Capillary electrophoretic chiral separation of hydroxychloroquine and its metabolites in the microsomal fraction of liver homogenates

2006 ◽  
Vol 27 (5-6) ◽  
pp. 1248-1254 ◽  
Author(s):  
Carmem Dickow Cardoso ◽  
Valquíria A. Polisel Jabor ◽  
Pierina Sueli Bonato
Keyword(s):  
Chiral Separation ◽  
Microsomal Fraction ◽  
Capillary Electrophoretic ◽  
Liver Homogenates

scholarly journals ANALYTICAL STUDY OF MICROSOMES AND ISOLATED SUBCELLULAR MEMBRANES FROM RAT LIVER

1974 ◽  
Vol 61 (1) ◽  
pp. 201-212 ◽  
Author(s):  
Alain Amar-Costesec ◽  
Henri Beaufay ◽  
Maurice Wibo ◽  
Denise Thinès-Sempoux ◽  
Ernest Feytmans ◽  
...  
Keyword(s):  
Analytical Study ◽  
Microsomal Fraction ◽  
Chemical Constituents ◽  
Microsomal Protein ◽  
High Yield ◽  
Marker Enzymes ◽  
Cytoplasmic Granules ◽  
Liver Homogenates ◽  
Nadph Cytochrome C Reductase ◽  
Phosphatase Alkaline

Liver homogenates have been submitted to quantitative fractionation by differential centrifugation. Three particulate fractions: N (nuclear), ML (large granules), and P (microsomes), and a final supernate (S) have been obtained. The biochemical composition of the microsomal fraction has been established from the assay and distribution pattern of 25 enzymatic and chemical constituents. These included marker enzymes for mitochondria (cytochrome oxidase), lysosomes (acid phosphatase and N-acetyl-ß-glucosaminidase), and peroxisomes (catalase). The microsomal preparations were characterized by a moderate contamination with large cytoplasmic granules (only 6.2% of microsomal protein) and by a high yield in microsomal components. Enzymes such as glucose 6-phosphatase, nucleoside diphosphatase, esterase, glucuronyltransferase, NADPH cytochrome c reductase, aminopyrine demethylase, and galactosyltransferase were recovered in the microsomes to the extent of 70% or more. Another typical behavior was shown by 5'-nucleotidase, alkaline phosphatase, alkaline phosphodiesterase I, and cholesterol, which exhibited a "nucleomicrosomal" distribution. Other complex distributions were obtained for several constituents recovered in significant amount in the microsomes and in the ML or in the S fraction.

scholarly journals Bile acids of snakes of the subfamily Viperinae and the biosynthesis of C-23-hydroxylated bile acids in liver homogenate fractions from the adder, Vipera berus (Linn.)

1976 ◽  
Vol 153 (2) ◽  
pp. 343-350 ◽  
Author(s):  
S Ikawa ◽  
A R Tammar
Keyword(s):  
Bile Acids ◽  
Microsomal Fraction ◽  
Liver Homogenate ◽  
Mitochondrial Fraction ◽  
Sodium Cholate ◽  
Vipera Berus ◽  
C 23 ◽  
Liver Homogenates ◽  
Generating System ◽  
Cholanic Acid

1. Analysis of bile salts of four snakes of the subfamily Viperinae showed that their bile acids consisted mainly of C-23-hydroxylated bile acids. 2. Incubations of 14C-labelled sodium cholate (3 α, 7 α, 12 α-trihydroxy-5 β-cholan-24-oate) and deoxycholate (3 α, 12 α-dihydroxy-5 β-cholan-24-oate) with whole and fractionated adder liver homogenates were carried out in the presence of molecular oxygen and NADPH or an NADPH-generating system. The formation of C-23-hydroxylated bile acids, namely bitocholic acid (3 α, 12 α, 23xi-trihydroxy-5 β-cholan-24-oic acid) and 3 α, 7 α, 12 α, 23 ξ-tetrahydroxy-cholanic acid (3 α, 7 α, 12 α, 23 ξ-tetrahydroxy-5 β-cholan-24-oic acid), was observed mainly in the microsomal fraction and partly in the mitochondrial fraction. 3. Biosynthetic pathways of C-23-hydroxylated bile acids are discussed.

scholarly journals The metabolism of phenylmercury by the rat

1972 ◽  
Vol 129 (4) ◽  
pp. 961-967 ◽  
Author(s):  
J. W. Daniel ◽  
J. C. Gage ◽  
P. A. Lefevre
Keyword(s):  
Small Proportion ◽  
Microsomal Fraction ◽  
Inorganic Mercury ◽  
Elemental Mercury ◽  
Microsomal Enzymes ◽  
Single Subcutaneous Dose ◽  
Subcutaneous Dose ◽  
Liver Homogenates

The metabolism of [U-14C]phenylmercury acetate was studied in the rat. After a single subcutaneous dose a small proportion is excreted unchanged in urine, and a larger amount in bile with some resorption from the gut. The greater part of the dose is broken down in the tissues to yield inorganic mercury which is excreted mainly in faeces, and conjugates of phenol and quinol are excreted in urine. In experiments in vitro phenylmercury is broken down by liver homogenates to release inorganic mercury and benzene; this reaction is effected by the soluble, but not the microsomal, fraction and does not require NADPH or NADH. No elemental mercury is formed under these conditions. It is probable that this reaction occurs in vivo and the benzene produced is rapidly converted into phenol and quinol by microsomal enzymes.

Depolymerization of polyadenylic acid by subcellular fractions of rat brain and liver

1969 ◽  
Vol 47 (3) ◽  
pp. 283-289 ◽  
Author(s):  
M. R. V. Murthy ◽  
A. D. Bharucha ◽  
C. Raynaud-Jammet
Keyword(s):  
Rat Brain ◽  
Microsomal Fraction ◽  
Soluble Fraction ◽  
Total Activity ◽  
Relative Increase ◽  
Nuclear Fraction ◽  
Polyadenylic Acid ◽  
Rat Brain And Liver ◽  
Liver Homogenates ◽  
Maximum Increment

Rat brain and liver homogenates depolymerized polyadenylic acid when added to a reaction mixture containing this polynucleotide. The activity in the homogenate declined progressively with the age of the tissues. This was reflected in a parallel reduction in the activity of the soluble fraction. In brain, the activity in the nuclear fraction also declined in the adult to half the level of the newborn. In contrast, liver nuclei had approximately the same activity at all stages of growth.With advancement in age, an increasingly greater proportion of the total activity of the tissues was contained in the nuclear fraction, while at the same time the proportion of activity in the soluble fraction decreased. The proportion of activity contained in the mitochondrial–microsomal fraction also increased with growth in brain, with the maximum increment in activity occurring after 8 weeks of age. In liver, there was actually a decrease of activity in this fraction during the same period. At all ages, the mitochondrial–microsomal fraction of brain contained a higher proportion of activity and the nuclear fraction of brain contained a lower proportion of activity compared to corresponding fractions of liver. The presence of polyadenylic acid degrading activity in these fractions and its relative increase with age may indicate a changing emphasis in the pattern of RNA metabolism during growth; for example, a higher rate of RNA synthesis in the young and a higher rate of RNA turnover in the adult.When the soluble fraction of rat brain was dialyzed, the polyadenylic acid degrading activity of this fraction was stimulated by the addition of inorganic orthophosphate. Brain and liver homogenates also mediated an ADP – inorganic phosphate exchange reaction which was highest in the newborn and decreased rapidly with age. These observations indicate that at least a part of the polyadenylic acid degrading activity in brain and liver extracts may be due to phosphorolytic action.

scholarly journals GOLGI FRACTIONS PREPARED FROM RAT LIVER HOMOGENATES

1973 ◽  
Vol 59 (1) ◽  
pp. 73-88 ◽  
Author(s):  
J. J. M. Bergeron ◽  
J. H. Ehrenreich ◽  
P. Siekevitz ◽  
G. E. Palade
Keyword(s):  
Endoplasmic Reticulum ◽  
Cytochrome C ◽  
Rat Liver ◽  
Microsomal Fraction ◽  
Companion Paper ◽  
Transferase Activity ◽  
Ethanol Treatment ◽  
Liver Homogenates ◽  
Cytochrome P 450 ◽  
Nadph Cytochrome C Reductase

The three Golgi fractions isolated from rat liver homogenates by the procedure given in the companion paper account for 6–7% of the protein of the total microsomal fraction used as starting preparation. The lightest, most homogeneous Golgi fraction (GF1) lacks typical "microsomal" activities, e.g., glucose-6-phosphatase, NADPH-cytochrome c-reductase, and cytochrome P-450. The heaviest, most heterogeneous fraction (GF3) is contaminated by endoplasmic reticulum membranes to the extent of ∼15% of its protein. The three fractions taken together account for nearly all the UDP-galactose: N-acetyl-glucosamine galactosyltransferase of the parent microsomal fraction, and for ∼70% of the activity of the original homogenate. Omission of the ethanol treatment of the animals reduces the recovery by half. The transferase activity is associated with the membranes of the Golgi elements, not with their content. Galactose is transferred not only to N-acetyl-glucosamine but also to an unidentified lipid-soluble component.

scholarly journals Molecular Modeling and Chiral Separation of Benzodiazepines by Capillary Electrophoresis Using Highly Sulfated Cyclodextrins

2018 ◽  
Vol 62 (2) ◽  
Author(s):  
María Gabriela Vargas Martinez ◽  
Guillermo Ramírez Galicia
Keyword(s):  
Capillary Electrophoresis ◽  
Chiral Separation ◽  
Theoretical Study ◽  
Equilibrium Constants ◽  
Fractional Factorial ◽  
Organic Modifier ◽  
Enantiomeric Resolution ◽  
Electrophoretic Method ◽  
Capillary Electrophoretic ◽  
Separation Conditions

<p>A capillary electrophoretic method for the chiral separation of the 3-chiral-1,4-benzodiazepines was developed. Enantiomeric resolution of oxazepam, lorazepam, temazepam, and lormetazepam was achieved using sulfated cyclodextrins (CD's) as chiral selectors. A 3-levels, 4-factors fractional factorial (34-2) design was applied to test 3 different CD's: heptakis-6-sulfato--cyclodextrin (HSCD), heptakis-(2,3-diacetyl-6-sulfato)--cyclodextrin (HDASCD), and heptakis-(2,3-dimethyl-6-sulfato)--cyclodextrin (HDMSCD). The CD type, its concentration, the pH of the electrolyte, and % organic modifier were tested as the factors in the experimental design. The highest resolution values were obtained using a 20 mM borate buffer, pH 9.0 with the addition of 5 % HSCD and 15 % methanol as an organic modifier. At these separation conditions, the equilibrium constants of the benzodiazepine-HSCD complex formation were calculated. A theoretical study of the interaction benzodiazepine-HSCD complex using semiempirical calculations is postulated.</p>

METABOLISM OF PORCINE, HUMAN AND SALMON CALCITONIN IN THE RAT

1972 ◽  
Vol 53 (3) ◽  
pp. 475-482 ◽  
Author(s):  
M. de LUISE ◽  
T. J. MARTIN ◽  
P. B. GREENBERG ◽  
V. MICHELANGELI
Keyword(s):  
Salmon Calcitonin ◽  
Microsomal Fraction ◽  
Rat Kidney ◽  
Mammalian Species ◽  
Rat Tissues ◽  
Human Calcitonin ◽  
Major Site ◽  
Major Activity ◽  
Liver Homogenates

SUMMARY Whereas the liver is the major site of accumulation of 125I-labelled porcine calcitonin soon after injection in the rat, both human and salmon calcitonin were rapidly taken up in rat kidney, with relatively insignificant amounts found in the liver. In-vitro studies of degradation of 125I-labelled calcitonins showed that human calcitonin was readily degraded by most rat tissues but the major activity was found in a kidney microsomal fraction, whereas the liver supernatant was most active towards pig calcitonin. Salmon calcitonin was resistant to breakdown by all tissues and fractions except the kidney microsomal fraction, which rapidly degraded it to trichloroacetic acid-soluble fragments. Liver homogenates from a number of mammalian and non-mammalian species degraded pig calcitonin but had little effect on salmon calcitonin. The results show that the kidney is the most important organ in the metabolism of human and salmon calcitonin in the rat, while confirming that the liver is mainly responsible for the metabolism of porcine calcitonin.

scholarly journals Differences in distribution of esterase between cell fractions of rat liver homogenates prepared in various media. Relevance to the lysosomal location of the enzyme in the intact cell

1971 ◽  
Vol 125 (2) ◽  
pp. 545-555 ◽  
Author(s):  
Patience C. Barrow ◽  
S. J. Holt
Keyword(s):  
Acid Phosphatase ◽  
Rat Liver ◽  
Microsomal Fraction ◽  
Intact Cell ◽  
Cellular Damage ◽  
Phase System ◽  
Two Phase ◽  
Liver Homogenates ◽  
Cell Fractions ◽  
Standard Fractionation

The distribution of esterase in subcellular fractions of rat liver homogenates was compared with that of the lysosomal enzyme acid phosphatase and the microsomal enzyme glucose 6-phosphatase. Most of the esterase from sucrose homogenate sediments with glucose 6-phosphatase and about 8% is recovered in the supernatant. However, up to 53% of the esterase can be washed from microtome sections of unfixed liver, in which less cellular damage would be expected than that caused by homogenization. About 40% of both esterase and acid phosphatase are recovered in the soluble fraction after homogenization in aqueous glycerol or in a two-phase system (Arcton 113–0.25m-sucrose), although glucose 6-phosphatase is still recovered in the microsomal fraction of such homogenates. The esterase of the microsomal fraction prepared from a sucrose homogenate is much more readily released by treatment with 0.26% deoxycholate than are other constituents of this fraction. The release of esterase from the microsomal fraction by the detergent and its concomitant release with acid phosphatase after homogenization in glycerol or the two-phase system suggests that a greater proportion of esterase may be present in lysosomes of the intact cell than is indicated by the results of standard fractionation procedures.

Capillary electrophoretic chiral separation of Cinchona alkaloids using a cyclodextrin selector

2008 ◽  
Vol 31 (6-7) ◽  
pp. 1130-1136 ◽  
Author(s):  
Dimitrios Tsimachidis ◽  
Petr Česla ◽  
Tomas Hájek ◽  
Georgios Theodoridis ◽  
Pavel Jandera
Keyword(s):  
Chiral Separation ◽  
Cinchona Alkaloids ◽  
Capillary Electrophoretic

Chiral Separation of Uncharged Pomalidomide Enantiomers Using Carboxymethyl-β-Cyclodextrin: A Validated Capillary Electrophoretic Method

Chirality ◽  
2015 ◽  
Vol 28 (3) ◽  
pp. 199-203 ◽  
Author(s):  
Zoltán-István Szabó ◽  
Levente Szőcs ◽  
Daniela-Lucia Muntean ◽  
Béla NoszáL ◽  
Gergő Tóth
Keyword(s):  
Chiral Separation ◽  
Electrophoretic Method ◽  
Capillary Electrophoretic
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