Narayana D B Anantha, Mukne Alka
Chief Scientific Officer, Ayurvidye Trust, Bengaluru, Karnataka, India.
Department of Pharmacognosy and Phytochemistry, Bombay College of Pharmacy, Mumbai, Maharashtra, India.
Indian J Pharmacol. 2025 Sep 1;57(5):302-307. doi: 10.4103/ijp.ijp_391_24. Epub 2025 Aug 22.
Acorus calamus L. (Sweet Flag), known in India as "Vacha," is widely used in traditional medicine, particularly for cognitive enhancement in infants. While traditionally considered safe, β-asarone - a key constituent - has shown potential genotoxicity in some in vitro studies. This study aimed to evaluate the mutagenic potential of Indian A. calamus rhizome, its extracts, and pure β-asarone using the Ames test in accordance with Organization for Economic Co-operation and Development guidelines.
Samples tested included pure β-asarone, dried rhizome powder, and extracts prepared in acidic and alkaline aqueous media. Marker compounds (β-asarone, α-asarone, and shyobunone) were quantified. The Ames test was conducted under GLP using Salmonella typhimurium strains TA97a, TA98, TA100, TA102, and TA1535 (Moltox Inc., USA). Each strain was tested with and without metabolic activation using Aroclor 1254-induced rat liver S9 homogenate. Positive controls included 2-aminoanthracene, sodium azide, mitomycin C, 9-aminoacridine, and 4-nitro-o-phenylenediamine. The vehicle was used as a negative control. Doses tested ranged from 39.06 to 5000 µg/plate.
No mutagenic response was observed in any of the tested samples across all five strains under either condition (±S9). Positive controls showed expected increases in revertant colonies. Revertant counts for test samples did not reach the threshold for a positive response (≥3 × control for TA1535, TA97a; ≥2× for TA98, TA100, and TA102).
Under the test conditions, A. calamus rhizome and β-asarone did not exhibit mutagenicity. Further studies, including chronic toxicity and carcinogenicity evaluations, are recommended to confirm safety across formulations.
菖蒲在印度被称为“瓦查”，在传统医学中广泛应用，尤其用于增强婴儿认知能力。虽然传统上认为其安全，但关键成分β-细辛醚在一些体外研究中显示出潜在的遗传毒性。本研究旨在根据经济合作与发展组织的指南，使用艾姆斯试验评估印度菖蒲根茎、其提取物及纯β-细辛醚的致突变潜力。
测试样品包括纯β-细辛醚、干燥根茎粉末以及在酸性和碱性水性介质中制备的提取物。对标记化合物（β-细辛醚、α-细辛醚和细辛酮）进行定量分析。使用鼠伤寒沙门氏菌菌株TA97a、TA98、TA100、TA102和TA1535（美国莫尔托克斯公司）在良好实验室规范（GLP）条件下进行艾姆斯试验。每个菌株在有和没有使用Aroclor 1254诱导的大鼠肝脏S9匀浆进行代谢活化的情况下进行测试。阳性对照包括2-氨基蒽、叠氮化钠、丝裂霉素C、9-氨基吖啶和4-硝基邻苯二胺。溶剂用作阴性对照。测试剂量范围为39.06至5000μg/平板。
在两种条件下（±S9），所有五个菌株的任何测试样品中均未观察到致突变反应。阳性对照显示回复突变菌落预期增加。测试样品的回复突变计数未达到阳性反应阈值（TA1535、TA97a为≥3×对照；TA98、TA100和TA102为≥2×）。
在测试条件下，菖蒲根茎和β-细辛醚未表现出致突变性。建议进行进一步研究，包括慢性毒性和致癌性评估，以确认不同制剂的安全性。
