Isoelectric focusing of apolipoproteins in immobilized pH gradients: improved determination of apolipoprotein E phenotypes

A method for isoelectric focusing of apolipoprotein E in an immobilized pH gradient with added carrier ampholytes has been developed. This method is an improvement over conventional isoelectric focusing of apolipoprotein E with respect to resolution, reproducibility, and simplicity. Since monosialo isoforms are resolved from the normally cofocusing asialo isoforms, unique patterns are obtained for all 6 common apolipoprotein E phenotypes. The method can also be applied to the screening of apolipoprotein A and C isoforms. Delipidated very low density lipoproteins (VLDL) have been used as the source of apolipoprotein E and C. Apolipoprotein A isoforms were focused directly from detergent-treated serum. Immunodetection of apolipoprotein E using capillary transfer was found to be compatible with the described method.     

Automated multiple-layer spotting for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of synthetic polymers utilizing ink-jet printing technology

Recently, a new multiple-layer matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) sample spotting technique for poly(ethylene glycol), offering improved analysis possibilities, was described. In this contribution the application of ink-jet printing to automated, multiple-layer MALDI-TOFMS sample preparation of synthetic polymers is presented, allowing accurate deposition of matrix, additive and analyte solutions. The new sample preparation technique was evaluated for poly(ethylene glycol) as well as poly(methyl methacrylate) standards, and optimized settings for both synthetic polymers have been obtained.     

The separation of palladium and platinum by ion flotation

Differences in the ion flotation properties of palladium(II) and platinum(IV) chloro complexes in aqueous solutions are used to achieve separations of these metals. The anionic chloro complex PtCl^2-₆ is floated selectively with cationic surfactants of the type, RNR'₃Br, from solutions of PdCl^2-₄ and various concentrations of hydrochloric acid. The palladium(II) does not float from solutions of ? 3.0 M HCl and the platinum(IV) floated from these solutions can be recovered free of palladium. However, the separation is incomplete as much of the platinum(IV) is also unfloated from these solutions. Quantitative separations are obtained by conversion of the palladium(II) to the cationic ammine, Pd(NH₃)₄²⁺ with aqueous ammonia prior to flotation. The anionic chloro complex of platinum(IV) is unaffected by the presence of ammonia and is floated quantitatively with the surfactant n-hexadecyltri-n-propylammonium bromide from 0.01 M ammonia solutions.     

Two-dimensional capillary gas chromatography without intermediate trapping. Electron capture detector quantitation of an amino alcohol (KABI 2128) in serum after trifluoroacetylation

A two- dimensional gas chromatograph based on the Deans switching principle is described. The unit comprises two separate ovens, each containing a fused silica capillary column. The columns are joined in a specially designed manifold permitting heart cuts to be performed without significant decrease in efficiency. The switching speed and the retention time stability of the system made it possible to perform heart cuts of only a few seconds' duration. The system has been used under isothermal conditions for the determination of an amino alcohol (KABI 2128) in the low ng/ml range after trifluoroacetylation and with electron capture detection. A much shorter clean-up procedure could be used in combination with the two-dimensional gas chromatograph as compared to a method using a single glass capillary column.     

Dispersion phenomena in reactors for flow analysis

Both coiled open tubular reactors and packed-bed reactors can be used in flow analysis. Band broadening and pressure drop in these reactors are discussed. Theoretical analysis shows that packed-bed reactors are to be preferred. It is shown that for a given residence time and equal band-broadening values the pressure drop over a packed-bed reactor is lower than over a coiled open tubular reactor. Rules for optimal design are given for coiled tubular reactors and packed-bed reactors. The application of both reactors is shown for the spectrophotometric determination of phosphate with a vanadomolybdate reagent yielding a yellow colour.     

On [Na(POCl3)4]+ Complex Ion Formation: Conductivity,Raman, and NMR studies in the system phosphoryl chloride–sodium tetrachloro aluminate and related compounds

The system POCl₃–NaAlCl₄ was investigated by measuring the conductivity and the Raman and NMR spectra (²⁷Al, ²³Na and ³¹P) as a function of the mol fraction x of NaAlCl₄ in POCl₃. Additionally, Raman spectra of POCl₃ solutions of NaFeCl₄, LiAlCl₄, LiFeCl₄, and KAlCl₄ were recorded. In solutions containing Li⁺ or Na⁺ ions a liquid to solid (or jelly) phase transition was observed under certain conditions, dependent on salt concentration and temperature. Observed changes in the Raman spectra of the electrolyte solutions in comparison to the pure solvent POCl₃ demonstrate the existence of interactions. Clearly, the POCl₃ eigenfrequencies and hence the molecules are pertubed. The formation of [M(POCl₃)₄]⁺ complexes (M = Li, Na) can be deduced from the Raman measurements. NMR investigations support this conclusion. For assigning of Raman spectra, (Li⁺, K⁺) cation and ([FeCl₄]^?, [SbCl₆]^?) anion substitutions were employed.     

Enantiomers of polarized alkenes: chromatographic enrichment and thermal interconversion

(+)- and (?)-$\text{1}$ as well as (+)- and (?)-$\text{2}$ were partially separated by liquid chromatography on triacetylcellulose; racemizations resulted in the barriers (Table 2) to rotation via planarized transition states.     

BRCA1 mutation screening using restriction endonuclease fingerprinting-single-strand conformation polymorphism in an automated capillary electrophoresis system

Efficient mutation scanning techniques are needed for the rapid detection of novel disease-associated mutations and rare-sequence variants of putative importance. The large size of the breast cancer 1 gene (BRCA1) and the many mutations found throughout its entire coding sequence make screening for mutations in this gene particularly challenging. We have developed a method for screening exon 11 of the BRCA1 gene based on restriction enzyme digestion of fluorescence-labeled polymerase chain reaction (PCR) products followed by single-strand conformation polymorphism (SSCP) using an automated capillary electrophoresis system, denoted capillary restriction endonuclease fingerprinting (REF)-SSCP electrophoresis. Using this strategy on a control set of samples, we were able to detect 17 of 18 known sequence alterations. The method was then applied to screen 73 Norwegian females with family histories of breast and/or ovarian cancer. A total of 172 sequence alterations were detected, including substitutions, insertions, and deletions. One novel substitution of unknown function was identified. Sequencing of all samples negative in the capillary REF-SSCP system gave no additional mutations confirming the high sensitivity of the described methodology. Capillary REF-SSCP electrophoresis appeared as a technically convenient technique, requiring amplification of fewer PCR fragments than traditional SSCP. The novel strategy allows high-throughput mutation scanning without radioactive labeling and polyacrylamide gel electrophoresis (PAGE).     

The solvent venting injection technique in capillary supercritical fluid chromatography

A solvent venting technique for injection of volumes up to 1 μl on 50 μm i.d. SFC columns has been compared to direct injection methods. The peak broadening and peak splitting observed with direct injection have been examined and found to be related to the starting pressure, the column temperature, and the sample solvent, in addition to the sample volume. The solvent venting technique removed peak splitting and improved the column efficiency. With a proper selection of experimental conditions, the sample recovery was 100%. The major part of the solvent was eluted in 15–20 s. Several applications have been demonstrated.     

Four levels of pattern recognition

Problems of pattern recognition in chemistry and other subjects can be divided conveniently into four different types depending on the level of scope of the problem.(1) Classification into one of a number of defined classes. As an example blood samples taken from persons known to be either controls or welders are considered. The problem is whether trace element concentrations in these samples contain information on whether or not a person is a welder.(2) Level 1 plus the possibility that an object is an outlier, i.e. does not belong to any of the defined classes. As an example, the üse of ¹³C-n.m.r. data to decide whether 2-substituted norbornanes have the exo or endo structure is discussed. (2A) Level 2, asymmetric. This situation occurs when one class does not have a systematic structure, but another class is homogeneous and can be described by a level 2 model. This occurs in the classification of materials or compounds as good or bad, active or inactive, and in binary classifications. As an example the use of trace element data to classify steel samples as having good or poor properties of strength is discussed.(3) Level 2 plus the ability to relate the variables measured to external properties of continuous character. As an example, the classification of a series of chemical compounds as β -receptor blockers, β -receptor stimulants, or neither, on the basis of their structural variables is discussed. In addition, relations between these structural variables and the measured biological activity are sought within each of the two classes.(4) Level 3 with the difference that several external property variables in the objects are measured. It may be desirable to use variables of the objects both for classification and for relations to several property variables: such examples are numerous in analytical chemistry.