Column chromatography isomer separation

Figure 12.12 Coupled SEC-RPLC separation of Plioflex rubber stock (a) SEC (b) RPLC ti ace of fraction 1, Wingstay 100 (Eive-peak pattern is representative of diarylphenylenedi-amine isomers) (c) RPLC ti ace of fraction 2, mixed disulfide and MBTS (2,2 -thiobis (ben-zothiazole)). Obtained under the same conditions as given for Eigure 12.11. Reprinted from Journal of Chromatography, 149, E. L. Johnson et al, Coupled column chromatography employing exclusion and a reversed phase. A potential general approach to sequential analysis , pp. 571-585, copyright 1978, with permission from Elsevier Science.

To this solution was added at one time the above-obtained ethyl acetate solution at -15°C, and the resulting mixture was stirred for 1 hour at -10°C to -15°C. The reaction mixture was cooied to -30°C, and water (80 ml) was added thereto. The aqueous layer was separated, adjusted to pH 4.5 with sodium bicarbonate and subjected to column chromatography on Diaion HP-20 resin (Mitsubishi Chemical Industries Ltd.) using 25% aqueous solution of isopropyl alcohol as an eluent. The eluate was lyophilized to give 7-[2-methoxyimino-2-(2-amino-1,3-thiazol-4-yl)acetamido] cephalosporanic acid (syn isomer) (1.8 g), MP 227°C (decomp.). 

Butylcyclohexanol has been prepared from />-/-butylphenol by reduction under a variety of conditions.3 4 Winstein and Holness5 prepared the pure trans alcohol from the commercial alcohol by repeated crystallization of the acid phthalate followed by saponification of the pure trans ester. Eliel and Ro 6 obtained 4-f-butylcyclohexanol containing 91% of the trans isomer by lithium aluminum hydride reduction of the ketone. Iliickel and Kurz 7 reduced />-/-butylphenol with platinum oxide in acetic acid and then separated the isomers by column chromatography. 

Careful chromatographic and detailed HNMR spectroscopic analysis of the products from the thermolyses of ethyl azidoformate in o-, m- and p-xylene revealed in all cases a mixture of 1 //-azepines.80 In o-xylene, only two of the four possible isomers were separated and characterized, namely, ethyl 4,5-dimethy 1-1 //-azepine-1 -carboxylate (9 %) and ethyl 3,4-dimethyl-l H-azepine-1-carboxylate (7 %). w-Xylene yielded a 2 3 mixture of ethyl 3,5-dimethyl-l//-azepine-1-carboxylate and ethyl 2,4-dimethyl-l//-azepine-l-carboxylate. The 2,4-dimethyl isomer (20 %) can be isolated from the mixture by removal of the 3,5-dimethyl isomer as its Diels-Alder cycloadduct with ethenetetracarbonitrile. p-Xylene gave a mixture of the two possible isomeric azepines which were partially separated by column chromatography. A pure sample of ethyl 2,5-dimethyl-1//-azepine-1-carboxylate (26%) was obtained from the mixture by selective decomposition of the 3,6-dimethyl isomer with refluxing alcoholic potassium hydroxide. 

The first reported example31,117 involved the diethyltetraphenyl-3//-azepines 18 and 19 which were obtained in 85% overall yield by the reaction of2,3-diethyl-2//-azirine with 2,3,4,5-tetraphenylcyclopentadienone (see Section 3.1.1.1.2.). The two isomeric azepines are separable by column chromatography (alumina or silica gel), and each isomer, on warming in xylene for three days, equilibrates to a 3 8 mixture of the 3//-azepines 18 and 19. 

Next we studied high temperature bromination of benzobarrelene at 150 C. NMR analysis indicated that the reaction mixture was very complex and consisted of at least ten products. After repeated column chromatography combined with fractional crystallization we have been able to separate 18 compounds (Scheme 6). Four of them were bromoalcohol compounds 18, 12, 22 and 2fl. After high temperature bromination we expected three isomeric non-rearranged products with benzobarrelene skeleton and isolated 22, 22, and 24 in yields of 34, 9.3, and 6.2 %, respectively. Because of the very close structural similarity we were not able to make a clear-cut differentiation between the stereochemistry of 22 and 24-Therefore, we carried out an X-ray analysis (ref. 9) of the isomer 22-... 

Grainger s group has developed an asymmetric route to (-)-cuparene (5-83) [22] using another photoinduced generation of a biradical (Scheme 5.16) [23]. Thus, irradiation of (S)-proline-derived 5-80 resulted initially in the formation of 5-81, which subsequently cyclizes in almost quantitative yield to afford a mixture of the four possible diastereomers 5-82a-d in an approximate 10 5 2 1 ratio. The two major isomers could be separated by column chromatography to provide 5-82a in 36% yield and the desired 5-82b in 55 %, which was converted into the natural product 5-83 in 24% yield over three steps. 

Unlike the behavior of 81, treatment of 2,3,5-tri-O-benzyl-D-ribofuranosyl bromide91 (110) with mercuric cyanide gave an ano-meric mixture of cyanides (111), which was reduced to an epimeric mixture of amines.92 Separation of this mixture by column chromatography gave the D-alio isomer (112), the D-altro isomer (113), and the glycal derivative (114). Compound 112 was, however, formed in moderate yield (18%), presumably because of a preponderance of the a anomer in the mixture of anomeric cyanides (111). Compound 112 was converted92 into the 1-ureido derivative (92) by treatment... 

As would be anticipated by the method of synthesis, two isomeric products were formed in a ratio of about 3 1 and these were easily separated by column chromatography. The compound formed in greater amount proved to be identical to th material isolated from the first preparation. A study of the C nmr spectra indicated that the more active isomer was the nicotinate 10 and this was based primarily on the coupling of the carboxyl carbon atom with the C-4 proton. 

Prbpy, 5-(2-MePr)bpy and 5-(2,2-Mc2Pr)bpy have been prepared and characterized. The mer-and /uc-isomers of each complex have been isolated by use of cation-exchange column chromatography as the steric requirements of the R group increase, the percentage of the /uc-isomer decreases. Enantiomers of [Ru(5-Prbpy)3] + were separated on SP Sephadex C-25. Electro-kinetic chromatography has been used to separate the enantiomers of [Ru(104)3] " anionic carboxymethyl-/ -cyclodextrin was employed as the chiral mobile phase additive. ... 

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