Analytical and Bioanalytical Chemistry

, Volume 408, Issue 30, pp 8701–8712 | Cite as

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

  • Sabine Neuberger
  • Kevin Jooß
  • Christian Ressel
  • Christian Neusüß
Research Paper
First Online:
Received:
Revised:
Accepted:
Part of the following topical collections:
  1. Fundamental Aspects of Electromigrative Separation Techniques

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.

Graphical Abstract

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

Keywords

Two-dimensional separation Mechanical valve Capillary zone electrophoresis-mass spectrometry Interference-free electrospray-ionization Interface 

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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Notes

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.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Sabine Neuberger
    • 1
  • Kevin Jooß
    • 1
  • Christian Ressel
    • 1
  • Christian Neusüß
    • 1
  1. 1.Faculty of ChemistryAalen UniversityAalenGermany

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