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<p><i>Morinda tinctoria</i> Roxb. (Rubiaceae) is an important herb traditionally used for the treatment of dyspepsia, stomatitis, diarrhea, fevers, and gout. The present work covers a new, accurate, and precise thin-layer chromatography (TLC)–densitometry method for the quality assurance of five therapeutically important biomarkers, namely, dehydromethoxygaertneroside (<b>1</b>), β-sitosterol-3-<i>O</i>-glucoside (<b>2</b>), 14-deoxy-11,12-didehydroandrographolide (<b>3</b>), rotundioic acid (<b>4</b>), and stigmasterol (<b>5</b>) from the leaves of <i>M. tinctoria</i>. All the five phytoconstituents are highly attributed with a variety of pharmacological activities. The precoated aluminum plates with silica gel 60 F<sub>254</sub> (10&#xa0;cm × 10&#xa0;cm), using toluene–diethyl ether–methanol–chloroform (5:0.5:1.5:3.5, <i>V/V</i>), were used for separation of markers. The identification of compounds (<b>1–5</b>) was achieved on TLC plate by derivatization using vanillin–sulfuric acid reagent and then heating for 3&#xa0;min at 125&#xa0;°C. The concentration range was 0.2–1.0&#xa0;μg/band with excellent correlation (0.9991–0.9997) for all the selected markers. The limit of detection and limit of quantification of the selected compounds varied between 20–33&#xa0;ng/band and 66–110&#xa0;ng/band, respectively. The validated method was precise, having recoveries vary from 93.09% to 107.69% for markers <b>1–5</b> in triplicate at different standard addition levels. The specificity was established through ultraviolet–visible spectra correlation and retention factor (<i>R</i><sub>F</sub>). Additionally, the greenness was evaluated using several metrics, including the Analytical GREEness (AGREE) metric, Complex Modified Green Analytical Procedure Index (ComplexMoGAPI), National Environmental Methods Index (NEMI), and Eco-Scale Assessment (ESA).</p> Graphical Abstract <p></p>

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Greenness assessment and validation of a thin-layer chromatography–densitometric method for the simultaneous estimation of five bioactive markers of Morinda tinctoria Roxb.

  • 期刊论文

  • 发表日期：

    2026年1月22日

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  • Priyanka Maurya,
  • Madhumita Srivastava,
  • Manju Singh,
  • Karuna Shanker

摘要

Morinda tinctoria Roxb. (Rubiaceae) is an important herb traditionally used for the treatment of dyspepsia, stomatitis, diarrhea, fevers, and gout. The present work covers a new, accurate, and precise thin-layer chromatography (TLC)–densitometry method for the quality assurance of five therapeutically important biomarkers, namely, dehydromethoxygaertneroside (1), β-sitosterol-3-O-glucoside (2), 14-deoxy-11,12-didehydroandrographolide (3), rotundioic acid (4), and stigmasterol (5) from the leaves of M. tinctoria. All the five phytoconstituents are highly attributed with a variety of pharmacological activities. The precoated aluminum plates with silica gel 60 F254 (10 cm × 10 cm), using toluene–diethyl ether–methanol–chloroform (5:0.5:1.5:3.5, V/V), were used for separation of markers. The identification of compounds (1–5) was achieved on TLC plate by derivatization using vanillin–sulfuric acid reagent and then heating for 3 min at 125 °C. The concentration range was 0.2–1.0 μg/band with excellent correlation (0.9991–0.9997) for all the selected markers. The limit of detection and limit of quantification of the selected compounds varied between 20–33 ng/band and 66–110 ng/band, respectively. The validated method was precise, having recoveries vary from 93.09% to 107.69% for markers 1–5 in triplicate at different standard addition levels. The specificity was established through ultraviolet–visible spectra correlation and retention factor (RF). Additionally, the greenness was evaluated using several metrics, including the Analytical GREEness (AGREE) metric, Complex Modified Green Analytical Procedure Index (ComplexMoGAPI), National Environmental Methods Index (NEMI), and Eco-Scale Assessment (ESA).

Graphical Abstract