Puthenvitil Smija Kakkuth, Perumal Palani
Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai 600025, India.
Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai 600025, India.
J Chromatogr B Analyt Technol Biomed Life Sci. 2025 Sep 1;1263:124707. doi: 10.1016/j.jchromb.2025.124707. Epub 2025 Jun 25.
The escalating demand for high-purity bioactive compounds in pharmaceutical research and quality control underscores the critical role of preparative High-Performance Liquid Chromatography. This study introduces a novel and optimized preparative HPLC protocol for the large-scale purification of thymol from Trachyspermum ammi (L.) essential oil. Addressing a notable knowledge gap, this is the first reported application of Prep-HPLC for thymol isolation from this specific botanical source, establishing a new, scalable protocol. An initial analytical HPLC method, optimized with Methanol:Water (60:40, v/v), was successfully scaled to a semi-preparative level. The optimized preparative method, utilizing Methanol:Water (90:10, v/v) as the mobile phase, achieved thymol purification with an impressive 98.87 % purity. The structural identity of the purified compound was unequivocally confirmed through comprehensive spectroscopic analyses, including GC-MS, FTIR, XRD, and NMR. Crucially, the purified thymol exhibited potent and broad-spectrum antifungal activity against both susceptible and multidrug-resistant strains of Candida albicans and Candida auris. Minimum inhibitory concentrations were determined to be as low as 0.125 μg/mL against multidrug-resistant C. albicans and 0.25 μg/mL against multidrug-resistant C. auris. This robust efficacy, particularly against challenging drug-resistant isolates, strongly positions the purified thymol as a promising candidate for further therapeutic development against candidiasis. This research significantly advances separation science and highlights the industrial potential of Prep-HPLC for high-value natural product isolation.
制药研究和质量控制中对高纯度生物活性化合物的需求不断增加，凸显了制备型高效液相色谱的关键作用。本研究介绍了一种新颖且优化的制备型高效液相色谱方案，用于从阿育吠陀（Trachyspermum ammi (L.)）精油中大规模纯化百里香酚。针对一个显著的知识空白，这是首次报道将制备型高效液相色谱应用于从这种特定植物来源中分离百里香酚，建立了一种新的、可扩展的方案。最初用甲醇:水（60:40，v/v）优化的分析型高效液相色谱方法成功扩展到半制备水平。优化后的制备方法以甲醇:水（90:10，v/v）作为流动相，实现了百里香酚的纯化，纯度达到令人印象深刻的98.87%。通过包括气相色谱 - 质谱联用（GC - MS）、傅里叶变换红外光谱（FTIR）、X射线衍射（XRD）和核磁共振（NMR）在内的综合光谱分析，明确确认了纯化化合物的结构身份。至关重要的是，纯化后的百里香酚对白色念珠菌和耳念珠菌的敏感菌株及多重耐药菌株均表现出强大的广谱抗真菌活性。对多重耐药白色念珠菌的最低抑菌浓度低至0.125μg/mL，对多重耐药耳念珠菌的最低抑菌浓度为0.25μg/mL。这种强大的功效，特别是对具有挑战性的耐药菌株，有力地将纯化后的百里香酚定位为进一步开发抗念珠菌病治疗药物的有前景候选物。本研究显著推动了分离科学的发展，并突出了制备型高效液相色谱在分离高价值天然产物方面的工业潜力。
