Elution profile showing resolution

The hydrophilic surface characteristics and the chemical nature of the polymer backbone in Toyopearl HW resins are the same as for packings in TSK-GEL PW HPLC columns. Consequently, Toyopearl HW packings are ideal scaleup resins for analytical separation methods developed with TSK-GEL HPLC columns. Eigure 4.44 shows a protein mixture first analyzed on TSK-GEL G3000 SWxl and TSK-GEL G3000 PWxl columns, then purified with the same mobile-phase conditions in a preparative Toyopearl HW-55 column. The elution profile and resolution remained similar from the analytical separation on the TSK-GEL G3000 PWxl column to the process-scale Toyopearl column. Scaleup from TSK-GEL PW columns can be direct and more predictable with Toyopearl HW resins. 

Fig. 2.145. Semipreparative HPLC separation of the subunits from P. cruentum B-PE. The elution profile monitored at 226 nm is shown. The identity of each peak was detemined from the visible absorption spectra, the known distribution and content of PEB (phycoeryhtrobihn) and PUB (phu-courobihn) and SDA-PAGe. Dotted line symbolizes the gradient in percentage of acetonitrile (for quantitative details see text). The order of subunit elution is y. a, p. The elution profile shows a partial resolution of at least three j- species. Reprinted with permission from R. Bermejo et al. [317].

Figure 16.15 shows the resulting chromatograms for the three glucan fractions obtained by previous preparative separation on Sephacryl S-200/S-1000 (Fig. 16.14). From the normalized fraction chromatograms, the elution profile of the initial mixture has been reconstructed by mixing 50% fraction 1, 40% fraction 2, and 10% fraction 3. Compared to the chromatogram of the preparative Sephacryl S-200/S-1000 system, separation with the TSK/ Superose system yields improved resolution in the low dp (high V, ) domain.

PAHs introduced in Section 34.1. A PCA applied on the transpose of this data matrix yields abstract chromatograms which are not the pure elution profiles. These PCs are not simple as they show several minima and/or maxima coinciding with the positions of the pure elution profiles (see Fig. 34.6). By a varimax rotation it is possible to transform these PCs into vectors with a larger simplicity (grouped variables and other variables near to zero). When the chromatographic resolution is fairly good, these simple vectors coincide with the pure factors, here the elution profiles of the species in the mixture (see Fig. 34.9). Several variants of the varimax rotation, which differ in the way the rotated vectors are normalized, have been reviewed by Forina et al. [2]. 

Figure 2. Elution profile of P. sajor-caju supernatant from DEAE-Bio-gel column, showing resolution of laccase and VAO peaks of activity.

For Th-FFF, column dispersion is well defined and can therefore be accounted for and removed from the elution profile [ 122,123]. In removing the effects of column dispersion, the smaller fractionating power of Th-FFF for M< 105 g/mol can be partially compensated. Similar findings were also published by Gunderson and Giddings [115] who found that, after dispersion corrections, Th-FFF shows a better resolution than SEC with the exception of low molar mass samples. 

Fig. 2 shows the fractionation of oligo (rA) with chain lengths of 3 to 37 nucleotides on TSK-DEAE 5PW. Similar results have been reported for Nucleogen DEAE 60 [16] and MONO-Q [17]. As an example for lEC separation of medium-sized nucleic acids, the elution profile of a crude plant RNA extract on Nucleogen DEAE 500 is shown in Fig. 3. A comparable resolution was obtained on TSK-DEAE 5PW although a slower elution was necessary [18]. The resin with the...

Fig. 3 shows the influence of temperature and isopropanol volume fraction in the term (a - Ha). There are examples of different situations for pairs 5-6 and 4—5, the (a - 1/a) term is markedly influenced by the isopropanol volume fraction the column temperature also exerts an important influence on this term for pairs 2-3 and 4-5. It is interesting to note that both parameters T and V) have important and similar effects on pairs 4-5 and 2-3 in this resolution term. For these cases, both parameters can be used to obtain a better resolution. Fig. 1C shows an elution profile under optimal elution conditions. 

Fig. 1. Purification of myristoylated Arfl. (A) Elution profile of Arfl proteins on Sephacryl S-100 26/60 column. The peak containing Arfl is labeled. Inset shows the expression of myristoylated Arfl in E. coli with NMT. Lane U, uninduced lane 1, induced., band containing Arfl. Proteins from the indicated elution fractions were separated by SDS-PAGE in the gel shown below the chromatogram. marks the band containing Arfl. (B) Resolution of myristoylated ARFI on Phenyl Sepharose HP column. The protein is loaded in the presence of 3 M NaCl and eluted with a descending sodium chloride gradient. In the SDS-PAGE fractionation of the proteins from the elution, shown below the chromatogram, the band containing Arfl is marked with. ...

In the top separation the peaks are only partially resolved. The bottom separation shows baseline resolution, i.e., the elution profile for the flrst-eluting peak reaches the system baseline before the second peak starting to elute. 

The stereochemistry of each enantiomer separated by the chiral HPLC has been studied after methanolysis of the epoxy ring. Examining the H NMR data of esters of the produced methoxyalcohols with (S)- and (R)-a-methoxy-a-(tri-fluoromethyl) phenylacetic acid by a modified Mosher s method [181], it has been indicated that the earlier eluting parent epoxides are (3S,4R)-, (6S,7R)-, and (9R,10S)-isomers (Table 7) [75, 76, 179]. The above three chiral HPLC columns show different resolution abilities but a different elution order is not observed. The resolution profile by the reversed-phase OJ-R column has been generalized with molecular shapes of the epoxy compounds considering the... 

Figure 1.4 Gradient elution of nine alkylphenones showing the effect of different gradient profiles on the peak resolution (a) linear, 0-100% acetonitrile (ACN), (b) linear, 30-100% ACN, (c) convex, 30-100% ACN, and (d) concave, 30-100% ACN. Curve numbers refer to profiles of Figure 1.3. (Adapted from Ref. 23 with permission.)...

Fig. 16.4 shows the capillary gas chromatograms obtained for a fraction. Fig. 16.4(b) shows the total ionisation chromatograms for the fraction obtained from the high resolution mass spectral data set. In general, the correlation between the total ionisation chromatogram profile and the flame ionisation detector profile is low due to both the differing relative detector responses and the consideration of scan cycle time (9.6s) with respect to chromatographic peak elution time ( 20s). 

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