A Novel Design of Simulated Moving Bed (SMB) Chromatography for Separation of Ketoprofen Enantiomer

Yoon, Tae-Ho,Chung, Bong-Hyun,Kim, In-Ho The Korean Society for Biotechnology and Bioengine 2004 Biotechnology and Bioprocess Engineering Vol.9 No.4

A simulated moving bed (SMB) chromatography system is a powerful tool for preparative scale separation, which can be applied to the separation of chiral compound. We have de-signed our own lab-scale SMB chromatography using 5 HPLC pumps, 6 stainless steel columns and 4 multi-position valves, to separate a racemic mixture of ketoprofen in to its enantiomers. Our design has the characteristics of the low cost for assembly for the SMB chromatography and easy repair of the unit, which differs from the designs suggested by other investigators. It is possible for the flow path through each column to be independently changed by computer control, using 4 multi-position rotary valves and 5 HPLC solvent delivery pumps. In order to prove the operability of our SMB system, attempts were made to separate the (S)-ketoprofen enantiomer from a ketoprofen racemic mixture. The operating parameters of the SMB chromatography were calculated for ketoprofen separation from a batch chromatography experiment as well as by the triangle theory. With a feed concentration of 1 mg/mL, (S)-ketoprofen was obtained with a purity of 96% under the calculated operating conditions.

Chiral Separation of Ketoprofen Racemate by using Chirex^(�) 3005 and Kromasil^(�) CHI-II Chiral Column

Yoon, Tae Ho,Kim, In Ho 한국화학공학회 2004 Korean Journal of Chemical Engineering Vol.21 No.2

Ketoprofen is a non-steroid anti-inflammatory drug(NSAID) that has analgesic, anti-inflammatory and antipyretic properties, of which pharmacological activity has been contributed mainly by the S-ketoprofen enantiomer. To gain highly purified S-ketoprofen enantiomer from ketoprofen racemate, suitable stationary phase was investigated by comparing R-type naphthyglucine and 3,5-dinitrobenzoic acid of Chirex_(ⓚ)3005 and O,O'-bis(4-tertbutyl-benzoyl)-N and N'-dially-L_tartardiamic of Kromasil_(ⓚ) CHI-II. S-ketoprofen enantiomer was successfully isolated by using the kromail column as well as the mobile phase of hexane/tert-buyl methyl ether/acetic(v/v)=60/40/0.1. Equilibrium constants of R-and S-ketoprofen were obtained by pulsed injection method(PIM) at various concentrations of ketoprofen racemate and loading sample amounts, which serve the basic infromation on overloading sample volume and amounts. A volume of 100㎕ containing 3.0㎎ of ketoprofen racemate to Kromasil column(250㎜×4.6㎜)resulted in 85% recovery of injected sample under an high concentration(30㎎/㎖)feed condition.

고성능 액체 크로마토그래피를 이용한 Pelubiprofen의 광학분할

김소영 ( Kim So Yeong ),이중기 ( Lee Jung Gi ),서성섭 ( Seo Seong Seob ),박태진 ( Park Tae Jin ),박달근 ( Park Dal Geun ) 한국공업화학회 2003 공업화학 Vol.14 No.6

Pelubiprofen, (±)-(E)-2-[4-(2-Oxo-cyclohexylidenemethyl)-phenyl] propionic acid는 비스테로이드 계열의 진통 및 소염제(NSAID)이다. 이 물질은 골 관절염, 류마티스성 관절염, 근골격성 통증, 수술 후 후유증, 치통 등과 같은 다양한 종류의 질병 치료에 약리활성을 보인다. 본 연구에서는 라세미 pelubiprofen을 HPLC를 이용하여 단일 enantiomer로 분리하기 위한 최적의 조건을 찾기 위한 것이다. 고정상으로는 Kromasil^(ⓡ)CHI-TBB (Eka, chem Sweden) 충진제를 이용하여 4.6×250 mm 크기의 칼럼에 직접 slurry packing을 한 것을 이용하였고, 최적의 이동상의 조성은 hexane/tert-butyl methyl ether/acetic acid의 비가 70/30/0.1 (v/v%)인 것으로 나타났다. 칼럼 효율에 영향을 주는 공정 변수를 조사하였는데 주입농도가 증가함에 따라서 분리도는 4.06에서 3.50으로 감소하였으나 이동상의 유속이 증가함에 따라 주입농도의 증가가 분리도에 미치는 영향은 다소 적은 것을 알 수 있었다. 최적의 분리도를 나타내는 조업조건은 이동상의 유속이 1 mL/min이고 주입부피가 5μL 일 때 4.11로 가장 높은 분리도를 나타내었다. Pelubiprofen, (±)-2-[4-(2-oxocyclohexylidenemethyl)phenyl]propionic acid, is a novel derivative of 2-arylpropionic acid non-steroidal anti-inflammatory drugs (profen NSAIDs). It has a wide variety of indications proposed: osteoarthritis, rheumatoid arthritis, musculoskeletal pain, post-operative trauma, low back pain, scapulohumeral periarthritis, neck-shoulder-arm syndrome and dental pain. In this study, we tried to determine the optimal separation conditions for chiral separation of pelubiprofen using HPLC(High Performance Liquid Chromatography). The Kromasil^(?) CHI-TBB(Eka chem, sweden) was used as a stationary phase and packed in a home-made 46×250 mm column. The mobile phase composition was determined by optimizing the solubility and resolution. It was found to be 70% of hexane, 30% of tert-butyl methyl ether and 0.1% of acetic acid. Process variables effecting column performance were investigated. Resolution decreased from 4.06 to 3.50 with increase of injection eoncentration, but the effect of injection concentration on the resolution was insignificant at higher flow rates. In the our experimental range studied, the highest resolution 4.11 was obtained at the mobile phase flow rate of 1 mL/min with an injection volume of 5 μL.

Determination of rabeprazole enantiomers in commercial tablets using immobilized cellulose-based stationary phase

김미리,유수경,Quoc Ky Truong,Xuan Lan Mai,정현규,강종성,김경호 대한약학회 2017 Archives of Pharmacal Research Vol.40 No.3

Rabeprazole is one of the latest proton-pumpinhibitors used for treatment of several gastrointestinaldisorders. For therapeutic applications, rabeprazole hasbeen administered as a mixture of R-(?) and S-(-) enantiomers. Owing to pharmacological and toxicological differencesbetween stereoisomers, chiral recognition has nowbecome an integral part of drug research and development. A simple and rapid liquid chromatographic method forenantioselective separation and determination of R-(?) andS-(-) enantiomers of rabeprazole in bulk drug and pharmaceuticalformulations was developed. Chiralpak IC(150 9 4.6 mm, 5 lm) column and lmobile phase containinghexane:ethanol:ethylenediamine (30:70:0.05 v/v) inan isocratic mode yielded baseline separation with resolutiongreater than 6.0 at 35 C. Effects of additives andn-hexane were evaluated. Optimized condition was validatedas per ICH guidelines. The method has good linearity,high sensitivity with LOD was 0.01 lg/mL andLOQ was 0.03 lg/mL for both enantiomers. Intra-dayprecision varied between 0.44 and 1.79% for S-(-) enantiomer,0.65 and 1.97% for R-(?) enantiomer. Relativestandard deviations of inter-day precision were less than1.81% for both enantiomers. The percentage recovery forboth enantiomers of rabeprazole ranged between 99.81 and101.95%, 98.82 and 101.36% in material and tablets,respectively. The method was successfully applied todetermine content of each enantiomer in commercialtablets.

Pharmacokinetic Behavior and Tissue Distribution of Verapamil and Its Enantiomers in Rats by HPLC

He, Langchong,Wang, Sicen The Pharmaceutical Society of Korea 2003 Archives of Pharmacal Research Vol.26 No.9

The differences in pharmacokinetic behavior and tissue distribution of verapamil and its enantiomers were investigated in rats. In high-performance liquid chromatographic method, an achiral ODS column (150 mm $\times$ 4.6 mm i.d.) with the mobile phase consisting of methanol-water (73:30, v/v) was used for the determination of the concentration for racemic verapamil, and a Chiralcel OJ column (250 mm$\times$4.6 mm i.d.) with the mixture of n-haxane-ethanol-triethylamine (85:15:0.2, v/v/v) as mobile phase was used to determine the concentrations of verapamil enantiomers. A fluorescence detector in the analytical system was set at excitation and emission wavelengths of 275 nm and 315 nm. The differences between enantiomers were apparent in the pharmacokinetics in rats. The area under the concentration-time curve (AUC) of S-(-) verapamil was higher than that of R-(+) verapamil. The half-distribution time ($T_{1/2(\alpha)}$) of S-(-) verapamil which distributing to tissue from blood was shorter than that of R-(+) verapamil, but the elimination half-time ($T_{1/2(\beta)}$) was longer in rat following oral administration of racemic verapamil. At 1.3 h after oral administration of racemic verapamil, however, there were no significant differences between enantiomers for the distributions in major tissues such as heart, cerebrum, cerebellum, liver, spleen and kidney.

Chiral Separation of Fluvastatin Enantiomers by Capillary Electrophoresis

Trung, Tran Quoc,Dung, Phan Thanh,Hoan, Nguyen Ngoc,Kim, Dae-Joong,Lee, Joo-Huyn,Kim, Kyeong-Ho 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.8

An analytical CE method was developed for the enantiomeric purity determination of fluvastatin enantiomers. Fluvastatin enantiomers were separated on an uncoated fused silica with 100 mM-borate solution containing 30 mg/mL of (2-hydroxypropyl)-$\beta$-cyclodextrin (HP-$\beta$-CD) as running buffer and fenoprofen as an internal standard. The linearity was observed within a $400-700\;{\mu}g/mL$ concentration range ($r^2{\geq}0.995$) for both fluvastatin enantiomers. The repeatability expressed as coefficient of variation (CV) of the method were 0.96 and 0.92% for (+)-3R, 5S and (-)-3S, 5R-fluvastatin, respectively. The limit of detection and quantification for both fluvastatin enantiomers were $1.5\;{\mu}g/mL$ and $2.5\;{\mu}g/mL$, respectively.

Chiral counter-current chromatography of gemifloxacin guided by capillary electrophoresis using (+)-(18-crown-6)-tetracarboxylic acid as a chiral selector

Kim, Eunsook,Koo, Yoon-Mo,Chung, Doo Soo Elsevier 2004 Journal of chromatography Vol.1045 No.1

<P><B>Abstract</B></P><P>(+)-(18-crown-6)-tetracarboxylic acid (18C6H<SUB>4</SUB>) has been known as a highly efficient chiral selector for resolving primary amine enantiomers in capillary electrophoresis (CE). We investigated the chiral separation of gemifloxacin using 18C6H<SUB>4</SUB> in analytical counter-current chromatography (CCC). The separation conditions for CE, including the binding constant, pH, and run buffer constituents, provided a helpful guideline for chiral CCC. A successful separation of gemifloxacin enantiomers could be achieved using a two-phase solvent system composed of 1-butanol-ethyl-acetate-bis(2-hydroxyethyl)aminotris(hydroxymethyl)methane acetate buffer with a small amount of 18C6H<SUB>4</SUB>. The hydrophobicity of the solvent system and the 18C6H<SUB>4</SUB> concentration were varied to optimize the chiral separation.</P>

Quantitative Analysis and Enantiomeric Separation of Ephedra Alkaloids in Ma Huang Related Products by HPLC-DAD and UPLC-MS/MS

Kyoung moon Han,황진우,이선희,박보름,김형일,백선영 한국생약학회 2022 Natural Product Sciences Vol.28 No.4

Ephedra is a genus of the Ephedraceae family and is found in temperate regions, such as Central Asia and Europe. Among the various ephedra species, Ma Huang (Ephedra herb) is derived from the aerial parts of Ephedra sinica Stapf, Ephedra equisetina Bunge, and Ephedra intermedia Schrenk & C.A. Mey. Ma Huang contains various ephedra alkaloids, including (-)-ephedrine, (+)-pseudoephedrine, (-)-norephedrine, (+)-norpseudoephedrine, (-)-methylephedrine, and (+)-methylpseudoephedrine, which are found naturally as single enantiomers, although they can be prepared as racemates. Although the use of Ma Huang in foods is prohibited in Korea, products containing Ma Huang can be imported, and so it is necessary to develop a suitable analytical technique for the detection of Ma Huang in foods. Herein, we report the development of analytical methods for the detection of ephedra alkaloids in products containing Ma Huang. Following sample purification by solid phase extraction, quantitative analysis was performed using ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS). Additionally, the enantiomers were successfully separated using HPLC-DAD. We successfully analyzed various food samples, where the ephedra alkaloids were qualitatively and quantitatively determined, and the enantiomers were separated. It is expected that these methods may contribute toward preventing the distribution of illegal products containing Ma Huang.

Liquid Chromatographic Enantiomer Separation of α-Amino Acid Esters as Nitrobenzoxadiazole Derivatives Using Polysaccharide-Derived Chiral Stationary Phases

Islam, Md. Fokhrul,Lee, Wonjae The Basic Science Institute Chosun University 2015 조선자연과학논문집 Vol.8 No.2

Liquid chromatographic enantiomer separation of ${\alpha}$-amino acid esters as nitrobenzoxadiazole (NBD) derivatives was performed using several chiral stationary phases (CSPs) based on polysaccharide derivatives under fluorescence detection. For enantiomer separation by normal HPLC, the non-aqueous derivatization method of ${\alpha}$-amino acid esters for NBD analytes was introduced. Among the six CSPs used in this study, the performance of Chiralpak IA was superior for enantiomer resolution of NBD derivatives of several ${\alpha}$-amino acid methyl esters. Also the convenient analytical method using polysaccharide-derived CSPs developed in this study was applied to determine the optical purity of ${\alpha}$-amino acids esters. It was investigated that the enantiomeric impurity levels of 0.02-1.73% were found after determination of enantiomeric purities of several commercially available L-amino acid methyl esters. It is expected to be quite useful for enantiomer separation of other ${\alpha}$-amino acid esters as NBD derivatives by normal HPLC.

Dual-mode of Reversed and Normal-phase Liquid Chromatographic Enantiomer Separation on Coated Cellulose Derived Chiral Stationary Phases

Jong Sung Jin,Kyung-Ah Lee,강종성,Young Koo Kang,Chae-Sun Baek,Wonjae Lee* 대한화학회 2006 Bulletin of the Korean Chemical Society Vol.27 No.8