Sequential Solvent Extraction of Urinary Corticosteroids and its Clinical Application

1967 ◽  
Vol 13 (10) ◽  
pp. 855-876 ◽  
Author(s):  
Habeeb Bacchus
Keyword(s):  
Carbon Tetrachloride ◽  
Ethyl Acetate ◽  
Clinical Laboratory ◽  
Ethyl Acetate Fraction ◽  
Polar Compounds ◽  
Subsequent Treatment ◽  
Salting Out ◽  
Adrenocortical Hyperplasia ◽  
The Difference ◽  
Hepatocellular Disease

Abstract A procedure which permits the measurement of corticosteroid metabolites in three solvent extracts of hydrolyzed urine is described. Carbon tetrachloride extracts the 11-desoxy, 17-hydroxycorticosteroids, and the 17-OH, 21-deoxycorticosteroids. These are quantitated by the Porter-Silber reaction and the method for 17-ketogenic steroids. The difference between 17-KGS and PSC is a measure of the total 17-OH, 21-deoxycorticosteroids. Subsequent treatment of the residual urine with methylene dichloride extracts compounds of the cortisol series, i.e., cortisol and cortisone in small amounts, THF and THE in larger amounts. These compounds are quantitated by either the Porter-Silber reaction or the 17-KGS procedure. Further treatment of the urine residue with ethyl acetate (employing the "salting out" procedure) results in the extraction of polar compounds including cortol, cortolone, 6 -hydroxycortisol, and its reduction product. This procedure was employed to study several clinical disorders. Congenital virilizing adrenocortical hyperplasia is characterized by a marked increase in the 17-KGS in the carbon tetrachloride extract; in adult-onset virilizing adrenocortical hyperplasia this increase is also significant, but not as marked. In adrenal hyperactivity (e.g., Cushing's syndrome, ACTH infusion) there are increases in all solvent fractions studied. Inhibition of 11- hydroxylase activity induced by SU 4885 (Metopirone) is accompanied by an increase in the PSC in the carbon tetrachloride fraction. In hepatocellular disease and in pregnancy, the most noteworthy changes are in the increase in polar compounds extracted in the ethyl acetate fraction. The method described is attractive in its simplicity and is readily applicable in the clinical laboratory.

scholarly journals A Polyphenol-Rich Fraction from Eugenia uniflora Exhibits Antioxidant and Hepatoprotective Activities In Vivo

2020 ◽  
Vol 13 (5) ◽  
pp. 84
Author(s):  
Mansour Sobeh ◽  
Marwa S. Hamza ◽  
Mohamed L. Ashour ◽  
Mona Elkhatieb ◽  
Mohamed A El Raey ◽  
...  
Keyword(s):  
Carbon Tetrachloride ◽  
Liver Injury ◽  
Ethyl Acetate ◽  
Acute Liver Injury ◽  
Antioxidant Activities ◽  
Biological Effects ◽  
Ethyl Acetate Fraction ◽  
Eugenia Uniflora ◽  
Quercetin Derivatives

Leaves from Eugenia uniflora, the red Brazilian cherry, have a high content of flavonoids that possess several biological effects such as anti-inflammatory, antioxidant, and antidiabetic activities. However, their influence on carbon tetrachloride (CCl4)-induced acute liver injury in rats has not been investigated. In the current study, a bioguided fractionation assay revealed that the ethyl acetate fraction (EAF) of Eugenia uniflora is the safest and most active fraction. LC-MS analysis of the ethyl acetate fraction revealed 22 secondary metabolites, mainly myricetin and quercetin derivatives. EAF did not show toxicity up to 2000 mg/kg, and exhibited antioxidant activities in vitro in DPPH assay with IC50 of 3.35 µg/mL. Additionally, EAF exhibited substantial antioxidant activities in vivo by counteracting the oxidative damage of the prooxidant juglone [80 µM] in Caenorhabditis elegans model organism and increased its survival rate in a dose-dependent fashion through the DAF-16/Foxo pathway. Furthermore, the hepatoprotective activity of EAF (200 mg/kg against carbon tetrachloride (CCl4) intoxicated male Wistar rats was assessed. EAF significantly inhibited CCl4-induced elevation of alanine aminotransferase (ALT), aspartate transaminase (AST), total bilirubin (TB), total cholesterol (TC), and triglycerides (TG), in the blood serum and prevented lipid peroxidation and restored superoxide dismutase (SOD) activity and glutathione (GSH) content in liver tissues. The observed hepatoprotective effects of EAF, which were supported by histopathological observations as pretreatment with EAF, effectively attenuated the CCl4-induced histopathological changes. In conclusion, EAF of Eugenia uniflora leaves has substantial hepatoprotective activities against CCl4 induced acute liver injury in rats due to its antioxidant activity.

Protective effects of ethyl acetate fraction ofLawsonia inermisfruits extract against carbon tetrachloride-induced oxidative damage in rat liver

2013 ◽  
Vol 32 (4) ◽  
pp. 694-706 ◽  
Author(s):  
Anis Ben Hsouna ◽  
Saoudi Mongi ◽  
Gérald Culioli ◽  
Yves Blache ◽  
Zohra Ghlissi ◽  
...  
Keyword(s):  
Carbon Tetrachloride ◽  
Oxidative Damage ◽  
Ethyl Acetate ◽  
Rat Liver ◽  
Ethyl Acetate Fraction ◽  
Protective Effects

scholarly journals Evaluation of the antioxidant potentials of extract and fractions of Vitex doniana fruit and identification of the bioactive metabolites using HPLC-DAD-MS analysis

2021 ◽  
Vol 17 (1) ◽  
pp. 160-175
Author(s):  
Keziah Uchechi Ajah ◽  
Ozioma Anne Asomugha ◽  
Chinazom Perpetua Ifedigbo ◽  
Kosoluchukwu Chidubem Umeh ◽  
Onyeka Chinwuba Obidiegwu ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Liquid Chromatography ◽  
Carbon Tetrachloride ◽  
Ethyl Acetate ◽  
Ethyl Acetate Fraction ◽  
Fruit Extract ◽  
Ms Analysis ◽  
Vitex Doniana

Isoflavones and flavonoids in general found in fruits and vegetable act as natural antioxidants. This study was aimed at screening the isoflavone-rich fractions of Vitex doniana fruit extract for antioxidant activity using in vitro and in vivo models. The ethyl acetate and butanol fractions were screened for in vitro antioxidant activity by DPPH free radical scavenging and FRAP (Fe3+ Reducing Antioxidant Power) assays using ascorbic acid as standard. In vivo antioxidant activity was also tested against carbon tetrachloride-induced toxicity in mice using carbon tetrachloride (4 mL/kg) and silymarin (100 mg/kg) as standard. The ethyl acetate fraction and butanol Vacuum Liquid Chromatography (VLC) sub-fractions were subjected to High Performance Liquid Chromatography–Diode Array Detector (HPLC-DAD) and Liquid Chromatography-Mass Spectrometry (LC-MS) analyses. The ethyl acetate and butanol fractions inhibited DPPH radicals with an IC50 of 897 µg/mL and 2809.38 µg/mL respectively. Both fractions also showed mild FRAP result with EC50 of 1401 µg/mL and 7051 µg/mL respectively. The fractions produced significant decrease (P < 0.05) in ALP levels only while ethyl acetate fraction caused a significant reduction in the value of only malondialdehyde. Both fractions also increased levels of superoxide dismutase (SOD). HPLC-DAD analysis led to the detection of cinnamic acid, protocatechuic acid and two isoflavones daidzein and genistein. Further LC-MS analysis confirmed the presence of the isoflavones tectorigenin, 5-O-methylgenistein, and 5-O-methyltectorigenin, among other flavonoids, peonidin, 5-methylpeonidin, methylmalvidin and kaempferol-3-O-(2''galloyl)-glucopyranoside. The detected isoflavones and other flavonoids may contribute to the observed mild in vitro and good in vivo antioxidant activity of V. doniana fruit extract and fractions.

scholarly journals The Hypoglycemic Effects of Ethanol Leaves Extract of Cymbopogon citratus and Its Fractions in Alloxan-Induced Diabetic Mice

2020 ◽  
pp. 1-11
Author(s):  
Joyce Temu ◽  
Haikael D. Martin ◽  
Elingarami Sauli
Keyword(s):  
Body Weight ◽  
Ethyl Acetate ◽  
Water Extract ◽  
Fine Powder ◽  
Ethyl Acetate Fraction ◽  
Negative Control ◽  
Hypoglycemic Activity ◽  
Cymbopogon Citratus ◽  
The Difference ◽  
Body Weight Dose

Varieties of plants including Cymbopogon citratus are traditionally used in controlling hyperglycemia by either stimulating insulin secretion, inhibition α- Glucosidase or α-amylase activity. Cymbopogon citratus leaves were shade dried, grinded into fine powder and then extracted by cold maceration using ethanol. Fractionation was done by VLC using dichloromethane, ethyl acetate and ethanol. OGTT was performed for both crude extract and fractions. Diabetes was induced in mice by intraperitoneal injection of freshly prepared alloxan monohydrate (170 mg/kilogram body weight). The mice were treated with ethyl acetate fraction once daily at 400 mg/kilogram body weight dose for the period of 20 days.FBG and weight were then recorded in days 1, 5, 10, 15 and 20 after six hours of fasting. Safety of crude water extract and ethyl acetate fractions were evaluated in mice by using Lorke’s method, followed by 5 days observation for their mortality and behavioral changes. Comparisons of results among groups were analyzed using One-way ANOVA. The difference between the means of the two population groups (each against negative control) was considered significant at p< 0.05. Results were expressed as mean ± SD. Both crude and ethyl acetate fractions from C. citratus showed significant hypoglycemic activity. Moreover, higher hypoglycemic activity was shown by ethyl acetate fraction (p = 0.004). No mortality was observed at 5000 mg/kilogram body weight dose but sleeping and tremor were observed at a 1000 -5000 mg/kilogram body weight dose. Good hypoglycemic and safety results from ethyl acetate fraction highly suggest that Cymbopogon citratus extracts are effective against insulin-dependent hyperglycemia, which may be contributed by the action of the present of alkaloids, saponins, antraquinone, phenol and tannins. Isolation and testing of the active ingredients from the C. citratus extract are thus warranted for use in developing pharmaceutical anti-hyperglycemic drugs from this herbal plant.

scholarly journals The Effect of Methanolic Extract and Ethyl Acetate Fraction of Kepel Fruit (Stelechocarpus burahol) to α-Glutathione S-Transferase Enzyme Concentration of Rat Liver and Blood Induced by CCl4

2020 ◽  
pp. 3
Author(s):  
Yusuf Rabbani ◽  
Claude Mona Airin ◽  
Sugeng Riyanto
Keyword(s):  
Free Radical ◽  
Carbon Tetrachloride ◽  
Ethyl Acetate ◽  
Methanolic Extract ◽  
Ethyl Acetate Fraction ◽  
Positive Control ◽  
Enzyme Concentration ◽  
The Body ◽  
Female Rats ◽  
Glutathione S Transferase

Degenerative diseases are mostly caused by high free radical concentrations in the body. Kepel fruit(Stelechocarpus burahol) is known to contain flavonoid compounds, a class of compounds that has free radicalscavenging activity in the body and could affect the activity of antioxidant enzymes. There have been reportsthat methanolic extract and ethyl acetate fractions of Kepel fruit have the lowest IC50 values, compared toextracts or fractions which are made using other organic solvents, on in vitro assays so that flavonoids aresuspected as responsible compounds to the free radical scavenging activity. This study aimed to identify thepresence of flavonoid content in methanolic extract and the fraction of ethyl acetate of Kepel fruit and also itseffect on rat alpha Glutathione S-Transferase (α-GST) enzyme concentration for the function to prevent hepaticcell damage due to carbon tetrachloride exposure. The identification of flavonoids content used a series ofcolor chemical reaction tests and thin layer chromatography that used silica gel 60F254 as the solid phase andmixture of ethyl acetate, formic acid, glacial acetic acid, and water (100: 11 : 11: 20 v/v) as the mobile phase.This study used 30 Sprague-Dawley female rats (age 2-3 months) that were grouped into 5 groups as solventcontrol (CMC-Na) group, negative control (CCl4) group, positive control (vitamin C) group, methanolic extractgroup, and ethyl acetate fraction group. All groups were induced with carbon tetrachloride (CCl4) compoundsexcept for the CMC-Na solvent control group. Blood sampling at 0 h, 24 h, and 72 h, while liver sampling at 24h and 72 h, with 3 samples each group. The analysis of rat α-GST enzyme concentrations used theEnzyme-linked Immunosorbent Assay (ELISA) method. Based on the phytochemical test, the methanolicextract and the ethyl acetate fraction of Kepel fruit have been identified contains flavonoids compound. Basedon the in vivo study, the positive control group's α-GST enzyme concentration was decreased, while thenegative control group's α-GST enzyme concentration was increased. Administration of methanolic extractand ethyl acetate fraction also decreased both liver and blood rat α-GST enzyme concentrations, althoughwithout significant correlation, and still could prevent the hepatic cell damage due to carbon tetrachlorideexposure.

Assessment of Antimicrobial Activity and total Polyphenol Content of Rosa multiflora and Rosa wichuraiana

2019 ◽  
Vol 10 (04) ◽  
pp. 646-650
Author(s):  
Hyeusoo Kim ◽  
Kyeong Won Yun
Keyword(s):  
Antimicrobial Activity ◽  
Ethyl Acetate ◽  
Antimicrobial Agents ◽  
Ethyl Acetate Fraction ◽  
Rosa Multiflora ◽  
Polyphenol Content ◽  
Total Polyphenol ◽  
Water Fraction ◽  
Total Polyphenol Content ◽  
Rosa Species

The fruit of Rosa multiflora has been used as traditional herbal medicine in Asian countries. The present investigation was undertaken to study the antimicrobial activity and total polyphenol content of hexane, ether, ethyl acetate, water fraction of methanol extract of fruit and flower from Rosa multiflora and Rosa wichuraiana. Antimicrobial activity of the mentioned fractions against 3 gram-positive and 4 gram-negative bacteria using disk diffussion method. The measurement of minimal inhibition concentration (MIC) showed that the ethyl acetate fraction of the two Rosa species is the most effective against the tested bacteria. The total polyphenol content of ethyl acetate fraction of the two Rosa species is higher than the other fractions. The results indicate the antimicrobial activity was related with the total polyphenol content and the fruit and flower of the two Rosa species can be considered as a natural source of antimicrobial agents.

Antioxidant Activity, Inhibition α-Glucosidase of Ethanol Extract of Strychnos nitida G. Don and Identification of Active Compounds

2019 ◽  
Vol 5 (3) ◽  
pp. 11-20
Author(s):  
Stefani Dhale Rale ◽  
Hasim Hasim ◽  
Syamsul Falah
Keyword(s):  
Antioxidant Activity ◽  
Ethyl Acetate ◽  
Ethanol Extract ◽  
Ethyl Acetate Fraction ◽  
Inhibition Activity ◽  
Active Compounds ◽  
Activity Inhibition ◽  
Activity Test ◽  
Ic50 Value ◽  
Treatment Of Diabetes

This study aims to find the treatment of diabetes using natural materials by exploring plants in the province of East Nusa Tenggara. his research was conducted out by extracting the Strychnos nitida G.Don stem using a method of maceration by ethanol 70%. Ethanol extract was then fractionated using n-hexane and ethyl acetate. Simplicia from maceration and fractionation results were then tested for antioxidant activity, α-glucosidase inhibition activity and identification of active compounds. The results showed that ethyl acetate fraction had the lowest IC50 value of 86.83 μg / ml. Results of the α-glucosidase activity test showed that ethyl acetate fraction and n-heksan fraction at 900 ppm had the highest percentage of inhibition of 34.23% and 33.89%. Identification using LCMS/MS method showed that ethyl acetate fraction consist of Benzenemethamine, N, N-dioctyl- as an antioxidantcompound and compound 24-methyl-5-cholestone-hexol as an antidiabetic compound. From the results of this study, we concluded that the extract of kayu ular Strychnos nitida G.Don stem has inhibition activity toward α-glucosidase enzyme.

Sign in / Sign up

Export Citation Format

Share Document