Analytical Methodologies and Stability Assessment of Bioactive Compounds in Complex Matrices

A special issue of Processes (ISSN 2227-9717). This special issue belongs to the section "Separation Processes".

Deadline for manuscript submissions: 30 June 2026 | Viewed by 454

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Department of Inorganic Chemistry and Pharmaceutical Analytics, Faculty of Pharmacy, Jagiellonian University Medical College, 9 Medyczna St., 30-688 Kraków, Poland
Interests: chromatographic techniques; methods validation; drug analysis; stability testing; interactions; drug impurities; remediation
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Dear Colleagues,

Analytical methods used to analyze bioactive compounds in complex matrices utilize specialized techniques such as high-performance liquid chromatography (HPLC), gas chromatography, and thin-layer chromatography, coupled with various detectors. The methodology is preceded by advanced sample preparation, including microextraction techniques (i.e., SPE, UASE). The complexity of matrices such as blood, plasma, urine, plant material, or food requires highly selective and sensitive analytical methods to avoid matrix interferences. Assessing the stability of compounds involves analyzing the impact of various environmental conditions on their degradation process over time. The obtained results allow us to identify potential degradation pathways, ensuring the authenticity and purity of the products. To ensure the reliability of the results, validation of the methods used is necessary, including checking linearity, precision, accuracy, and limits of detection and quantification. An important condition for modern analytical procedures is also the issue of developing sustainable and environmentally friendly methods (‘Green Chemistry’), using, for example, ecological solvents for extraction.

This Special Issue may constitute a review of works that meet the above assumptions and will allow for the characterization of active substances, including origin, production methods, extraction methods and chemical and/or microbiological composition, using validated analytical methods. We invite authors of papers from various scientific disciplines for whom the value of the finished product (drug, dietary supplement, food, etc.) is important from the point of view of the declared and expected health effect.

Dr. Malgorzata Starek
Guest Editor

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Keywords

  • analytical techniques
  • chromatography
  • extraction
  • matrix effects
  • biological material
  • drugs
  • dietary supplements
  • compound stability
  • degradation
  • method validation

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Published Papers (1 paper)

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Research

19 pages, 968 KB  
Article
Process Development and Validation of Reverse-Phase High-Performance Liquid Chromatography Method for Simultaneous Quantification of Quercetin, Thymoquinone, and Pterostilbene
by Ushasi Das, Sanchita Mandal, Ketan Ranch and Sudarshan Singh
Processes 2026, 14(3), 428; https://doi.org/10.3390/pr14030428 - 26 Jan 2026
Viewed by 344
Abstract
The simultaneous HPLC method for quantifying Quercetin (Que), Thymoquinone (Thy), and Pterostilbene (Pte) aims at the precise measurement of these polyphenols alone or in complex mixtures, targeting their therapeutic potential in disorders such as diabetes and epilepsy. The method focuses on quantifying Que, [...] Read more.
The simultaneous HPLC method for quantifying Quercetin (Que), Thymoquinone (Thy), and Pterostilbene (Pte) aims at the precise measurement of these polyphenols alone or in complex mixtures, targeting their therapeutic potential in disorders such as diabetes and epilepsy. The method focuses on quantifying Que, Thy, and Pte, utilizing optimized reversed-phase HPLC conditions as per ICH Q2(R1) standards. Key validation aspects include linearity, specificity, precision, and accuracy, ensuring compliance for quality control in nanomedicine and nutraceuticals, and the method’s applications support pharmacokinetic studies and stability testing, contributing to personalized medicine and addressing pharmaco-resistance. The HPLC method development and validation were performed on a phenyl column using the mobile phase consisting of solvent A (0.1% orthophosphoric acid in HPLC water) and solvent B (acetonitrile) at a ratio of 55:45 in an isocratic elution mode at a flow rate of 1 mL/min and at a column temperature of 35 °C. Ultraviolet detection was measured at 254 nm. Moreover, the method was validated for accuracy, precision, linearity, specificity, and sensitivity. The retention time for tested Que, Thy, and Pte was observed at 4.15 min, 8.70 min, and 10.75 min, respectively. Limits of detection for Que, Thy, and Pte were 1.55 μg/mL, 2.40 μg/mL, and 70.79 µg/mL, whereas limits of quantification were 4.69 μg/mL, 7.28 μg/mL, and 214.52 µg/mL, respectively. Linearity and correlation coefficients for Que, Thy, and Pte were found in the range of 50–250 μg/mL (0.9999), 50–250 μg/mL (0.9999), and 620–3100 μg/mL (0.9996), respectively. A reasonable level of accuracy was indicated by the tested method suggesting extremely high recovery levels (98–102%). The separation of tested compounds was achieved within 11 min. The developed and validated RP-HPLC–UV method was successfully applied for the identification and quantification of Que, Thy, and Pte for their combined estimation in complex formulations. From the validation study, it was found that the tested method is specific, accurate, precise, reliable, and reproducible. Full article
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