peak elution volume (time)in column chromatography, V̄R, t̄R

We have devised a method to fractionate low density lipoprotein (LDL) into subspecies by means of column chromatography. DEAE-agarose columns, 2.6 × 60 cm, were loaded with LDL (25–45 mg LDL protein) and eluted with a 0.045–0.13 M NaCl gradient. The LDL eluted over a volume of 900 ml. Specific portions of the eluted LDL, reapplied to a column identical with the original, reelute at about the same point. Altering the NaCl concentration of the elution fluid changed the elution volume. The cholesterol-protein ratio of the LDL subfractions was progressively lower in fractions eluting at higher NaCl concentrations. These results indicate the LDL is not a homogenous lipoprotein species but consists of subfractions which differ in at least charge and cholesterol content.

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Purification and properties of bovine pituitary follitropin

Biochemical Journal ◽

10.1042/bj1590651 ◽

1976 ◽

Vol 159 (3) ◽

pp. 651-659 ◽

Cited By ~ 19

Author(s):

K W Cheng

Keyword(s):

Amino Acid ◽

Column Chromatography ◽

Gel Filtration ◽

Deae Cellulose ◽

Elution Volume ◽

Purification And Properties ◽

Receptor Assay ◽

Ion Exchange Column ◽

Pituitary Glands ◽

Radioligand Receptor Assay

A reproducible procedure was developed for the purification of follitropin from frozen bovine pituitary glands. The method involved precipitation with (NH4)2SO4 and acetone, followed by ion-exchange column chromatography on CM-cellulose and DEAE-cellulose and gel filtration on Sephadex G-100. A specific radioligand-receptor assay for follitropin was used to locate the activity in eluates after column chromatography and gel filtration. The potency of the highly purified bovine follitropin as measured by Steelman-Pohley bioassay was 164 times that of NIH-FSH-S1 standard preparation. They yield of bovine follitropin was 2.9 mg/kg of frozen pituitary glands. Electrophoretically, bovine follitropin was more acidic in nature and migrated further towards the anode than lutropin and thyrotropin. The elution volume of bovine follitropin by gel filtration on Sephadex G-100 was very similar to that of bovine lutropin. The amino acid composition of bovine follitropin was similar to that of sheep and human follitropin, being rich in lysine, aspartic acid, threonine, serine, glutamic acid and half-cystine.

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Heparin Excretion in Intact and Hepatectomized Rats

Thrombosis and Haemostasis ◽

10.1055/s-0038-1650152 ◽

1981 ◽

Vol 45 (02) ◽

pp. 146-149

Author(s):

Ray Losito ◽

Harry Gattiker ◽

Ginette Bilodeau

Keyword(s):

Column Chromatography ◽

Normal Values ◽

Anticoagulant Activity ◽

Biliary Fistula ◽

Kinetics Of ◽

Intact Rats

SummaryMetabolism and kinetics of 3H-heparin were compared in intact and hepatectomized rats. Rats were divided into three groups: 1) intact rats with biliary fistulas and cystostomies 2) intact rats with only cystostomies and 3) hepatectomized rats with cystostomies. Radioactivity in blood, bile and urine besides anticoagulant activity in blood and urine were examined. In addition, column chromatography of urine was used to isolate possible metabolites. Seventy percent and 80% of the radioactive dose was found in the urine of intact rats at 24 hr and 48 hr. Close to 5% of the radioactivity was found in bile or rats with a biliary fistula after 48 hr. The APTT declined to near normal values at 1 hr whether rats had a biliary fistula or not. In contrast, only 25 % of the radioactivity could be excreted into the urine of hepatectomized rats in 24 hr; the APTT did not decline as fast and at 5 hr, it was still 100 seconds. Only one radioactive component could be isolated on chromatography from all urines of these animals and appears to be similar to the original heparin. Thus, the liver has an important role to play in regulating the anticoagulant effects and excretion of heparin.

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VANILLINSÄURE ALS ENDPRODUKT DES STOFFWECHSELS VON ADRENALIN UND NORADRENALIN. II.

Acta Endocrinologica ◽

10.1530/acta.0.0440101 ◽

1963 ◽

Vol 44 (1) ◽

pp. 101-106 ◽

Cited By ~ 4

Author(s):

Wilhelm Dirscherl ◽

Helmut Thomas

Keyword(s):

Rat Liver ◽

Paper Chromatography ◽

Column Chromatography ◽

Vanillic Acid ◽

Hippuric Acid ◽

Single Spot ◽

Solvent Systems ◽

Perfusion Experiment ◽

Kinetics Of

ABSTRACT Perfusion of rat liver with vanillic acid yielded only one metabolite. In paper chromatography with three different solvent systems, the substance showed the same RF-values as vanillyolglycine (3-methoxy-4-hydroxyhippuric acid) and in mixed chromatograms there was only one single spot. After separation by column chromatography, the UV- and IRspectra of the reaction product were identical with those of 3-methoxy4-hydroxy-hippuric acid. During the perfusion experiment, the kinetics of the conjugation were investigated.

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DIE ZUSAMMENSETZUNG DER JODHALTIGEN VERBINDUNGEN UND IHRE BEZIEHUNG ZUM JODUMSATZ IN EUTHYREOTEN STRUMEN

Acta Endocrinologica ◽

10.1530/acta.0.0410584 ◽

1962 ◽

Vol 41 (4) ◽

pp. 584-594 ◽

Cited By ~ 2

Author(s):

Dankwart Reinwein ◽

Erich Klein

Keyword(s):

Column Chromatography ◽

Iodine Content ◽

High Plasma ◽

Secretion Rate ◽

Total Iodine ◽

Iodine Compound ◽

Hormonal Secretion ◽

Carrier Free ◽

Iodine Metabolism ◽

Diffuse Goiter

ABSTRACT The iodine metabolism was investigated in 29 patients with euthyroid non-endemic diffuse goiter. 1 to 14 days before thyroidectomy the patients received carrier-free 131I. The chemical iodine fractions (PBI, BEI and iodide) of the thyroid and the labelled iodine compound were analyzed by means of paper- and column chromatography. In one gland the total iodine content varied only by ± 19.6% of the average, the relative shares of PBI, BEI and iodide as well as that of the iodoamino acids being equal. Monoiodotyrosine, diiodotyrosine and thyroxine were found in the thyroid homogenate without hydrolysis. The homogenate after hydrolysis contained more iodotyrosines at the expense of iodothyronines than do normal glands. 17 goiters with normal 131I-uptake showed a high total iodine content (14.2 ± 5.0 mg) whilst 9 goiters with an increased 131I-uptake had low values (3.58 ± 0.6 mg). The opposite was found for the relative shares of BEI with the chromatographically isolated iodothyronines thyroxine, triiodothyronine and an unidentified iodine compound. Goiters with »high plasma PB131I« were characterized by a faster transfer of 131I into the more heavily iodinated compounds than is found in glands with a normal hormonal secretion rate. The highest values for the iodothyronines were found in goiters with increased 131I-uptake together with a high hormonal secretion rate. From this study it appears that the changes in the iodine-poor glands are due to a defective exo- or endogenous iodine supply. The observed alterations in iodine-rich glands are probably induced by a faulty iodine utilization characterized by an incomplete iodotyrosyl-coupling defect.

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Protein Solvent Interaction: Transition of Protein-solvent Interaction Concept from Basic Research into Solvent Manipulation of Chromatography

Current Protein and Peptide Science ◽

10.2174/1389203718666171024121529 ◽

2018 ◽

Vol 20 (1) ◽

pp. 34-39 ◽

Cited By ~ 1

Author(s):

Tsutomu Arakawa ◽

Yoshiko Kita

Keyword(s):

Column Chromatography ◽

Rapid Development ◽

Academic Research ◽

Basic Research ◽

Protein Stabilization ◽

Special Issue ◽

Solvent Interaction ◽

Protein Formulations ◽

Solvent Additive ◽

Downstream Processes

Previously, we have reviewed in this journal (Arakawa, T., Kita, Y., Curr. Protein Pept. Sci., 15, 608-620, 2014) the interaction of arginine with proteins and various applications of this solvent additive in the area of protein formulations and downstream processes. In this special issue, we expand the concept of protein-solvent interaction into the analysis of the effects of solvent additives on various column chromatography, including mixed-mode chromatography. Earlier in our research, we have studied the interactions of such a variety of solvent additives as sugars, salts, amino acids, polymers and organic solvents with a variety of proteins, which resulted in mechanistic understanding on their protein stabilization and precipitation effects, the latter known as Hofmeister series. While such a study was then a pure academic research, rapid development of genetic engineering technologies and resultant biotechnologies made it a valuable knowledge in fully utilizing solvent additives in manipulation of protein solution, including column chromatography.

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A Cytotoxic Natural Product from Patrinia villosa Juss

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520619666190416101014 ◽

2019 ◽

Vol 19 (11) ◽

pp. 1399-1404 ◽

Cited By ~ 1

Author(s):

Yangcheng Liu ◽

Wei Liu ◽

Changlan Chen ◽

Zheng Xiang ◽

Hongwei Liu

Keyword(s):

Cell Cycle ◽

Cell Lines ◽

Silica Gel ◽

Column Chromatography ◽

Bioactive Compound ◽

Antitumor Agent ◽

2D Nmr ◽

Preparative Hplc ◽

Dependent Manner ◽

2D Nmr Spectra

Background and Purpose:: Patrinia villosa Juss is an important Chinese herbal medicine widely used for thousands of years, but few reports on the ingredients of the herb have been presented. In this study, we aim to isolate the bioactive compound from the plant. Material and Methods:: The air-dried leaves of P. villosa (15kg) were extracted three times with 70% EtOH under reflux. The condensed extract was suspended in H2O and partitioned with light petroleum, dichloromethane and n-BuOH. The dichloromethane portion was then subjected to normal-phase silica gel column chromatography, ODS silica gel column chromatography and semi-preparative HPLC to yield compound 1. Cytotoxicities of 1 were assayed on HepG2, A549 and A2780 cell lines. The mechanism of apoptosis and cell cycle on A549 was confirmed subsequently. Results: A new impecylone (Impecylone A) was isolated from the leaves of Patrinia villosa Juss, and its structures were established using 1D, 2D-NMR spectra and HR-ESI-MS. Impecylone A could selectivity inhibit HepG2 and A549 cell lines. The compound could induce apoptosis of A549 and arrest the cell cycle at G2/M phase in a dose-dependent manner. Conclusion: Impecylone A is a novel compound from Patrinia villosa Juss and could be a potential antitumor agent especially in the cell lines of A549.

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Isolation of Sinapic Acid from Habenaria intermedia D. Don: A New Chemical Marker for the Identification of Adulteration and Substitution

Current Traditional Medicine ◽

10.2174/2215083804666181030101709 ◽

2020 ◽

Vol 6 (4) ◽

pp. 380-387

Author(s):

Jaswinder Kaur Virk ◽

Vikas Gupta ◽

Mukesh Maithani ◽

Ravindra K. Rawal ◽

Sanjiv Kumar ◽

...

Keyword(s):

Column Chromatography ◽

Sinapic Acid ◽

Phytochemical Screening ◽

Chemical Marker ◽

Reference Compound ◽

Crystalline Powder ◽

Chemical Test ◽

Herbal Plants ◽

Single Compound ◽

Layer Chromatography

Background: Vriddhi is one of the Rasayana herbs in Ayurveda broadly used in vitality, strengthening Ayurvedic formulations. To fulfill steeply increased demand and declined supply, tubers have been collected in destructive manner resulting in reduced plant population and pushing the plant in Red list of IUCN endangered species. However, manufacturers are using substitutes and other substandard drugs leading to adulteration which puts the importance of therapeutically rich herbal plants at stake. Lack of chemical markers is the main inability of regulatory authorities for not taking any action against this adulteration. Objective: Isolation of chemical marker of plant that can be used as a reference compound for identification of unauthorized substitution. Methods: Preliminary phytochemical screening of methanolic and toluene extract of H. intermedia D. Don was done using standard methods followed by column chromatography for the isolation of phytoconstituents. A total of 3004 fractions were collected with Thin Layer Chromatography (TLC) profiling and different fractions were pooled. A single compound was isolated and confirmed by chemical test, melting point, spectral analysis and compared with the literature. Results: Phytochemical screening of extracts shows the presence of alkaloids, carbohydrates, steroids, terpenoids, flavonoids, tannins and phenolics. A pure white crystalline powder was isolated by column chromatography which was characterized as 3,5-dimethoxy-4- hydroxycinnamic acid (Sinapic acid) with the help of IR and Mass spectroscopy. Conclusion: This is the first report of Sinapic acid as a novel compound from Vriddhi, Habenaria genus and Orchidaceae family. It can be used as a marker for the identification of unauthorized substitution and adulteration claiming the use of Vriddhi.

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Phytoceuticals in aniseeds: a practical approach for isolation and identification

International Journal of Bioassays ◽

10.21746/ijbio.2016.01.002 ◽

2016 ◽

Vol 5 (01) ◽

pp. 4701

Author(s):

Shobha R. I. ◽

Andallu B.

Keyword(s):

Chronic Diseases ◽

Column Chromatography ◽

High Performance ◽

Methanolic Extract ◽

Analytical Techniques ◽

Therapeutic Agents ◽

Systematic Research ◽

Isolation And Identification ◽

Traditional System ◽

Pharmacologically Active

Nature has provided mankind with a broad and structurally diverse array of pharmacologically active chemical compounds, phytoceuticals, which have proved to be indispensable for the cure of chronic diseases or as lead structures for novel therapeutic agents. Almost, 70% of modern medicines in India have been developed from plants used in the traditional system of medicine. With the technological advances and the development of more sophisticated isolation and analytical techniques, there is great scope for further systematic research to screen and isolate many more phytoceuticals which might be more effective/as effective as synthetic drugs and thereby assess their potential in protecting against chronic diseases. The present investigation is aimed at isolation and identification of phytoceuticals in a spice, aniseed (Pimpinella anisum L), therapeutically less-exploited and widely used only for culinary purpose, using analytical techniques viz. column chromatography, high performance thin layer chromatography (HPTLC.) and nuclear magnetic resonance spectrometry. Among all the solvent fractions of methanolic extract tested, methanolic extract and ethyl acetate fraction possessed highest amounts of bioactive compounds viz. phenolics, flavonoids, flavonols which was confirmed by qualitative, quantitative and HPTLC. analyses. Spectral analysis using NMR. of one of the sub-fractions of aniseeds obtained by column chromatography, revealed the presence of a glycosylated flavone, luteolin-6C-glucoside in aniseeds. The present study revealed aniseeds to be a great source of bioactive phytoceuticals which can be novel candidates for development of new therapeutic agents.

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Antioxidant Activity of the Isoflavonoids from the Roots of Maackia amurensis

Natural Product Communications ◽

10.1177/1934578x1300800511 ◽

2013 ◽

Vol 8 (5) ◽

pp. 1934578X1300800 ◽

Cited By ~ 1

Author(s):

Nadezda I. Kulesh ◽

Sergey A. Fedoreyev ◽

Marina V. Veselova ◽

Natalia P. Mischenko ◽

Vladimir A. Denisenko ◽

...

Keyword(s):

Oxidative Stress ◽

Antioxidant Activity ◽

Glutathione Peroxidase ◽

Column Chromatography ◽

Thiobarbituric Acid ◽

Formalin Injection ◽

Thiobarbituric Acid Reactive Substances ◽

H Nmr ◽

Maackia Amurensis

Seven isoflavonoids, including a new glycoside, (6a R,11a R)-medicarpin-3- O-gentiobioside (6), were isolated from the roots of Maackia amurensis using repeated column chromatography on a Toyopearl HW-50F sorbent and identified by HPLC–PDA–MS, 1H NMR, 13C, 1H–1H COSY, HSQC NMR and HMBC NMR analyses as daidzin (1), genistein-7- O-gentiobioside (2), pseudobaptigenin-7- O-gentiobioside (3), formononetin-7- O-gentiobioside (4), (6a R,11a R)-maackiain-3- O-gentiobioside (5), and 5- O-methylgenistein-7- O-gentiobioside (7). In the model of oxidative stress induced by formalin injection, the isolated isoflavone and pterocarpan glucosides 1-7 were shown to reduce the formation of malondialdehyde (MDA) and other thiobarbituric acid reactive substances (TBARS), as well as glutathione peroxidase (GPO) activity in rats.

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