C. L. Gopu Poona College of Pharmacy, Bharati Vidyapeeth University, Erandwane, Pune, 411038, Maharashtra, India

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S. S. Gilda Poona College of Pharmacy, Bharati Vidyapeeth University, Erandwane, Pune, 411038, Maharashtra, India

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A. R. Paradkar Poona College of Pharmacy, Bharati Vidyapeeth University, Erandwane, Pune, 411038, Maharashtra, India

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K. R. Mahadik Poona College of Pharmacy, Bharati Vidyapeeth University, Erandwane, Pune, 411038, Maharashtra, India

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A simple, sensitive, and reproducible high-performance thin-layer chromatographic method has been established for simultaneous analysis of trigonelline and 4-hydroxyisoleucine from fenugreek seeds (Trigonella foenum-graceum). Chromatography was performed on silica gel GF254 with n-butanol-methanol-acetic acid-water 4:1.5:1:1 (υ/υ) as mobile phase. Densitometric scanning of the plates at 266 nm was used for analysis of trigonelline, and plates were scanned at 395 nm after derivatisation with ninhydrin reagent for analysis of 4-hydroxyisoleucine. The method was validated for specificity, precision (intraday and interday), accuracy, and robustness. Response was a linear function of the concentration of standard solutions in the range 100 to 1000 ng per band and 50 to 500 ng per band, respectively, with correlation coefficients of 0.9992 ± 0.04 and 0.9986 ± 0.06, for trigonelline and 4-hydroxyisoleucine. The accuracy of the method was checked by determination of recovery at three different levels; average percentage recovery was 99.39 ± 0.34 for trigonelline and 99.03 ± 0.26 for 4-hydroxyisoleucine. The method was found to be simple, precise, specific, sensitive, and accurate and can be used for routine quality control of fenugreek seeds and also for quantitative analysis of these marker compounds in processed fenugreek and its marketed preparations.

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Senior editors

Editor(s)-in-Chief: Kowalska, Teresa (1946-2023)

Editor(s)-in-Chief: Sajewicz, Mieczyslaw, University of Silesia, Katowice, Poland


  • Danica Agbaba, University of Belgrade, Belgrade, Serbia
  • Łukasz Komsta, Medical University of Lublin, Lublin, Poland
  • Ivana Stanimirova-Daszykowska, University of Silesia, Katowice, Poland
  • Monika Waksmundzka-Hajnos, Medical University of Lublin, Lublin, Poland

Editorial Board

  • Ravi Bhushan, The Indian Institute of Technology, Roorkee, India
  • Jacek Bojarski, Jagiellonian University, Kraków, Poland
  • Bezhan Chankvetadze, State University of Tbilisi, Tbilisi, Georgia
  • Michał Daszykowski, University of Silesia, Katowice, Poland
  • Tadeusz H. Dzido, Medical University of Lublin, Lublin, Poland
  • Attila Felinger, University of Pécs, Pécs, Hungary
  • Kazimierz Glowniak, Medical University of Lublin, Lublin, Poland
  • Bronisław Glód, Siedlce University of Natural Sciences and Humanities, Siedlce, Poland
  • Anna Gumieniczek, Medical University of Lublin, Lublin, Poland
  • Urszula Hubicka, Jagiellonian University, Kraków, Poland
  • Krzysztof Kaczmarski, Rzeszow University of Technology, Rzeszów, Poland
  • Huba Kalász, Semmelweis University, Budapest, Hungary
  • Katarina Karljiković Rajić, University of Belgrade, Belgrade, Serbia
  • Imre Klebovich, Semmelweis University, Budapest, Hungary
  • Angelika Koch, Private Pharmacy, Hamburg, Germany
  • Piotr Kus, Univerity of Silesia, Katowice, Poland
  • Debby Mangelings, Free University of Brussels, Brussels, Belgium
  • Emil Mincsovics, Corvinus University of Budapest, Budapest, Hungary
  • Ágnes M. Móricz, Centre for Agricultural Research, Budapest, Hungary
  • Gertrud Morlock, Giessen University, Giessen, Germany
  • Anna Petruczynik, Medical University of Lublin, Lublin, Poland
  • Robert Skibiński, Medical University of Lublin, Lublin, Poland
  • Bernd Spangenberg, Offenburg University of Applied Sciences, Germany
  • Tomasz Tuzimski, Medical University of Lublin, Lublin, Poland
  • Yvan Vander Heyden, Free University of Brussels, Brussels, Belgium
  • Adam Voelkel, Poznań University of Technology, Poznań, Poland
  • Brata Walczak, University of Silesia, Katowice, Poland
  • Wiesław Wasiak, Adam Mickiewicz University, Poznań, Poland
  • Igor G. Zenkevich, St. Petersburg State University, St. Petersburg, Russian Federation


KOWALSKA, TERESA (1946-2023)


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Acta Chromatographica
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