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PubMed1996

Affinity countercurrent chromatography using a ligand in the stationary phase.

In countercurrent chromatography (CCC), an addition of a ligand to the liquid stationary phase remarkably improved both retention time and peak resolution of the analytes: various amino acid derivatives were separated by N-dodecanoyl-L-proline-3,5-dimethylanilide, while polar catecholamines and dipeptides were separated by bis-(2-ethylhexyl)phosphoric acid. By selecting an appropriate ligand and dissolving it in the liquid stationary phase, the present CCC technique can perform a variety of separations comparable to chiral chromatography, ion chromatography, and affinity chromatography. Leakage of the ligand from the column can be entirely eliminated by introducing a small volume of a ligand-free stationary phase at the end of the column as an absorbent. The method further facilitates application of pH-zone-refining CCC and can increase the sample loading capacity over 10 times for a given column.

Amino Acids↗
PubMed1998

On-line microporous membrane liquid-liquid extraction for sample pretreatment combined with capillary gas chromatography applied to local anaesthetics in blood plasma.

A new automated procedure for analyzing complex samples has been developed utilizing microporous membrane liquid-liquid extraction (MMLLE) combined with capillary gas chromatography. Some local anaesthetics were used as model compounds in aqueous solution as well as in blood plasma. The MMLLE procedure was performed in a flow system with the sample fed to the donor side of the hydrophobic microporous membrane and with an organic solvent (hexane) in the pores and as the acceptor solution. The analytes in a small volume of sample (< 1 mL) were extracted into the organic acceptor phase which was transferred into the gas chromatographic system by utilizing a loop-type interface compatible with large-volume (300 microL) injection. High selectivity and low carry-over effects were obtained with the system. The detection limits were 0.5-1 ng/mL using 0.5 mL of human plasma, and the precision was approximately 5%. The effects of pH, flow rates, and adsorption of the analytes were evaluated.

Adsorption↗
PubMed1999

On-line microextraction of metal traces for subsequent determination by plasma atomic emission spectrometry using pH peak focusing countercurrent chromatography.

Metal ions were highly efficiently enriched by pH peak focusing high-speed countercurrent chromatography. The peak intensity for a 10-mL standard sample in the effluent stream was increased over 100-fold compared to conventional plasma atomic emission spectrometry. Ca, Cd, Cu, Mg, Mn, Ni, and Zn are chromatographically extracted in a basic organic stationary phase containing a complex-forming reagent such as bis(2-ethylhexyl) phosphoric acid. After the sample solution is introduced into the column, metal ions remain around the sharp pH border formed between acidic and basic zones, moving toward the column outlet. Enriched metal ions are finally eluted with the sharp pH border as a highly concentrated peak into a volume of less than 100 microL. We evaluated this method for concentration efficiency in trace determination in tap water using different column diameters.

Countercurrent Distribution↗
PubMed1976

Nucleotide sequence of phenylalanine transfer ribonucleic acid from pea (Pisum sativum, Alaska).

Phenylalanine transfer ribonucleic acid from peas (Pisum sativum, Alaska) was completely digested with beef pancreatic ribonuclease (RNase I) and with ribonuclease T1. The resulting oligonucleotides were compared with those from the corresponding hydrolyses of phenylalanine transfer ribonucleic acid from wheat germ. The structures of both ribonucleic acids appeared to be identical. This report is the first to show that identical structures for the same specific acceptor transfer ribonucleic acid are present in two different plant species.

Base Sequence↗
PubMed1976

Membrane surface properties other than charge involved in cell separation by partition in polymer, aqueous two-phase systems.

When aqueous solutions of dextran and of poly-(ethylene glycol) are mixed above certain concentrations, immiscible, liquid two-phase systems are obtained which are useful for separating cells by partition. Some salts partition unequally between the phases, giving rise to an electrostatic potential difference between them. Partition of cells has therefore been thought to depend predominantly on membrane charge. We now report two instances in which membrane charge either does not determine or is not the main determinant of cell partition. (A) Cell partition coefficients in phase systems approaching the critical point (the component concentrations below which a homogeneous solution occurs) increase, even in phase systems in which the phase potential difference is practically zero. Furthermore, in such systems, the partition coefficient of (human) erythrocytes is not reduced by complete removal of sialic acid. (B) Rat and mouse erythrocytes have sizable partition coefficients in a phase system away from the critical point with no potential difference between the phases. Cell surface interaction with the polymers is probably responsible for cell partition in these cases. Partition studies on erythrocytes from nine mammalian species in phases near the critical point with and without electrostatic potential differences reveal major species-specific differences in the membrane charge/noncharge components. A correlation has been found, in phases near the critical point that have essentially no electrostatic potential difference, between partition coefficient and the ratio of poly/monounsaturated fatty acids in the membranes of red cells from different species. Our present results thus provide parameters for the separation of cells by partition in addition to or instead of membrane charge depending on the polymer and salt composition and concentration selected.

Animals↗