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

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.

Long-term stability of gentamicin sulfate-ethylenediaminetetraacetic acid disodium salt (EDTA-Na2) solution for catheter locks

“…As reported in previous study assaying EDTA-Na 2 in pharmaceutical formulation, EDTA-Cu 2+ , as ionic analyte, showed minimal retention (Fig. 2C, < 1 min) through reversed-phase by using liquid chromatography with ion-pairing and elution gradient [42], and co-elution of nitrilotriacetic acid-Cu 2+ complex (Fig. 2B).

Fig.

…”

Encapsulation of Zn-DTPA into poly lactic-co-glycolic acid nanoparticles via a modified double emulsion method for extended release into lung fluid

“…Then, 0.4 µL of the dispersed nanoparticles was transferred into 1.5 mL centrifugation tubes. At appropriate time intervals, the dispersed NPs were centrifuged, and 20 µL of the supernatant was transfer into a vial, and diluted with 100 µL of iron (III) chloride solution at 5 mg/mL and 880 µL of 0.1% formic acid to obtain 1 mL as a total volume [43]. Samples were complexed to Fe-DTPA 2-due to the highest affinity of DTPA 5towards the Fe3+ metal ion compared to various metal ions such as Ca 2+ , Cd 2+ , Co 2+ , Cu 2+ , Mn 2+ and Zn 2+ that are present in the lung fluid [44].…”

Prospects for ion chromatography in quality assessment of biologicals

“…[1] и за короткое время из способа детектирования небольшого числа ионов превратился в самостоятельный вид анализа [2]. Ион-парная ВЭЖХ, ион-эксклюзионная ВЭЖХ, ионообменная ВЭЖХ с последующим УФ-или флуориметрическим детектированием продуктов разделения применяется при оценке качества биологических лекарственных препаратов (БЛП) достаточно давно [3][4][5][6][7][8][9][10][11][12][13][14][15][16]. Также в последние годы в нормативной документации на БЛП представлены методики ионной хроматографии с кондуктометрическим или амперометрическим детектированием.…”