Isolation and purification of recombinant human plasminogen Kringle 5 by liquid chromatography and ammonium sulfate salting-out

2013 ◽  
Vol 28 (7) ◽  
pp. 957-965 ◽  
Author(s):  
Liujiao Bian ◽  
Xu Ji ◽  
Wei Hu
Keyword(s):  
Liquid Chromatography ◽  
Ammonium Sulfate ◽  
Salting Out ◽  
Isolation And Purification ◽  
Kringle 5 ◽  
Human Plasminogen

scholarly journals DEVELOPMENT OF RECOMBINANT PEPTIDE TECHNOLOGY WITH ANTIMICROBIAL PROPERTIES OF A BROAD ACTION SPECTRUM

2017 ◽  
pp. 3-14
Author(s):  
Alexander Prosekov ◽  
Alexander Prosekov ◽  
Olga Babich ◽  
Olga Babich ◽  
Irina Milenteva ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Ammonium Sulfate ◽  
Action Spectrum ◽  
Antimicrobial Properties ◽  
Hplc Method ◽  
Salting Out ◽  
Antimicrobial Spectrum ◽  
Recombinant Peptide ◽  
Isolation And Purification ◽  
Protein Nature

Bacteriocins are antibacterial, mainly complex, substances of protein nature. The promising strains producing bacteriocins used in the food industry are lactic acid microorganisms. This study examines the development of a technology for the production of a recombinant peptide with broad-spectrum antimicrobial properties. An important step is the isolation and purification of the recombinant peptide. It has been proved that the highest antimicrobial activity is manifested by a recombinant peptide isolated by a method based on salting out with ammonium sulfate. During the purification of the recombinant bacteriocin preparation, three kinds of columns were used. In the purification process, the volume of bacteriocin produced decreases 3-fold, while the RU/mL increases 3-fold, and RU/mg increases 6-fold. Purification allows the use of a smaller amount of recombinant bacteriocin in technologies with greater efficacy. Based on the results of determining the molecular weight and purity of the recombinant bacteriocin, it was found that the molecular weight of the recombinant bacteriocin having the amino acid sequence: KYYGNGVTCCKHSCSVDXGKASSCIINNGAMAXATGGH GGNHCCGMSRYIQGIPDFLRGYLHGISSANKHKKGRL, is 13 kDa. A technology for the preparation of a broad-action antimicrobial spectrum peptide has been developed. The process of production of antimicrobial peptide includes such stages as: cultivation of the recombinant strain of Escherichia coli BL21DE3; separation of biomass from the nutrient medium; precipitation of bacteriocins by ammonium sulfate; centrifugation; washing the precipitate; centrifugation at 4200 rpm and separation of the preparation; purification of bacteriocins by HPLC method; packing in bags of polymeric and combined materials; storage at a temperature of 18±2°C for 12 months.

Extraction, Cleanup, and Quantitative Determination of Aflatoxins in Corn

1980 ◽  
Vol 63 (3) ◽  
pp. 631-633 ◽  
Author(s):  
James E Thean ◽  
David R Lorenz ◽  
David M Wilson ◽  
Kathleen Rodgers ◽  
Richard C Gueldner
Keyword(s):  
High Pressure ◽  
Liquid Chromatography ◽  
Quantitative Determination ◽  
Ammonium Sulfate ◽  
Silica Gel ◽  
Normal Phase ◽  
Aqueous Methanol ◽  
Normal Phase Hplc ◽  
Water Saturated

Abstract A method is proposed for extraction and cleanup of corn samples for the quantitation of 4 aflatoxins by high pressure liquid chromatography (HPLC). After aqueous methanol extraction, ammonium sulfate treatment, and partition of aflatoxins into chloroform, sample extracts are partially purified on Sep-Pak cartridges or small columns packed with HPLC grade silica; cleanup requires only 13 mL solvent/sample. Aflatoxins B1, B2, G1, and G2 in the purified extract are resolved in ca 10 min by normal phase HPLC on a microparticulate (5 μm) silica gel column with a 50% water-saturated chloroform-cyclohexaneacetonitrile- ethanol solvent, and are measured by ultraviolet fluorescence in a silica gel-packed flowcell. Recoveries of added aflatoxins B1, B2, G1, and G2 were 84–118 % at levels of 1.5–125 μg/kg

High Performance Liquid Chromatography Coupled with Radioactivity Detection: A Powerful Tool for Determining Drug Metabolite Profiles in Biological Fluids

1986 ◽  
Vol 41 (1-2) ◽  
pp. 115-125 ◽  
Author(s):  
K.-O. Vollmer ◽  
W. Klemisch ◽  
A. von Hodenberg
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Biological Fluids ◽  
Gradient Elution ◽  
Reversed Phase ◽  
Isolation And Purification ◽  
Carbon 14 ◽  
Tracer Techniques ◽  
Metabolite Profiles

Abstract High performance liquid chromatography coupled with continuous radioactivity detection rep­resents an advancement in drug metabolism research. Using radioactive substances labelled in biologically stable positions, all metabolites can be specifically detected by radioactivity measure­ment. Thus no clean-up of biological fluids is required prior to HPLC. This can prevent artefact formation from unstable metabolites, reduces recovery problems and facilitates quantitation. Separation of highly polar and unpolar metabolites is possible in a single chromatographic run using gradient elution and reversed phase materials. This technique is also well-suited for prepara­tive isolation and purification of metabolites for subsequent structure elucidation. Various metabolite profiles of drugs labelled with carbon-14 or tritium are shown. Metabolites of the following drugs are presented: norfenefrine, etozolin, thymoxamine, naloxone, and levobunolol. We review the general methodology and report our experience with this technique. In principle, this technique may be useful for all biological systems in which tracer techniques are applied.

scholarly journals Salting-out assisted liquid–liquid extraction coupled with high-performance liquid chromatography for the determination of vitamin D3 in milk samples

2019 ◽  
Vol 6 (8) ◽  
pp. 190952 ◽  
Author(s):  
Nur Hidayah Sazali ◽  
Anas Alshishani ◽  
Bahruddin Saad ◽  
Ker Yin Chew ◽  
Moi Me Chong ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Vitamin D3 ◽  
High Performance ◽  
Liquid Extraction ◽  
Sample Treatment ◽  
Salting Out ◽  
Milk Samples ◽  
Liquid Liquid Extraction

In this study, salting-out assisted liquid–liquid extraction (SALLE) as a simple and efficient extraction technique followed by high-performance liquid chromatography (HPLC) was employed for the determination of vitamin D3 in milk samples. The sample treatment is based on the use of water-miscible acetonitrile as the extractant and acetonitrile phase separation under high-salt conditions. Under the optimum conditions, acetonitrile and ammonium sulfate were used as the extraction solvent and salting-out agent, respectively. The vitamin D3 extract was separated using Hypersil ODS (250x i.d 4.6 mm, 5 µm) HPLC column that was coupled with diode array detector. Vitamin D2 was used as internal standard (IS) to offset any variations in chromatographic conditions. The vitamin D3 and the IS were eluted in 18 min. Good linearity ( r 2 > 0.99) was obtained within the range of 25–600 ng g −1 with the limit of detection of 15 ng g −1 and limit of quantification of 25 ng g −1 . The validated method was applied for the determination of vitamin D3 in milk samples. The recoveries for spiked samples were from 94.4 to 113.5%.

Salting-out induced liquid–liquid microextraction based on the system of acetonitrile/magnesium sulfate for trace-level quantitative analysis of fluoroquinolones in water, food and biological matrices by high-performance liquid chromatography with a fluorescence detector

2014 ◽  
Vol 6 (17) ◽  
pp. 6973-6980 ◽  
Author(s):  
Dongli Du ◽  
Guozhong Dong ◽  
Yuanyuan Wu ◽  
Jingjing Wang ◽  
Ming Gao ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Quantitative Analysis ◽  
High Performance ◽  
Biological Matrices ◽  
Fluorescence Detector ◽  
Trace Level ◽  
Salting Out ◽  
Liquid Microextraction

A convenient, economical SILLME method coupled with HPLC/FLD for sample preparation, extraction and trace-level quantitative determination of fluoroquinolones is reported.

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