Eluents phosphate buffer

Eluent Phosphate buffer (10 mM) Phosphate buffer Phosphate buffer... 

Figure 10.3. Size exclusion chromatogram obtained by UV and OC detection (SEC/UV-DOC) for a river sample (river Neckar, Heidelberg, 22.03.2007, DOC = 2.3 mg liter 1, chromatographable organic carbon (COC = 75%). Column TSK HW50S (250 x 20mm). Eluent phosphate buffer, 26.8mmol liter 1, UV (254nm) and organic carbon (OC) detection, exclusion volume V0, permeation volume Vp).

Fig. 1 Dependence of k on adrenaline (squares) and L-tyrosine hydrazide (circles), on mobile-phase concentration of 1-hexane-sulfonate. Column Synergi Hydro-RP (Phenomenex) 150 x 4.6 mm ID, particle size 4 pm, and bonded phase coverage 4.05 pmol/m. Eluent phosphate buffer 37.10 mM KH2PO4 and 4.29 mM Na2HP04 calculated to provide a pH of 6.0. After addition of the desired amount of sodium 1-hexanesulfonate, NaCl was added so that the total sodium concentration was 50 mM (constant ionic strength). Experimental data were fitted by Eq. 8.

Three different types of columns packed with gels of different pore sizes are available. Columns should be selected that are suitable for the molecular weight range of specific samples, as each type has a different exclusion limit (Fig. 6.41, page 215). Bovine serum albumin (BSA), myoglobin, and lysozyme show good peak shapes using only 100 mM of sodium phosphate buffer as an eluent. There is no need to add any salt to the eluent to reduce the ionic interaction between protein and gel. 

FIGURE 7.13 Preparative separation of various proteins on Fractogel EMD BioSEC (S). The length of the column was 1000 mm and the inner diameter 100 mm. The flow rate was 6.2 ml/min with 20 sodium phosphate buffer (pH 7.2) containing 0.3 M NaCI as the eluent. The injected standard proteins can be used to create a calibration curve. 

FIGURE 7.17 Separation of a complex mixture on Fractogel EMD BioSEC (S) with a column dimension of 1000 X 50 mm (Superformance glass column). The sample contained ferritin (I), immunoglobulin G (2), transferrin (3), ovalbumin (4), myoglobin (5), aprotinin (6), and vitamin B, (7). Five milliliters of the mixture was injected onto the column at a flow rate of 3 ml/min (eluent 20 mAI sodium phosphate buffer, 0.1 M NaCI, pH 7.2). 

Use of 10 pm LiChrosorb RP18 column and binary eluent of methanol and aqueous 0.1 M phosphate buffer (pH 4.0) according to suitable gradient elution program in less than 20-min run time with satisfactory precision sensitivity of spectrophotometric detection optimized, achieving for all additives considered detection limits ranging from 0.1 to 3.0 mg/1, below maximum permitted levels Simultaneous separation (20 min) of 14 synthetic colors using uncoated fused silica capillary column operated at 25 kV and elution with 18% acetonitrile and 82% 0.05 M sodium deoxycholate in borate-phosphate buffer (pH 7.8), recovery of all colors better than 82%... 

To test the quality of some synthetic dyes according to standardized procedures, a screening is recommended based on TLC analysis on silica plates 60 F 254 using elutions with an ethyl acetate pyridine water 25 25 20 (v v v) mixture. To determine purity and secondary dyes (components or by-products of a dye that are not allowed to be present), successive TLC separations are recommended or, for more accurate answers, HPLC-DAD using RP-18 columns and eluents like acetonitrile and phosphate buffer."... 

An alternative system proved to be both simpler and more user friendly (Unger et al., 2004 Machtejevas et al., 2006). Thus far we have used this configuration to analyze human plasma, sputum, urine, cerebrospinal fluid, and rat plasma. For each particular analysis we set up an analytical system based on a simple but specific strategy (Figure 9.5). The analysis concept is based on an online sample preparation and a two-dimensional LC system preseparating the majority of the matrix components from the analytes that are retained on a RAM-SCX column followed by a solvent switch and transfer of the trapped peptides. The SCX elution used five salt steps created by mixing 20 mM phosphate buffer (pH 2.5) (eluent Al) and 20 mM phosphate buffer with 1.5 M sodium chloride (eluent Bl) in the following proportions 85/15 70/30 65/45 45/55 0/100 with at the constant 0.1 mL/min flow rate. Desorption of the... 

The retention indices, measured on the alkyl aryl ketone scale, of a set of column test compounds (toluene, nitrobenzene, p-cresol, 2-phenyl ethanol, and IV-methylaniline) were used to determine the changes in selectivity of a series of ternary eluents prepared from methanol/0.02M phosphate buffer pH 7 (60 40), acetonitrile/0.02 M phosphate buffer pH 7 (50 50) and tetrahydrofuran/0.02 M phosphate buffer pH 7 (25 65). The analyses were carried out on a Spherisorb ODS reversed-phase column. The selectivity changes were often nonlinear between the binary composition [83]. 

Figure 3.18 Adenosine phosphates in blood on vinyl polymer column. Conditions column, Asahipak GS320 (vinyl alcohol copolymer gel), 50 cm x 7.6 mm i.d. eluent, 0.1 M sodium phosphate buffer containing 3 M sodium chloride pH 7.0 flow rate, 1.0 ml min-, detection, UV 260 nm. Peaks 1, haemoglobin 2, adenosine triphosphate 3, adenosine diphosphate and 4, adenosine monophosphate.

Figure 4.8 Cation-exchange liquid chromatography of basic proteins. Column, Asahipak ES502C eluent, 20 min linear gradient of sodium chloride from 0 to 500 mM in 50 mM sodium phosphate buffer pH 7.0 flow rate, 1 ml min-1 temperature, 30 °C detection, UV 280 nm. Peaks 1, myoglobin from horse skeletal muscle (Mr 17 500, pi 6.8-7.3) 2, ribonuclease from bovine pancreas (Mr 13 700, pi 9.5-9.6) 3, a-chymotrypsinogen A from bovine pancreas (Mr 257 000, pi 9.5) and 4, lysozyme from egg white (Mr 14 300, pi 11.0-11.4). (Reproduced by permission from Asahikasei data)...

Figure 4.11 Direct injection analysis of haemoglobin in red blood cells. Column, Asahipak ES-502C eluent, 32 min linear gradient from 25% 30 mM sodium phosphate buffer to 65% 30 mM sodium phosphate containing 300 mM sodium chloride pH 5.5 flow rate, 1.0 ml min-1 detection, 425 nm. Samples. A, normal subject and B, diabetic patient.

Figure 4.20 Calibration curves for size-exclusion liquid chromatography. Column, TSK GEL G3000SW, 120 cm x 7.5 mm i.d. eluent, 0.2 m sodium phosphate buffer pH 6.8 flow rate, 1.0 ml min-1. Standards 1, protein-, 2, dextran, and 3, polyethylene glycol.

Apply the hybridoma culture supernatant to the protein A column and monitor the eluent for protein at 280 nm. Wash the column with 30-40 mL of phosphate buffer, pH 8.0 (see Note 4). 

Fio. 47. Retention factors of pteroyl-oliso-yL-glutamates as a ftinction of the eluent pH. The solid lines were calculated as diKussed in Bush et at. (204). Data were obtained for folic acid (FA) and oligo yglutainates containing one (Ft Glu,), three (Ft GIU ). hve (Ft GtU ), and seven (Ft Glur) utamyl residues. The data were obtained on 5 Partisil ODS-2 with 0.1 M phosphate buffers containing 696 (v/v) acetonitrile as eluent, 4S C, with detection at 254 nra. Reprinted with permission from Bush rf a/. (204). 

Fig. 42., Flota of the retention foctors of monoprotic acid versus the pH of the eluent, rhr iliilii wcrciohliiiiicil on l

Fig. 5A-C. Gel-filtration chromatograms on a column (column size, 10 mm (I.D.) x 200 mm flow rate, 2.0 cm min ) of Sephadex G-50 with aqueous phosphate buffer (10 mmol dm ) at pH 7.0 as eluent A cage-type cyclophane 8 alone (1.0 x 10" mol) B hybrid assembly formed with 7 (1.0 X 10" mol) and 9 (2.0 x 10" mol) C hybrid assembly formed with 8 (1.0 x 10" mol) and 9 (2.0 X 10" mol)...

Figure 5. Lignins dissolved on heating spruce wood meal with phosphate buffer, pH 6.8, for 6 h at 150°C. Column Sephadex G-50. Eluent 0.5 M NaOH.

Red-cell (dark adapted) Blepharisma japonicum were cultured in Pisa, in the dark, at 23 °C, in the presence of the Enterobacter aerogenes bacterium as food supply [7]. Blue-cell (light adapted) Blepharisma japonicum were produced by in vivo photoconversion of blepharismin into oxyblepharismin under a low intensity cold white lamp (below 10 W/m2). Blue cells were washed, collected by low speed centrifugation and resuspended in a 20-mM sodium cholate solution. The chromoprotein was obtained by FPLC chromatography of this preparation, on a hydroxyapatite column. The applied eluent was a phosphate buffer (pH 7.4), first 0.05 M and then 0.2 M. This ionic strength step affects the affinity of the biomolecules with the hydroxyapatite [8]. 

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