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Two-Dimensional Chromatographic Isolation of High Purity Erinacine A from Hericium erinaceus

Studies
Author/s: Asst.Prof. Andrej Gregori, PhD, Katerina Naumoska, Alen Albreht
Publish date: 13.05.2026

Medicinal mushrooms, which have been used for centuries in traditional Chinese medicinal mushrooms, which have been used for centuries in traditional Chinese medicine, offer a viable alternative to modern medicine by providing effective solutions for diseases that cannot be  treated with conventional therapies.

Hericium erinaceus (Bull.: Fr.) Pers., commonly known as Lion’s Mane, is a well known medicinal mushroom that has attracted great interest from academia and the industry as it contains a wide range of health-promoting compounds, with erinacine A  being particularly noteworthy.

A simple and robust two-dimensional chromatographic fractionation protocol for the isolation of the neuroprotective compound erinacine A from Hericium erinaceus is proposed.

This production platform yielded 19.4 mg of erinacine A from approximately 130 g of mushroom material, with a chromatographic purity of 97.4%. The procedure includes extraction, concentration, fractionation, purification, and characterisation of the bioactive compound.

The crude H. erinaceus extract was fractionated in the first dimension by normal-phase chromatography, and the fraction containing erinacine A was further purifies in the second dimension by semi-preparative reversed-phase chromatography. This strategy utilises the orthogonality of the two chromatographic modes of effectively eliminate difficult impurities, including structural isomers and analogues of erinacine A.

Complementary analytical approaches such as: high-performance thin-layer chromatography (HPTLC) and high-performance liquid chromatography with ultraviolet and tandem mass spectrometric detection (HPLC-UV-MS/MS) were employed to unambiguously confirm erinacine A in the isolated fractions, while HPLC with a charged aerosol detector (CAD) was used to determine its purity.

Given the limited commercial availability and the high price of erinacine A, the described method offers a straightforward and cost-effective alternative to obtain this valuable compound for further research and applications.

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