scholarly journals Production of Fucoxanthin from Phaeodactylum tricornutum Using High Performance Countercurrent Chromatography Retaining Its FOXO3 Nuclear Translocation-Inducing Effect

Marine Drugs ◽  
2021 ◽  
Vol 19 (9) ◽  
pp. 517
Author(s):  
Daniela Bárcenas-Pérez ◽  
Antonín Střížek ◽  
Pavel Hrouzek ◽  
Jiří Kopecký ◽  
Marta Barradas ◽  
...  
Keyword(s):  
Phaeodactylum Tricornutum ◽  
High Performance ◽  
Nuclear Translocation ◽  
Solvent System ◽  
Biologically Active ◽  
Countercurrent Chromatography ◽  
Reverse Phase ◽  
General Process ◽  
Lower Phase ◽  
Isolation Methods

Phaeodactylum tricornutum is a rich source of fucoxanthin, a carotenoid with several health benefits. In the present study, high performance countercurrent chromatography (HPCCC) was used to isolate fucoxanthin from an extract of P. tricornutum. A multiple sequential injection HPCCC method was developed combining two elution modes (reverse phase and extrusion). The lower phase of a biphasic solvent system (n-heptane, ethyl acetate, ethanol and water, ratio 5/5/6/3, v/v/v/v) was used as the mobile phase, while the upper phase was the stationary phase. Ten consecutive sample injections (240 mg of extract each) were performed leading to the separation of 38 mg fucoxanthin with purity of 97% and a recovery of 98%. The process throughput was 0.189 g/h, while the efficiency per gram of fucoxanthin was 0.003 g/h. Environmental risk and general process evaluation factors were used for assessment of the developed separation method and compared with existing fucoxanthin liquid-liquid isolation methods. The isolated fucoxanthin retained its well-described ability to induce nuclear translocation of transcription factor FOXO3. Overall, the developed isolation method may represent a useful model to produce biologically active fucoxanthin from diatom biomass.

scholarly journals Separation of the Glycosylated Carotenoid Myxoxanthophyll from Synechocystis Salina by HPCCC and Evaluation of Its Antioxidant, Tyrosinase Inhibitory and Immune-Stimulating Properties

Separations ◽  
2020 ◽  
Vol 7 (4) ◽  
pp. 73
Author(s):  
Michaela Nováková ◽  
Tereza Fábryová ◽  
Doris Vokurková ◽  
Iva Dolečková ◽  
Jiří Kopecký ◽  
...  
Keyword(s):  
High Performance ◽  
Large Scale ◽  
Solvent System ◽  
Inhibitory Effect ◽  
Scale Production ◽  
Unicellular Cyanobacterium ◽  
Lower Phase ◽  
Large Scale Production ◽  
Global Demand ◽  
Potential Benefits

Global demand for natural pigments has increased in the past few years. Myxoxanthophyll, a glycosylated monocyclic carotenoid, is a pigment that occurs naturally in cyanobacteria but no scalable isolation process has been developed to obtain it from its natural source to date. In this study, myxoxanthophyll was isolated from unicellular cyanobacterium Synechocystis salina (S. salina) using high-performance countercurrent chromatography (HPCCC), where the lower phase of the biphasic solvent system composed of n-heptane–ethanol–water (2:4:4, v/v/v) was used as a mobile phase, whereas its upper phase was the stationary phase. For the HPCCC isolation, a multi-injection method was developed, and four consecutive sample injections (70 mg each) were performed, obtaining, in total, 20 mg of myxoxanthophyll, which was finally purified with high-performance liquid chromatography (HPLC). Overall, a final myxoxanthophyll yield of 15 mg (98% purity) was obtained. The target pigment showed a weak antioxidant and tyrosinase inhibitory effect, and exhibited immune-stimulating properties by activating human granulocytes. The results presented here form a basis for the large-scale production of myxoxanthophyll, and show the potential benefits of this pigment for human health.

Distribution and Expression of Adrenomedullin in Human Gastrointestinal Tissue

1998 ◽  
Vol 35 (5) ◽  
pp. 643-648 ◽  
Author(s):  
Michitaka Kitani ◽  
Junichiro Sakata ◽  
Yujiro Asada ◽  
Kazuo Kitamura ◽  
Tanenao Eto
Keyword(s):  
High Performance ◽  
Northern Blot Analysis ◽  
Immunohistochemical Study ◽  
Gel Filtration ◽  
Biologically Active ◽  
Gastrointestinal Hormone ◽  
Reverse Phase ◽  
Gastrointestinal Tissue ◽  
Biologically Active Peptide

Adrenomedullin (AM) is a biologically active peptide recently isolated from phaeochromocytoma. We report here the distribution and characterization of immunoreactive AM and gene expression of AM in human gastrointestinal tissue. Using a sensitive radioimmunoassay system for the peptide, immunoreactive human AM was detected in the stomach, duodenum, jejunum, ileum and colon. The AM concentration of these tissues was about 0.4–0.8 pmol/g wet tissue. Reverse phase and gel filtration high-performance liquid chromatographies showed that most of the immunoreactive AM in stomach and jejunum was identical to authentic human AM. By northern blot analysis, human AM mRNA was found to be expressed ubiquitously in the human gastrointestinal tissues. Furthermore, an immunohistochemical study revealed that immunoreactive AM cells were present in the gastrointestinal glands. These results suggest that AM may play some role as a gastrointestinal hormone.

Isolation and Purification of Aloin a and Aloin B by High-Speed Counter-Current Chromatography

2013 ◽  
Vol 634-638 ◽  
pp. 1241-1246 ◽  
Author(s):  
Shi Rong Tang ◽  
Shang Long Chen ◽  
Sang Sang Lu ◽  
Xin Yang Hu
Keyword(s):  
High Speed ◽  
High Performance ◽  
Volume Ratio ◽  
Solvent System ◽  
Two Phase ◽  
Lower Phase ◽  
Isolation And Purification ◽  
Counter Current ◽  
Chloroform Methanol ◽  
Crude Methanol Extract

High-speed counter-current chromatography(HSCCC) was successfully used for isolation and purification of Aloin A and Aloin B from the crude methanol extract of Aloe with a two-phase solvent system composed of chloroform–methanol–n-butylalcohol-water at an optimized volume ratio of 4:3:1:2 (v/v/v/v). The lower phase was used as the mobile phase in the head to tail elution mode. The preparative HSCCC separation was performed on 180 mg of the crude extract yielding pure Aloin A(18mg) and Aloin B(16mg) at purities of 95.2% and 96.8%, respectively, as determined by high performance liquid chromatography (HPLC). HSCCC is a powerful technique for isolation and separation of chemical composition from aloe.

Isolation and Purification of Ginkgo Flavonoids by High-Speed Counter-Current Chromatography

2013 ◽  
Vol 781-784 ◽  
pp. 741-745
Author(s):  
Sang Sang Lu ◽  
Hui Song ◽  
Jing Zhi Miao ◽  
Shi Rong Tang
Keyword(s):  
Mobile Phase ◽  
High Speed ◽  
High Performance ◽  
Leaf Extract ◽  
Volume Ratio ◽  
Solvent System ◽  
Two Phase ◽  
Lower Phase ◽  
Isolation And Purification ◽  
Counter Current

High-speed counter-current chromatography (HSCCC) was successfully used for isolation and purification of Ginkgo flavonoids from the Ginkgo biloba L. leaf extract (GBE) with a two-phase solvent system composed of n-hexaneethyl acetatemethanolwater at an optimized volume ratio of 4:6:5:5(v/v/v/v). The lower phase was used as the mobile phase in the head to tail elution mode. The preparative HSCCC separation was performed on 200 mg of GBE yielding pure Quercetin (22mg), Kaempferol (15mg) and Isorhamnetin (4mg) at purities of 96.6%, 92.3% and 93.6%, respectively, as determined by high performance liquid chromatography (HPLC). HSCCC is a powerful technique for isolation and separation of chemical composition from GBE.

High Performance Liquid Chromatographic Determination of Subsidiary Colors in FD&C Red No. 3

1982 ◽  
Vol 65 (5) ◽  
pp. 1080-1085
Author(s):  
Robert J Calvey ◽  
Allen L Goldberg
Keyword(s):  
High Performance ◽  
Chromatographic Determination ◽  
Total Sample ◽  
Solvent System ◽  
High Performance Liquid Chromatographic ◽  
Phase Method ◽  
Reverse Phase ◽  
Liquid Chromatographic Determination ◽  
Liquid Chromatographic

Abstract Two rapid, sensitive, reproducible methods that use a Zorbax C-8 reverse phase column and high performance liquid chromatography are described for the determination of the subsidiary colors in FD&C Red No. 3. With the first method, 8 subsidiary colors (fluorescein, 2'-iodofluorescein, 4'-iodofluorescein, 2',5'-diiodofluorescein, 2',7'-diiodofluorescein, 4',- 5'-diiodofluorescein, 2',4',7'-triiodofluorescein, and 2',4',5'-triiodofluorescein) are eluted in a reproducible pattern by increasing the organic nature of a buffered mobile phase. Method 1 is capable of quantitating all the subsidiary colors except 4'-iodofluorescein and 4',5'-diiodofluorescein. If these 2 subsidiary colors are seen, the sample must be run again by method 2, which uses a different program and solvent system to quantitate them. The average recoveries for the 6 subsidiary colors quantitatively determined in FD&C Red No. 3 by method 1 ranged from 96 to 98%. The average recoveries for 4'-iodofluorescein and 4',5'-diiodofluorescein, quantitatively determined in FD&C Red No. 3 by method 2, were 101 and 103%, respectively. The amounts of the 6 subsidiary colors recovered by method 1 were 0.04-7.4% by weight of the total sample. The amounts of the 2 subsidiary colors recovered by method 2 were 0.13-2.6% by weight of the total sample.

scholarly journals Methodology for Porphyrin Isolation by High-Performance Countercurrent Chromatography

2014 ◽  
Vol 9 (6) ◽  
pp. 1934578X1400900
Author(s):  
Amaro C. Ramos ◽  
Fernanda S. Neves ◽  
Maria Raquel G. Vega ◽  
Edmilson J. Maria ◽  
Rodrigo R. Oliveira’
Keyword(s):  
Methyl Ester ◽  
High Performance ◽  
Solvent System ◽  
Distribution Coefficients ◽  
Countercurrent Chromatography ◽  
Pheophorbide A ◽  
Resolution Factor ◽  
Enhanced Solubility ◽  
Versatile Technique ◽  
Heterocyclic Structure

Countercurrent chromatography is a versatile technique for the isolation of a wide variety of plant substances. However, little attention has been devoted to the application of this technique for the isolation of porphyrins. This class of compounds are of great importance in the medical area and in photocatalysis due to their heterocyclic structure, composed of four modified pyrrol subunits interconnected on their α carbon atoms by methinic bridges. The methanol extract of Gallesia integrifolia was partitioned using different solvents; the dichloromethane fraction was then submitted to countercurrent chromatography. The solvent system composed of n-hexane, ethyl acetate, methanol, and water (1:2.5:2.5:1) was chosen to perform the chromatographic analysis due to the enhanced solubility and the best distribution coefficients of the target compounds. Two porphyrins were isolated by this method and identified as 132-hydroxypheophorbide a methyl ester and pheophorbide a, methyl ester. The solvent system proposed provided good distribution coefficients for both substances (1.27 and 1.87, respectively), and a high resolution factor.

Box-Behnken Design for Extraction Optimization Followed by High Performance Countercurrent Chromatography: Production of a Flavonoid-enriched Fraction from Achyrocline Satureioides

Planta Medica ◽  
2019 ◽  
Vol 86 (02) ◽  
pp. 151-159 ◽  
Author(s):  
Vanessa Pittol ◽  
George González Ortega ◽  
Eduarda Doneda ◽  
Sara Elis Bianchi ◽  
Marí Castro Santos ◽  
...  
Keyword(s):  
High Performance ◽  
Extraction Procedure ◽  
Solvent System ◽  
Biological Properties ◽  
Extraction Yield ◽  
Quadratic Term ◽  
Solvent Ratio ◽  
Countercurrent Chromatography ◽  
Box Behnken Design ◽  
Achyrocline Satureioides

AbstractThe biological properties of Achyrocline satureioides have been mostly ascribed to its major flavonoids quercetin (QCT), luteolin (LUT), and 3-O-methylquercetin (3OMQ). The present study aimed to optimize the extraction by dynamic maceration of the major phenolic compounds in order to obtain in a subsequent step a flavonoid-enriched fraction (FEF) using high performance countercurrent chromatography (HPCCC). A 3-level Box-Behnken design (BBD) was applied to maximize the extraction of the substances, using the plant : solvent ratio (X1 ), extraction time (X2 ), and ethanol concentration (X3 ) as factors. One-step HPCCC semipreparative separation with a solvent system composed of hexane : ethyl acetate : methanol : water (0.9 : 0.9 : 0.8 : 1.0, v/v) was employed to obtain the FEF. The second-order polynomial model was able to fit the experimental data adequately. The linear and quadratic terms of X3 were the most significant factors that affected all the responses. The positive linear term of X3 indicated a substantial increase in extraction yield, while the negative quadratic term showed a nonlinear tendency. Linear terms of X1 suggested a tendency to solvent saturation, except for QCT. The terms of X2 did not affect the responses substantially. The HPCCC method was found to be efficient and rapid for separating the FEF with 71% (w/w) flavonoid content. Overall, the developed extraction procedure coupled with HPCCC proved to be efficient for obtaining an enriched fraction with a very high content of flavonoids from A. satureioides.

scholarly journals Laguncularia racemosa Phenolics Profiling by Three-Phase Solvent System Step-Gradient Using High-Performance Countercurrent Chromatography with Off-Line Electrospray Mass-Spectrometry Detection

Molecules ◽  
2021 ◽  
Vol 26 (8) ◽  
pp. 2284
Author(s):  
Fernanda das Neves Costa ◽  
Gerold Jerz ◽  
Peter Hewitson ◽  
Fabiana de Souza Figueiredo ◽  
Svetlana Ignatova
Keyword(s):  
Mass Spectrometry ◽  
High Performance ◽  
Gradient Elution ◽  
Solvent System ◽  
Laguncularia Racemosa ◽  
Phase System ◽  
Ionization Mass ◽  
Countercurrent Chromatography ◽  
Esi Ms ◽  
Three Phase

The detailed metabolite profiling of Laguncularia racemosa was accomplished by high-performance countercurrent chromatography (HPCCC) using the three-phase system n-hexane–tert-butyl methyl ether–acetonitrile–water 2:3:3:2 (v/v/v/v) in step-gradient elution mode. The gradient elution was adjusted to the chemical complexity of the L. racemosa ethyl acetate partition and strongly improved the polarity range of chromatography. The three-phase solvent system was chosen for the gradient to avoid equilibrium problems when changing mobile phase compositions encountered between the gradient steps. The tentative recognition of metabolites including the identification of novel ones was possible due to the off-line injection of fractions to electrospray ionization mass spectrometry (ESI-MS/MS) in the sequence of recovery. The off-line hyphenation profiling experiment of HPCCC and ESI-MS projected the preparative elution by selected single ion traces in the negative ionization mode. Co-elution effects were monitored and MS/MS fragmentation data of more than 100 substances were used for structural characterization and identification. The metabolite profile in the L. racemosa extract comprised flavonoids, hydrolysable tannins, condensed tannins and low molecular weight polyphenols.

Rapid scaleup of high performance countercurrent chromatography from bench to kilogram

Planta Medica ◽  
2012 ◽  
Vol 78 (11) ◽  
Author(s):  
GH Harris ◽  
C De Amicis ◽  
NA Edwards ◽  
MB Giles ◽  
P Hewitson ◽  
...  
Keyword(s):  
High Performance ◽  
Countercurrent Chromatography

Preparation and Characterization of NH2-Terminal Fibrinogen Bβ Fragments from N-DSK of Human Fibrinogen

1984 ◽  
Vol 51 (01) ◽  
pp. 016-021 ◽  
Author(s):  
S Birken ◽  
G Agosto ◽  
B Lahiri ◽  
R Canfield
Keyword(s):  
High Performance ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Reverse Phase ◽  
Human Fibrinogen ◽  
Human Volunteers ◽  
Cnbr Cleavage ◽  
Two Peaks

SummaryIn order to investigate the early release of NH2-terminal plasmic fragments from the Bβ chain of fibrinogen, substantial quantities of Bβ 1-42 and Bβ 1-21 are required as immunogens, as radioimmunoassay standards and for infusion into human volunteers to determine the half-lives of these peptides. Towards this end methods that employ selective proteolytic cleavage of these fragments from fibrinogen have been developed. Both the N-DSK fragment, produced by CNBr cleavage of fibrinogen, and Bβ 1-118 were employed as substrates for plasmin with the finding of higher yields from N-DSK. Bβ 1-42 and Bβ 1-21 were purified by gel filtration and ion-exchange chromatography on SP-Sephadex using volatile buffers. When the purified preparation of Bβ 1-42 was chromatographed on reverse-phase high performance liquid chromatography, two peaks of identical amino acid composition were separated, presumably due either to pyroglutamate or to amide differences.

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