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Fluid migration through the tail of the Dalkon shield intrauterine device.

X-ray radiographs were used to monitor the migration of Renouve -dip radiopaque tracer through the multifilament tail of Dalkon Shield intrauterine devices (IUDs). The movement of the tracer was more rapid and migrated a greater distance through the long end of the tail than through the short end of the tail. The double knot serves as at least a temporary barrier to the migration of tracer from the long end of the tail, but this barrier was circumvented when drops of Renouve -dip radiotracer were placed at both ends of the tail. Migration of Renouve -dip radiotracer was prevented when the end of the Dalkon Shield tail was fused by heating. No migration of this tracer occurred on the tails of commonly used IUDs which possessed monofilament tails, thus demonstrating that fluid migration occurs within and not on the surface of the multifilament tail.

Capillary Action↗

Negative chemotaxis does not control quail neural crest cell dispersion.

Negative chemotaxis has been proposed to direct dispersion of amphibian neural crest cells away from the neural tube (V. C. Twitty, 1949, Growth 13(Suppl. 9), 133-161). We have reexamined this hypothesis using quail neural crest and do not find evidence for it. When pigmented or freshly isolated neural crest cells are covered by glass shards to prevent diffusion of a "putative" chemotactic agent away from the cells and into the medium, we find a decrease in density of cells beneath the coverslip as did Twitty and Niu (1948, J. Exp. Zool. 108, 405-437). Unlike those investigators, however, we find the covered cells move slower than uncovered cells and that the decrease in density can be attributed to cessation of cell division and increased cell death in older cultures, rather than directed migration away from each other. In cell systems where negative chemotaxis has been demonstrated, a "no man's land" forms between two confronted explants (Oldfield, 1963, Exp. Cell Res. 30, 125-138). No such cell-free space forms between confronted neural crest explants, even if the explants are closely covered to prevent diffusion of the negative chemotactic material. If crest cell aggregates are drawn into capillary tubes to allow accumulation of the putative material, the cells disperse farther, the wider the capillary tube bore. This is contrary to what would be expected if dispersion depended on accumulation of this material. Also, no difference in dispersion is noted between cells in the center of the tubes versus cells near the mouth of the tubes where the tube medium is freely exchanging with external fresh medium. Alternative hypotheses for directionality of crest migration in vivo are discussed.

Animals↗

Stereoisomeric purity determination of captopril by capillary gas chromatography.

The GC method developed for the stereoisomeric purity determination of captopril is based on the combined information derived from the analyses of the captopril sample on two GC systems, one with a chiral and the other with an achiral column. The limit of detection has been determined to be 0.02% (w/w) for (R,S) or (S,R) and 0.03% for (R,R), with corresponding minimum quantifiable levels of 0.08% and 0.09%.

Capillary Action↗

Optimization of resolution in capillary zone electrophoresis: combined effect of applied voltage and buffer concentration.

Expressions are formulated for the prediction of solute migration time and resolution as a function applied voltage and buffer concentration in capillary zone electrophoresis. The resolution equation assumes that solute diffusion is the only operative zone-broadening mechanism. A resolution surface in applied voltage and buffer concentration space is presented featuring isochrones that are used to predict the behavior of resolution under constant analysis time. In the resolution-voltage planes the resolution increases continuously with increasing voltage. At the high-voltage border, the resolution decreases continuously with increasing concentration, however, at the low-voltage border the resolution passes through a shallow maximum as the buffer concentration is increased. At constant analysis time, resolution is optimized by simultaneously increasing the voltage and the buffer concentration. In comparison, this theoretical approach, which predicts resolution from solute migration times only, gives values that are consistently about 40-50% higher than experimentally determined resolution.

Acetates↗

Determination of antihistamines in pharmaceuticals by capillary electrophoresis.

Capillary electrophoresis with on-column UV detection at 214 nm was used to separate a group of nine antihistamines. All these compounds were satisfactorily separated within ca. 6 min using a mixed carrier system containing sodium dodecyl sulphate with beta-cyclodextrin and tetrabutylammonium hydrogensulphate as modifiers. The application of the method to the determination of the amount of antihistamines present in commercial pharmaceutical samples was demonstrated. In addition, the migration behaviour of the antihistamines in the mixed carrier system was examined.

Capillary Action↗

Separation of dideoxyribonucleosides in trace amounts by automated liquid chromatography and capillary electrophoresis.

No satisfactory high-performance liquid chromatographic (HPLC) method is currently available for the separation of the major dideoxyribonucleosides (ddNs) and their derivatives. A method involving HPLC has been developed for the separation of five major ddNs [ddA, ddC, ddI, azT and 2',3'-dideoxy-2',3'-didehydrothymidine (d4T)]. Elution of the common and modified components of DNA was also examined under the selected separation conditions of HPLC. The elution characteristics of these compounds were studied using serum plasma samples spiked with ddN derivatives. In addition, capillary electrophoresis (CE) was investigated for the separation of ddNs and their derivatives. Picomolar amounts of the five major ddNs and the metabolic product of azT [5'-O-glucuronide-3'-azido-3'-deoxythymidine (Glo-azT)] were satisfactorily resolved in 10 min by using a modification of CE. The spectral properties of the ddNs were characterized under different pH conditions and compared with those of their parent deoxyribonucleosides (dNs) because these compounds are commonly measured in HPLC by their spectral properties. The spectra of ddC and ddT derivatives resemble very closely those of dC and dT, but those of ddA and ddI differ to some extent from their parent dNs. The HPLC method was extensively examined for satisfactory resolutions of these compounds. For example, an isocratic elution method, although simple, failed to resolve these compounds and ion-pair chromatography did not offer any advantage. Gradient elution involving buffered solutions and increasing amounts of an organic modifier yielded satisfactory results. Methanol appeared to be the organic modifier of choice. A reversed-phase matrix with smaller than octadecyl alkyl chains did not produce the necessary interactions. Uniform spherical beads of smaller diameter produced superior resolutions. The separation of these compounds on three commercially available columns is discussed. The separation of human plasma samples spiked with dideoxynucleoside derivatives by HPLC was accomplished in ca. 16 min. The presence of the dNs did not interfere in their separations.

Capillary Action↗

Determination of sulphonamides in pork meat extracts by capillary zone electrophoresis.

For sixteen sulphonamides the effective mobility was measured as a function of pH and from the effective mobilities determined in two different electrolyte systems the pK value and mobility at infinite dilution were calculated. A pH of 7.0 was found to be the optimum pH for the separation for both standard mixtures and mixtures of sulphonamides dissolved in pork meat extracts. For the determination of the sulphonamides in pork meat only a very simple pretreatment consisting of extraction with acetonitrile and centrifugation is suitable, as the matrix effects at pH 7.0 do not affect the separation. Calibration graphs for five sulphonamides were constructed, and regression coefficients of at least 0.999 were obtained. The limit of detection for the method varies from 2 to 9 ppm for a pressure injection time of 10 s (injection volume ca. 18 nl) using a Polymicro Technology capillary of length 116.45 cm, distance between injection and detection 109.75 cm and I.D. 50 microns.

Animals↗

Enzymophoresis of nucleic acids by tandem capillary enzyme reactor-capillary zone electrophoresis.

Enzymophoresis with coupled heterogeneous capillary enzyme reactor-capillary zone electrophoresis was developed and evaluated in the area of nucleic acids. Ribonuclease T1, hexokinase and adenosine deaminase were successfully immobilized on the inner walls of short fused-silica capillaries through glutaraldehyde attachment. These open-tubular capillary enzyme reactors were quite stable for a prolonged period of use under operation conditions normally used in capillary zone electrophoresis. The capillary enzyme reactors coupled in series with capillary zone electrophoresis served as peak locator on the electropherogram, improved the system selectivity, and facilitated the quantitative determination of the analytes with good accuracy. Also, they allowed the on-line digestion and mapping of minute amounts of transfer ribonucleic acids, and the simultaneous synthesis and separation of nanogram quantities of oligonucleotides.

Adenosine Deaminase↗

Determination of aminoglycoside antibiotics in pharmaceuticals by capillary zone electrophoresis with indirect UV detection coupled with micellar electrokinetic capillary chromatography.

Aminoglycoside antibiotics can be determined by capillary zone electrophoresis (CZE) with indirect UV detection in the anionic mode with a reversed electroosmotic flow (EOF) by addition of FC 135 to the background electrolyte. The effective mobilities of thirteen aminoglycoside antibiotics were determined as a function of pH. Applying CZE with indirect UV detection in the anionic mode and reversed EOF coupled with micellar electrokinetic capillary chromatography with the cationic surfactant cetyltrimethylammonium bromide, both neutral and charged antibiotics can be determined in combined pharmaceuticals. As an example, neomycin and hydrocortisone were determined in Otosporin eardrops.

Aminoglycosides↗

Supercritical fluid chromatography of Fusarium mycotoxins.

Capillary- and packed-column supercritical fluid chromatography has been used for the separation of Fusarium mycotoxins of various structure types such as the trichothecenes including deoxynivalenol and its acetylated derivatives and T-2 toxin, as well as butenolide, culmorin, sambucinol and zearalenone. The effect of modifier concentration and column temperature and pressure was also studied. Retention indices based on alkylphenones were determined for these mycotoxins on two of the capillary columns employed.

Capillary Action↗

Determination of propionate in bread using capillary zone electrophoresis.

A method for the determination of propionate in bread is described. The propionate was extracted from the bread with a repeated extraction procedure and measured using capillary zone electrophoresis in the indirect UV mode applying a background electrolyte of 0.005 M Tris adjusted at pH 4.6 by adding benzoic acid. Using laboratory-baked bread containing known amounts of sodium propionate, recoveries of ca. 95% could be established, validating the method.

Bread↗

Comparison of flame ionization and inductively coupled plasma mass spectrometry for the detection of organometallics separated by capillary supercritical fluid chromatography.

Organotin compounds are separated by capillary supercritical fluid chromatography (SFC) and a comparison of the detection by flame ionization (FID) and inductively coupled plasma mass spectrometry (ICP-MS) is presented. Resolution, detection limits, linear dynamic range and reproducibility are the parameters compared between SFC-FID and SFC-ICP-MS, for the detection of tri- and tetraorganotin compounds. The resolution obtained in the SFC-FID system is not always observed in SFC-ICP-MS. Degradation in resolution is due to fluctuations in transfer line temperature. Baseline resolution for the organotins considered is achieved in both systems by using a longer column. Detection limits (DLs) are calculated as 3 sigma/S, where sigma is the standard deviation of the blank signal and S is the slope of the calibration curve. Detection limits of 10.3, 12.5, 12.0 and 9.0 pg are obtained for tetrabutyltin, tributyltin chloride, triphenyltin chloride and tetraphenyltin, respectively, using SFC-FID. An improvement in detection limits of one order of magnitude is achieved by SFC-ICP-MS for the same organotins (0.26, 0.80, 0.57 and 0.20 pg, respectively). The relative standard deviations using SFC-FID for five 50-nl injections, containing 0.5 ng Sn, ranged from 3.2 to 6.4%. Using SFC-ICP-MS, five replicate injections of 0.05 ng Sn give R.S.D.s from 1.3 to 3.4%.

Capillary Action↗

Capillary zone electrophoresis and packed capillary column liquid chromatographic analysis of recombinant human interleukin-4.

Capillary zone electrophoresis (CZE) and packed capillary column liquid chromatography (micro-LC) have been applied to the analysis of the recombinant human protein interleukin-4 (rhIL-4). Separations for both the parent protein and its enzymatic digest were developed for the purpose of characterizing protein purity and identity. CZE separations of the intact protein were investigated over the pH range of 4.5 to 8.0 using uncoated fused silica capillaries. Gradient reversed-phase micro-LC was performed using 0.32 mm packed capillary columns at flow-rates of 5-6 microliters/min. Emphasis was placed on the ability of these methods to separate close structural variants and degradation products of the protein. Peptide mapping of the tryptic digest of rhIL-4 using a combination of CZE and micro-LC provided complimentary high resolution methods for establishing protein identity. Reproducible separations were achieved using sub-picomol amounts of sample. The advantages and problems encountered with these two techniques for characterizing rhIL-4 were assessed.

Capillary Action↗

Measurement of vitamin C by capillary electrophoresis in biological fluids and fruit beverages using a stereoisomer as an internal standard.

Ascorbic acid (or vitamin C) is an important component of many biological systems and various physiological roles have been described for it. A rapid and simple capillary electrophoresis method for ascorbic acid measurements in biological fluids as well as in beverages was developed. A stereoisomer of ascorbic acid, isoascorbic acid, not normally found in nature, was used as the internal standard for this assay. The analysis was performed in a 30 cm x 75 microns I.D. fused-silica capillary with 100 mM tricine buffer, pH 8.8, and measured by UV absorbance at 254 nm. The method was sensitive to 1.6 micrograms/ml and linear to 480.1 micrograms/ml. Within-run R.S.D. was 3.2% (93.5 +/- 3.0 micrograms/ml, mean +/- S.D., n = 18) and run-to-run R.S.D. was 3.3% (35.6 +/- 1.2 micrograms/ml, mean +/- S.D., n = 10) and 1.9% (149.4 +/- 2.8 micrograms/ml, mean +/- S.D., n = 10). Average spiked recovery from human plasma samples was 98.0%. The technique has been demonstrated to be suitable for assay of vitamin C in biological samples and some fruit juices.

Ascorbic Acid↗

Separation of transfer RNA and 5S ribosomal RNA using capillary electrophoresis.

Capillary gel electrophoresis and capillary electrophoresis using entangled polymer solutions was investigated for their applicability for the separation of low-molecular-mass RNAs (transfer RNA and 5S ribosomal RNA), with a size range of 70-135 nucleotides, from bacteria. Cross-linked polyacrylamide gel-filled capillaries (3 and 5%) were used for capillary gel electrophoresis. Good resolution was obtained using gel-filled capillaries only for small tRNAs with lengths to 79 nucleotides, larger tRNAs and 5S rRNA could not be resolved using this method. Buffers containing sieving additives were employed to improve separations of RNA by capillary electrophoresis using entangled polymer solutions. The use of linear sieving polymers in buffers resolved 5S rRNA and tRNAs, even when they possessed only different secondary structure or small differences in length (1-5 nucleotides).

Buffers↗

Sequencing of antisense DNA analogues by capillary gel electrophoresis with laser-induced fluorescence detection.

A method using capillary gel electrophoresis with laser-induced fluorescence detection is described which permits complete sequence determination of antisense DNA analogues of unknown sequence. This method, originally created as a tool to confirm the sequence of antisense oligonucleotides being developed as therapeutic drugs, utilizes data collected under a range of experimental conditions described by the Ogston model as applied to gel electrophoresis. A linear relationship independent of experimental conditions between the relative electrophoretic migration time and the oligonucleotide base number was observed and is shown to be consistent with a simplified version of this model and can be used to facilitate the sequence determination.

Base Sequence↗

Analysis of metallothionein isoforms by capillary electrophoresis: optimisation of protein separation conditions using micellar electrokinetic capillary chromatography.

Capillary electrophoresis (CE) techniques have been successfully applied to the separation of metallothionein (MT) isoforms and have proved to be rapid, practical and economical. Study of a variety of different electrolytes and capillaries has shown that electrolyte buffer composition and capillary wall surface modifications can have considerable influence on isoform separation and resolution. Ionic surfactants such as sodium dodecyl sulphate (SDS) form micelles at elevated concentrations and the partitioning of molecules between the hydrophobic micelle phase and the aqueous phase and their resulting migration in an electric field is the basis of the technique known as micellar electrokinetic capillary chromatography (MECC). In the present work, we have used sheep and rabbit MT to optimise MECC conditions for analysis of MT isoforms. Capillaries of 57 cm gave much better separations than shorter columns although analysis times were increased to about 12 min. Changing the buffer and SDS concentration or the pH affected the selectivity of isoform separation and up to 5 isoforms in sheep MT and 6 in rabbit MT were completely or partially resolved. Comparing different diameter capillaries we conclude that 25 microns I.D. columns give better separations than 50 or 75 microns I.D. columns although sensitivity is reduced by a factor of about 3 and 5, respectively. Using our MECC conditions, columns coated with C1 or C18 hydrophobic material were not found to be useful in improving MT separation or resolution although further evaluation of these columns is in progress.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗