Solvent Extraction

Aqueous Ethanol Extraction: For red pigments produced by Monascus purpureus, using 60% ethanol at pH 6, 30°C, with an extraction time of 16 hours and an agitation rate of 180 rpm yielded optimal results [1].

Methanol and Acetone Extraction: For astaxanthin from Xanthophyllomyces dendrorhous, a combination of ethanol, methanol, and acetone was effective. Microwave treatment to destroy cell walls, followed by extraction at 30°C over 24 hours with 10% ethanol was optimal [2] .

Aqueous Two-Phase Extraction

Ionic Liquids: Using ammonium- or imidazolium-based ionic liquids for extracting polyketide-based pigments from fungi like Talaromyces albobiverticillius and Emericella purpurea showed promising results. Tetrabutylammonium bromide ([N4444]Br-) was particularly effective [3] .

Physical Disruption Techniques

Microwave-Assisted Extraction (MAE): Enhances the efficiency of pigment extraction when combined with deep eutectic solvents (DES. Optimal conditions include temperatures between 20-90°C, extraction times of 5-70 minutes, and ultrasound frequencies of 18-80 kHz [4] [5] .

Chromatography Techniques

Centrifugal Partition Chromatography: For anthraquinones from Cortinarius sanguineus, a biphasic solvent system comprising chloroform/ethyl acetate/methanol/water/acetic acid (3:1:3:2:1, v/v/v/v/v) was effective [6] .

Column Chromatography: Used for purifying pigments from seaweeds and fungi, often combined with thin-layer chromatography (TLC) and UV-visible spectrophotometry for characterisation [7] [8] .

Modern Extraction Methods

Supercritical Fluid Extraction: Offers high efficiency and development potential for extracting pigments from various sources [9] .

Ultrasound-Assisted Extraction (UAE): Combined with DES, UAE provides higher performance compared to conventional methods like maceration and soxhlet extraction [5] .

Specific Case Studies

Monascus purpureus: Optimal pigment production in submerged fermentation conditions at 30°C and pH 6.5, using rice and peptone as carbon and nitrogen sources respectively [10] .

Fusarium chlamydosporum: Homogenization technique followed by purification using TLC, column chromatography, and spectrophotometry for extracting fusarubin [8] .

e.g.

Aqueous Ethanol Extraction, 60% ethanol, pH 6, 30°C, 16h, 180 rpm Monascus purpureus, Red Pigments [1]

Methanol/Acetone Extraction: Microwave treatment, 30°C, 24h, 10% ethanol Xanthophyllomyces dendrorhous, Astaxanthin [2]

Ionic Liquids Tetrabutylammonium bromide ([N4444]Br-) Talaromyces albobiverticillius, Polyketide Pigments [3]

Centrifugal Partition Chromatography Chloroform/ethyl acetate/methanol/water/acetic acid Cortinarius sanguineus, Anthraquinones [6]

Supercritical Fluid Extraction Various conditions Multiple species Various pigments [9]

UAE with DES 20-90°C, 5-70 min, 18-80 kHz Multiple species Various pigments [5]

These methodologies should provide a comprehensive guide to effectively extract fungal pigments. Adjustments may be necessary based on specific fungal strains and pigment types.

References

1. Recovery of Red Pigments from Monascus purpureus FTC 5357 by Extraction of Fermented Solids: Operational Conditions and Kinetics

Daud, N.F.S., Said, F.M., Chisti, Y., Yasin, N.H.M.

Brazilian Archives of Biology and Technology, 2021

https://www.scopus.com/pages/publications/85116392130

2. An efficient method for the extraction of astaxanthin from the red yeast Xanthophyllomyces dendrorhous

Choi, S.-K., Kim, J.-H., Park, Y.-S., (...), Chang, H.-I.

Journal of Microbiology and Biotechnology, 2007

https://www.scopus.com/pages/publications/34250206672

3. Aqueous two-phase system extraction of polyketide-based fungal pigments using ammonium-or imidazolium-based ionic liquids for detection purpose: A case study

Lebeau, J., Petit, T., Fouillaud, M., (...), Caro, Y.

Journal of Fungi, 2020

https://www.scopus.com/pages/publications/85098225723

4. Sustainable green extraction of anthocyanins and carotenoids using deep eutectic solvents (DES): A review of recent developments

Osamede Airouyuwa, J., Sivapragasam, N., Ali Redha, A., Maqsood, S.

Food Chemistry, 2024

https://www.scopus.com/pages/publications/85188998001

5. Application of emerging technologies for extraction of pigments using green solvents: case of deep eutectic solvents combined with ultrasound-assisted extraction technique

Zannou, O., Oussou, K.F., Mohammed, S., (...), Esatbeyoglu, T.

Biomass Conversion and Biorefinery, 2025

https://www.scopus.com/pages/publications/105003128532

6. A convenient separation strategy for fungal anthraquinones by centrifugal partition chromatography

Hammerle, F., Zwerger, M., Höck, A., (...), Siewert, B.

Journal of Separation Science, 2022

https://www.scopus.com/pages/publications/85122870880

7. Seaweed Calliblepharis jubata and Fucus vesiculosus Pigments: Anti-Dermatophytic Activity

Gomes, L., Cotas, J., Fernandes, C., (...), Pereira, L.

Applied Sciences (Switzerland), 2024

https://www.scopus.com/pages/publications/85192460507

8. Production of a Polyketide Pigment by Fusarium chlamydosporum

Thomas, T.A., Tirumale, S.

Science, 2022

https://www.scopus.com/pages/publications/85132411167

9. Characteristics and extraction technology of natural pigments from animals and plants

Zheng, H., Zhang, H., Zhang, Z.-H.

Forest Research, 2003

https://www.scopus.com/pages/publications/1942540465

10. Isolation and optimization of Monascus ruber OMNRC45 for red pigment production and evaluation of the pigment as a food colorant

Darwesh, O.M., Matter, I.A., Almoallim, H.S., (...), Oh, Y.-K.

Applied Sciences (Switzerland), 2020

https://www.scopus.com/pages/publications/85098503975