Chromatographic purification, analysis

The biosynthesis, occurrence, and biological activities of piperidine alkaloids arising from lysine have been studied Punica granatum. Piper species, and Lobelia inflata). The isolation process, chromatographic purification/analysis, and structure elucidation of the significant alkaloids are discussed. 

HF-induced elimination (5). To a solution of aqueous HF (4 drops, 50%) in MeCN (8 ml) was added a solution of the /3-hydroxysilane (lmmol) in MeCN (2ml), and the mixture was stirred at room temperature until t.l.c. analysis indicated completion (5-20min). The reaction mixture was then partitioned between pentane (50 ml) and saturated sodium hydrogen carbonate solution (10ml). The aqueous layer was extracted thoroughly with pentane (3 x 50 ml), and the combined organic extracts were washed with brine and dried. Concentration followed by chromatographic purification gave the product alkenes. 

Londo, T., Lynch, R, Kehoe, T., Meys, M., and Gordon, N., Accelerated recombinant protein purification process development. Automated, robotics-based intergration of chromatographic purification and analysis, /. Chromatogr. A, 798, 73, 1998. 

Figure 9.17. Overview of the manufacture of Ceprotin. As the active ingredient is derived directly from pooled human plasma, particular emphasis is placed upon ensuring that the finished product is pathogen-free. Precautions entail the incorporation of two independent viral inactivation steps and high-resolution chromatographic purification. Additionally, extensive screening of plasma pool source material for blood-borne pathogens is undertaken. Viral screening is undertaken using a combination of immunoassay and PCR analysis for the presence of viral nucleic acid...

To a 1-4 ml plasma sample is added 10 ng of deu-terated internal standard (A1-THC-d3) dissolved in 50 yl ethanol. The extraction procedure and the liquid chromatographic purification on Sephadex LH-20 columns are described in detail in our previous studies (1-3). The purified sample was dissolved in absolute ethanol (10 yl) and kept cold (-20°C) and dark until analysis. This solution can be subjected to mass fragmentography directly or after silylation. 

To a solution of 1 mmol of (-)-/V-bcnzyloxycarbonylnorcphcdrinc in 10 mL of dry benzene is added 0.3 mmol of pyridinium tosylate and 1.1 mmol of methyl 4 4-dimethoxy-2-butenoate. The mixture is refluxed for 12 h with a bypassed dropping funnel filled with 4 A molecular sieves placed between the flask and the reflux condenser. The mixture is cooled, 10 mL of Et20 arc added, the filtrate washed with 5% aq NaHCOj and sat. aq NaCl and finally dried over Na2S04. The mixture is then filtered and the solvent evaporated in vacuo. GC analysis of the crude product gives a d.r. of 94 6. Chromatographic purification gives the 2-alkenyloxazolidine 1 in 88% yield. [a]D —81.6 (c = 1, CHCf,). 

The A,0-dimethyl hydroxamate (3 mmol) is dissolved in 25 mL of anhydrous THE The reaction mixture is cooled down to 0°C and LiAlHt (6 mmol, 228 mg) is added portionwise. After stirring the reaction for 30 min, ethyl acetate (70 mL) is added first, followed by a 1 M potassium hydrogen sulfate solution (50 mL), and the mixture is stirred for 30 min. The organic layer is then collected, washed with 1 M potassium hydrogen sulfate (2 X 50 mL) and a saturated NaCl solution, and dried over magnesium sulfate. The solvent is removed in vacuo to give the A/-protected a-amino aldehyde. The a-amino aldehyde obtained by this method can be used without further purification as the thin-layer chromatographic (TLC) analysis shows only... 

These compounds, whose general formula is AraPoXa, have been detected among the /3-decay products of Ara °BiX2. For X = F, compounds which Ar is 2,5-dimethylphenyl, jc-tolyl, jc-anisyl, jc-methoxyphenyl, and a-naphthyl have been claimed to be indentified by chromatographic analysis. Another method of preparation has also been reported. This involves the treatment of AraPo with AraBiFa and subsequent chromatographic purification. Similar compounds have been reported in which X is chloride or bromide. 

Dissolve the antibody to be conjugated in 0.1 M sodium phosphate, 0.15 M NaCl, pH 7.5, at a concentration of 10 mg/ml. If the antibody contains oligomers (as evidenced by nondenaturing electrophoresis or HPLC gel filtration analysis), then the monomeric IgG form should be isolated by gel filtration using a column of Sephacryl S-200HR. If no oligomers are present, then omit the chromatographic purification. 

Falcarinol, falcarindiol, and myristicin contents of carrots, Daucus carota L., were determined by a sequence of dichloromethane extraction, column chromatographic purification, and gas-liquid chromatographic analysis. High Color 9, Long Imperator 58, Danvers 126, and Spartan Bonus varieties were grown in Wisconsin (1979-1982), Florida (1980-1982), California (1980-1982), Arizona... 

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