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Gradient conditions

Amino acid R in Fig. 26-1 3-Letter code 1-Letter code [Pg.551]

This gradient system has been adapted for the analysis of coastal tind interstitial waters where compounds derived from amino acid degradation such as P-alaavae, taurine or amino butyric acids may occur in addition to the standard amino acids given in Table 26-1. For less complex samples such as, e.g., hydrolysates the gradient run time may be abbreviated and a linear gradient employed. It should, however, be noted that under these conditions glycine and threonine are usually not separated. [Pg.552]


At the solid walls, the boundary conditions state that the velocity is zero (i.e. no slip). Also at the walls, the temperature is either fixed or a zero-gradient condition is applied. At the surface of the spinning disk the gas moves with the disk velocity and it has the disk temperature, which is constant. The inlet fiow is considered a plug fiow of fixed temperature, and the outlet is modeled by a zero gradient condition on all dependent variables, except pressure, which is determined from the solution. [Pg.338]

Relationships between lipophilicity and retention parameters obtained by RPLC methods using isocratic or gradient condition are reviewed. Advantages and limitations of the two approaches are also pointed out, and general guidelines to determine partition coefficients in 1-octanol-water are proposed. Finally, more recent literature data on Hpophilicity determination by capillary electrophoresis of neutral compounds and neutral forms of ionizable compounds are compiled. Quotation is restricted to key references for every method presented - an exhaustive listing is only given for the last few years. [Pg.332]

Snyder, L. R., Dolan, J. W., and Cox, G. B., Preparative high-performance liquid chromatography under gradient conditions. III. Craig simulations for heavily overloaded separations, J. Chromatogr., 484, 437, 1989. [Pg.126]

A typical HPLC separation using a 15-cm column of 15,000 theoretical plates produces peak capacity (Giddings, 1991) of about 80-100 under isocratic conditions and up to 150 under gradient conditions in 1 h(Eq. 7.3, n peak capacity, A number of theoretical plates of a column, and fR and t retention time of the last and the first peak of the chromatogram, respectively). An increase in the number of separated peaks per unit time can be achieved by increased separation speed made possible by monolithic silica columns (Deng et al., 2002 Volmer et al., 2002). This has also been shown for peptides and proteins (Minakuchi et al., 1998 Leinweber et al., 2003). [Pg.158]

A simple and rapid RP-HPLC method was developed for the determination of retinoid in galenicals. Commercial preparations were diluted, filered and used for separation. Measurements were carried out in an ODS column (150 X 4.6 mm i.d. particle size 3 /xm). Solvents A and B were methanol-10 mM ammonium acetate (75 25, v/v) and methanol-THF (84 16, v/v), respectively. The flow rate was 0.8ml/min. Gradient conditions were 0-25 min, 0 per cent B 35 min, 100 per cent B, isocratic for 10 min. Typical chromatograms are shown in Fig. 2.37. The repeatability of peak area ranged between 0.48 -3.2 per cent for UV-DAD and 0.57 - 3.1 per cent for fluorescence detection. The reproducibility varied between 0.26 - 4.6 per cent. It was found that the method is precise, selective, sensitive and linear, therefore, it can be employed for the routine quality control of this class of drags [85],... [Pg.132]

The anthocyanin profile of the flowers of Vanda (Orchidaceae) was investigated with a similar technique. Flowers (2 kg) were extracted with 101 of methanol-acetic acid-water (9 l 10,v/v) at ambient temperature for 24 h. The extract was purified by column chromatography, paper chromatography, TLC and preparative RP-HPLC. Analytical HPLC was carried out in an ODS column (250 X 4.6 mm, i.d.) at 40°C. Gradient conditions were from 40 per cent to 85 per cent B in 30 min (solvent A 1.5 per cent H3P04 in water solvent B 1.5 per cent H3P04, 20 per cent acetic acid and 25 per cent ACN in water). The flow rate was 1 ml/min and analytes were detected at 530 nm. The chemical structures of acylated anthocyanins present in the flowers are compiled in Table 2.90. The relative concentrations of anthocyanins in the flower extracts are listed in Table 2.91. It can be concluded from the results that the complex separation and identification methods (TLC, HPLC, UV-vis and II NMR spectroscopy, FAB-MS) allow the separation, quantitative determination and identification of anthocyanins in orchid flowers [262],... [Pg.276]

Normal-phase HPLC has also found application in the analysis of pigments in marine sediments and water-column particulate matter. Sediments were extracted twice with methanol and twice with dichloromethane. The combined extracts were washed with water, concentrated under vacuum and redissolved in acetone. Nomal-phase separation was performed with gradient elution solvents A and B being hexane-N,N-disopropylethylamine (99.5 0.5, v/v) and hexane-2-propanol (60 40, v/v), respectively. Gradient conditions were 100 per cent A, in 0 min 50 per cent A, in 10 min 0 per cent A in 15 min isocratic, 20 min. Preparative RP-HPLC was carried out in an ODS column (100 X 4.6 mm i.d. particle size 3 jum). Solvent A was methanol-aqueous 0.5 N ammonium acetate (75 25, v/v), solvent B methanol-acetone (20 80, v/v). The gradient was as follows 0 min, 60 per cent A 40 per cent A over 2 min 0 per cent A over 28 min isocratic, 30 min. The same column and mobile phase components were applied for the analytical separation of solutes. The chemical structure and retention time of the major pigments are compiled in Table 2.96. [Pg.287]

Another RP-HPLC technique has been applied for the determination of synthetic food dyes in soft drinks with a minimal clean-up. Separation of dyes was obtained in an ODS column (150 x 4 mm i.d. particle size 5 pm). Solvents A and B were methanol and 40 mM aqueous ammonium acetate (pH = 5), respectively. Gradient conditions were 0-3 min, 10 per cent A 3-5 min, to 25 per cent A 5-8 min, 25 per cent A 8-18 min, to 75 per cent A 18-20 min, 75 per cent A. The flow rate was 1 ml/min and dyes were detected at 414 nm. The separation of synthetic dyes achieved by the method is shown in Fig. 3.35. The concentrations of dyes found in commercial samples are compiled in Table 3.21. The quantification limit depended markedly on the type of dye, being the highest for E-104 (4.0 mg/1) and the lowest for E-102 and E-110 (1.0 mg/1). The detection limit ranged from 0.3 mg/1 (E-102 and E-110) to 1.0 mg/ml (E-104 and E-124). It was suggested that the method can be applied for the screening of food colourants in quality control laboratories [113]. [Pg.421]

Fig. 3.89. Separation of the six anionic dyes under gradient conditions in 5 pm Hypersil ODS, 100 X 3 mm i.d. column. Eluent (first solution) 25 mM TBAN03, 25 mM acetate buffer pH 4.7 (second solution) methanol. Flow rate, 0.7 ml/min injection volume, 20 pV, gradient indicated in the Figure sample solution, mixture of anionic dyes, concentration of each 25 pg/ml Absorption detection at 500 nm. Reprinted with permission from R. M. Seifar et al. [150]. Fig. 3.89. Separation of the six anionic dyes under gradient conditions in 5 pm Hypersil ODS, 100 X 3 mm i.d. column. Eluent (first solution) 25 mM TBAN03, 25 mM acetate buffer pH 4.7 (second solution) methanol. Flow rate, 0.7 ml/min injection volume, 20 pV, gradient indicated in the Figure sample solution, mixture of anionic dyes, concentration of each 25 pg/ml Absorption detection at 500 nm. Reprinted with permission from R. M. Seifar et al. [150].

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Boundary conditions gradient elution

Box 25-3 Choosing Gradient Conditions

Broad gradient conditions, HPLC

Conditional gradient-correlation matrix

Gradient chromatography isocratic conditions from

Gradient elution chromatography under nonlinear conditions

Gradient elution mode isocratic condition

Gradient polymer elution chromatography critical conditions

Latitude-conditioned gradients

Retention differences under gradient conditions

Zero-gradient condition

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