Biomedical subjects

K Albert

Publications and source records attributed to K Albert.

At least 37 records · Page 2Linked to original sources

PubMed2000

Application of HPLC-NMR coupling using C30 phase in the separation and identification of flavonoids of taxonomic relevance.

The on-line separation and identification of two important taxonomic markers for plant species of the Paepalanthus genus, the flavonoids 6-methoxykaempferol-3-O-beta-D-glucopyranoside and 6-methoxykaempferol-3-O-beta-D-6"(p-coumaroyl)glucopyranoside, has been performed with an HPLC-NMR coupling using C30 phase. 1D spectra have been recorded in the stopped-flow mode for the two predominant chromatographic peaks. This is the first application of HPLC-NMR coupling using C30 phase to a taxonomic problem. The technique drastically reduces the required amount of sampling for structure determination.

Chromatography, High Pressure Liquid↗

PubMed1999

n-Alkyl fluorenyl phases in chromatography. II. Dynamic behavior and high-performance liquid chromatography applications.

The dynamic behavior of two n-hexyl fluorenyl phases (fluorene-6A [3a(Tn)(Qm)y], fluorene-6B [3b(Tn)(Qm)y]) and three n-decyl fluorenyl phases (fluorene-10A [4a(Tn)(Qm)y], fluorene-10B [4b(Tn)(Qm)y], and fluorene-10C [4c(M1)(Qm)y]) is investigated by solid-state nuclear magnetic resonance (NMR) spectroscopy using the dipolar filter technique with both 13C and 1H detection. These results are compared with those from other dynamic measurements, like the relaxation times in the rotating frame (T1pH) and the variation of the contact time (T(CH)). Additionally, another type of a fluorenyl phase [5a(Tn)(Qm)y], which has an aromatic moiety connected to the silica gel by amido couplings, was also investigated by the dipolar filter method. The solid-state NMR dynamic measurements indicate an increased mobility of the n-alkyl fluorenyl phases compared to the amido coupled fluorenyl phase. The lower the ligand density of the studied n-alkyl fluorenyl phases, the higher their mobility. The separation behavior of the respective phases in high-performance liquid chromatography was investigated with samples containing polycyclic aromatic hydrocarbons and nitro explosives. Depending on the amount of the chemically bound aromatic moiety and the length of their n-alkyl spacer groups, pi-pi interactions with the solute molecules are involved in the separation process and cause it to proceed at a different rate. Therefore, n-alkyl fluorenyl phases can be classified as mixed-mode phases.

Chromatography, High Pressure Liquid↗

PubMed1999

Liquid chromatography-nuclear magnetic resonance spectroscopy.

A general overview of the experimental set-up for performing analytical-scale and nanoliter-scale liquid chromatography-1H nuclear magnetic resonance spectroscopy (LC-1H-NMR) experiments is given. The high power of combining LC with 1H-NMR spectroscopy is demonstrated by two examples, where NMR acquisition was performed either in the continuous-flow mode on the analytical scale or in the stopped-flow mode on the nanoliter scale. Current developments employing the on-line coupling of capillary as well as supercritical fluid separation methods with 1H-NMR spectroscopy together with LC-13C-NMR spectroscopy are discussed.

Chromatography, Liquid↗

PubMed1999

Separation and identification of various carotenoids by C30 reversed-phase high-performance liquid chromatography coupled to UV and atmospheric pressure chemical ionization mass spectrometric detection.

In this paper the application of on-line HPLC-UV-APCI (atmospheric pressure chemical ionization) mass spectrometry (MS) coupling for the separation and determination of different carotenoids as well as cis/trans isomers of beta-carotene is reported. All HPLC separations were carried out under RP conditions on self-synthesized polymeric C30 phases. The analysis of a carotenoid mixture containing astaxanthin, canthaxanthin, zeaxanthin, echinenone and beta-carotene by HPLC-APCI-MS was achieved by scanning the mass range from m/z 200 to 700. For the characterization of a sample containing cis/trans isomers of beta-carotene as well as their oxidation products, a photodiode-array UV-visible absorbance detector was used in addition between the column and the mass spectrometer for structural elucidation of the geometrical isomers. The detection limit for beta-carotene in positive-ion APCI-MS was determined to be 1 pmol. In addition, an extract of non-polar substances in vegetable juice has been analyzed by HPLC-APCI-MS. The included carotenoids could be identified by their masses and their retention times.

Atmospheric Pressure↗

PubMed1999

Gradient elution capillary electrochromatography and hyphenation with nuclear magnetic resonance.

Coupling of gradient capillary electrochromatography (gradient CEC) and capillary zone electrophoresis (CZE) with nuclear magnetic resonance spectroscopy (NMR) was performed using a recently developed capillary NMR interface. This technique was applied for the analysis of pharmaceuticals and food. An analgesic was investigated using isocratic and gradient continuous-flow CEC-NMR. Comparison of the results demonstrated the superiority of gradient CEC over isocratic CEC. Aspartame and caffeine, both ingredients of soft beverages, were separated and analyzed by continuous flow CZE-NMR. The order of elution could be reversed by altering the pH. This reversal led to an increased sample concentration in the NMR detection cell, thus allowing the acquisition of a totally correlated spectroscopy (TOCSY) two-dimensional (2-D) spectrum of the synthetic peptide aspartame.

Acetaminophen↗

PubMed1999

Hyphenation of high performance liquid chromatography with nuclear magnetic resonance spectroscopy for the characterization of beta-carotene isomers employing a C30 stationary phase.

The hyphenation of HPLC together with NMR spectroscopy proves advantageous for the structure elucidation of oxidation- and UV-sensitive compounds such as beta-carotene isomers. In the closed-loop HPLC-NMR system, degradation or isomerization of separated compounds is largely hindered. With the help of 3-microm C30 stationary phases a better separation efficiency towards the different beta-carotene cis/trans isomers could be obtained in comparison to a 5-microm material, resulting in sharper peaks and a better resolution of all compounds. This effect greatly facilitated the structure determination of the isomers by HPLC-NMR coupling. Due to the introduction of a superior stationary phase, the structure of seven cis-isomers of beta-carotene could thereby be determined employing the stopped-flow HPLC-1H-NMR mode.

Chromatography, High Pressure Liquid↗

PubMed1998

Reversed-phase high-performance liquid chromatographic identification of lutein and zeaxanthin stereoisomers in bovine retina using a C30 bonded phase.

An efficient reversed-phase high-performance liquid chromatographic assay with ultraviolet detection at 450 nm for simultaneous determination of lutein and zeaxanthin stereoisomers in bovine retina is described. The procedure involves rapid and careful one-step hexane extraction of the carotenoids from the homogenized liquid and enrichment by on-line solid phase extraction on a polystyrene cartridge. The substances were eluted at a flow rate of 1 ml/min with acetone-water (85:15, v/v) on a C30 reversed-phase column. C30 phases exhibit superior shape selectivity for the separation of carotenoid stereoisomers compared with conventional C18 phases. For comparison a mixture of iodine-isomerized lutein and zeaxanthin standards was separated.

Animals↗

PubMed1998

Silver-plated vitamins: a method of detecting tocopherols and carotenoids in LC/ESI-MS coupling.

In LC/MS, nonpolar substances in the majority of cases cannot be ionized by standard electrospray ionization (ESI) because they obviously lack a site for protonation or deprotonation. The ionization of carotenoids and tocopherols can be greatly enhanced by the addition of silver ions. The Ag(+)-carotenoid and Ag(+)-tocopherol adducts thus formed render these substances amenable to MS. alpha-, beta-, gamma-, and delta-tocopherol, alpha-tocopherol acetate, and the various isomers of lycopene and beta-carotene were separated by C30 RP-HPLC and could be identified by online ESI-MS. A mixture of six different carotenoids was analyzed by scanning the mass range from m/z 500 to 800. The mass spectra of the peaks revealed that all carotenoids and most tocopherols were partially oxidized to radical cations. The detection limit for canthaxanthin was approximately 500 fmol while that of beta-carotene was below 300 fmol. An increase in sensitivity in the MRM mode can be attained by monitoring ions formed by loss of elemental silver from the adducts in the CID cell. Dichloromethane extracts of tomato, carrot, and vegetable juices, a vitamin drink, and a commercial infant food product were analyzed by LC/MS. After postcolumn argentation, from the mass-selective extracts of the TIC, the carotenoids and tocopherols present could be identified by their masses and their retention times. For all studies, a silver perchlorate solution with an overall concentration of 50 micrograms/mL was used.

Antioxidants↗

PubMed1998

n-alkyl fluorenyl phases in chromatography. I. Synthesis and characterization.

A new class of silica gel-bound fluorene phases is described. The compounds are synthesized via reaction of fluorenyl lithium with omega-alkenyl bromides leading to 9-(5'-hexenyl)-9H- and 9-(9'-decenyl)-9H-fluorene (1 and 2), followed by hydrosilation reactions with different hydrosilanes. The resulting omega-functionalized silylalkyl fluorenes 3a(T0), 3b(T0), 4a(T0), 4b(T0) and 4c(M0) (Scheme 1) react with surface silanol groups of silica gel to generate the new fluorene phases 3a(Tn)(Qm)y, 3b(Tn)(Qm)y, 4a(Tn)(Qm)y, 4b(Tn)(Qm)y and 4c(M1)(Qm)y. The phases are characterized by employing 1H, 13C and 29Si solid-state nuclear magnetic resonance spectroscopy. Their applicability in high-performance liquid chromatography is proved by the Sander and Wise test (SRM 869). In contrast to conventional n-alkyl phases, pi-pi interactions are additionally involved in the separation process and, therefore, the retention times of the polycyclic aromatic hydrocarbons sample molecules depend on the ligand densities of the applied fluorene phases.

Chromatography, High Pressure Liquid↗

PubMed1998

Shape selectivity of C30 phases for RP-HPLC separation of tocopherol isomers and correlation with MAS NMR data from suspended stationary phases.

Vitamin E (tocopherol) acts in various organisms as the main free radical scavenger. This capacity, which is enhanced by the synergetic effect of vitamin C and carotenes, points to a possible application as anti-tumor agent in chemotherapy. There are several isomeric forms, namely alpha-, beta-, gamma-, and delta-tocopherol, having different antioxidative abilities, with alpha-tocopherol being the most biologically active. Using methanol as eluent and a C30 stationary phase, we achieved complete separation of alpha-, beta-, gamma-, and delta-tocopherol and alpha-tocopherol acetate by RP-HPLC within 14 min. Detection was performed by UV and 1H NMR spectroscopy. The advantage of NMR is the possibility of structural identification of chromatographic peaks. Also, coeluting peaks are easily recognized. The enhanced shape recognition of the C30 phase has been attributed to the high order of the alkyl chains of the stationary phase. This has, so far, been proven by solid-state NMR spectroscopy. We now introduce the technique of 13C MAS NMR spectroscopy of suspended stationary phases. The resulting NMR spectra reveal that, in the presence of weak eluents, like methanol, the overall high order of the C30 chains is slightly altered, whereas in stronger eluents, like MTBE, the alkyl chains possess a higher mobility.

Chromatography, High Pressure Liquid↗

PubMed1998

Directly coupled CZE-NMR and CEC-NMR spectroscopy for metabolite analysis: paracetamol metabolites in human urine.

Direct coupling of NMR spectroscopic detection with both capillary zone electrophoresis (CZE) and capillary electrochromatography (CEC) was applied to the separation of metabolites of the drug paracetamol in an extract of human urine. Continuous-flow CZE-NMR and CEC-NMR allowed the detection of the major metabolites, the glucuronide and sulfate conjugates of the drug and the endogenous material hippurate. Identification of these substances was achieved by examination of individual rows of the NMR chromatogram and this also gave estimates of the detection limits. For CEC-NMR, spectra were also obtained in the stopped-flow mode including a two-dimensional TOCSY NMR experiment which afforded confirmatory evidence for paracetamol glucuronide. Characterisation of drug metabolites using NMR spectroscopy is therefore possible with nanolitre sample volumes.

Acetaminophen↗

PubMed1997

Stationary interphases with extended alkyl chains: a comparative study on chain order by solid-state NMR spectroscopy.

Stationary interphases with long n-alkyl chains (n = 18, 22, 30, 34) have been examined by solid-state NMR spectroscopy. The determination of the silane functionality and the degree of cross-linking of silane ligands on the silica surface was performed by 29Si CP/MAS NMR spectroscopy. High-speed 1H MAS and 13C CP/MAS NMR spectroscopy were utilized to assess alkyl chain order and mobility of the different bonded phases. For this purpose, 1H NMR line widths and 13C chemical shifts have been evaluated. It is shown that stationary phase order and rigidity increase with alkyl chain length. In addition, the temperature-dependent trans/gauche conformational change occurs at higher temperatures for a polymeric C34 phase compared with a C30 sorbent. This behaviour is discussed in the context of previously reported chromatographic (HPLC) shape selectivity differences.

Carbon Isotopes↗

PubMed1996

1H MAS NMR spectroscopy of chemically modified silica gels: a fast method to characterize stationary interphases for chromatography.

Chemically modified silica gels used as stationary phases in chromatography have been investigated by means of solid-state 1H magic angle spinning (MAS) NMR spectroscopy. Since the organosilanes are bonded to the surface of the silica gel, their protons are diluted and possess a higher mobility in comparison to protons in pure organic solids. Thereby the usually strong homonuclear dipole-dipole interactions among the protons are reduced and it is possible to obtain well-resolved 1H NMR spectra of the organic interphases with MAS-only techniques. Effects of temperature and magnetic field strength on the resolution of the spectra are examined as well as the dependence of T1 and T1pH relaxation times on temperature and spinning speed.

Chemical Phenomena↗

PubMed1995

Detection of psychoactive drugs using 19F MR spectroscopy.

In vivo 19F resonance spectroscopy measurements of trifluorinated neuroleptics (fluphenazine and trifluoperazine) and later trifluorinated antidepressants (fluoxetine and fluvoxamine) began with animal experiments in 1983. Using rats which have been treated with high oral doses of fluphenazine over a period of three weeks in the beginning of these experiments the measurement time was very long (up to 10 h). The application of better techniques using surface coils led to marked improvement of the signal noise ratio and measurement times in animal experiments could be reduced to minutes. These results encouraged us and other groups to perform experiments in humans to detect and try to estimate brain levels of trifluorinated neuroleptics and antidepressants. The present data of several research groups demonstrate that 19F MR spectroscopy has the potential of becoming a valuable tool for monitoring drug levels at the site of action. The extension of the animal studies to humans might facilitate a better treatment of schizophrenic and depressive patients.

Animals↗

PubMed1995

Solid state NMR and extraction studies on "phenyl"-bonded stationary phases used for solid phase extraction.

The solid phase extraction (SPE) and elution of [14C]-propranolol from aqueous buffer has been studied for a range of phenyl-bonded SPE materials. Differences were noted in the recovery of the analyte using methanol-water eluents depending upon the manufacturer and whether or not phase had been endcapped. Efficient recoveries of [14C]-propranolol were only achieved when triethylamine was added to the eluting solvent as a competing base. Solid state cross polarisation/magic angle spinning (CP/MAS) NMR spectroscopy was used to characterise the phases further, which revealed differences in endcapping between materials as well as differences in the type and extent of cross-linking.

Magnetic Resonance Spectroscopy↗

PubMed1994

Characterization of wear in composite material orthopaedic implants. Part I: The composite trunnion/ceramic head interface.

Carbon fiber reinforced polyetheretherketone (C/PEEK) composite materials are being investigated as an alternative to metal in the femoral component of a total hip arthroplasty. Wear is among the issues that must be addressed before introducing a new orthopaedic implant material. This study examines the generation of wear debris when zirconia femoral heads are mechanically attached to C/PEEK trunnions and loaded under simulated physiological conditions. Mechanical testing was performed on a trunnion/head assembly loaded from 445 to 4450N at an angle of 39 degrees to the long axis of the trunnion. The trunnions were tested at a frequency of 20 Hz for 10 million cycles. After completion of the fatigue test, solution from the test assembly was characterized by laser scattering and by SEM image analysis to determine the size, shape, total number, and identify of the particles. In addition, the peak load to pull the head from the trunnion was measured. The total number of particles generated during the test was in the range of 10(5) as indicated by both laser scattering and (SEM) image analysis. Both carbon fiber and PEEK particles were found in an average proportion of about 1:13, respectively. The carbon fiber particle size average was 153 microns and the PEEK particle size average was 2.2 microns. The zirconia heads remained well attached to the C/PEEK trunnions as indicated by a mean peak distraction force of 1942 +/- 116N.

Benzophenones↗

PubMed1994

Characterization of wear in composite material orthopaedic implants. Part II: The implant/bone interface.

Carbon fiber/PEEK polymer (C/PEEK) composite materials are being developed for use as orthopaedic implant materials. Wear is an issue of increasing importance in orthopaedic implants; particulate debris generated by the wearing of biomaterials may be a causal factor leading to osteolysis and implant loosening. Therefore, numerical and experimental studies were completed to characterize the wear of C/PEEK composite materials in comparison to current orthopaedic implant materials. Finite element analyses (FEA) of a composite material hip stem implanted in a femur and loaded at 890 N determined that peak contact stresses will occur at the proximal-medial and distal regions of the implant. These contact stresses were found to be below 1.0 MPa over most of the implant surface; however the peak stress in the proximal-medial region was 1.8 MPa and higher still at the distal portion of the stem. In vivo forces result in contact stress values up to 9.0 MPa. The composite implant exhibited 10-40% lower contact stresses in the distal region compared to a titanium-alloy implant of identical design. Composite material wear samples were slid against porous hydroxylapatite (HA) to simulate the stem/bone interface. An identical series of experiments was run for comparison to a current orthopaedic implant material--Ti6A14V titanium alloy. Two domains of motion were studied; a composite ring-on-HA disc large amplitude sliding wear test; and a composite pin-on-HA disc small amplitude fretting regimen. Nominal contact pressures during testing were 1.4 MPa and 7.6 MPa for sliding and fretting tests, respectively. Fretting and sliding abrasive wear tests resulted in the composite material exhibiting a lower wear rate than the titanium-alloy. The magnitude of the difference was greatly dependent on the contact pressures, sliding amplitudes, and counterface material properties.

Alloys↗

PubMed1993

13C NMR investigation of synovial fluids.

13C NMR spectra of synovial fluids from 20 patients suffering from rheumatic diseases have been recorded. Structural changes in hyaluronic acid, the main carrier of the viscoelastic properties of synovial fluid, could be observed in the NMR spectra of the native biological fluid. By comparing these spectra with those of purified hyaluronic acid, a rough estimation of the degree of depolymerization of synovial hyaluronic acid was possible. The patients with rheumatoid arthritis appeared to have a lower degree of polymerization compared to patients with osteoarthrosis. Thus, 13C NMR spectroscopy provides useful information about biophysical properties of synovial fluid.

Arthritis, Rheumatoid↗