HPLC Determination of Dimethyl-4,4'-Dimethoxy-5,6,5',6' Dimethylene Dioxybiphenyl-2,2'-Dicarboxylate (DDB) and Co-Adminstered Diuretics as an Extemporaneous Mixture in Human Plasma


  • University of Alexandria, El-Messalah, Department of Pharmaceutical Analytical Chemistry, Alexandria, 21521, Egypt


Background: Diuretics are co-adminstered with hepatoprotectant drugs for the treatment of oedema and ascitis in patients suffering from hepatic cirrhosis.This work presents an accurate and simple method for the in vivo analysis of Dimethyl-4,4'-dimethoxy-5,6,5',6'-dimethylenedioxybiphenyl-2,2'-dicarboxylate (DDB), a hepatoprotectant drug, with hydrochlorothiazide (HCT) and amiloride hydrochloride (AM) diuretics in human plasma using HPLC. Methodology/Principal Findings: The separation was achieved on Spheri-5 RP-8 (5μ) and Spheri-5 RP-18 (5μ) columns (220 X 4.6 mm i.d.) using 0.02 M disodium hydrogen phosphate, 0.12% w/v sodium laurylsulfate, 0.1% v/v triethylamine adjusted to pH 4 and acetonitrile in a ratio 65:35 v/v as mobile phase. The separation was carried out at ambient temperature with a flow rate of 1.0 mL min-1. Quantitation was achieved with UV detection at 278 nm based on peak area with linear calibration curves at concentration ranges 0.03- 30 μg mL -1 for HCT and 0.05- 30 μg mL -1 for both AM amd DDB. Conclusions: The proposed method was successfully applied to the determination of the investigated drugs in human plasma sample obtained from a healthy male volunteer four hours after single oral dose administration of the three drugs. The proposed method was validated in terms of linearity, accuracy, precision, limits of detection and quantitation and other aspects of analytical validation.


Bifendate, Amiloride Hydrochloride, Hydrochlorothiazide, Co-Administered, Human Plasma, HPLC.

Subject Discipline

Pharmacy and Pharmacology

Full Text:


X. S. Li, B. L. Zhao, G. T. Liu, and W. J. Xiu, Scavenging effects on active oxygen radicals by Schizandrins with different structures and configurations, Free Radic. Biol. Med. 1990; 9(2), 99-104.

P. Y. Chin, H. Y. Leung, M. K. Poo, and K. M. Ko, Effects of Schisandrin B Enantiomers on Cellular Glutathione and Menadione Toxicity in AML12 Hepatocytes, Pharmacology 2006; 77(2), 63-70.

G. M. Hadad, Validated stability-indicating HPLC method for the determination of dimethyl-4,4_dimethoxy-5,6,5_,6_-dimethylene dioxybiphenyl-2,2_-dicarboxylate (DDB) and its degradation products, J. Pharm. Biomed. Anal. 2008; 47, 695–703.

C. K. Jeong, S. B. Kim, S. J. Choi, D. H. Sohn, G. I. Ko, and H. S. Lee, Rapid microbore liquid chromatographic analysis of biphenyldimethyl dicarboxylate in human plasma with on-line column switching, J. Chromatogr. B: Biomed. Appl. 2000; 738(1), 175-179.

L. Zhou, L. Gu, Y. Wang, and J. Liang, Determination of bifendate in human plasma by HPLC–MS and bioequivalence on bifendate pills in healthy volunteers, J. Pharm. Biomed. Anal. 2006; 40(4), 10251030.

Z. Chena, J. Zhua, H. Chenb, and Y. Xiaoa, A simple HPLC method for the determination of bifendate: Application to a pharmacokinetic study of bifendate liposome, J. Chromatogr. B 2007; 857(2), 246-250

T. M. Huang, Z. He, B. Yang, L. P. Shao, X. W. Zheng, and G. L. Duan, Simultaneous determination of captopril and hydrochlorothiazide in human plasma by reverse-phase HPLC from linear gradient elution, J. Pharm. Biomed. Anal. 2006; 41, 644-648.

N. Sultana, M. S. Arayne, S. S. Ali, and S. Sajid, Simultaneous determination of olmesartan medoxomil and irbesartan and hydrochlorothiazide in pharmaceutical formulations and human serum using high performance liquid chromatography, Chinese Journal of Chromatography 2008; 26(5), 544-549.

A. Dominquiz-Vidal, P. Ortega-Barrales, and A. Molina-Diaz, Fast flow-injection fluorimetric determination of amiloride by using a solid sensing zone, Talanta 2002; 56(6), 1005-1013.

A. C. Moffat, M. D. Osselton, and B. Widdop, Clarke’s Analysis of Drugs and Poisons, thirrd edition, London, 2004.

Y. Lee, C. K. Shim, and S. S. Chung, Determination of Dimethyl-4,4'-Dimethoxy-5,6,5',6'-Dimethylene Dioxybiphenyl-2,2'-Dicarboxylate in Human Serum by High Performance Liquid Chromatography Anal.

Lett. 1999; 32(5), 945-954.

The United States Pharmacopeia, Thirty-second edition, The National Formulary, Twenty-seventh edition (2009), United States Pharmacopeial Convention, Inc., p. 1095.


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