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Ion-pairing HPLC methods to determine EDTA and DTPA in small molecule and biological pharmaceutical formulations

Authors: George Wang, Frank P.Tomasella

Abstract:

Ion-pairing high-performance liquid chromatography–ultraviolet (HPLC–UV) methods were developed to determine two commonly used chelating agents, ethylenediaminetetraacetic acid (EDTA) in Abilify® (a small molecule drug with aripiprazole as the active pharmaceutical ingredient) oral solution and diethylenetriaminepentaacetic acid (DTPA) in Yervoy® (a monoclonal antibody drug with ipilimumab as the active pharmaceutical ingredient) intravenous formulation. Since the analytes, EDTA and DTPA, do not contain chromophores, transition metal ions (Cu2+, Fe3+) which generate highly stable metallocomplexes with the chelating agents were added into the sample preparation to enhance UV detection. The use of metallocomplexes with ion-pairing chromatography provides the ability to achieve the desired sensitivity and selectivity in the development of the method. Specifically, the sample preparation involving metallocomplex formation allowed sensitive UV detection. Copper was utilized for the determination of EDTA and iron was utilized for the determination of DTPA. In the case of EDTA, a gradient mobile phase separated the components of the formulation from the analyte. In the method for DTPA, the active drug substance, ipilimumab, was eluted in the void. In addition, the optimization of the concentration of the ion-pairing reagent was discussed as a means of enhancing the retention of the aminopolycarboxylic acids (APCAs) including EDTA and DTPA and the specificity of the method. The analytical method development was designed based on the chromatographic properties of the analytes, the nature of the sample matrix and the intended purpose of the method. Validation data were presented for the two methods. Finally, both methods were successfully utilized in determining the fate of the chelates.

Keywords

EDTA; DTPA; Ion-pairing HPLC; Aripiprazole; Ipilimumab

Citation: George Wang, Frank P.Tomasella Ion-pairing Hplc Methods To Determine Edta And Dtpa In Small Molecule And Biological Pharmaceutical Formulations doi:10.1016/j.jpha.2016.01.002

Received: 6 October 2015, Revised: 15 January 2016, Accepted: 20 January 2016, Available online: 21 January 2016

Copyright: © 2016 Xi'an Jiaotong University. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)

Conclusion

In conclusion, two fit for purpose, ion-pairing reversed-phase HPLC methods were developed to determine EDTA in Abilify® oral solution and DTPA in Yervoy® intravenous solution drug products. TBA was added into the mobile phase to form ion pairs with metallocomplexes of EDTA and DTPA to enhance retention on reversed-phase HPLC columns. The analytical method utilized gradient and isocratic mobile phases in the determination of an APCA in a small molecule and a biological formulation, respectively. The use of metallocomplexes with ion-pairing chromatography provides the ability to achieve the desired sensitivity and selectivity in the development of the method.

Specifically, the sample preparation involving metallocomplex formation allows UV detection with high sensitivity. Copper (II) was utilized for the determination of EDTA and iron (III) was utilized for the determination of DTPA. For the determination of EDTA, a gradient mobile phase separated the components of the formulation from the analyte. In the case of DTPA, the active drug substance, ipilimumab, was eluted in the void. To the best of our knowledge, there is no report on analytical HPLC method for APCAs by direct injections of monoclonal drug formulations. In addition, the optimization of the concentration of the ion-pairing reagent was discussed as a means of enhancing the retention of the APCAs and the specificity of the method. The analytical method development was designed based on the chromatographic properties of the analytes, the nature of the sample matrix and the intended purpose of the method.

Acknowledgments

The authors wish to acknowledge the following that provided the needed supplies or stimulating conversation: Jonathan D. Basch, Jacob Bongers, Mi Jin, David K. Lloyd and Rao V. Mantri, Bristol-Myers Squibb Company New Brunswick, NJ, USA.

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