboxylic groups of macromolecules permit to bond LM to side groups of synthetic and natural polymers and copolymers of acrylic and vinylbenzoic acids hydrolyzed copolymers of maleic anhydride , carboxymethylcellulose comb-like alkyl acrylates , polyglutamic acid and other synthetic polypeptides as well as proteins, such as invertase or ribonuclease l... 

Investigations have shown that 9-anthrylcarbene generated by thermal or photoiytic decomposition of 9-anthryldiazomethane in a polymer solution, can be inserted into the C—H bonds of the mactomolecules . As a result, anthracene groups are covalently bonded to the macromolecules (Table 1 LMn). 

For example, the single-stage preparation of the polymers and copolymers of styrene with LM of the 9-anthrylmethyloxycarbonyl structure (Table 1 LMi) by copolymerization is accompanied by homolytic side reactions (Scheme 1). However, such labeled polymers can be obtained in two stages by copolymerization of the main monomer with the labeling amount of acrylic, methacrylic or vinylbenzoic acids and subsequent anthrylmethylation of the carboxylic groups of the polymer using 9-anthryldiazomethane. 

The treatment of a copolymer of methyl methacrylate with a labeling amount of methacrylic acid units by 9-anthryldiazomethane, lO-methyl-P-anthryWiaro-methane or lO-phenyl-9-anthryldiazomethane serves as an example of the preparation of polymer (copolymer) samples of the same structure but with different LM by a combined method ° l... 

Anthryldiazomethane ADAM Carboxyl (and other acidic groups)... 

The following fluorescent alkylating agents have been introduced as pre-chromatographic reagents 9-bromo-methylacridine [480], 3-bromomethyl-6,7-dimethoxy-l-methyl-2(lH)-quinoxalinone [481], naphthacyl bromide (2-bromoacetonaphthone) [482, 483], p-(anthroyloxy)-phenacyl bromide (panacyl bromide) [484], 1-bromo-acetylpyrene [485], 9-chloromethy[anthracene [486], and 9-anthryldiazomethane [487, 488]. 

A method for the determination of arachidonic acid metabolites with 9-anthryldiazomethane has been reported [489]. 

Seven fatty acids ( ,4.0, C,6 o, C,g., C,g.o, C g., C,g.2, and 20 4) were extracted from cardiac tissue and analyzed as their 9-anthryldiazomethane derivatives [1061]. Excellent peak shsqjes and baseline resolution were obtained on a C,g column (A = 365 nm) using a 93/7 acetonitrile/water mobile phase. The total analysis time was 60 min. The authors noted that small changes in the mobile phase composition or minor deviations from a flow rate of 0.6 mL/min destroyed the resolution between the Ci4 o, C15.1, Cjg.j, and 20 4 compounds. The detection limits were reported as 15 flnol injected (S/N = 4). Linear working curves are shown for up to 400pmol injected. 

Okadaic acid and pectenotoxin-2 (P2) were extracted from phytoplankton, derivatized with 9-anthryldiazomethane, and separated in 18 min on a C g column... 

Nimura and Kinoshita [96] developed a derivatization technique for the analysis of fatty acids that is based on 9-anthryldiazomethane (ADAM). The latter is very stable in solution and reacts with fatty acids at room temperature without a catalyst to yield strongly fluorescent esters, which can be chromatographed on an ODS phase with acetonitrile/water mixtures. The derivatization of fatty acid is carried out with a 0.1% methanohc solution of 9-anthryldiazomethane, which is prepared via oxidation of 9-anthraldehyde-hydrazone following a procedure by Nakaya et al. [97]. The fluorescence and excitation spectra of fatty acid-methyl-anthracene esters exhibit maxima at 412 and 365 nm, respectively. As an example, Figure 8.62 shows the separation of ADAM derivatives, a mixture of saturated and unsaturated long-chain fatty acids, which can be detected down to the lowest picomole range. 

Biotin and its analogues can be discriminated by their structural dilference using reversed phase or anion exchange HPLC (Bowers-Komro et al. 1986 Chastain et al. 1985 Livaniou et al. 2000) with no further derivitization. However, fluorescent derivatizations with 4-bromomethyl-7-methoxycumarin (Br-Mmc), 9-anthryldiazomethane (ADAM), 1-pyrenyldiazomethane (PDAM), thiamine, o-phthalaldehyde (OPA) or 3-mercaptopropionic acid (3-MPA) have usually been attempted to obtain better detection limit (Nojiri et al. 1998 Yokoyama and Kinoshita 1991). 

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