Resolution of racemic metoprolol, propranolol, carvedilol, bisoprolol, salbutamol, and labetalol, commonly used β-blockers, into their enantiomers has been achieved by TLC on silica gel plates impregnated with optically pure L
-Glu and L
-Asp. Acetonitrile-methanol-water-dichloromethane and acetonitrile-methanol-water-glacial acetic acid mobile phases in different proportions enabled successful separation. The spots were detected with iodine vapor. The detection limits were 0.23, 0.1, 0.27, 0.25, 0.2, and 0.2 μg for each enantiomer of metoprolol, propranolol, carvedilol, bisoprolol, salbutamol, and labetalol, respectively.
A simple and rapid method has been established for indirect separation of the enantiomers of (R,S)-metoprolol and (R,S)-carvedilol by reversed-phase TLC. Beta blockers derivatized with 1-fluoro-2,4-dinitrophenyl-5-l-alanine amide (Marfey’s reagent, FDNP-l-Ala-NH2) and its six structural variants (FDNP-l-Phe-NH2, FDNP-l-Val-NH2, FDNP-l-Pro-NH2, FDNP-l-Leu-NH2, FDNP-l-Met-NH2, and FDNP-d-Phg-NH2) were spotted on precoated plates. (R,S)- Metoprolol and (R,S)-carvedilol were isolated from pharmaceutical dosage forms and purified. The diastereomers were separated most effectively by use of mobile phases containing acetonitrile and triethylamine-phosphate buffer (50 mM, pH 5.5). The results obtained by use of Marfey’s reagent were compared with those obtained by use of the other variants. The effects of buffer concentration, pH, and concentration of organic modifier were studied.
Direct resolution of enantiomers of (±)-bupropion (BUP) was achieved by thin-layer chromatography on silica gel plates impregnated with optically pure l-Glu as chiral selector. The solvent system acetonitrile-methanol-dichloromethane-water (5.6:1:2.2:1, v/v) was successful in resolving the enantiomers. Spots were detected by use of iodine vapor. The detection limit was 0.2 μg for each enantiomer of BUP. The effects of concentration of chiral selector, temperature, and pH on enantiomeric resolution were examined. The separation of BUP enantiomers was also investigated by high-performance liquid chromatography (HPLC) on a chlorinated methylated cellulose-based stationary phase. Reversed phase HPLC was successful using binary mixture of aqueous ammonium formate and methanol for separation of enantiomeric pair with detection at 230 nm. The factors influencing HPLC separation were also investigated.
Resolution of the enantiomers of racemic atenolol, metoprolol, propranolol, and labetalol, commonly used β-blockers, has been achieved by TLC on silica gel plates using vancomycin as chiral impregnating reagent or as chiral mobile phase additive. With vancomycin as impregnating agent, successful resolution of the enantiomers of atenolol, metoprolol, propranolol, and labetalol was achieved by use of the mobile phases acetonitrile-methanol-water-dichloromethane 7:1:1:1 (
), acetonitrile-methanol-water 6:1:1 (
), acetonitrile-methanol-water-dichloromethane-glacial acetic acid 7:1:1:1:0.5 (
), and acetonitrile-methanol-water 15:1:1 (
), respectively. With vancomycin as mobile phase additive, successful resolution of the enantiomers of metoprolol, propranolol, and labetalol was achieved by use of the mobile phases acetonitrile-methanol-0.56 mM aqueous vancomycin (pH 5.5) 6:1:1 (
), acetonitrile-methanol-0.56 mM aqueous vancomycin (pH 5.5) 15:1:2 (
), and acetonitrile-methanol-0.56 mM aqueous vancomycin (pH 5.5)-dichloromethane 9:1:1.5:1 (
), respectively. Spots were detected by use of iodine vapor. The detection limits were 1.3, 1.2, 1.5, and 1.4 μg for each enantiomer of atenolol, metoprolol, propranolol, and labetalol, respectively.
Authors:Sonika Batra, Manisha Singh, and Ravi Bhushan
The present paper deals with direct enantioresolution of (±)-bupropion using thin-layer chromatography and different Cu(II)-l-amino acid complexes as chiral ligand exchange reagent (LER). Cu(II) acetate and four l-amino acids (viz., l-proline, l-histidine, l-phenylalanine, and l-tryptophan) were used for the preparation of LER. Four different approaches were adopted for impregnating/loading the plate with the LER. In the present work, plate impregnation was achieved (a) by mixing LER with silica gel slurry, (b) by developing plain plates with solutions of the Cu complexes, (c) using a solution of Cu(II) acetate as mobile phase additive for the thin-layer chromatography (TLC) plates impregnated with one of the l-amino acids, and (d) by using the LER as mobile phase additive. Spots were located using iodine vapor. The results obtained with all the approaches have been compared in terms of resolution. Effect of concentration of Cu(II) acetate and chiral selector has also been studied.
Authors:Ravi Bhushan, Deepak Gupta, and Aishwarya Jain
A simple and sensitive method for separation and quantitative determination of antidiabetic drugs in pharmaceutical preparations has been established and validated. Commercial formulations of five antidiabetic drugs (metformin, pioglitazone, rosiglitazone, glibenclamide, and gliclazide) were chosen for the studies. The compounds were extracted, isolated, purified, recrystallized, and characterized by measurement of melting point,
, and IR. Quantitative determination was performed by HPLC, TLC, and column chromatography supplemented with UV spectrophotometry. Two of the combinations, metformin + pioglitazone and metformin + gliclazide, were separated by open-column chromatography. Detection was by UV spectrophotometry in HPLC and by use of iodine vapor in TLC.
Ketorolac (Ket) is a potent non-narcotic analgesic drug (among the nonsteroidal anti-inflammatory drugs). The physiological activity of Ket resides with (S)-(−)-Ket while the drug is marketed and administered as a racemic mixture. Therefore, it is desirable that the pharmacokinetics is measured and quantified for enantiomers individually and not as a total drug. The present paper is focused on relevant literature on LC enantioseparation of (RS)-Ket along with bioassay, pharmacokinetic and clinical studies within the discipline of analytical chemistry. HPLC and Thin layer chromatography (TLC) methods using both direct and indirect approaches are discussed. The methods provide chirality recognition even in the absence of pure enantiomers. Besides, a brief discussion on resolution by crystallization and enzymatic methods is included. The most interesting aspects include establishment of structure and molecular asymmetry of diastereomeric derivatives using LC-MS, proton nuclear magnetic resonance spectrometry, and by drawing conformations in three dimensional views by using certain software. A brief discussion has also been provided on the recovery of native enantiomers by TLC.
Authors:Dipshi Singh, Poonam Malik, and Ravi Bhushan
The scientific development in the area of enantioseparation during the last few decades has centered on the production of new chiral stationary phases (CSPs) and new chiral derivatizing reagents (CDRs) for use in liquid chromatography, and in particular high-performance liquid chromatography (HPLC) only. Both CSPs and CDRs have several limitations which, in general, are ignored. Little attention has been paid to thin-layer chromatography (TLC) despite its many advantages compared to HPLC in pharmaceutical and drug analysis and the areas of natural products chemistry and organic synthesis, particularly enantioselective synthesis in purification of the product prior to establishing enantiomeric ratio by different method(s). TLC provides a rapid, easy, aff ordable, and simple approach in all these situations. The demonstrated capability and effi- ciency of TLC in direct resolution of the racemate clearly establish its superiority, and the methodology should allow its application in the resolution of several other racemates, irrespective of the functional group, in a very short time along with the recovery of native enantiomers (for further use).
Authors:Hariom Nagar, Jürgen Martens, and Ravi Bhushan
Enantioresolution of three active pharmaceutical ingredients (APIs), namely, atenolol, betaxolol, and orciprenaline, marketed as racemic mixture, has been achieved in a direct mode using (S)-glutamic acid as chiral additive in thin-layer chromatography. Two different approaches were adopted: (1) (S)-glutamic acid was mixed in the silica gel slurry for making thin-layer plates, or (2) it was added in the mobile phase and plain plates without any chiral additive were used. Both (1) and (2) were capable of separating enantiomers of all the three racemates, but different combinations and proportions of solvents were found successful in the two cases. Good resolution was achieved in both cases, and the results are compared for these two sets of studies among themselves and with other literature reports. Iodine was used to locate the spots of the corresponding enantiomers. The detection limits for each enantiomer were found in the range of 1.4–1.9 μg (per spot).