Biomedical subjects

R Gonzalo-Lumbreras

Publications and source records attributed to R Gonzalo-Lumbreras.

14 recordsLinked to original sources

PubMed2005

HPLC method development for testosterone propionate and cipionate in oil-based injectables.

Two isocratic liquid chromatographic methods for the determination of testosterone propionate (TP) and cipionate (TC) in oil-based injectables using methyltestosterone and bolasterone as internal standards, respectively, have been developed and validated. Mobile phases 57% water:acetonitrile 43% (v:v) and 54% water:acetonitrile 46% (v:v) were used for TP and TC, respectively. For both methods, a bonded-silica Luna CN (250 mm x 4.6 mm i.d., 5 microm) (25 degrees C) column, a flow-rate 1 ml min(-1) and UV absorbance detection at 245 nm were used and two separations up to base line were achieved. Prior to HPLC analysis, sample preparation was required, including extraction of TP and TC from oil-based injectables using the surfactant sodium dodecyl sulphate.

Chromatography, High Pressure Liquid↗

PubMed2004

Method development for betamethasone and dexamethasone by micellar liquid chromatography using cetyl trimethyl ammonium bromide and validation in tablets. Application to cocktails.

An isocratic liquid chromatographic method for the determination of betamethasone (BM) and dexamethasone (DM) using methylprednisolone (MPL) as internal standard and micellar mobile phases consisting of cetyl trimethyl ammonium bromide (CTAB) and organic modifiers such as propanol, butanol and pentanol has been developed. The effect of organic modifiers, surfactant concentration, temperature and flow-rate on the separation has been studied. Method validation for dexametasone or bethametasone in tablets was carried out using a mobile phase 0.24% pentanol and 32.5 mM CTAB, a flow-rate of 0.5 ml min(-1), an Hypersil C18 column (60 degrees C), and UV detection at 243 nm. The recoveries for BM and DM found in the accuracy test were 99 +/- 3 and 101 +/- 2, respectively. Repeatability and intermediate precision expressed as R.S.D. were lower than 5% for both compounds. The proposed method was applied to cocktails containing both compounds.

Betamethasone↗

PubMed2003

Optimization of the separation of a complex mixture of natural and synthetic anabolic steroids by micellar liquid chromatography.

A systematic optimization of the HPLC separation of a complex mixture containing natural and synthetic anabolic steroids by micellar liquid chromatography using a Hypersil (150 mm x 3.0 mm i.d., 5 microm) C18 column and UV detection at 245 nm (exception is made for oxymetolone and danazol which were monitorized at 280 nm) has been carried out. The isocratic micellar mobile phases (from binary to quaternary) consisted of sodium dodecyl sulphate and organic modifiers such as acetonitrile, tetrahydrofuran, propanol, butanol or pentanol. The effect of the organic modifiers, surfactant concentration, temperature, ionic strength and flow-rate on the separation has been studied. A micellar mobile phase 5% propanol and 40 mM surfactant allowed the separation of 12 steroids out of 14 tested in about 20 min. A bivariant optimization method for the micellar mobile phase propanol-surfactant corroborated the above results.

Anabolic Agents↗

PubMed2003

Method development for corticosteroids and anabolic steroids by micellar liquid chromatography.

A systematic optimization of the HPLC separation of a complex mixture containing urinary steroids (anabolics and corticoids), boldenone and bolasterone (synthetic anabolics) by micellar liquid chromatography has been carried out. The isocratic micellar mobile phases (from binary to quaternary) consisted of sodium dodecyl sulphate and organic modifiers such as acetonitrile, tetrahydrofuran, propanol, butanol or pentanol. The effect of the organic modifiers, surfactant concentration, temperature, ionic strength and flow-rate on the separation has been studied. A micellar mobile phase made of 5% propanol and 40 mM surfactant allowed the separation of 13 steroids in about 23 min. A bivariant optimization method for the micellar mobile phase surfactant-propanol corroborated the above results. The separations obtained show good perspectives for future developments.

Adrenal Cortex Hormones↗

PubMed2003

Conventional and micellar liquid chromatography method development for danazol and validation in capsules.

Two isocratic liquid chromatographic methods (conventional and micellar) for the determination of danazol (DZ) in capsules using canrenone (CAN) as internal standard have been developed and validated. In conventional liquid chromatography a mobile phase 35% water:acetonitrile 65%, v:v, a flow-rate 1 ml min(-1) and a C18 Hypersil ODS (250 x 4.6 mm, 5 microm) column (25 degrees C) were used. In micellar liquid chromatography (MLC) the conditions were: mobile phase 40 mM sodium dodecyl sulfate:2% pentanol, flow-rate 0.5 ml min(-1) and C18 Hypersil ODS (150 x 3.0 mm, 5 microm) column (60 degrees C). For both methods. UV absorbance detection at 280 nm was used and a separation up to base line was achieved. Prior to HPLC analysis a simple sample preparation was required. The recoveries found in the accuracy test were 99 +/- 10 and 101 +/- 8%, in conventional liquid chromatography (CLC) and MLC, respectively. Repeatability and intermediate precision expressed as R.S.D. were lower than 5% for both methods. Detection limits obtained were 2.4 and 3.0 ng g(-1) in CLC and CLM, respectively.

Canrenone↗

PubMed2003

Optimization and validation of conventional and micellar LC methods for the analysis of methyltestosterone in sugar-coated pills.

Two isocratic liquid chromatographic methods (conventional and micellar) for the determination of methyltestosterone in sugar-coated pills using fluoxymesterone as internal standard have been developed and validated. In conventional liquid chromatography a mobile phase 45% water:acetonitrile 55% (v:v), a flow-rate 1 mlmin(-1) and a C(18) Hypersil ODS (250 x 4.6 mm, 5 microm) column (25 degrees C) were used. In micellar liquid chromatography the conditions were: mobile phase 40 mM sodium dodecyl sulfate: 10% propanol, flow-rate 0.5 mlmin(-1) and C(18) Hypersil ODS (150 x 3.0 mm, 5 microm) column (60 degrees C). For both methods, UV absorbance detection at 245 nm was used and a separation up to base line was achieved. Prior to HPLC analysis a simple sample preparation was required.

Anabolic Agents↗

PubMed2001

Solvent and solid-phase extraction of natural and synthetic anabolic steroids in human urine.

Liquid-liquid (using dichloromethane) and liquid-solid extraction processes (using disposable C18 cartridges) were applied to human urine samples spiked with 15 androgenic anabolic steroids (natural and synthetic). The extraction recoveries were assessed from different HPLC separations of anabolic steroids using water-acetonitrile mobile phase, and using calibration graphs obtained by injection into HPLC of standard samples of these compounds before and after extraction. The procedures, including sample preconcentration, showed extraction efficiencies over 90% which were independent on a wide range of concentrations tested. Solid phase extraction yielded poor results for oximetolone, danazol and dehydroepiandrosterone. For real urine samples, hydrolysis using beta-glucuronidase and washing using sodium hydroxide before and after solvent extraction, respectively, is recommended.

Acetonitriles↗

PubMed2000

High-performance liquid chromatographic optimization study for the separation of natural and synthetic anabolic steroids. Application to urine and pharmaceutical samples.

An HPLC separation of a complex mixture containing 14 androgenic anabolic steroids (natural and synthetic) for screening purposes has been carried out. The applied optimization method involved the use of binary, ternary and quaternary mobile phases containing acetonitrile, methanol or tetrahydrofuran as organic modifiers. The effect of different reversed-phase packings and temperature on the separation using acetonitrile as organic modifier was studied. The optimum separation was achieved by using a water-acetonitrile (55:45, v:v) mobile phase and a Hypersil ODS (250 mm x 4.6 mm) 5 microm column (30 degrees C) in about 38 min, allowing the separation of 14 out of 14 compounds tested (when danazol is excluded, 13 out of 14 were separated in 23 min). Calibration graphs were obtained using bolasterone, methyltestosterone and canrenone as internal standards. Detection limits were in the range 0.012-0.11 microg ml(-1). The optimized separation was applied for monitoring the norethindrone acetate hydrolysis from tablets and to the analysis, after liquid-liquid extraction, of urine samples spiked with steroids.

Anabolic Agents↗

PubMed2000

Optimization of the high-performance liquid chromatographic separation of a complex mixture containing urinary steroids, boldenone and bolasterone: application to urine samples.

An HPLC separation of a complex mixture containing 13 urinary anabolics and corticoids, and boldenone and bolasterone (synthetic anabolics) has been carried out. The applied optimization method involved the use of binary, ternary and quaternary mobile phases containing acetonitrile, methanol or tetrahydrofuran as organic modifiers. The effect of different reversed-phase packings and temperature on the separation was studied. The optimum separation was achieved by using a water-acetonitrile (60:40, v/v) mobile phase in reversed-phase HPLC at 30 degrees C, allowing the separation of all the analytes in about 24 min. Calibration graphs were obtained using bolasterone or methyltestosterone as internal standards. Detection limits were in the range 0.012-0.107 microg ml(-1). The optimized separation was applied to the analysis, after liquid-liquid extraction, of human urine samples spiked with steroids.

Adrenal Cortex Hormones↗

PubMed1995

Simultaneous determination of cortisol and cortisone in urine by reversed-phase high-performance liquid chromatography. Clinical and doping control applications.

A reversed-phase high-performance liquid chromatography (HPLC) method for the simultaneous determination of cortisol and cortisone in human urine samples using methylprednisolone as the internal standard is described. The method involves the systematic use of isocratic mobile phases of water and methanol, acetonitrile or tetrahydrofuran and a reversed-phase Hypersil C18 column. A water-acetonitrile mixture used as the mobile phase proved to be the most adequate one for analyzing urine samples purified by solvent extraction. The proposed method is sensitive, reproducible and selective. It was applied to the determination of cortisol and cortisone in several human urine samples: healthy subjects, sportsmen before and/or after stress for doping control purposes, and patients with Cushing's syndrome.

Calibration↗

PubMed1994

Extraction and high-performance liquid chromatographic separation of deflazacort and its metabolite 21-hydroxydeflazacort. Application to urine samples.

Two HPLC methods for the separation of a mixture of corticoids including the oxazolinic corticoid deflazacort and its metabolite 21-hydroxydeflazacort using different water-tetrahydrofuran mobile phases were developed. Both separations allowed the detection and determination of fifteen out of sixteen corticoids using different C18 columns. Extraction data for deflazacort and its metabolite using different extraction procedures are also reported. These separation conditions were applied to urine samples from two male volunteers administered Dezacor, with both doping control and clinical purposes.

Adrenal Cortex Hormones↗

PubMed1994

Solvent and solid-phase extraction of natural and synthetic corticoids in human urine.

Optimization of the main variables that affect solvent and solid-phase extraction processes, using disposable C18 cartridges and the non-ionic polymeric resin Serdolit AD-2, of human urine containing natural and synthetic corticoids is described. The data were obtained from different HPLC separations of these compounds using calibration graphs obtained before and after extraction of these compounds. The procedures, including sample preconcentration, showed efficiencies over 90%. NaCl was used to avoid emulsion formation in solvent extraction. The results achieved using solvent and solid-phase extraction are discussed.

Adrenal Cortex Hormones↗

PubMedDate not supplied

Development and method validation for testosterone and epitestosterone in human urine samples by liquid chromatography applications.

An isocratic high-performance liquid chromatographic method for the determination of testosterone (T) and epitestosterone (ET) in human urine using liquid-liquid or solid-phase extraction (SPE) is developed and validated. The optimum separation is achieved using a Hypersil C(18) column, water-acetonitrile (57:43, v/v) as the mobile phase and UV-absorbance detection at 245 nm. The recoveries obtained for T and ET in liquid-liquid and SPE demonstrate that these procedures are interchangeable. Quantitation limits for T and ET are 8.6 and 5.4 ng/mL using solvent extraction and 7.3 and 5.7 ng/mL using SPE, respectively. The proposed method is used to evaluate the urinary T, ET, and the T/ET ratio for a healthy male population using liquid-liquid extraction, and the T and ET excretion profile for nine healthy men using SPE.

Adult↗

PubMedDate not supplied

Method development for cortisol and cortisone by micellar liquid chromatography using sodium dodecyl sulphate: application to urine samples of rugby players.

The chromatographic behavior of cortisol and cortisone using a micellar medium of sodium dodecyl sulphate (SDS) as surfactant, a Hypersil C18 (150- x 3.2-mm i.d., 5 microm) column, a flow rate of 0.5 mL/min, and UV absorbance detection at 245 nm is described. The effect of several organic modifiers and the surfactant concentration on the separation is studied. A mobile phase of 18 mM SDS and 8.3% tetrahydrofuran allows for the separation of cortisol and cortisone up to baseline. These results are also achieved by applying a bivariant optimization method. The proposed method is sensitive, reproducible, and selective. In addition, it is less expensive than conventional high-performance liquid chromatography methods for cortisol and cortisone. The method is applied to the determination of cortisol and cortisone in urine samples of rugby players before and after stress for doping control purposes.

Chromatography, Liquid↗