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Quantification of ascorbic acid and acetylsalicylic acid in effervescent tablets by CZE-UV and identification of related degradation products by heart-cut CZE-CZE-MS

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Abstract

Capillary electrophoresis is commonly applied for the analysis of pharmaceutical products due to its high separation efficiency and selectivity. For this purpose, electrospray-ionization-(ESI)-interfering additives or electrolytes are often required, which complicates the identification of impurities and degradation products by mass spectrometry (MS). Here, a capillary zone electrophoresis (CZE) method with ultraviolet (UV) absorption detection for the simultaneous determination and quantification of ascorbic acid and acetylsalicylic acid in effervescent tablets was developed. Related degradation products were identified via CZE-CZE-MS. Systematic optimization yielded 100 mM tricine (pH = 8.8) as appropriate background electrolyte, resulting in baseline separation of ascorbic acid, acetylsalicylic acid, and related anionic UV-active degradation products. The CZE-UV method was successfully validated regarding the guidelines of the Food and Drug Administration. The validated method was applied to trace the degradation rate of the active pharmaceutical ingredients at defined ambient conditions. A heart-cut CZE-CZE-MS approach, including a 4-port-nL-valve, was performed for the identification of the observed degradation products. This 2D setup enables a precise cutting of accurate sample volumes (20 nL) and the independent operation of two physically separated CZE dimensions, which is especially beneficial regarding MS detection. Hence, the ESI-interfering tricine electrolyte components were separated from the analytes in a second electrophoretic dimension prior to ESI-MS detection. The degradation products were identified as salicylic acid and mono- and diacetylated ascorbic acid. This setup is expected to be generally applicable for the mass spectrometric characterization of CZE separated analytes in highly ESI-interfering electrolyte systems.

A CZE-UV method for the quantification of effervescent tablet ingredients and degradation products was developed and validated. In order to identify unknown degradation products separated in the CZE-UV, a 2D heart-cut approach was performed applying a mechanical 4-port-valve. The unknown substances were transferred from the 1st to the 2nd dimension followed by the separation of ESI-interfering tricine from the analytes prior to mass spectrometric detection

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Abbreviations

AA:

Ascorbic acid

APIs:

Active pharmaceutical ingredients

ASA:

Acetylsalicylic acid

BGE:

Background electrolyte

CE:

Capillary electrophoresis

CEC:

Capillary electrochromatography

CIEF:

Capillary isoelectric focusing

CZE:

Capillary zone electrophoresis

DHAA:

Dehydroascorbic acid

DKG:

2,3-Diketogulonic acid

EOF:

Electroosmotic flow

ESI:

Electrospray-ionization

HPLC:

High-performance liquid chromatography

IS:

Internal standard

LOD:

Limit of detection

MS:

Mass spectrometry

NSA:

2-Naphthalenesulfonic acid

PVA:

Poly(vinyl alcohol)

RSD:

Relative standard deviation

S/N:

Signal-to-noise

SA:

Salicylic acid

SL:

Sheath liquid

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Acknowledgments

The authors acknowledge the partners within the MIME project: J&M Analytic AG, LabCognition Analytical Software GmbH & Co. KG, Glatt GmbH Systemtechnik, Pharma Test AG, Bundeskriminalamt Wiesbaden (BKA) and University of Münster and the Federal Ministry of Education and Research (BMBF) for financial support in the course of the announcement “Sicherung der Warenketten” within the scope of the program “Forschung für die zivile Sicherheit” of the Federal Government (FKZ 13 N12009). We thank Norbert Schaschke and Jens Hühner from the University of Aalen for helpful suggestions.

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    1. Faculty of Chemistry, Aalen University, Beethovenstraße 1, 73430, Aalen, Germany

      Sabine Neuberger, Kevin Jooß, Christian Ressel & Christian Neusüß

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    1. Sabine Neuberger
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    2. Kevin Jooß
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    3. Christian Ressel
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    4. Christian Neusüß
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    Correspondence to Christian Neusüß.

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    Published in the topical collection Fundamental Aspects of Electromigrative Separation Techniques with guest editors Carolin Huhn and Pablo A. Kler.

    Sabine Neuberger and Kevin Jooß contributed equally to this work.

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    Neuberger, S., Jooß, K., Ressel, C. et al. Quantification of ascorbic acid and acetylsalicylic acid in effervescent tablets by CZE-UV and identification of related degradation products by heart-cut CZE-CZE-MS. Anal Bioanal Chem 408, 8701–8712 (2016). https://doi.org/10.1007/s00216-016-9734-2

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