Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “CHROMATOGRAPHY”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,495 records · Page 83Linked to original sources
PubMed1984

Trace analysis of organic compounds by high-performance liquid chromatography with photoionization detection.

Photoionization detection (PID) has now been successfully interfaced with modern high-performance liquid chromatography (HPLC) to provide a new "hyphenated" technique, HPLC-PID. This method is compatible with reversed-phase HPLC solvents and conditions, as well as with certain organic solvents used in normal-phase liquid chromatography. The HPLC-PID interface consists of a variable-ratio eluent splitter, operated inside a heated oven, which vaporizes both analytes and mobile phase prior to introduction into the detector. Depending on the energy (eV) of the photoionization detector lamp, most of the solvents commonly used in reversed-phase liquid chromatography are compatible with long-term, on-line, real-time, continuous photoionization detector operation. Virtually all organic classes now detectable by gas chromatography-PID can also be detected by HPLC-PID, but with somewhat poorer detection limits. In certain instances, minimum detection limits are in the 5-500 ng/injection range. The HPLC-PID system appears ideal for aromatic or aliphatic amines, substituted hydrocarbons, and certain other classes of organic compounds. Various applications of HPLC-PID, under reversed-phase conditions are illustrated and discussed.

Chromatography, Gas↗
PubMed1983

Bovine viral diarrhea virus: purification of surface proteins in detergent-containing buffers by fast protein liquid chromatography.

Bovine viral diarrhea virus was purified by lectin chromatography. The glycoprotein peplomers were dissociated from the virion by treatment with detergent. By a second lectin gel chromatography the glycoconjugates containing terminal galactose were prepared. In combination with lectin affinity chromatography, ion-exchange chromatography on Mono-Q in the presence of the low-UV-absorbing detergent Berol 172 proved to be a powerful technique both for analytical and preparative applications.

Chromatography, Affinity↗
PubMed1977

Application of quantitative high-performance thin-layer chromatography in the antibiotic industry.

For the in-process control of antibiotic fermentations and for routine assays of samples for scaling up and for pure products, quantitative high-performance thin-layer chromatography (HPTLC) can be used with advantage. Rapid chromatography on high-performance layers, combined with an automatic spraying device for exact derivatization on the plate and precise computation of the calibration line within an automatic measurement and evaluation, represents a new, inexpensive analysis system. There are only 2 min of labour time (one fifth of that required in thin-layer chromatography) required for one sample and the total analysis time varies from 3 to 9 min (one third to one quarter of that required in thin-layer chromatography) based on one plate with 12 samples. The 95% confidence limits (N = 10) range between 0.5 and 3.0%.

Anti-Bacterial Agents↗
PubMed1990

Fractionation of basic nuclear proteins of human sperm by zinc chelate affinity chromatography.

Immobilized metal affinity chromatography was investigated for the fractionation of basic nuclear proteins of human sperm. Human sperm nuclei essentially contain two classes of protamines: a protamine of type P1 (HPl), rich in cysteine but with only one histidine, and three protamines of type P2 (HP2, HP3, HP4), rich in cysteine and histidine (nine in protamine HP2), potential ligands for transition metal ions. The critical conditions for metal affinity chromatography were defined: choice of metal, protein material and buffer, type of elution and sample loading. Chromatography of nuclear proteins, without histones and with cysteine residues alkylated by iodoacetamide, was optimum on zinc Chelating Sepharose in a Tris-acetate buffer and elution with an increasing concentration gradient of imidazole. Under these conditions, the two classes of protamines were completely separated. The intermediate basic proteins were further purified by reversed-phase high-performance liquid chromatography. Heterogeneity of binding to zinc of protamine HP1 was demonstrated. The proposed method is simple and reproducible and the recovery of proteins is high. It may be applied to study the expression and function of P1 and P2 protamines, e.g., in the case of infertile men.

Buffers↗
PubMed1989

The separation of pancreatic procarboxypeptidases by high-performance liquid chromatography and chromatofocusing.

Different experimental conditions and chromatographic supports have been selected for the most efficient and rapid purification of procarboxypeptidases from porcine and human pancreas by different high-performance liquid chromatography (HPLC) variants (anion exchange, reversed phase and gel filtration). Anion-exchange chromatography was found to be the most capable and permitted the isolation, in a single step, of three different porcine procarboxypeptidases (2A + 1B forms) and five different human procarboxypeptidases (2B + 3A forms) in a native and pure state from whole pancreas extracts. Other pancreatic proproteases are also cleanly isolated in the same step. Reversed-phase chromatography under mild conditions separated porcine or human procarboxypeptidases A from other pancreatic proteins in a very short time but was unable further to subfractionate the same proteins. The sequential use of gel filtration (or anion-exchange) and reversed-phase HPLC chromatography permitted, in a simple way, the isolation and dissociation of the strongly bound components of the binary complexes between procarboxypeptidases A and proproteinase E in either porcine or human pancreas extracts. Chromatofocusing on a fast protein liquid chromatographic support was also found to be a very efficient technique, showing a slightly lower capability to separate procarboxypeptidases than anion-exchange HPLC though in a much shorter time and in larger quantities.

Animals↗
PubMed2002

Composition analysis of poly(ethylene glycol)-poly(L-lactide) diblock copolymer studied by two-dimensional column chromatography.

We used two-dimensional column chromatography to analyze the composition of a sample of presumably a diblock copolymer of poly(ethylene glycol) (PEG) and poly(L-lactide) synthesized from monomethoxy-terminated PEG. The first dimension of the separation is phase fluctuation chromatography to prepare fractions that contain various components of the copolymer in different ratios. The second dimension is size-exclusion chromatography, NMR, and HPLC at the critical condition of PEG. The PEG initiator has small amounts of diol-terminated dimeric components. We found that the copolymer sample contains a triblock copolymer and low-molecular-mass components in addition to the main part of the diblock copolymer. The SEC chromatograms show that the main part consists of two components with distinct peak lengths for the PLLA block. The low-molecular-mass components have a broad distribution in chemical composition. Phase fluctuation chromatography enriched the triblock copolymer and the diblock copolymer with the longer PLLA block in early fractions when the column was packed with carboxymethyl-modified porous silica. When the porous medium was PLLA-grafted silica, size exclusion dominated, but the low-molecular-mass components were separated according to their chemical composition.

Chromatography, Gel↗
PubMed1997

Inductively coupled plasma mass spectrometric detection for chromatography and capillary electrophoresis.

Inductively coupled plasma mass spectrometry (ICP-MS) is now a well established detection technique for liquid chromatography, gas chromatography, supercritical fluid chromatography and capillary electrophoresis. A review of the literature with particular regard to ICP-MS as a chromatographic and capillary electrophoretic detector is presented. The various modes of chromatography and capillary electrophoresis are discussed and practical descriptions for hyphenating the techniques with the ICP mass spectrometer are given. Sample introduction systems and data acquisition methods are reviewed along with the numerous applications of ICP-MS as a chromatographic detector. In addition, alternative plasma sources, such as the atmospheric and reduced pressure helium microwave-induced plasmas for chromatographic detection are described.

Chromatography, Gas↗
PubMed1997

Single-step purification of bispecific monoclonal antibodies for immunotherapeutic use by hydrophobic interaction chromatography.

A method for large scale production and single-step purification of bispecific antibodies is described. Hybrid-hybridomas were grown in hollow-fibre bioreactors with an average yield of 8 to 12 g of immunoglobulin per month. Bispecific antibodies were purified from the bioreactor supernatant by hydrophobic interaction chromatography which resolves bispecific antibodies, monospecific immunoglobulins, and culture medium supplements in one single chromatographic step. Proteins were analyzed by ELISA, SDS-PAGE, isoelectric focussing, indirect fluorescence staining, CTL-stimulation and T-cell proliferation assays. Finally, antibody preparations were checked for the presence of endotoxin and mouse DNA. Our results suggest that functional bispecific antibodies for use in therapeutic applications can be batch purified from bioreactor harvest by hydrophobic interaction chromatography in a single step. Compared to other methods such as affinity chromatography (protein A/G), ion-exchange or hydroxyapatite chromatography, our protocol offers a substantial reduction in labor time, cost, protein loss, and risk of contamination.

Animals↗
PubMed1999

Application of immunoaffinity chromatography for detection of tetrodotoxin from urine samples of poisoned patients.

Immunoaffinity chromatography using the monoclonal antibody (Tl-1) specific for tetrodotoxin (TTX) has been developed for isolating TTX from urine samples. By combining immunoaffinity chromatography with fluorometric high performance liquid chromatography (HPLC), it has become possible to detect a small amount of TTX in urine samples. The detection limit of TTX in urine was 2 ng/ml. By this combined method, TTX was detected in all the urine samples that were collected from poisoned patients during the week following TTX ingestion. The combination of immunoaffinity chromatography with HPLC was very useful in detecting TTX from the urine samples of poisoned patients for diagnosis of TTX-food poisoning.

Antibodies, Monoclonal↗
PubMed1982

Anion-exchange high-performance liquid chromatography of human serum apolipoproteins.

The rapid separation of seven urea-soluble apolipoprotein species from delipidated human serum very low density lipoproteins (VLDL) and high density lipoproteins (HDL) has been achieved by high-performance liquid chromatography on an anion-exchange column of Syn-Chropak AX 300. Effluent chromatographic peaks were detected by absorbance at 280 nm in a flow-through cell. Peaks corresponding to apolipoproteins AI1, AI2, AII, CI, CII, CIII1, and CIII2 were identified by amino acid analysis, gel electrophoresis, and isoelectric focusing. Maximum efficient loading of semipreparative columns (250 X 9.0 mm) was established to be ca. 20 mg HDL apolipoprotein. Minimum detectable protein was shown to be ca. 1 microgram on an analytical-scale column (300 X 4.5 mm). Chromatographic resolution is comparable to that of conventional DEAE-cellulose column chromatography. The ratio of apoAI1 to apoAI2 was considerably greater in high-performance liquid chromatography, suggesting that the variants seen in conventional chromatography and isoelectric focusing are in part artifactual.

Amino Acids↗
PubMed1989

Catecholamine measurements in urine by high-performance liquid chromatography with amperometric detection--comparison with an autoanalyser fluorescence method.

In order to validate different methods of measuring urinary catecholamines (norepinephrine, epinephrine and dopamine) in humans, methods based on separation of catecholamines using reversed-phase or cation-exchange high-performance liquid chromatography with electrochemical detection were compared with an autoanalyser-based fluorescence method. Different methods for pre-chromatography sample purification were also studied. For measurements of urinary catecholamines, the reversed-phase-based chromatographic techniques studied were found to give less reliable results than cation-exchange chromatography, even if one of them (Clin Rep Urine Catecholamine Kit) gave almost as precise estimates. The autoanalyser technique yielded good results. It is concluded that cation-exchange chromatography with an appropriate sample work-up procedure (a combination of organic solvent extraction and alumina adsorption) is a reliable and accurate method for analyses of urinary catecholamines.

Acetates↗
PubMed1989

Determination of methandrostenolone and its metabolites in equine plasma and urine by coupled-column liquid chromatography with ultraviolet detection and confirmation by tandem mass spectrometry.

Monitoring steroid use requires an understanding of the metabolism in the species in question and development of sensitive methods for screening of the steroid or its metabolites in urine. Qualitative information for confirmation of methandrostenolone and identification of its metabolites was primarily obtained by coupled-column high-performance liquid chromatography-tandem mass spectrometry. The steroids and a sulphuric acid conjugate were isolated and identified by their daughter ion mass spectra in the urine of both man and the horse following administration of methandrostenolone. Spontaneous hydrolysis of methandrostenolone sulphate gave 17-epimethandrostenolone and several dehydration products. This reaction had a half-life of 16 min in equine urine at 27 degrees C. Mono- and dihydroxylated metabolites were also identified. Several screening methods were evaluated for detection and confirmation of methandrostenolone use including thin-layer chromatography and high-performance liquid chromatography. Coupled-column liquid chromatography was used for automated clean-up of analytes difficult to isolate by manual methods. The recovery of methandrostenolone was 101 +/- 3.3% (mean +/- S.D.) at 6.5 ng/ml and both methandrostenolone and 17-epimethandrostenolone were quantified in urine by ultraviolet detection up to six days after a 250-mg intramuscular dose to a horse. The utility of on-line tandem mass spectrometry for confirmation of suspected metabolites is also shown.

Animals↗
PubMed1986

Purification of human plasma lipid transfer protein using fast protein liquid chromatography.

A system for the isolation of human plasma lipid transfer protein (LTP) has been devised using a combination of conventional and high-performance ion-exchange chromatography. Following initial purification by ammonium sulphate precipitation, ultracentrifugation, hydrophobic interaction and cation-exchange chromatography, appropriate fractions were further purified using the Pharmacia fast protein liquid chromatography system. Using this method of purification, human plasma LTP has been purified more rapidly and with greater recovery than with conventional column chromatography. Whereas two forms of LTP were previously reported from the authors' laboratory [LTP-I, molecular mass (Mr) 69,000 and LTP-II, Mr 55,000], with an improved chromatographic system only one form of LTP (LTP-I) has been isolated. This suggests that LTP-II may have been a fragment of LTP-I, produced during the previously used lengthy purification process.

Buffers↗
PubMed1981

Identification of two Thormählen-positive compounds from melanotic urine by gas chromatography-mass spectrometry.

The isolation of two Thormählen-positive compounds from the urine of a patient with malignant melanoma and the elucidation of their structure by gas chromatography-mass spectometry is described. The compounds were isolated using a poly-N-vinylpyrrolidone column and separated by preparative thin-layer chromatography. After elution they were analyzed by gas chromatography and gas chromatography-mass spectrometry as their trimethylsilyl derivatives and after hydrolysis also as their tert.-butyldimethylsilyl derivatives. The results showed the main Thormählen-positive compound A to be the glucuronide of 5-hydroxy-6-methoxyindole, whereas the minor compound AX appeared to be the glucuronide of its isomer 6-hydroxy-5-methoxyindole.

Chromatography, Thin Layer↗
PubMed2002

Analysis of alpha-amylase-derived pyridylamino-dextran sulfate oligomers by the combination of size-exclusion and reversed-phase high-performance liquid chromatography.

In a previous study, we reported a novel method for the separation and quantification of a strong negatively charged material, dextran sulfate sodium (DSS), using fluorometric labeling with 2-aminopyridine and size-exclusion high-performance liquid chromatography. In the present study, we developed a method for the separation of pyridylamino-DSS (PA-DSS) using reversed-phase high-performance liquid chromatography (RPLC). In vitro enzymatic degradation of the PA-DSS was carried out using alpha-amylase. In RPLC, depolymerized PA-DSS was eluted on the basis of molecular mass (in the order pentamer, trimer, dimer, and monomer of PA-DSS) and separations were more sharply than in size-exclusion chromatography. The combination of RPLC and size-exclusion chromatography also separated depolymerized PA-DSS as effectively as RPLC alone.

Chromatography, Gel↗
PubMed2005

Enantioselective chromatography in drug discovery.

Molecular chirality is a fundamental consideration in drug discovery, one necessary to understand and describe biological targets as well as to design effective pharmaceutical agents. Enantioselective chromatography has played an increasing role not only as an analytical tool for chiral analyses, but also as a preparative technique to obtain pure enantiomers from racemates quickly from a wide diversity of chemical structures. Different enantioselective chromatography techniques are reviewed here, with particular emphasis on the most widespread high performance liquid chromatography (HPLC) and the rapidly emerging supercritical fluid chromatography (SFC) techniques. This review focuses on the dramatic advances in the chiral stationary phases (CSPs) that have made HPLC and SFC indispensable techniques for drug discovery today. In addition, screening strategies for rapid method development and considerations for laboratory-scale preparative separation are discussed and recent achievements are highlighted.

Amylose↗
PubMed1991

Theory for electrostatic interaction chromatography of proteins.

A simple theoretical framework for the effect of the eluting salt concentration on the retention factor of proteins in ion-exchange chromatography under conditions of linear elution is described. It is based on the solution of the linearized Poisson-Boltzmann equation for two oppositely charged planar surfaces in contact with a salt solution. The theory predicts a linear relation between the logarithmic retention factor and the reciprocal square root of the ionic strength of the eluent in the salt concentration range used in linear elution chromatography. A large body of retention data obtained in ion-exchange chromatography of proteins over a wide range of experimental conditions was plotted as ln k' vs 1/square root of I, where k' and I are the retention factor and ionic strength, respectively. The plots are linear or nearly so, as predicted for a moderate salt concentration range by the theory. From the slope of such plots the characteristic charges of the proteins were estimated by using only fundamental physicochemical constants. The chromatographically measured protein charges compare well to those obtained from titrimetric experiments at the same pH, although certain deviations are noted. The theoretical approach presented here offers a more realistic treatment of the ion-exchange chromatography of proteins than the stoichiometric displacement model and can serve as a convenient framework for the analysis of retention data.

Chromatography↗
PubMed1990

Affinity chromatography purification of angiotensin II receptor using photoactivable biotinylated probes.

We have developed biotinylated photoactivable probes that are suitable for covalent labeling of angiotensin II (AII) receptors and the subsequent purification of covalent complexes through immobilized avidin or streptavidin. One of these probes, biotin-NH(CH2)2SS(CH2)2CO-[Ala1,Phe(4N3)8]AII, which contains a cleavable disulfide bridge in its spacer arm and which displays, in its radioiodinated form, very high affinity for AII receptors (Kd approximately 1 nM), proved to be suitable for indirect affinity chromatography of rat liver receptor with facilitated recovery from avidin gels by use of reducing agents. This constituted the central step of an efficient partial purification scheme involving hydroxylapatite chromatography, streptavidin chromatography, and thiopropyl-Sepharose chromatography. SDS-PAGE analysis and autoradiography established the identity of the purified entity (molecular weight 65K) as the AII receptor. Possible ways of completing purification to homogeneity and extrapolation of the protocols to a preparative scale are discussed, as well as the potential contribution of our new probes to the study of the structural properties of angiotensin receptors.

Amino Acid Sequence↗