Computer-aided multichannel detection in high-performance liquid chromatography

Summary The development of a computer-aided rapid-scanning detector for HPLC based on the linear photodiode array UV-visible spectrophotometer is described. Chomatograms monitored at up to three wavelengths, with automatic capture of spectra for eluted peaks and post-run processing of spectral data to generate log₁₀ (A) spectra, second derivative and fourth derivative spectra, are described. Methods are reported for the analysis of forensic samples of diacetylmorphine (heroin) in the presence of the degradation products and potential contaminants caffeine, papaverine, 6-acetylcodeine, thebaine, 6-acetyl-morphine, procaine and morphine separated by HPLC. The novel use of second and higher derivative spectra for the qualitative characterisation of drugs and contaminants separated by HPLC is described.Presented at the 14th International Symposium on Chromatography London, September, 1982     

205Tl NMR methods for the characterization of monovalent cation binding to nucleic acids

Monovalent cations play an important role in many biological functions. The guanine rich sequence, d(G4T4G4), requires monovalent cations for formation of the G-quadruplex, d(G4T4G4)2. This requirement can be satisfied by thallium (Tl+), a potassium (K+) surrogate. To verify that the structure of d(G4T4G4)2 in the presence of Tl+ is similar to the K+-form of the G-quadruplex, the solution structure of the Tl+-form of d(G4T4G4)2 was determined. The 10 lowest energy structures have an all atom RMSD of 0.76 +/- 0.16 A. Comparison of this structure to the identical G-quadruplex formed in the presence of K+ validates the isomorphous nature of Tl+ and K+. Using a 1H-205Tl spin-echo difference experiment we show that, in the Tl+-form of d(G4T4G4)2, small scalar couplings (<1 Hz) exist between 205Tl and protons in the G-quadruplex. These data comprise the first 1H-205Tl scalar couplings observed in a biological system and have the potential to provide important constraints for structure determination. These experiments can be applied to any system in which the substituted Tl+ cations are in slow exchange with the bulk ions in solution.     

Studies on alkaline protease produced by Bacillus sp. NG312

An alkalophilic hyperproducer of alkaline protease, Bacillus sp. NG312, was isolated, and the enzyme showed maximum activity at pH 11.0 and 60°C. The temperature optimum was increased by 10°C in presence of Ca²⁺. The crudeenzyme was found to have half-life of 11 d at 37°C and maximum stability at pH 9.0–10.0. It also exhibited very good stability in presence of detergent components and some locally available commericial detergent powders.     

Electron impact and chemical ionization mass spectra of 2,2-diphenyl-3-arylcyclobutanone oximes. Formation of ion m/z 167

The electron impact (EI) and chemical ionization (CI) spectra of 2,2-diphenyl-3-aryl cyclobutanone oximes (1–5) are reported. Formation of diphenylmethyl cation at m/z 167 is a major fragmentation process in both EI and CI spectra. Labelling studies established that the hydrogen involved in this rearrangement transfers from the NOH group and not from cyclobutane ring positions. The [M + 3]⁺ ions are formed under CI conditions as a result of C?N double bond reduction. An interesting secondary kinetic isotope effect is observed in the formation of ion e at m/z 183 in both EI and CI spectra. Other characteristic fragmentation pathways occurring in the EI and CI spectra of these compounds are outlined.     

Thermal decomposition of cesium hexanitratouranium(IV)

As cesium hexanitratouranium(IV), Cs₂U(NO₃)₆, has the same Cs:U stoichiometry as that of Cs₂UO₄, thermal decomposition of this nitrato complex in air and nitrogen was studied in detail as a possible alternate method of preparing pure Cs₂UO₄. The volatility of cesium nitrate, which is one of the intermediate products, changed this Cs:U ratio during thermal decomposition. Hence, only Cs₂U₂O₇ was obtained on heating the sample to 775 K or higher. A scheme for the thermal decomposition of Cs₂U(NO₃)₆ is given by combining the observed TG, XRD and IR data.     

A COMPARISON OF MAMMALIAN CELL SENSITIVITY TO EITHER 254 nm OR ARTIFICIAL "SOLAR" SIMULATED RADIATION

The sensitivity of six mammalian cell strains to either germicidal (254 nm) or artificial "solar" simulated radiation was tested. The solar simulator used had an output similar, in some respects, to natural sunlight. Cellular capacity for Herpes simplex virus production was used as the assay procedure. The tested cells were a strain of African green monkey kidney cells and five human skin fibroblast cell strains. The latter included a "normal" cell strain, and four photosensitive cell strains; three of which were strains of xeroderma pigmentosum cells, and one strain of Bloom's syndrome cells. When comparing the D¹⁰ values, the different cell strains varied by a factor of six in response to germicidal radiation, but only by a factor of two to artificial "solar" simulated radiation. The relative sensitivity of the cells to either type of radiation also varied from 1.7 to 10.9. Large variations in response occurred even among the xeroderma pigmentosum cell strains. These responses suggest that mammalian cell sensitivity to 254 nm radiation may not be a true indicator of a cell's responses to natural sunlight.     

High-performance liquid chromatography systems for the separation of benzodiazepines and their metabolites

The high-performance liquid chromatographic (HPLC) retention characteristics of 21 benzodiazepine drugs and some of their metabolites have been examined on both silica and ODS-silica packing materials. Four HPLC systems have been considered and retention data are presented for the drugs on these systems. The correlation of retention data on the systems is considered with reference to the problem of identifying unknown benzodiazepines.