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<p>Perillae folium (PF) is widely used in both food and medicine, with considerable variation in its volatile oil composition, which can be categorized into distinct chemotypes that significantly impact quality. This study aimed to establish a rapid and efficient method for screening and identifying antimicrobial compounds in PF volatile oils by integrating high-performance thin-layer chromatography (HPTLC), bioautography, and mass spectrometry (MS). The method enables effect-directed analysis by combining on-plate bioautography following HPTLC separation with MS-based compound identification. Three PF chemotypes, perillaldehyde type (PA), perilla ketone type (PK), and mixed type (PAPK), were characterized by HPTLC, revealing distinct chromatographic profiles. Bioactivity assays using <i>Vibrio fischeri</i> and <i>Bacillus subtilis</i> were performed directly on the HPTLC plates. Numerous volatile compounds showed strong antibacterial activity against <i>V. fischeri</i>, while a few also inhibited <i>B. subtilis</i>. HPTLC‒MS identified eight antimicrobial compounds across the three chemotypes. Among them, caryophyllene oxide, perilla ketone, perillaldehyde, sageone, and eugenol were the major contributors to antimicrobial activity. In addition, <i>α</i>-asarone and caryophyllene oxide displayed broad-spectrum activity against both bacterial strains. ImageJ software was used to quantify inhibition bands, indicating that the antibacterial potency followed the order: PK &gt; PAPK &gt; PA. In summary, the established HPTLC‒bioautography‒MS method enables rapid separation and identification of antimicrobial compounds in PF volatile oils. The results reveal significant differences in bioactive compounds among chemotypes of PF, leading to varying antimicrobial effects. This study provides data supporting PF quality evaluation and also presents a practical strategy for screening bioactive compounds in other medicinal plants.</p>

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Identification of antibacterial compounds in various Perillae folium chemotypes by high-performance thin-layer chromatography‒bioassay‒mass spectrometry

  • 期刊论文

  • 发表日期：

    2025年7月15日

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  • Zihao Si,
  • Mooseob Kim,
  • Lihua Gu,
  • Linnan Li,
  • Li Yang,
  • Zhengtao Wang

摘要

Perillae folium (PF) is widely used in both food and medicine, with considerable variation in its volatile oil composition, which can be categorized into distinct chemotypes that significantly impact quality. This study aimed to establish a rapid and efficient method for screening and identifying antimicrobial compounds in PF volatile oils by integrating high-performance thin-layer chromatography (HPTLC), bioautography, and mass spectrometry (MS). The method enables effect-directed analysis by combining on-plate bioautography following HPTLC separation with MS-based compound identification. Three PF chemotypes, perillaldehyde type (PA), perilla ketone type (PK), and mixed type (PAPK), were characterized by HPTLC, revealing distinct chromatographic profiles. Bioactivity assays using Vibrio fischeri and Bacillus subtilis were performed directly on the HPTLC plates. Numerous volatile compounds showed strong antibacterial activity against V. fischeri, while a few also inhibited B. subtilis. HPTLC‒MS identified eight antimicrobial compounds across the three chemotypes. Among them, caryophyllene oxide, perilla ketone, perillaldehyde, sageone, and eugenol were the major contributors to antimicrobial activity. In addition, α-asarone and caryophyllene oxide displayed broad-spectrum activity against both bacterial strains. ImageJ software was used to quantify inhibition bands, indicating that the antibacterial potency followed the order: PK > PAPK > PA. In summary, the established HPTLC‒bioautography‒MS method enables rapid separation and identification of antimicrobial compounds in PF volatile oils. The results reveal significant differences in bioactive compounds among chemotypes of PF, leading to varying antimicrobial effects. This study provides data supporting PF quality evaluation and also presents a practical strategy for screening bioactive compounds in other medicinal plants.