The influence of synthetic conditions on the stability of methotrexate-monoclonal antibody conjugates determined by reversed phase high performance liquid chromatography.

Methotrexate (MTX) has been convalently attached to an IgG-type monoclonal antibody (791T/36) directed to tumour-associated antigen gp72. Conjugates were synthesized by the active ester method using MTX N-succinimidyl ester at various pH values (7.5-10.5). Following purification by gel filtration, high performance liquid chromatography was used to assess the free drug or its derivatives in samples of MTX-791T/36 conjugates previously treated (or not) with hydroxylamine. Quantitative analysis, performed on a reversed phase column (pore size 300 A) with isocratic acetonitrile-sodium acetate buffer (pH 4.8) as mobile phase, indicated no detectable amount of free methotrexate in hydroxylamine-treated conjugates even six months after their preparation. Similar observations were made with conjugates, whose synthesis were performed at pH greater than or equal to 10. In contrast, the presence of increasing amounts of drug/metabolite could be demonstrated in samples produced at lower pH values. Based on these findings, the pH-dependent kinetics of MTX release has been determined and used to design conditions under which stable MTX-791T/36 conjugates could be prepared without hydroxylamine reaction.     

The use of naphthalene derivatives in inorganic analysis : The nitronaphthylamines as new reagents for mercuric mercury and ceric cerium

The use of 4-nitro-1-naphthylamme as a spot test for mercuric mercury and 1-nitro-2-naph-thylamine as a specific spot test for ceric cerium have been observed In each case, the reactions appear to be suitable for general qualitative inorganic analysis The ceric cerium test was specific whereas in the case of mercuric mercury. Ce⁺⁴ was the only on of 51 tested which interfered.     

Negative ion mass spectra of metal complexes: Secondary electron capture and CS2 elimination from nickel (ii) dithiocarbamates

Negative ion mass spectra of series of bis-(N, N-dithiocarbamato)nickel(II) complexes of formula [NiS₂C·NR¹R²]₂ (where·NR¹R² ? ·NEt₂ ·NPr₂, ·NBu₂, pyrrolidinyl, piperidyl, morpholinyl, and ·NEtPh) have been obtained by secondary electron capture. Intense molecular anions are given for all compounds, with most fragments originating from these ions. Metastable data indicate that CS₂ is eliminated from all molecular anions.     

Radiation-Induced Copolymerization of Styrene and Cellulose at Low Dose Rates

A study has been made of the radiation-induced grafting of styrene to cellulose in the presence of cobalt 40 gamma irradiation at low total doses and dose rates. For copolymerization, What mans 41 filter paper was immersed in solutions of styrene in methanol at various monomer concentrations. The dose rates were varied from 140 to 3100 rads/hr while the total dose varied from lo³ to 2 × 10⁵ rads.

In the presence of oxygen the grafting results showed considerable scatter, but a statistical analysis revealed that at all concentrations a linear relationship existed between total dose and graft %. Three factors contributing to the variance about the regression line were examined. Residual oxygen in the solution, while not important at high dose rates, was shown markedly to contribute to the variance in the range examined. A linear dose-rate effect involving a decrease in graft with increasing dose rate was demonstrated. A LET effect also contributed to the over-all variance. Preliminary results indicate the presence of a maximum which may be a Trommsdorff effect.