A sensitive and precise RP-HPLC method has been developed for the simultaneous estimation of clidinium bromide (CDB) and chlordiazepoxide (CDZ) in pure and pharmaceutical formulations. The separation was achieved on a Nucleodur C8 ( ？mm i.d., 5？μm particle size) column at 25°C. CH3CN-MeOH-NH4OAc 0.1M (30？:？40？:？30, v/v/v) was used as the mobile phase at a flow rate of 1.0？mL？min？1 and detector wavelength at 218？nm. Almotriptan (ALT) was used as internal standard. The validation of the proposed method was carried out for linearity, accuracy, precision, LOD, LOQ, and robustness. The method showed good linearity in the ranges of 2.5–300.0 and 3.0–500.0？μg？mL？1 for CDB and CDZ, respectively. The percentage recovery obtained for CDB and CDZ was 100.40–103.38 and 99.98–105.59%, respectively. LOD and LOQ were 0.088 and 0.294？μg？mL？1 for CDB and 0.121 and 0.403？μg？mL？1 for CDZ, respectively. The proposed method was successfully applied to the determination of CDB and CDZ in combined dosage forms and the results tallied well with the label claim. 1. Introduction Chlordiazepoxide (7-chloro-N-methyl-5-phenyl-3H-1, 4-benzodiazepine-2-amina-4-oxide) is used as an anxiolytic, sedative, hypnotic, anticonvulsant, and/or skeletal muscle relaxant. The drug may inhibit monosynaptic and polysynaptic reflexes by acting as an inhibitory neuronal transmitter or by blocking excitatory synaptic transmission. The drug may also directly depress motor nerve and muscle function [1, 2]. Clidinium bromide (3-[(hydroxy-diphenylacetyl)-oxy]-1-methyl-1-azoniabicylo-[2.2.2] octane bromide is an anticholinergic drug which may help symptoms of cramping and abdominal stomach pain by decreasing stomach acid and slowing the intestines. It is commonly prescribed in combination with chlordiazepoxide by the name of clidinium-c [3]. The United States Pharmacopeia (USP) stated the nonaqueous titration method for the assay of clidinium bromide and chlordiazepoxide [4]. Few methods for the determination of clidinium bromide and chlordiazepoxide in combined dosage forms including HPLC [5–7], derivative spectrophotometry [8, 9], spectrophotometry using multivariate calibration techniques [10], and capillary SFC [11] have been reported. Literature survey revealed that some analytical methods have been used for the individual estimation of clidinium bromide and chlordiazepoxide. Capillary electrophoresis [12] and kinetic spectrophotometric [13] methods for clidinium bromide have been described. Chlordiazepoxide has been determined either alone or with other compounds in pharmaceutical formulations using
References
[1]
G. K. Mcevory, Ed., AHFS Drug Information, American Society of Hospital Pharmacists, 1990.
[2]
J. Gasparic and J. Zimak, “Analysis of the 1, 4-benzodiazepines by methods based on hydrolysis,” Journal of Pharmaceutical and Biomedical Analysis, vol. 1, no. 3, pp. 259–279, 1983.
[3]
B. C. Rudy and B. Z. Senkowski, “Clidinium Bromide,” in Analytical Profiles of Drug Substances, K. Florey, Ed., vol. 2, pp. 145–161, Academic Press, New York, NY, USA, 1973.
[4]
United States Pharmacopoeia, United States Pharmacopoeia Convention, Rockville, Md, USA, 34th edition, 2011.
[5]
A. Pathak, P. Rai, and S. J. Rajput, “Stability-indicating HPLC method for simultaneous determination of clidinium bromide and chlordiazepoxide in combined dosage forms,” Journal of Chromatographic Science, vol. 48, no. 3, pp. 235–239, 2010.
[6]
S. M. Yuen and G. Lehr, “Liquid chromatographic determination of clidinium bromide and clidinium bromide-chlordiazepoxide hydrochloride combinations in capsules,” Journal of the Association of Official Analytical Chemists, vol. 74, no. 3, pp. 461–464, 1991.
[7]
I. M. Jalal, S. I. Sa'sa, A. Hussein, and H. S. Khalil, “Reverse-phase high-performance liquid chromatographic determination of clidinium bromide and chlordiazepoxide in tablet formulations,” Analytical Letters, vol. 20, no. 4, pp. 635–655, 1987.
[8]
M. I. Toral, P. Richter, N. Lara, P. Jaque, C. Soto, and M. Saavedra, “Simultaneous determination of chlordiazepoxide and clidinium bromide in pharmaceutical formulations by derivative spectrophotometry,” International Journal of Pharmaceutics, vol. 189, no. 1, pp. 67–74, 1999.
[9]
S. A. ？zkan, N. Erk, and Z. Sentürk, “Simultaneous determination of two-component mixtures in pharmaceutical formulations containing chlordiazepoxide by ratio spectra derivative spectrophotometry,” Analytical Letters, vol. 32, no. 3, pp. 497–520, 1999.
[10]
M. R. Khoshayand, H. Abdollahi, A. Moeini, A. Shamsaie, A. Ghaffari, and S. Abbasian, “Simultaneous spectrophotometric determination of chlordiazepoxide and clidinium using multivariate calibration techniques,” Drug Testing and Analysis, vol. 2, no. 9, pp. 430–435, 2010.
[11]
N. K. Jagota and J. T. Stewart, “Separation of chlordiazepoxide and selected chlordiazepoxide mixtures using capillary SFC,” Journal of Liquid Chromatography, vol. 16, no. 2, pp. 291–305, 1993.
[12]
B. Nickerson, “The determination of a degradation product in clidinium bromide drug substance by capillary electrophoresis with indirect UV detection,” Journal of Pharmaceutical and Biomedical Analysis, vol. 15, no. 7, pp. 965–971, 1997.
[13]
A. Sheibani, M. R. Shishehbore, and Z. T. Ardakani, “Kinetic spectrophotometric determination of bromide in clidinium-c drug,” Chinese Chemical Letters, vol. 22, no. 5, pp. 595–598, 2011.
[14]
S. E. Roberts and M. F. Delaney, “Determination of chlordiazepoxide, its hydrochloride and related impurities in pharmaceutical formulations by reversed-phase high-performance liquid chromatography,” Journal of Chromatography, vol. 283, pp. 265–272, 1984.
[15]
J. B. Zagar, F. J. Van Lenten, and G. P. Chrekian, “High pressure liquid chromatographic separation and quantitation of chlordiazepoxide.HCl and two of its related compounds,” Journal of the Association of Official Analytical Chemists, vol. 61, no. 3, pp. 678–682, 1978.
[16]
D. Burke and H. Sokoloff, “Simultaneous high-performance liquid chromatographic determination of chlordiazepoxide and amitriptyline hydrochloride in two-component tablet formulations,” Journal of Pharmaceutical Sciences, vol. 69, no. 2, pp. 138–140, 1980.
[17]
R. T. Sane, D. P. Gangal, R. V. Tendolkar, R. M. Kothurkar, and K. D. Ladage, “Simultaneous high performance liquid chromatographic determination of amitriptyline hydrochloride and chlordiazepoxide from pharmaceutical preparations,” Indian Journal of Pharmaceutical Sciences, vol. 51, no. 2, pp. 68–70, 1989.
[18]
M. A. Abuirjeie and M. E. Abdel-Hamid, “Simultaneous high-performance liquid chromatographic and first-derivative spectrophotometric determination of amitriptyline hydrochloride and chlordiazepoxide in capsules,” Analytical Letters, vol. 22, no. 4, pp. 951–962, 1989.
[19]
R. S. Haggag, R. A. Shaalan, and T. S. Belal, “Validated HPLC determination of the two fixed dose combinations (chlordiazepoxide hydrochloride and mebeverine hydrochloride; carvedilol and hydrochlorothiazide) in their tablets,” Journal of AOAC International, vol. 93, no. 4, pp. 1192–1200, 2010.
[20]
S. K. Patel and N. J. Patel, “Simultaneous RP-HPLC estimation of trifluoperazine hydrochloride and chlordiazepoxide in tablet dosage forms,” Indian Journal of Pharmaceutical Sciences, vol. 71, no. 5, pp. 545–547, 2009.
[21]
B. Q. Che, “Determination of five components in compound hypotensive tablet by HPLC,” Yaoxue Xuebao, vol. 39, no. 8, pp. 618–620, 2004.
[22]
A. Zevzikoviene, A. Zevzikovas, and A. Bertulis, “Determination of diazepine derivatives: alprazolam, medazepam, chlordiazepoxid mixture by high performance liquid chromatography,” Medicina, vol. 39, pp. 37–41, 2003.
[23]
S. K. Patel and N. J. Patel, “Simultaneous determination of imipramine hydrochloride and chlordiazepoxide in pharmaceutical preparations by spectrophotometric, rp-hplc, and hptlc methods,” Journal of AOAC International, vol. 93, no. 3, pp. 904–910, 2010.
[24]
M. I. Walash, M. Rizk, and A. El-Brashy, “Spectrophotometric determination of chlordiazepoxide and nitrazepam,” Talanta, vol. 35, no. 11, pp. 895–898, 1988.
[25]
D. J. White, J. T. Stewart, and I. L. Honigberg, “Quantitative analysis of chlordiazepoxide hydrochloride and related compounds in drug substance and tablet dosage form by HPTLC and scanning densitometry,” Journal of Planar Chromatography Modern, vol. 4, no. 4, pp. 330–332, 1991.
[26]
G. B. El-Hefnawey, I. S. El-Hallag, E. M. Ghoneim, and M. M. Ghoneim, “Voltammetric behavior and quantification of the sedative-hypnotic drug chlordiazepoxide in bulk form, pharmaceutical formulation and human serum at a mercury electrode,” Journal of Pharmaceutical and Biomedical Analysis, vol. 34, no. 1, pp. 75–86, 2004.
[27]
Y. M. Issa, N. T. Abdel-Ghani, A. F. Shoukry, and H. M. Ahmed, “New conventional coated-wire ion-selective electrodes for flow-injection potentiometric determination of chlordiazepoxide,” Analytical Sciences, vol. 21, no. 9, pp. 1037–1042, 2005.
[28]
D. J. Garretty, K. Wolff, and A. W. M. Hay, “Micro-extraction of chlordiazepoxide and its primary metabolites, desmethylchlordiazepoxide and demoxepam, from plasma and their measurement by liquid chromatography,” Annals of Clinical Biochemistry, vol. 35, no. 4, pp. 528–533, 1998.
[29]
J. H. Riddick, “An ultraviolet and visible spectrophotometric assay method for chlordiazepoxide,” Clinical Biochemistry, vol. 6, no. 3, pp. 189–199, 1973.
[30]
G. L. Long and J. D. Winefordner, “Limit of detection: a closer look at the IUPAC definition,” Analytical Chemistry A, vol. 55, no. 7, pp. 712–724, 1983.
