A Comparative Evaluation of Polystyrene Divinylbenzene Copolymer HPLC Columns on the Chromatographic Performance of the Compendial Method for Doxycycline Hyclate Capsules: Implications for Method Implementation of a Medical Countermeasure Medication

Firat Yerlikaya, Adil Mohammad, Patrick J. Faustino, Mansoor A. Khan, Saeed R. Khan

Division of Product Quality Research, Office of Testing and Research, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, Food and Drug Administration, Silver Spring, MD, USA.

DOI: 10.4236/jasmi.2015.53003                

Abstract

The purpose of this study was to evaluate the impact of polystyrene divinylbenzene copolymer HPLC columns on the chromatographic performance of the USP compendial method for doxycycline hyclate. The compendial method was implemented based on the assessment of the chromatographic performance of six USP defined L21 polystyrene divinylbenzene HPLC columns. Modifications to the method were based on USP <621> for chromatography. The method was validated for the determination of doxycycline hyclate and its impurities in commercially available drug products. A number of different polystyrene-divinylbenzene columns were tested and failed to provide selectivity for the resolution of doxycycline and its impurities. Separation was optimally achieved on an Agilent PLPR-S column (250 × 4.6 mm, 8 μm) by using an Agilent 1260 series HPLC system. Doxycycline hyclate and its impurities were eluted isocratically at a flow rate of 1 mL/min with mobile phase and detected at 270 nm. The column temperature was maintained at 60^oC. The method was validated according to USP category I requirements for Assay. Validation acceptance criteria were met in all cases. The analytical range for doxycycline hyclate was 50 - 250 μg/mL and the linearity was r² > 0.999 over three days. The method was determined to be specific. Both accuracy (95.1% - 102.4%) and precision (0.50% - 4.8%) were established across the analytical range for low, intermediate and high QC concentrations. Method applicability was demonstrated by analyzing marketed products of doxycycline hyclate, in which results showed potency meeting USP acceptance criteria. In conclusion, this study described the remarkable differences in selectivity that were encountered during the implementation phase for the compendial methods for doxycycline and its impurities in marketed products and it could be used in the future to assss the product quality of doxycycline hyclate capsules stored in the National stockpiles.

Keywords

Doxycycline, 4-Epidoxycycline, 6-Epidoxycycline, Metacycline, Polystyrene-Divinylbenzene

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Jantratid, E., Strauch, S., Becker, C., Dressman, J.B., Amidon, G.L., Junginger, H.E., Kopp, S., Midha, K.K., Shah, V.P., Stavchansky, S. and Barends, D.M. (2010) Biowaiver Monographs for Immediate Release Solid Oral Dosage Forms: Doxycycline Hyclate. Journal of Pharmaceutical Sciences, 99, 1639-1653.
[2]	Shariati, S., Yamini, Y. and Esrafili, A. (2009) Carrier Mediated Hollow Fiber Liquid Phase Microextraction Combined with HPLC-UV for Preconcentration and Determination of Some Tetracycline Antibiotics. Journal of Chromatography B, 877, 393-400. http://dx.doi.org/10.1016/j.jchromb.2008.12.042
[3]	Mack, G.D. and Ashworth, R.B. (1978) A High Performance Liquid Chromatographic System for the Analysis of Tetracycline Drug Standards, Analogs, Degradation Products and Other Impurities. Journal of Chromatographic Science, 16, 93-101. http://dx.doi.org/10.1093/chromsci/16.3.93
[4]	Oka, H., Ikai, Y., Kawamura, N., Uno, K., Yamada, M., Harada, K. and Suzuki, M. (1987) Improvement of Chemical Analysis of Antibiotics: XII. Simultaneous Analysis of Seven Tetracyclines in Honey. Journal of Chromatography A, 400, 253-261. http://dx.doi.org/10.1016/S0021-9673(01)81619-8
[5]	Oka, H., Ikai, Y., Kawamura, N., Uno, K., Yamada, M., Harada, K., Uchiyama, M., Asukabe, H. and Suzuki, M. (1987) Improvement of Chemical Analysis of Antibiotics. X. Determination of Eight Tetracyclines Using Thin-Layer and High-Performance Liquid Chromatography. Journal of Chromatography, 393, 285-296.
[6]	Ding, X.J. and Mou, S.F. (2000) Ion Chromatographic Analysis of Tetracyclines Using Polymeric Column and Acidic Eluent. Journal of Chromatography A, 897, 205-214. http://dx.doi.org/10.1016/S0021-9673(00)00779-2
[7]	Seth, P. and Stamm, A. (1986) Quantitative Estimation and Separation of Doxycycline HCl and Its Related Products. Drug Development and Industrial Pharmacy, 12, 1469-1475. http://dx.doi.org/10.3109/03639048609065871
[8]	Pesek, J.J. and Matyska, M.T. (1996) Separation of Tetracyclines by High-Performance Capillary Electrophoresis and Capillary Electrochromatography. Journal of Chromatography A, 736, 313-320. http://dx.doi.org/10.1016/0021-9673(95)01316-4
[9]	Kazemifard, A.G. and Moore, D.E. (1997) Evaluation of Amperometric Detection for the Liquid-Chromatographic Determination of Tetracycline Antibiotics and Their Common Contaminants in Pharmaceutical Formulations. Journal of Pharmaceutical and Biomedical Analysis, 16, 689-696. http://dx.doi.org/10.1016/S0731-7085(97)00089-7
[10]	Zhao, F.N., Zhang, X.Z. and Gan, Y.R. (2004) Determination of Tetracyclines in Ovine Milk by High-Performance Liquid Chromatography with a Coulometric Electrode Array System. Journal of Chromatography A, 1055, 109-114. http://dx.doi.org/10.1016/j.chroma.2004.08.131
[11]	Skulason, S., Ingolfsson, E. and Kristmundsdottir, T. (2003) Development of a Simple HPLC Method for Separation of Doxycycline and Its Degradation Products. Journal of Pharmaceutical and Biomedical Analysis, 33, 667-672. http://dx.doi.org/10.1016/S0731-7085(03)00316-9
[12]	Yekkala, R., Diana, J., Adams, E., Roets, E. and Hoogmartens, J. (2003) Development of an Improved Liquid Chromatographic Method for the Analysis of Doxycycline. Chromatographia, 58, 313-316.
[13]	Kogawa, A.C. and Salgado, H.R.N. (2012) Doxycycline Hyclate: A Review of Properties, Applications and Analytical Methods. International Journal of Life Science and Pharma Research, 2, 11-25.
[14]	Hoogmartens, J., Khan, N.H., Vanderhaeghe, H., Vanderleeden, A.L., Oosterbaan, M., Veldtulp, G.L., Plugge, W., Vandervlies, C., Mialanne, D., Melamed, R. and Miller, J.H.M. (1989) A Collaborative Study of the Analysis of Doxycycline Hyclate by High-Performance Liquid Chromatography on Polystyrene-Divinylbenzene Packing Materials. Journal of Pharmaceutical and Biomedical Analysis, 7, 601-610. http://dx.doi.org/10.1016/0731-7085(89)80226-2
[15]	USP37-NF32 (2014) United States Pharmacopeia 37: National Formulary 32.
[16]	EP8.0 (2014) European Pharmacopoeia 8.0.
[17]	BP (2014) British Pharmacopoeia 2014.
[18]	Dihuidi, K., Kucharski, M.J., Roets, E., Hoogmartens, J. and Vanderhaeghe, H. (1985) Quantitative-Analysis of Doxycycline and Related Substances by High-Performance Liquid Chromatography. Journal of Chromatography A, 325, 413-424. http://dx.doi.org/10.1016/S0021-9673(00)96051-5
[19]	Young, J.E., Matyska, M.T., Azad, A.K., Yoc, S.E. and Pesek, J.J. (2013) Separation Differences among Phenyl Hydride, Udc Cholesterol, and Bidentate C8 Stationary Phases for Stability Indicating Methods of Tetracyclines. Journal of Liquid Chromatography & Related Technologies, 36, 926-942.
[20]	De Leenheer, A.P. and Nelis, H.J. (1977) Reversed-Phase High-Performance Liquid Chromatography of Doxycycline. Journal of Chromatography A, 140, 293-299. http://dx.doi.org/10.1016/S0021-9673(00)93594-5
[21]	Pilorz, K. and Choma, I. (2004) Isocratic Reversed-Phase High-Performance Liquid Chromatographic Separation of Tetracyclines and Flumequine Controlled by a Chaotropic Effect. Journal of Chromatography A, 1031, 303-305. http://dx.doi.org/10.1016/j.chroma.2003.12.024
[22]	Naidong, W., Verresen, K., Busson, R., Roets, E. and Hoogmartens, J. (1991) Isolation of Doxycycline, 6-Epidoxycy- cline and 2-Acetyl-2- Decarboxamidometacycline from Commercial Metacycline by Preparative Column Liquid Chro- matography on Silica Gel. Journal of Chromatography A, 586, 67-72. http://dx.doi.org/10.1016/0021-9673(91)80026-D
[23]	Tylova, T., Kamenik, Z., Flieger, M. and Olsovska, J. (2011) Comparison of LC Columns Packed with 2.6 mu m Core-Shell and Sub-2 mu m Porous Particles for Gradient Separation of Antibiotics. Chromatographia, 74, 19-27. http://dx.doi.org/10.1007/s10337-011-2021-7
[24]	Waters (2014) Waters Corporation. http://www.waters.com/webassets/cms/support/docs/WAT044491.pdf
[25]	Monser, L. and Darghouth, F. (2000) Rapid Liquid Chromatographic Method for Simultaneous Determination of Tetracyclines Antibiotics and 6-Epi-Doxycycline in Pharmaceutical Products Using Porous Graphitic Carbon Column. Journal of Pharmaceutical and Biomedical Analysis, 23, 353-362. http://dx.doi.org/10.1016/S0731-7085(00)00329-0