CHAIN MAPPING BY CTA-SAX AND SIZE EXCLUSION/ESI-MS OF 5 SAMPLES

通过 CTA-SAX 和尺寸排除/ESI-MS 对 5 个样品进行链图谱

• 批准号: 8168905
• 负责人: Parastoo Azadi
• 金额: $ 0.17万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-10 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168905
• 关键词: Acids; Blood capillaries; Computer Retrieval of Information on Scientific Projects Database; Detection; Exclusion; Freeze Drying; Funding; Gases; Gel Chromatography; Grant; Hydrogen; Infusion procedures; Inorganic Sulfates; Institution; Ions; Low-Molecular-Weight Heparin; Maps; Mass Spectrum Analysis; Research; Research Personnel; Resources; Sampling; Series; Sodium Chloride; Solutions; Solvents; Source; Spectrometry, Mass, Electrospray Ionization; System; Temperature; Time; United States National Institutes of Health; Unspecified or Sulfate Ion Sulfates; Water; ammonium acetate; aqueous; capillary; detector; instrument; voltage

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. CTA SAX-HPLC The samples were dissolved in water (20 mg/mL) and 10 ¿L was injected onto a Phenomenex Gemini-NX C18 column (3 ¿m, 250x4.6 mm), coated with cetyltrimethylammonium hydrogen sulfate. Analytes were detected by their UV absorbance at 232 nm. Separation was effected by a salt gradient (Table 1) using the following system: Solvent A: Water (brought to pH 3.0 by addition of conc. methylsulfonic acid); Solvent B: 2.5 M methylsulfonic acid, pH 2.5. Preparative Gel Filtration Twenty-five ¿L of a 100 mg/mL aqueous solution of LMWH was injected onto two TSKGel G2000SW columns (7.8 mm ID X 30 cm, 5 ¿m), connected in series and equipped with a TSKGel guard column (6.0 mm ID X 4.0 cm, 7 ¿m), using 0.25 M ammonium acetate, pH 6.0 as eluent at a flow rate of 0.5 mL/min. Detection was by UV at 232 nm.. The fractions containing tetrasaccharide (41.8-42.9 min) were freeze-dried 3 times from water, redissolved in 25 ¿L water, and repurified on the same column. The tetrasaccharide containing fractions were freeze-dried 3 times from water. ESI Mass Spectrometry Mass spectrometry was performed on a Thermo Scientific LTQ Orbitrap Discovery instrument in ESI mode under direct infusion. Infusion rate was 5 ¿L/min, capillary temperature was 300 ¿C, capillary voltage -13 V, spray voltage -3.2 kV, and sheath gas flow rate was 18. Both the ion trap and the Orbitrap FT detector were used to acquire mass spectra.

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