Racemization detection

In the flavour extract of apricots, racemic dihydroactinidiolide (DHA) was found as the first natural racemate detected by enantio-MDGC analysis [16]. The absolute configurations and the optical activities have been reported to be (R)-(-) and (S)- +) enantiomers, respectively [17, 18]. 

An hplc assay was developed suitable for the analysis of enantiomers of ketoprofen (KT), a 2-arylpropionic acid nonsteroidal antiinflammatory dmg (NSAID), in plasma and urine (59). Following the addition of racemic fenprofen as internal standard (IS), plasma containing the KT enantiomers and IS was extracted by Hquid-Hquid extraction at an acidic pH. After evaporation of the organic layer, the dmg and IS were reconstituted in the mobile phase and injected onto the hplc column. The enantiomers were separated at ambient temperature on a commercially available 250 x 4.6 mm amylose carbamate-packed chiral column (chiral AD) with hexane—isopropyl alcohol—trifluoroacetic acid (80 19.9 0.1) as the mobile phase pumped at 1.0 mL/min. The enantiomers of KT were quantified by uv detection with the wavelength set at 254 nm. The assay allows direct quantitation of KT enantiomers in clinical studies in human plasma and urine after adrninistration of therapeutic doses. 

The reaction of 5-methoxy-2(5//)-furanone 168 with amines was also studied (89T6799). The conjugated addition of ethanolamine to the furanone 168 gave the racemic amino lactone 275 (R = CH2CH20H). Similarly, piperazine reacted with two equivalents of 168 to provide the diadduct 276 as a single diastereomer (no traces of the other isomer were detected). With tryptamine, the reaction was nearly quantitative with the the formation the tran -adduct 277 (R = tryptophanyl) (Scheme 72) (89T6799). 

Fig. 3-9. Preparative HPLC of 100 mg of the test racemate 8 in a single 2 mL injection using a 250 x 4.6 mm i.d. column containing (5)-Glu-(5)-Leu-DNB CSP. Conditions mobile phase ethyl acetate, flowrate 2.0 mL min , UV detection at 380 nm. Injection 2 mL of 50 mg mL racemate solution. Fractions collected before and after the indicated cut point were 98.4 % ee and 97 % ee pure, respectively. (Reprinted with permission from ref. [86]. Copyright 1999, American Chemical Society.)...

If some fields may be empty in the sublevels, all the fields in the main level are required for each entry. A new chiral separation record can be added in CHIRBASE solely if the authors correctly identify both sample and CSP. Since the beginning of the project, our policy has been to contact the authors of all publications containing incomplete, ambiguous or inconsistent data and to ask for additional information. Providing the separations with unique case numbers helps us considerably in this essential task, and also facilitates avoiding redundancies in the database. When chiral separations are reported for the second time in a new publication with exactly the same chromatographic conditions, this is stated in a footnote added in the field comments . In this field, miscellaneous information that cannot appear elsewhere are listed (detection limit, description of a reported chromatogram, racemization study, mobile phase limitations, etc.). 

The mixture of free amino acids is reacted with OPA (Fig. 7-8) and a thiol compound. When an achiral thiol compound is used, a racemic isoindole derivative results. These derivatives from different amino acids can be used to enhance the sensitivity of fluorescence detection. Figure 7-9 shows the separation of 15 amino acids after derivatization with OPA and mercaptothiol the racemic amino acids may be separated on a reversed-phase column. If the thiol compound is unichiral, the amino acid enantiomers may be separated as the resultant diastereomeric isoindole compound in the same system. Figure 7-10 shows the separation of the same set of amino acids after derivatization with the unichiral thiol compound Wisobutyryl-L-cysteine (IBLC). 

Derivatization of a racemic compound with an achiral group may play an important role in the analysis of a chiral compound (Fig. 7-15). In the case of substances with low or no UV-activity, the compounds can be rendered detectable by introducing an UV-absorbing or fluorescent group. If the racemate itself shows selectivity on a chiral stationary phase (CSP), this method can be applied to reduce the limit of detection. Examples have been reported in the literature, especially for the derivatization of amino acids which are difficult to detect using UV detection. Different derivatization strategies can be applied (Fig. 7-16). 

The submitters report obtaining the product in 99% yield. The enantiomeric excess of the Mosher ester of 3 was measured to be 98% using a Chiralcel OD column (40% 2-propanol/hexane). This optical purity measurement substantiated the optical purity assessment made by 111 NMR studies of 3 and racemic 3 prepared using a different method3. Addition of the chiral shift reagent tris[3-(heptafluoropropylhydroxymethylene)-(+)-camphorato]europium (III) resulted in clear resolution of the respective aromatic proton signals for the two enantiomers, which was demonstrated with the racemate. Under similar conditions, NMR analysis of 3 showed that within the detectable limits of the experiment (ca. <3%), there was none of the disfavored enantiomer. 

Child, A.M., Gillard, R.D. and Pollard, A.M. 1993 Microbially-induced promotion of amino acid racemization in bone isolation of the microorganisms and the detection of their enzymes. Journal of Archaeological Science 20 159-168. 

The high simple diastereoselectivities observed running the [4-1-3] cycloadditions raised the question concerning the induction of chirality. Preliminary experiments involving chiral menthyloxy Fischer carbenes 169 (R = (-)-men-thyl) resulted in the formation of the diastereomeric lactim ethers 173-1 and 173-2 in a 7 3 ratio, which could be separated by means of a crystallization. A final acidic hydrolysis gave the enantiomerically pure e-caprolactams 175 and ent-175 and the acyclic esters, respectively. No signs of racemization have been detected,Eqs. (18,19) [39b]. 

Searching for a method of synthesis of enantiopure lamivudine 1, the compound having a monothioacetal stereogenic centre, Rayner et al. investigated a lipase-catalysed hydrolysis of various racemic a-acetoxysulfides 2. They found out that the reaction was both chemoselective (only the acetate group was hydrolysed with no detectable hydrolysis of the other ester moieties) and stereoselective. As a result of the kinetic resolution, enantiomerically enriched unreacted starting compounds were obtained. However, the hydrolysis products 3 were lost due to decomposition." In this way, the product yields could not exceed 50% (Equation 1). The product 2 (R = CH2CH(OEt)2) was finally transformed into lamivudine 1 and its 4-epimer. ... 

When the natural product source contains racemic mixtures (of isomeric forms), then clearly the assignment of signal value to either or both variants of a compound needs to be determined. Alterations in receptor detection of chirality can change sensitivity to the geometrically alternate compound over the range 101 to 106. Of the lactones passed into urine and deposited on tarsal hairs of Black-tailed deer (Odocoileus... 

In practise, if using one of these reagents to follow the course of a chiral separation, it is essential to determine whether resolution is possible, by performing a test experiment either on a sample of racemate, or at least a sample known to contain significant quantities of both enantiomers. Once useable resolution has been established, the technique can be used to monitor solutions of unknown enantiomer ratios with reasonable accuracy, down to normal NMR detection limits. 

New analyses of material from the interior of the Orgeuil and Ivona meteorites show the presence of P-alanine, glycine and y-amino-n-butyric acid as the main components (0.6-2.0ppm) traces of other amino acids were also detected. The amino acids were present as racemic mixtures, i.e., d/l = 1, so that an extraterrestrial origin can be assumed (Ehrenfreund et al., 2001). 

Fig. 14A-C Chromatography of the racemic monoepoxy derivatives (I—III) of Z3,Z6,Z9-18 on chiral HPLC columns A Chiralpak AD B Chiralpak AS C Chiralcel OJ-R. The solvent system for the former two normal-phase columns is 0.1% 2-propanol in n-hexane (0.45 ml/min), and that of the third column is 15% water in MeOH (0.45 ml/min). Homo-conjugated dienes, epo3,Z6,Z9-18 H (I) and Z3,Z6,epo9-18 H (III), were detected by UV (215 nm), and Z3,epo6,Z9-18 H (II) was detected by RID. The earlier eluting isomers have a 3S,4R, 6S,7R, or 9R,10S configuration...

The pyrroline-iV-oxide 411 lost enantiomeric purity in the deprotection step. The THP protecting group could be deprotected under very mild conditions using Amberlyst 15 in methanol. However, the mixture was obtained in low yield accompanied by partial or total racemization as indicated by variation of specific rotation. Racemization also occurred during purification by silica gel chromatography or recrystallization. The lack of configurational stability of the nitrone 411 may be explained with the occurrence of a fast (not detectable by NMR), nitrone-hydroxyenamine tautomerism (Scheme 91). 

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