Highly sensitive assay for tyrosine hydroxylase activity by high-performance liquid chromatography.

A highly sensitive assay for tyrosine hydroxylase (TH) activity by high-performance liquid chromatography (HPLC) with amperometric detection was devised based on the rapid isolation of enzymatically formed DOPA by a double-column procedure, the columns fitted together sequentially (the top column of Amberlite CG-50 and the bottom column of aluminium oxide). DOPA was adsorbed on the second aluminium oxide column, then eluted with 0.5 M hydrochloric acid, and assayed by HPLC with amperometric detection. D-Tyrosine was used for the control. alpha-Methyldopa was added to the incubation mixture as an internal standard after incubation. This assay was more sensitive than radioassays and 5 pmol of DOPA formed enzymatically could be measured in the presence of saturating concentrations of tyrosine and 6-methyltetrahydropterin. The TH activity in 2 mg of human putamen could be easily measured, and this method was found to be particularly suitable for the assay of TH activity in a small number of nuclei from animal and human brain.     

Synthesis of methylpyridine derivatives—XXXIIIChem. Pharm. Bull.25, 203 (1977).: Phosphonylation and chlorination of methylpyridine and 3-nitro-methylpyridine derivatives

Novel phosphonylation of the active Me group of pyridine derivatives is described.Reaction of 2 (and 4) -methylpyridine (3 and 5) with PCl₅ in POCl₃ gave 2 (and 4) -trichloromethylpyridine (4 and 6). Similarly, reaction of 4 (and 6) methyl-3-nitropyridine (7 and 9) afforded the corresponding trichloromethyl derivative (8 and 10). However, the similar reaction of 2-methyl-3-nitropyridine (11) gave dichloro-(3-nitro-2-pyridyl)-methylphosphonic dichloride (12).Mechanisms of the formation of these products are discussed.     

Magnetic circular dichroism and anomalous fluorescence spectra of [2,2] (2,6)azulenophanes

The electronic absorption spectra of syn-[2,2] (2,6) azulenophane (1) and anti-[2,2] (2,6)azulenophane (2) were investigated via magnetic circular dichroism spectra. The anomalous fluorescence spectra were also observed. From the assignment of the absorption spectra, it was concluded that molecules 1 and 2 emitted the fluorescence from the third excited singlet state.     

Purification of enzymes by high-speed gel filtration on TSK-GEL SW columns

The purification of enzymes was investigated by high-speed gel filtration on TSK-GEL G3000SWG columns packed with porous silica gel deactivated by chemically bonded hydrophilic compounds. Crude β-galactosidase from bacterial cells and commercial urease were purified ca. 15-fold in a single gel filtration. These enzymes were eluted within an hour from the column and the recoveries of enzymatic activity were almost 100% although the operation was carried out at room temperature (22°). Samples up to 100 mg could be applied to the column without loss of separation efficiency.     

Base-induced rearrangement of 1-trimethylsilylpropargyl alcohol. Generation of lithiated trimethylsiloxyallenes and their reactions

Under the effect of butyllithium, 1-trimethylsilylpropargyl alcohol undergoes rearrangement to afford the corresponding trimethylsiloxyallene or its lithiated one, which can be used for the preparation of α,β-unsaturated ketone derivatives.     

Syntheses of 3-substituted furans via lithium di(3-furyl)cuprate—the application of lithium di(3-furyl)cuprate to the total synthesis of dendrolasin

One step syntheses of 3-substituted furans by the reactions of a new reagent, lithium di(3-furyl)cuprate $\text{1}$, with various substrates and the application of $\text{1}$ to a total synthesis of dendrolasin are described.     

Novel rearrangement of twisted 2,4-di-alkyl 2-trans-4-trans pentadienals: An approach to cyclopentenones based on molecular mechanical consideration

Rearrangement of 2,4-di-$\text{trans}$-pentadienals $\text{IVa-d}$ was described as a consequence of the successive alkyl substitutions on the C-2 and C-4 positions. ¹H-FT nmr spectroscopy suggested that the reaction involves ketene $\text{VIII}$ as an intermediate.