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Separations
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Determination and Quantification of Pharmaceuticals Using Chromatography Methods
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Determination and Quantification of Pharmaceuticals Using Chromatography Methods

A special issue of Separations (ISSN 2297-8739). This special issue belongs to the section "Analysis of Natural Products and Pharmaceuticals".

Deadline for manuscript submissions: 10 May 2026 | Viewed by 3929

Special Issue Editors

Dr. Karol Wróblewski

E-Mail Website
Guest Editor
Institute of Medical Sciences, College of Medical Sciences, University of Rzeszów, 35-959 Rzeszów, Poland
Interests: liquid chromatography; mass spectrometry; biomedical and pharmaceutical analysis; sample preparation methods; drug development; clinical trials
Special Issues, Collections and Topics in MDPI journals
Prof. Dr. Małgorzata Szultka-Młyńska

E-Mail Website
Guest Editor
Department of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University, Gagarin 7, 87-100 Torun, Poland
Interests: bioanalytics; chromatography; in vitro; in vivo; mass spectrometry; metabolomics; microextraction; sample preparation techniques

Special Issue Information

Dear Colleagues,

Various chromatographic techniques are used in the analysis of pharmaceuticals such as high-performance thin-layer chromatography (HPTLC), high-performance liquid chromatography (HPLC), ultra-high-performance liquid chromatography (UHPLC), and gas chromatography (GC). The ongoing evolution of chromatographic methods includes advances in components such as the types of stationary phases, detection systems, data handling systems, and sample preparation methods. Modern detection techniques such as diode array detection (DAD), fluorescence detection (FLD), mass spectrometry (MS), or tandem mass spectrometry (MS/MS) play an important role in the development of sensitive and selective methods for quantitative analysis.

This Special Issue on the “Determination and Quantification of Pharmaceuticals Using Chromatography Methods” will cover a wide range of topics, including, but not limited to the following: sample preparation methods, the development and validation of a stability-indicating method for the estimation of drugs in bulk and pharmaceutical formulations, stability testing in pharmaceutical development, the determination and characterization of degradation products of drugs, and the chromatographic analysis of natural products.

Dr. Karol Wróblewski
Prof. Dr. Małgorzata Szultka-Młyńska
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 250 words) can be sent to the Editorial Office for assessment.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Separations is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • chromatography
  • method validation
  • drug analysis
  • pharmaceutical analysis
  • analytical method development
  • pharmaceutical formulations
  • mass spectrometry

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Published Papers (3 papers)

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Research

11 pages, 800 KB  
Article
Determination of Paracetamol and Orphenadrine Citrate in Tablets via a Novel RP-HPLC Method: Development Following Box–Behnken Design, Validation, Robustness Testing, and Greenness Assessment
by Dimitris Valatsos, Kostas Gkountanas and Yannis Dotsikas
Separations 2026, 13(2), 58; https://doi.org/10.3390/separations13020058 - 6 Feb 2026
Viewed by 399
Abstract
Paracetamol (PAR) and orphenadrine citrate (ORPH) are two active substances commonly used in combination medicinal products, due to the analgesic effect of paracetamol and the muscle relaxant effect of orphenadrine, with a therapeutic indication of mild to moderate acute musculoskeletal pain. The aim [...] Read more.
Paracetamol (PAR) and orphenadrine citrate (ORPH) are two active substances commonly used in combination medicinal products, due to the analgesic effect of paracetamol and the muscle relaxant effect of orphenadrine, with a therapeutic indication of mild to moderate acute musculoskeletal pain. The aim of this work is to develop and validate an isocratic HPLC method for the simultaneous determination of PAR and ORPH in tablet formulation. Preliminary experiments showed that an analytical column with a chemically bound phenyl phase was required. A Box–Behnken design (BBD) was utilized to optimize the analytical method for two key responses, PAR asymmetry factor (AsymPAR) and ORPH capacity factor (kORPH), with three numerical factors: percentage of ACN in mobile phase (A); pH (B); and salt concentration in the aqueous solution (C). The optimized method consists of a Pinnacle DB Biphenyl (250 × 4.6 mm) 5 µm column, and a mobile phase of 37%/63% v/v ACN-NaH2PO4·H2O in 29 mM aqueous solution, pH = 2.5. The flow rate was set to 1.5 mL/min and detection occurred at 215 nm. After the optimization process the following chromatographic conditions were selected and the method was validated for various ICH parameters covering system suitability, specificity, linearity (R2 = 1.00), precision (%RSD ≤ 2), accuracy (98% ≤ %Recovery ≤ 102%), and robustness. Finally, the environmental friendliness of the novel method was assessed by using the Analytical GREEnness (AGREE) metric tool, obtaining a score of 0.67. Full article
(This article belongs to the Special Issue Determination and Quantification of Pharmaceuticals Using Chromatography Methods)
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20 pages, 2917 KB  
Article
Volatile Organic Compound Profiling of Traditional Multi-Herbal Prescriptions for Chemical Differentiation and Ethnopharmacological Insights
by Sumin Seo, Unyong Kim, Jiyu Kim, Chohee Jeong and Sang Beom Han
Separations 2026, 13(1), 8; https://doi.org/10.3390/separations13010008 - 24 Dec 2025
Viewed by 652
Abstract
Traditional herbal prescriptions composed of multiple botanicals remain central to ethnopharmacological practice; however, their chemical basis and classification remain poorly understood. Non-volatile compound analyses of herbal medicines are well established, but comparative studies focusing on volatile organic compounds (VOCs) across multi-herbal prescriptions are [...] Read more.
Traditional herbal prescriptions composed of multiple botanicals remain central to ethnopharmacological practice; however, their chemical basis and classification remain poorly understood. Non-volatile compound analyses of herbal medicines are well established, but comparative studies focusing on volatile organic compounds (VOCs) across multi-herbal prescriptions are scarce. To enhance the chemical understanding of traditional formulations and clarify prescription-level characteristics, this study applied headspace solid-phase microextraction coupled with gas chromatography–mass spectrometry (HS-SPME–GC–MS) to characterize VOC-based chemical signatures in 30 prescriptions composed of 76 herbal ingredients. Multivariate analyses such as principal component analysis, partial least squares discriminant analysis (PLS-DA), and orthogonal PLS-DA (OPLS-DA) enabled systematic differentiation of various prescriptions and identified 25 discriminant VOCs, 9 of which were common among multiple therapeutic categories. These shared compounds, such as 5-hydroxymethylfurfural (5-HMF) and 4H-pyran-4-one derivatives, reflect recurrent chemical patterns associated with broad-spectrum applications, whereas category-specific volatiles (including isopsoralen, senkyunolide, and fenipentol) delineated therapeutic boundaries, even among prescriptions with overlapping botanicals. Capturing both shared and distinct volatile signatures clarified ambiguous boundaries between categories such as cold, inflammation, or diabetes versus kidney disorder prescriptions, thereby linking chemical patterns with ethnopharmacological indications. Together, these findings highlight VOC profiling as a valuable diagnostic and interpretive tool that bridges traditional categorization systems with modern chemical analysis, offering a robust framework for future pharmacological and mechanistic investigations. Such an approach not only substantiates traditional categorization but also provides a practical basis for quality control and pharmacological evaluation of multi-herbal formulations. Full article
(This article belongs to the Special Issue Determination and Quantification of Pharmaceuticals Using Chromatography Methods)
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19 pages, 3465 KB  
Article
Metabolic Profiling and Pharmacokinetics Characterization of Yinhua Pinggan Granules with High-Performance Liquid Chromatography Combined with High-Resolution Mass Spectrometry
by Ningning Gu, Haofang Wan, Imranjan Yalkun, Yu He, Yihang Lu, Chang Li and Haitong Wan
Separations 2025, 12(5), 113; https://doi.org/10.3390/separations12050113 - 28 Apr 2025
Cited by 1 | Viewed by 2265
Abstract
Yinhua Pinggan Granules (YPG) is a patented traditional Chinese medicine (TCM) compound prescription, with wide clinical application against cold, cough, and relevant diseases. However, the chemical profiles of YPG in vivo are still unknown, hindering further pharmacological and quality control (QC) researches. This [...] Read more.
Yinhua Pinggan Granules (YPG) is a patented traditional Chinese medicine (TCM) compound prescription, with wide clinical application against cold, cough, and relevant diseases. However, the chemical profiles of YPG in vivo are still unknown, hindering further pharmacological and quality control (QC) researches. This study presents an ultra-high-performance liquid chromatography coupled with high-resolution orbitrap mass spectrometry (UHPLC-MS)-based method. Using the Compound Discoverer platform and a self-built ‘in-house’ compound database, the metabolic profiles and pharmacokinetics characters of YPG were investigated. Consequently, a total of 230 compounds (including 39 prototype components and 191 metabolites) were tentatively identified, in which the parent compounds were mainly flavonoids, alkaloids, and terpenoids, and the main metabolic pathways of metabolites include hydration, dehydration, and oxidation. The serum concentration of seven major representative compounds, including quinic acid, chlorogenic acid, amygdalin, 3′-methoxypuerarin, puerarin, glycyrrhizic acid, and polydatin, were also measured, to elucidate their pharmacokinetics behaviors in vivo. The pharmacokinetic study showed that the seven representative compounds were quantified in rat plasma within 5 min post-administration, with Tmax of less than 2 h, followed by a gradual decline in concentration over a 10 h period. The method demonstrated excellent linearity (R2 > 0.998), precision, and recovery (RSD < 15%). As the first systematic characterization of YPG’ s in vivo components and metabolites using UHPLC-MS, this study may contribute to comprehensively elucidate the metabolic profiles of the major components in YPG, and provide a critical foundation for further investigation on the QC and bioactivity research of YPG. Full article
(This article belongs to the Special Issue Determination and Quantification of Pharmaceuticals Using Chromatography Methods)
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