Biomedical subjects

F J Rupérez

Publications and source records attributed to F J Rupérez.

12 recordsLinked to original sources

PubMed2005

Tandem column for the simultaneous determination of arginine, ibuprofen and related impurities by liquid chromatography.

Ibuprofen arginate is a rapidly absorbed salt designed to promote more rapid onset of analgesia than commercially available forms of ibuprofen. Ibuprofen and arginine have very different polarities and this becomes in a chromatographic problem, further complicated with the determination of related compounds, which is necessary in stability assays of the pharmaceutical forms. The common solution is the employment of two separate methods, but this is time consuming. A LC method has been developed to determinate both compounds and related impurities in one run. Ibuprofen, arginine and three ibuprofen related impurities (B, E and J) have been baseline separated with isocratic conditions at pH 3.0 and run time under 20 min by employing a tandem combination of two different stationary phases: first a ZORBAX SB-C18 column from Agilent (250 mm x 4.6 mm and 5 microm) and downstream a SUPELCOSIL LC-NH2 column from Supelco (150 mm x 4.6 mm and 3 microm). The octadecyldiisobutylsilane column provides the separation of ibuprofen and its impurities by a hydrophobic mechanism, whereas aminopropyl column offers selective retention of arginine by dipolar interaction mechanism. Method has been successfully validated following ICH guidelines and it has been demonstrated to be reliable for arginine, ibuprofen and related impurities determination in sachets of two different dosages as pharmaceutical forms. Moreover, stress test has proved the selectivity of the method for degradation products, such as those that can emerge throughout long-term stability assays.

Arginine↗

PubMed2005

Capillary electrophoresis of glutathione to monitor oxidative stress and response to antioxidant treatments in an animal model.

Glutathione plays a central role in metabolism and antioxidant defence. Several factors can influence the analytical efficiency and rapidity of the quantitative determination of glutathione. Procedures in sample pre-treatment have been compared in order to minimize analytical errors. Capillary electrophoresis has been chosen as a more adequate technique for obtaining a rapid and simple method for glutathione and glutathione disulfide determination in the blood and liver of the rat. The methods, once optimised, have been validated and applied for monitoring the oxidative stress in an animal model, such as the rat made diabetic by streptozotocin injection, when the animals are treated with antioxidants and compared with the corresponding controls.

Animals↗

PubMed2005

LC methods for acyclovir and related impurities determination.

Acyclovir, guanine, and impurity A have been baseline separated with isocratic conditions at pH=3.0 and run time under 15 min by employing a SB-CN column from Agilent (150 mm x 4.6mm and 3.5 microm). Moreover, when run time was increased to 40 min six impurities (guanine, impurities A, F, G, Vir 3/4 and N(7)) plus acyclovir were separated in the same conditions. The mobile phase consisted of buffer A/acetonitrile 96:4 (v/v), being buffer A:25 mM H(3)PO(4) (Milli-Q H(2)O) brought to pH 3.0 with KOH. The same column provided separation for all the seven impurities described in pharmacopoeia, including impurity C, which coeluted with acyclovir in the previous conditions with a mobile phase prepared with 25 mM phosphoric acid (pH=1.8)/acetonitrile 96:4 (v/v). The method has been validated following ICH guidelines and it has demonstrated to be reliable for acyclovir and its impurities determination.

Acyclovir↗

PubMed2004

Direct liquid chromatography method for retinol, alpha- and gamma-tocopherols in rat plasma.

An HPLC method for Vitamins A and E in rat plasma has been developed. The main goals of the method are the small amount of sample, 50 microl, and the direct extraction of analytes in one step with acetone, which is a solvent compatible with the reverse-phase mobile phases. Recoveries, as compared with classical and more tedious methods, were near 100%. The method employs a Supelco Discovery C18 column and methanol/water (95:5, v/v) as mobile phase. After being developed, the method was validated following ICH guidelines, with UV, fluorescence and electrochemical detectors. It proved to be selective, lineal, accurate and precise. This method greatly simplifies sample treatment and that is a critical point when working with a large number of samples.

Animals↗

PubMed2003

Capillary electrophoresis determination of loratadine and related impurities.

While HPLC has traditionally been the method of choice for purity determination of pharmaceutical substances, capillary electrophoresis (CE) offers a different selectivity and hence it is a complementary technique to HPLC. Loratadine, an antihistamine, could include in its raw material seven impurities that ought to be separated, identified and quantified for drug development and quality control. As a complementary tool for undoubtful identification, a CE method has been developed. The separation was carried out with an uncoated fused-silica capillary (57 cm x 50 microm ID) and was operated at 20 kV potential. Temperature was maintained at 25 degrees C. The final separation buffer was prepared with 100 mM H(3)PO(4) made up to pH 2.5 with NaOH and with 10% acetonitrile added (v/v). Impurities can be detected at the 0.1% level of the active and validation parameters for linearity accuracy and precision are adequate for all the analytes and that permits to consider the method reliable and suitable for application to long-term stability and purity studies.

Drug Contamination↗

PubMed2003

Poly(ethyleneglycol) column for the determination of acetaminophen, phenylephrine and chlorpheniramine in pharmaceutical formulations.

New polar reversed-phase stationary phases in HPLC provide specific selectivities which can help to solve traditional chromatographic problems related to the development of chromatographic methods with widely different retention times for the sample components. One such case is the analysis of pharmaceutical formulations against the common cold. Acetaminophen, phenylephrine and chlorpheniramine, compounds with different polarities, are frequently associated in these drugs. An isocratic and rapid HPLC method for the simultaneous determination of the three compounds, acetaminophen, phenylephrine and chlorpheniramine, in capsules as pharmaceutical formulations, including the separation of impurities (4-aminophenol and 4-chloracetanilide) and excipients, has been developed and validated. The final chromatographic conditions employed a Supelco Discovery HS PEG column poly(ethyleneglycol) 15x0.46 cm, 5 microm. The mobile phase was 20 mM phosphate buffer, pH 7.0-acetonitrile (90:10, v/v) at a flow-rate of 1 ml/min. UV detection was performed at 215 nm for all the compounds except acetaminophen, which was measured at 310 nm. Validation parameters permit us to consider this method suitable.

Acetaminophen↗

PubMed2002

LC determination of loratadine and related impurities.

Loratadine, an antihistamine, could include in its raw material seven impurities that ought to be separated identified and quantified for drug development and quality control. A HPLC method employing a SymmetryShield RP8 column has been developed and validated for loratadine and related compounds measurement, the last ones under the 0.1% level. The mobile phase consisted of methanol-buffer A (65:35, v/v), being buffer A: H(3)PO(4) 10 mM (H(2)O) brought up to pH 7.00 with triethylamine. UV detection was performed at 244 nm. Validation parameters for linearity, accuracy and precision are in agreement with ICH guidelines for all the analytes and that permits to consider the method reliable and suitable for application to long-term stability and purity studies.

Chromatography, High Pressure Liquid↗

PubMed2001

Chromatographic analysis of alpha-tocopherol and related compounds in various matrices.

Tocopherols and tocotrienols (Vitamin E) are part of a group of "minor components" of main interest, present in the unsaponifiable fraction of many samples. Their importance in biological, metabolical and nutritional studies makes determination of tocopherols and related compounds of major interest. Present work critically reviews the different ways to perform sample pre-treatment and analysis of these compounds, related to the matrices, other analytes to be measured, sensitivity, and simplicity. The review includes well referenced tables that provide in-depth summaries of methodology for the chromatographic analysis of alpha-tocopherol and related compounds in foods, pharmaceuticals, plants, animal tissues and other matrices.

Chromatography, Gas↗

PubMed2001

Capillary electrophoresis for evaluating orange juice authenticity: a study on Spanish oranges.

Fruit juices have very distinct organic acid profiles that can be used as fingerprints for establishing possible adulteration. Recently, our group developed and validated a capillary electrophoresis method using UV detection for determining citric, isocitric, tartaric, and malic acids in natural and commercial orange juices. Sample treatment consisted of only dilution and centrifugation or filtration. This method has been applied to evaluate these acids and their ratios in 63 samples of Navelina, the most common variety of Spanish oranges, over a three month period. This evaluation has been conducted to establish ranges of acid concentrations and to compare them with those found in commercial juices. The more reliable parameter, because of the lower variability in fresh samples, was found to be the citrate/isocitrate ratio with a value of 113 (RSD = 10%). Only one of nine ramdonly selected commercial juices presented values within the range of those of the population of just-pressed Navelina orange juice. Moreover, three of them had measurable tartrate values, which is not a natural component of orange juice, showing mixtures with cheaper fruits.

Beverages↗

PubMed2000

Low arachidonic acid rather than alpha-tocopherol is responsible for the delayed postnatal development in offspring of rats fed fish oil instead of olive oil during pregnancy and lactation.

This study was designed to compare in rats the effects of dietary fish oil and olive oil during pregnancy and lactation on offspring development, fatty acid profile and vitamin E concentration. From d 0 of pregnancy, female Sprague-Dawley rats were divided into two groups that were fed purified diets that differed only in their nonvitamin lipid components. One diet contained 10 g fish oil/100 g diet (FOD), whereas the other contained 10 g olive oil/100 g diet (OOD). At d 20 of gestation, maternal adipose tissue fatty acid profile did not differ between rats fed the two diets, whereas both maternal and fetal plasma and liver arachidonic acid (AA) contents were proportionally lower and eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid contents were higher in the FOD group than in the OOD group. alpha-Tocopherol concentration was lower in maternal and fetal plasma, liver and brain in the FOD group than in the OOD group. The postnatal increase in body weight and length was less and body and psychomotor maturation indices were delayed in pups from FOD-fed dams compared with those from OOD-fed dams. This difference was maintained when pups were cross-fostered at birth, with the delay in postnatal development present in the pups suckling dams fed FOD during lactation. At age 21 d, pups suckling dams fed FOD had lower AA and higher EPA and DHA concentrations in brain phospholipids. Although alpha-tocopherol in plasma and liver was lower in pups suckling dams fed FOD rather than OOD, brain alpha-tocopherol concentrations did not differ. Milk yield and milk alpha-tocopherol and AA concentrations were lower and EPA and DHA were higher in the milk of dams fed FOD compared with those fed OOD. Postnatal development indices and the proportion of plasma, liver and brain AA concentrations, although not plasma, liver and brain alpha-tocopherol concentrations, recovered to the values found in dams fed OOD when the FOD was supplemented with gamma-linolenic acid. However, postnatal development indices were not recovered when the FOD was supplemented with sufficient exogenous vitamin E to increase plasma and liver alpha-tocopherol concentrations above those in dams fed OOD. Thus, although feeding FOD during pregnancy and lactation decreases both alpha-tocopherol and AA concentrations, the latter deficiency rather than the former seems to be responsible for delayed postnatal development of rat pups.

Adipose Tissue↗

PubMed1999

Determination of alpha-tocopherol and alpha-tocopherol acetate in diets of experimental animals. Study of stability in the diets.

A simple method is described which permits, avoiding saponification, alpha-tocopherol and alpha-tocopheryl acetate measurement in semi-synthetic diets for experimental animals by HPLC, with both UV and fluorescence detection. Phenyldodecane was chosen as internal standard with remarkable performances, and EDTA and BHT were added to prevent oxidation in aqueous and non-aqueous phases respectively. The mobile phase was methanol-water (94:6 v/v) at a flow-rate of 2 ml/min. Samples were homogenized and extracted twice with n-hexane by probe sonication. Extracts were evaporated to dryness and redissolved with chloroform-methanol (1:1, v/v). Validation parameters were studied between 25 ng and 6 micrograms for alpha-tocopherol and between 3 and 24.2 micrograms for alpha-tocopheryl acetate, which corresponds to the range of values in the existing diets. Results had correlation coefficients > 0.99; recoveries > 85%; R.S.D. < 6%, so the method is adequate to control vitamin E intake in animals as well as vitamin E stability in food during storage.

Animal Feed↗

PubMed1998

Simplified method for vitamin E determination in rat adipose tissue and mammary glands by high-performance liquid chromatography.

A method for vitamin E (alpha-tocopherol) measurement in rat adipose tissue and mammary gland has been developed and validated. Tissues were homogenized in ethanol-water (1:1) and extracted with n-hexane. Vitamin K1 was used as internal standard. Separation was performed by HPLC with methanol-water (96.5:3.5) as eluent in a Nucleosil C18 column (15 x 0.46 cm) at 40 degrees C. Detection was by fluorescence with excitation at 295 nm and emission at 350 nm for vitamin E and at 330 and 440 nm for vitamin K1. Standards and tissue extracts were checked for linearity giving correlation coefficients over 0.99 in a range of concentrations from 0.56 to 4.51 nmol/g in adipose tissue and from 2.18 to 17.4 nmol/g in mammary gland tissue. Intra-assay precision (R.S.D.) varied between 3 and 4%, whereas inter-assay precision was between 8 and 9%. Recoveries ranged between 95 +/- 3% and 98 +/- 11% for the two tissues, respectively. Vitamin E was measured in rats that had previously received one oral dose of this vitamin. Whereas vitamin E content in adipose tissue did not differ between late-pregnant and virgin rats, it was significantly higher in mammary gland of pregnant rats, and this difference could be related to the enhanced lipoprotein lipase activity in this group.

Adipose Tissue↗