Column-switching High-performance Liquid Chromatography for On-line Simultaneous Determination and Resolution of Enantiomers of Verapamil and Its Metabolites

Overview

Journal Pharm Res

Specialties Pharmacology
Pharmacy

Date 1991 Aug 1

PMID 1924170

Citations 3

Authors

Y Oda

N Asakawa

T Kajima

Y Yoshida

T Sato

Affiliations

Soon will be listed here.

Abstract

An on-line simultaneous assay for the enantiomers of verapamil (VA) and its three metabolites in plasma was developed with column-switching HPLC. This system consists of an ovomucoid protein chiral stationary phase coupled to an achiral reversed-phase column via a dilution tube and a trapping column. The reversed-phase column was used to separate and quantitate VA, its metabolites, and internal standard, without interference from plasma components. Then each of the eluates containing VA and its metabolites was selectively switched into a sampling loop, and the samples were transferred successively to the trapping column after dilution with a new mobile phase. After concentration on the trapping column, each sample was passed to the ovomucoid column, where the enantiomers were resolved and quantitated. The mobile phases for both HPLC columns were independently optimized and the diffusion of the sample during column switching was minimized. This method was shown to be efficient and reliable.

Citing Articles

Development and Validation of a Chiral Liquid Chromatographic Assay for Enantiomeric Separation and Quantification of Verapamil in Rat Plasma: Stereoselective Pharmacokinetic Application.

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Chromatographic Studies of Protein-Based Chiral Separations.

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References

Armstrong D, Ward T, Armstrong R, Beesley T . Separation of drug stereoisomers by the formation of beta-cyclodextrin inclusion complexes. Science. 1986; 232(4754):1132-5. DOI: 10.1126/science.3704640. View

Wainer I, Stiffin R . Direct resolution of the stereoisomers of leucovorin and 5-methyltetrahydrofolate using a bovine serum albumin high-performance liquid chromatographic chiral stationary phase coupled to an achiral phenyl column. J Chromatogr. 1988; 424(1):158-62. DOI: 10.1016/s0378-4347(00)81088-9. View

Echizen H, Vogelgesang B, Eichelbaum M . Effects of d,l-verapamil on atrioventricular conduction in relation to its stereoselective first-pass metabolism. Clin Pharmacol Ther. 1985; 38(1):71-6. DOI: 10.1038/clpt.1985.137. View

Kuwada M, TATEYAMA T, Tsutsumi J . Simultaneous determination of verapamil and its seven metabolites by high-performance liquid chromatography. J Chromatogr. 1981; 222(3):507-11. DOI: 10.1016/s0378-4347(00)84156-0. View

Cole S, Flanagan R, Johnston A, Holt D . Rapid high-performance liquid chromatographic method for the measurement of verapamil and norverapamil in blood plasma or serum. J Chromatogr. 1981; 218:621-9. DOI: 10.1016/s0021-9673(00)82087-7. View

Ellrodt G, Chew C, Singh B . Therapeutic implications of slow-channel blockade in cardiocirculatory disorders. Circulation. 1980; 62(4):669-79. DOI: 10.1161/01.cir.62.4.669. View

Echizen H, Brecht T, Niedergesass S, Vogelgesang B, Eichelbaum M . The effect of dextro-, levo-, and racemic verapamil on atrioventricular conduction in humans. Am Heart J. 1985; 109(2):210-7. DOI: 10.1016/0002-8703(85)90585-x. View

SCHAMROTH L, Krikler D, Garrett C . Immediate effects of intravenous verapamil in cardiac arrhythmias. Br Med J. 1972; 1(5801):660-2. PMC: 1787795. DOI: 10.1136/bmj.1.5801.660. View

Kapur P, Law T, Watson E . Simultaneous quantitation of verapamil, norverapamil, and N-dealkylated metabolites in human plasma following oral administration. J Chromatogr. 1985; 337(1):160-5. DOI: 10.1016/0378-4347(85)80024-4. View

10.

Eichelbaum M, Mikus G, Vogelgesang B . Pharmacokinetics of (+)-, (-)- and (+/-)-verapamil after intravenous administration. Br J Clin Pharmacol. 1984; 17(4):453-8. PMC: 1463390. DOI: 10.1111/j.1365-2125.1984.tb02371.x. View

11.

Harapat S, Kates R . High-performance liquid chromatographic analysis of verapamil. II. Simultaneous quantitation of verapamil and its active metabolite, norverapamil. J Chromatogr. 1980; 181(3-4):484-9. View

12.

Vogelgesang B, Echizen H, Schmidt E, Eichelbaum M . Stereoselective first-pass metabolism of highly cleared drugs: studies of the bioavailability of L- and D-verapamil examined with a stable isotope technique. Br J Clin Pharmacol. 1984; 18(5):733-40. PMC: 1463564. DOI: 10.1111/j.1365-2125.1984.tb02536.x. View

13.

Chu Y, Wainer I . Determination of the enantiomers of verapamil and norverapamil in serum using coupled achiral-chiral high-performance liquid chromatography. J Chromatogr. 1989; 497:191-200. DOI: 10.1016/0378-4347(89)80018-0. View

14.

Schill G, Wainer I, Barkan S . Chiral separations of cationic and anionic drugs on an alpha 1-acid glycoprotein-bonded stationary phase (EnantioPac). II. Influence of mobile phase additives and pH on chiral resolution and retention. J Chromatogr. 1986; 365:73-88. DOI: 10.1016/s0021-9673(01)81544-2. View

15.

Chu Y, Wainer I . The measurement of warfarin enantiomers in serum using coupled achiral/chiral, high-performance liquid chromatography (HPLC). Pharm Res. 1988; 5(10):680-3. DOI: 10.1023/a:1015991324316. View

16.

Hasegawa J, Fujita T, Hayashi Y, Iwamoto K, Watanabe J . pKa determination of verapamil by liquid-liquid partition. J Pharm Sci. 1984; 73(4):442-5. DOI: 10.1002/jps.2600730405. View

17.

Pieper J, Rutledge D . Determination of verapamil and its primary metabolites in serum by ion-pair adsorption high-performance liquid chromatography. J Chromatogr Sci. 1988; 26(9):473-7. DOI: 10.1093/chromsci/26.9.473. View

18.

Singh B, Ellrodt G, Peter C . Verapamil: a review of its pharmacological properties and therapeutic use. Drugs. 1978; 15(3):169-97. DOI: 10.2165/00003495-197815030-00001. View

19.

Kacprowicz A, Fullinfaw R, Bury R . High-performance liquid chromatographic measurement of verapamil in plasma using a diol column. J Chromatogr. 1985; 337(2):412-5. DOI: 10.1016/0378-4347(85)80056-6. View

20.

Miwa T, Miyakawa T, Kayano M, Miyake Y . Application of an ovomucoid-conjugated column for the optical resolution of some pharmaceutically important compounds. J Chromatogr. 1987; 408:316-22. DOI: 10.1016/s0021-9673(01)81816-1. View