High performance liquid chromatography buffers

While the apparent molecular weight was about 47,000 g/mol or daltons (Da) by mobUify on SDS-PAGE, separate analysis by sedimentation equilibrium measurements and capillary high-performance liquid chromatography (HPLC) in SDS buffer gave values near 23,000 Da. 

Chloroform, sodium chloride, anhydrous sodium sulfate, sulfuric acid (97%), hydrochloric acid (36%), sodium bicarbonate, trifluoroacetic acid, tris(hydro-xymethyl)aminomethane (Tris), special grade Water, high-performance liquid chromatography grade 0.1 M Phosphate buffer solution (pH 7.0)... 

High performance liquid chromatography-mass spectrometric methods Nitin et al. [75] developed and validated a sensitive and selective liquid chromatography-tandem mass spectrometric method (LC MS MS) for the simultaneous estimation of bulaquine and its metabolites primaquine in monkey plasma. The mobile phase consisted of acetonitrile ammonium acetate buffer (20 mM, pH 6) (50 50, v/v) at a flow rate of 1 mL/min. The chromatographic separations were achieved on two Spheri cyano columns (5 pm, 30 cm x 4.6 mm), connected in... 

A new hydroxynitrile lyase (HNL) was isolated from the seed of Japanese apricot Prunus mume). It accepts benzaldehyde and a large number of unnatural substrates for the addition of HCN to produce the corresponding (7 )-cyanohydrins in excellent optical and chemical yields. A new high-performance liquid chromatography (HPLC)-based enantioselective assay technique was developed for the enzyme, which promotes the addition of KCN to benzaldehyde in a buffered solution (pH 4.0). Asymmetric synthesis of (7 )-cyanohydrins by a new HNL is described (Figure 8.4). ... 

In the separation of biomolecules, sample preparation almost always involves the use of one or more pretreatment techniques. With high-performance liquid chromatography (HPLC), no one sample preparation technique can be appHed to all biological samples. Several techiques may be used to prepare the sample for injection. For example, complex samples require some form of preffactionation before analysis, samples that are too dilute for detection require concentration before analysis, samples in an inappropriate or incompatible solvent require buffer exchange before analysis, and samples that contain particulates require filtration before injection into the analytical instrument. 

Determination of brinzolamide and its 3 principal metabolites (the N-desethyl, A -desmethoxypropyl and O-desmethyl analogs) in whole blood and plasma from clinical and pre-clinical studies was performed using high performance liquid chromatography (HPLC) with UV detection. After addition of a known amount of internal standard (AL-5138, the 4-methoxybutyl analog of brinzolamide), the sample was acidified with 50 mM sodium phosphate buffer, pH 3.0 and extracted with ethyl acetate. 

A method for the determination of personal exposure to benzidine-based dyes has been developed. This procedure involved the reduction of benzidine-based dye filter samples to free benzidine with neutral buffered sodium hydrosulfite solution. The benzidine-containing reduction solution was then analyzed by high performance liquid chromatography. The reduction was found to be quantitative by visible-spectrum analysis. This reduction and analysis method was evaluated with four benzidine-based dyes over the range from 12 to 300 micrograms per sample. Precision for the reduction and analysis of the four dyes falls within % coefficient of variation. This method can differentiate between benzidine-and benzidine congener-based dyes. Results are reported in terms of free benzidine. 

Evaporate a 5- ll aliquot to dryness under a vacuum. Redissolve the residue in a suitable buffer prior to analysis by an amino acid analyzer or into the appropriate eluant if high performance liquid chromatography (HPLC) analysis is used. 

Development of fast, accurate, and reproducible high-performance liquid chromatography (HPLC) methods has offset the use of traditional open-column and TLC methods in modern chlorophyll separation and analysis. A number of normal and reversed-phase methods have been developed for analysis of chlorophyll derivatives in food samples (unit F4.4), with octadecyl-bonded stationary phase (C]8) techniques predominating in the literature (Schwartz and Lorenzo, 1990). Inclusion of buffer salts such as ammonium acetate in the mobile phase is often useful, as this provides a proton equilibrium suitable for ionizable chlorophyllides and pheophorbides (Almela et al., 2000). 

JS Hamada. Separation and molecular mass distribution of rice proteins by size-exclusion high-performance liquid chromatography in a dissociating buffer. J Chromatogr A 734 195-203, 1996. 

A high-performance, liquid chromatography (h.p.l.c.) assay (reversed phase, C lg column, methanol - water- acetate buffer) for isosorbide dinitrate along with the two mononitrates in pharmaceutical formulations has been described.45 A similar method can be found in Ref. 44. 

For small molecule analytes (see Note 6) for which a radiotracer form is available, sequentially load a known quantity of tracer dissolved in buffer and determine the amount of analyte in the eluant. When the radioactivity not retained by the immunoaffinity column plateaus, the column binding sites are saturated. Wash the column, and elute the retained radioactivity. The mass of analyte in the eluted volume is the apparent column capacity. In many instances a radio-labeled analyte may not be available. In such cases, high-performance liquid chromatography, UV spectroscopy, or any other analytical tool capable of selectively quantifying the analyte may be used to determine column capacity. 

The use of high performance liquid chromatography (HPLC) on-line or off-line is an essential feature for peptide mapping to integrate the removal of buffers and salts (purification) and the separation of analytes (preconcentration) with mass spectrometry. With on-line LC/MS approaches, low flow rates (<100pL/min) have been demonstrated to provide maximum sensitivity with ESI techniques for the analysis of proteins. In the work performed by Arnott and... 

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