LC/MS for the degradation profiling of cough–cold products under forced conditions

doi:10.1016/j.jpba.2004.03.011

Journal of Pharmaceutical and Biomedical Analysis

Volume 35, Issue 5, 3 September 2004, Pages 1035-1045

https://doi.org/10.1016/j.jpba.2004.03.011 Get rights and content

Abstract

Heat, acid, base, UV radiation and oxidation stress methods were applied to study the stability of cough–cold products containing acetaminophen, phenylephrine or phenylpropanolamine hydrochloride and chlorpheniramine maleate. Liquid chromatography coupled with mass spectrometry was used to analyze the degraded samples and obtain molecular weights information. Different volatile buffers (ammonium bicarbonate and ammonium acetate) were assayed in LC/MS methods and retention times of the analytes were compared with those obtained in HPLC with UV detection employing a conventional sodium phosphate buffer to establish the possibility of results transference between the two systems.

Introduction

Pharmaceutical formulations against the common cold use to contain acetaminophen, phenylephrine or phenylpropanolamine hydrochloride and chlorpheniramine maleate. Acetaminophen presents known impurities commercially available such as 4-aminophenol, 4-chloracetanilide and 4-nitrophenol, but this is not the case with the other compounds. To establish the stability characteristics and degradation pathways of these compounds, forced degradation studies (stress testing) were carried out on these drug substances under heat, acid, base, UV radiation and oxidation stress methods, as recommended the ICH guideline Q1A(R2) on stability testing of new drug substances and products [1]. Forced degradation has to demonstrate specificity when developing stability-indicating methods and for this reason it should be performed prior to implementation of stability studies to assure analytical methods are stability-indicating. With that purpose, it is necessary documented evidence that analytical procedures are validated and suitable for the detection and quantisation of degradation products, as emphasizes the ICH guideline Q3B(R) entitled “impurities in new drug products” [2]. It is also required that analytical methods should be validated to demonstrate that impurities unique to the new drug substance do not interfere with, or are separated from, specified and unspecified degradation products in the drug products.

The techniques employed for the analysis of stability samples can be titrimetric, spectophotometric and chromatographic methods. A critical review of these methods is shown in the work of Bakshi and Singh [3]. Within the chromatographic methods, HPLC is the most employed due to its high-resolution capacity, sensitivity and specificity, and because a lot of compounds can be analysed using this technique. Nevertheless, some times the information that supplies the HPLC is not enough for identity confirmation of known and unknown degradation products and their selective determination. For this purpose, hyphenated LC/MS technique that supplies molecular weight and fragmentation information is an indispensable tool of analysis. But in many cases, method development is required because HPLC/UV methods are not easily transferred to LC/MS due to many of the HPLC applications require non-volatile buffer solutions for optimal separation that the LC and MS interface does not tolerate.

The objective of this study was the transference of a HPLC method with a phosphate buffer to the LC/MS system for the determination of the impurity profiling of the components of a cough–cold formulation, employing volatile buffers like ammonium bicarbonate and ammonium acetate.

Section snippets

Chemicals

Standards of actives (acetaminophen, phenylephrine, phenylpropanolamine and chlorpheniramine maleate) and impurities (4-aminophenol, 4-chloracetanilide and 4-nitrophenol) as well as capsules, and excipients of the specialities were kindly provided by CINFA, S.A. (Pamplona, Spain). NaOH and H₂O₂ were from Panreac (Barcelona, Spain), H₃PO₄, NH₄COOCH₃ and CH₃CN (HPLC) from Merck (Darmstadt, Germany), NH₄HCO₃ was from Sigma–Aldrich (Steinheim, Germany), HCl was from Carlo Erba and water was

LC/MS method optimization

In the previous work, we developed and validated a HPLC method for the simultaneous determination of acetaminophen, phenylephrine and chlorpheniramine in pharmaceutical formulations such as capsules and sachets. It included the separation of impurities, excipients and phenylpropanolamine, which can be contained in some formulations instead of phenylephrine. The method was a gradient elution in a Symmetry Shield RP8 [4]. The run lasted for 20 min. The method was employed for stability assays, but

Conclusions

Two LC/MS methods using ammonium bicarbonate and ammonium acetate as volatile buffers were developed and applied to the degradation profile estimation of cough–cold products under stress forced conditions. Ammonium acetate permitted us the results transference between the HPLC method with UV detection and a conventional sodium phosphate buffer, and the LC/MS method with an electrospray ionization interface, used to obtain molecular weights information. Thus, when a degradation product appears

Acknowledgements

We would like to thank Lilly laboratories for the technical support and Cinfa laboratories for providing the test materials.

References (20)

M. Bakshi et al.
J. Pharm. Biomed. Anal.
(2002)
A. Marı́n et al.
J. Pharm. Biomed. Anal.
(2002)
A. Garcı́a et al.
J. Chromatogr. B.
(2003)
D.V. McCalley
J. Chromatogr. A.
(2003)
S. Espinosa et al.
J. Chromatogr. A.
(2002)
A. Espada et al.
J. Chromatogr. A.
(2003)
A. Espada et al.
J. Chromatogr. A.
(2004)
J.J. Kirkland et al.
J. Chromatogr. A.
(1995)
ICH topic Q1A(R2). Stability testing of new drug substances and products, 2003. Available on:...
ICH topic Q3B(R). Impurities in new drug products, 2003. Available on:...

There are more references available in the full text version of this article.

Cited by (40)

Raman imaging studies on the stability of Paracetamol tablets under different storage conditions
2023, Vibrational Spectroscopy
Citation Excerpt :
The compound 4-AP is reported to have significant nephrotoxicity and teratogenic effects [21,22], and it has been limited to the low level of 50 ppm (0.005 % w/w) in pharmaceutical formulations by the European and US Pharmacopoeias, by employing a manual colourimetric limit test [23,24]. Other techniques have been used for determining 4-AP in APAP tablets, such as spectrophotometry,[25] high-performance liquid chromatography (HPLC) [26–28], liquid chromatography coupled with mass spectroscopy (LC/MS) [29], capillary electrophoresis [30,31], flow injection analysis [24], and micellar electrokinetic chromatography [32,33]. However, most of the procedures mentioned above are time-consuming, expensive and require specialized labour.
The applicability of Raman imaging for pharmaceutics production ranges from the characterization of pharmaceutical formulations, kinetic processes in drug delivery to the rapid detection and identification of counterfeit drugs/contaminants. Acetaminophen (Paracetamol, APAP) is an analgesic and antipyretic drug and one of the most consumed medicines worldwide. On the other hand, the compound 4-aminophenol (4-AP) is a hydrolytic product of APAP with nephrotoxicity and teratogenic effects. In this work, we have explored for the first time Raman imaging methods to characterize the main components of commercial APAP tablets (APAP-tablets) and to inquire about the potential of this optical technique to identify 4-AP in APAP tablets, which have been previously spiked with such contaminant. The laboratorial treated APAP-tablet samples were subjected to different temperature, humidity and sunlight exposure conditions, mimicking storage conditions, and then the Raman spectra and images were collected to monitor changes that might occur in those conditions. Although the lower limit of detection of 4-AP in APAP-tablets is above the minimum levels established by Pharmacopoeias (0.005 %), this research demonstrates that Raman imaging still allows the detection of small amounts of the contaminant, thus opening perspectives for exploring this technique for characterizing APAP products.
Choices of chromatographic methods as stability indicating assays for pharmaceutical products: A review
2021, Heliyon
Stability indicating assay describes a technique which is used to analyse the stability of drug substance or active pharmaceutical ingredient (API) in bulk drug and pharmaceutical products. Stability indicating assay must be properly validated as per ICH guidelines. The important components in a stability indicating assay include sensitivity, specificity, accuracy, reliability, reproducibility and robustness. A validated assay is able to measure the concentration changes of drug substance/API with time and make reliable estimation of the quantity of the degradation impurities. The drug substance is separated and resolved from the impurities. Pros and cons of HPLC, GC, HPTLC, CE and SFC were discussed and reviewed. Stability indicating assay may consist of the combination of chromatographic separation and spectroscopic detection techniques. Hyphenated system could demonstrate parallel quantitative and qualitative analysis of drug substances and impurities. Examples are HPLC-DAD, HPLC-FL, GC-MS, LC-MS and LC-NMR. The analytes in the samples are separated in the chromatography while the impurities are chemically characterised by the spectroscopy in the system. In this review, various chromatographic methods which had been employed as stability indicating assays for drug substance and pharmaceutical formulation were systematically reviewed, and the application of hyphenated techniques in impurities characterisation and identification were also discussed with supporting literatures.
Peak-tracking algorithm for use in comprehensive two-dimensional liquid chromatography – Application to monoclonal-antibody peptides
2021, Journal of Chromatography A
Citation Excerpt :
When utilizing such software, it is of utmost importance to track as many compounds as possible to obtain the most realistic predictions of separations of the analysed sample. In addition to its use for method optimization, peak tracking can also be used for impurity profiling [21–23]. When comparing chromatograms of different samples, compounds that cannot be tracked may be impurities in specific samples, which may be highly interesting for many applications.
A peak-tracking algorithm was developed for use in comprehensive two-dimensional liquid chromatography coupled to mass spectrometry. Chromatographic peaks were tracked across two different chromatograms, utilizing the available spectral information, the statistical moments of the peaks and the relative retention times in both dimensions. The algorithm consists of three branches. In the pre-processing branch, system peaks are removed based on mass spectra compared to low intensity regions and search windows are applied, relative to the retention times in each dimension, to reduce the required computational power by elimination unlikely pairs. In the comparison branch, similarity between the spectral information and statistical moments of peaks within the search windows is calculated. Lastly, in the evaluation branch extracted-ion-current chromatograms are utilized to assess the validity of the pairing results. The algorithm was applied to peptide retention data recorded under varying chromatographic conditions for use in retention modelling as part of method optimization tools. Moreover, the algorithm was applied to complex peptide mixtures obtained from enzymatic digestion of monoclonal antibodies. The algorithm yielded no false positives. However, due to limitations in the peak-detection algorithm, cross-pairing within the same peaks occurred and six trace compounds remained falsely unpaired.
Development of a new liquid chromatographic method for the simultaneous separation of ciprofloxacin degradation products with 5-hydroxymethyl-furfural, impurity of D-glucose, in an intravenous solution for perfusion
2019, Annales Pharmaceutiques Francaises
Citation Excerpt :
Until nowadays, liquid chromatography (LC) has been a major used technique for their determination [33,34] coupled to ultraviolet (UV) [35], fluorescence (FL) [36], or mass spectrometry (MS) [3,37] detectors. The particular advantage of LC relative to the other widely available techniques is its high-resolution capacity, sensitivity, specificity and ability to establish the quantification limits of impurities [38]. Several references about the determination of FQs in biological fluids, by using HPLC have been reported [39,40], for example, ofloxacin [41,42], levofloxacin [43,44], lomefloxacin [45,46], ciprofloxacin [47–50].
The place occupied by fluoroquinolones is very important clinically. Ciprofloxacin has been the most widely prescribed one which exhibits good activity against Pseudomonas aeruginosa and Acinetobacter. D-glucose is frequently used as an excipient in most formulations for perfusion solutions. Since there is much interest in pharmaceutical quality control of such formulations, separation of impurities from the main drug substances and accurate assay quantification, and since there is no reference or monograph until nowadays that has been reported for the simultaneous separation of ciprofloxacin degradation products along with 5-hydroxymethyl-furfural (5-HMF), impurity of D-glucose, classified as a high toxic substance, thus our aim of this work is to develop a new simple, sensitive and stability indicating method allowing this separation by high-performance liquid chromatography. We have started from the chromatographic conditions recommended by the British Pharmacopoeia, and by optimizing the nature of the stationary phase, the composition of the mobile phase and the injection volume. After optimisation, the retained chromatographic conditions have enabled the separation of all impurities with good resolution factor greater than 1.5 for each pair of peaks and with good symmetry peak shape. The developed method was validated according to the International Conference of Harmonisation (ICH) guidelines for specificity, detection and quantification limits, and then it was applied to stability study of the formulation subjected to different ICH prescribed stress conditions. The 5-HMF was checked to be the impurity issued from D-glucose hydrolysis by high temperature mainly after autoclaving of pharmaceuticals. The developed method was proved to be simple, specific with very low limit of quantification. Hence, it can be considered as a method for stability indicating and routine quality control analysis in pharmaceutical industries.
La place occupée par les fluoroquinolones est très importante sur le plan clinique. La ciprofloxacine a été la plus largement prescrite et présente une bonne activité contre Pseudomonas aeruginosa et Acinetobacter. Le D-glucose est fréquemment utilisé comme excipient dans la plupart des formulations pour solutions de perfusion. Comme le contrôle de la qualité pharmaceutique de ces formulations suscite beaucoup d’intérêt, il convient de séparer les impuretés des principales substances médicamenteuses et de quantifier les dosages avec précision, et comme il n’existe jusqu’à présent aucune référence ou monographie concernant la séparation simultanée des produits de dégradation de la ciprofloxacine avec le 5-hydroxyle-méthyle-furfural (5-HMF), impureté de D-glucose, classée substance hautement toxique, notre objectif est de développer une nouvelle méthode simple, spécifique et indicatrice de stabilité permettant cette séparation par chromatographie en phase liquide à haute performance. Nous sommes partis des conditions chromatographiques recommandées par la pharmacopée britannique et en optimisant la nature de la phase stationnaire, la composition de la phase mobile et le volume d’injection. Après optimisation, les conditions chromatographiques retenues ont permis la séparation de toutes les impuretés avec un bon facteur de résolution supérieur à 1,5 pour chaque paire de pics et avec une bonne symétrie des pics. La méthode développée a été validée conformément aux directives de la conférence internationale d’harmonisation (ICH) pour la spécificité, les limites de détection et de quantification, puis appliquée à l’étude de stabilité de la formulation soumise aux différentes conditions de stress recommandées par l’ICH. Il a été vérifié que le 5-HMF est l’impureté issue de l’hydrolyse du D-glucose obtenue à forte température, principalement après autoclavage de produits pharmaceutiques. La méthode développée s’est avérée simple, spécifique et avec une limite de quantification très basse. Par conséquent, elle peut être considérée comme une méthode indicatrice de stabilité et de contrôle en routine de la qualité de ces formulations dans les industries pharmaceutiques.
Catalytic reduction of Nitrophenols using silver nanoparticles-supported activated carbon derived from agro-waste
2018, Journal of Environmental Chemical Engineering
Citation Excerpt :
The next most abundant ion in the spectrum is at m/z 80 attributed to pyridinium cation formed by rearrangement. The ion at m/z 53 was attributed to loss of HCN from pyridinium cation providing evidence towards the formation of aminophenol [34]. The absence of peak at m/z 139 due to nitrophenol further confirms the formation of nitrophenol.
Immobilization of Ag nanoparticles (Agnps) on solid supports has been considered as the most efficient way to prevent their aggregation and use them as catalysts. Agrowaste derived carbons provide enormous benefits such as environmentally friendliness, economic viability and porosity when used as supports for Agnps. The present study was conducted to investigate the use of palm shell agrowaste derived catbon(Agnp-PSAC) as a support for Ag nanoparticles. The prepared Agnp-PSAC was characterized using Transmission electron Microscopy (TEM), Fourier-transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy with Energy Dispersive Spetroscopy (SEM-EDAX), Raman and UV–visible spectroscopic techniques. The catalytic potential of the prepared Agnp-PSAC was investigated. Nitrophenols such as 4-nitrophenol (4-NP), 2,4-dinitrophenol (DNP) and 2,4,6 tri nitrophenol (TNP) were used as substrates. The supported Agnps could be reused upto four cycles. The reduction followed pseudo-first-order kinetics. A value added application of palm shell derived activated carbon is thus proposed.
Rapid assay of the comparative degradation of acetaminophen in binary and ternary combinations
2014, Arabian Journal of Chemistry
Citation Excerpt :
The quality and the stability of pharmaceutical drugs are of fundamental significance for patient’s safety and rapid health recovery. Different environmental factors such as humidity, temperature and UV (ultraviolet) light influence the stability (Marin and Barbas, 2004) of these drugs, since these elements produce various degradants. In consequence, the presence of degradant metabolites profoundly influences the efficacy of these medicines.
The study is intended to monitor the comparative degradation rates of acetaminophen in binary and ternary combinations by UV–vis spectroscopy. The drugs were exposed to UV-rays in blister packing. The exposition time was 24, 48 and 72 h for both shorter and longer wavelengths. The problem of overlapping UV bands of aspirin and caffeine with acetaminophen was solved by extracting them in diethylether, therefore, we developed a straightforward, rapid and accurate assay method for measuring acetaminophen concentration in binary and ternary mixtures and to monitor its degradation.

View all citing articles on Scopus

View full text

LC/MS for the degradation profiling of cough–cold products under forced conditions

Abstract

Introduction

Section snippets

Chemicals

LC/MS method optimization

Conclusions

Acknowledgements

J. Pharm. Biomed. Anal.

J. Pharm. Biomed. Anal.

J. Chromatogr. B.

J. Chromatogr. A.

J. Chromatogr. A.

J. Chromatogr. A.

J. Chromatogr. A.

J. Chromatogr. A.