As a rare edible and medicinal fungus, black truffle (Tuber melanosporum) not only has a unique flavor, but is also rich in high-value natural active ingredients such as polysaccharides and L-ergothioneineineineineineineineine. Modern research shows that these ingredients have various physiological functions such as antioxidant, anti-inflammatory, anti-aging, and immune regulation, which makes black truffles show great application potential in the fields of cosmetics and functional foods.

However, its raw material development has long faced two core bottlenecks:

1. Unclear ingredients: Although black truffles are dubbed “rare”, the specific contents of their core functional ingredients (such as polysaccharides and L-ergothioneineineineineineineineine) are unclear, resulting in a lack of accurate data support for product efficacy claims and difficulty in maintaining constant quality;
2. Origin based on experience: The selection of raw materials mostly relies on traditional experience or the reputation of the origin. There is a lack of scientific ingredient data as a basis for selection, resulting in the inability to achieve precise selection of materials for specific effects, and the value of raw materials cannot be maximized.

In order to solve the above pain points, Landivine Research Institute innovatively applied liquid chromatography-tandem mass spectrometry (LC-MS/MS) precise quantification method to the systematic evaluation of black truffle raw materials. Related papers have been officially published in academic journals.

By establishing a highly sensitive and highly reproducible detection method, this study achieved for the first time an accurate “portrait” of the L-ergothioneineineineineineineineine and polysaccharide content in black truffles from different origins such as Liaoning and Yunnan, providing a solid scientific basis for the precise development and application of raw materials.

Technological breakthrough

In order to achieve accurate quantification of the core components of black truffles, this study used liquid chromatography-tandem mass spectrometry (LC-MS/MS) and phenol-sulfuric acid method to systematically analyze the L-ergothioneineineineineineineineine and polysaccharide contents of black truffles from Anshan, Liaoning, Kunming, Yunnan, and Gongshan, Yunnan.

In detecting the content of L-ergothioneineineineineineineineine in black truffles, the LC-MS/MS method used in this study shows the following core advantages:

1. High sensitivity and wide linear range: This method has a good linear relationship within the concentration range of 0.01~1.0μg/mL, and the lower limit of quantification is as low as 0.01μg/mL. This shows that this method has strong applicability and stability;
2. Excellent precision and accuracy: Methodological examination results show that, except for L-ergothioneineineineineineineineine mass concentration of 10ng/mL, the relative standard deviation of all tested concentrations is <3%, and the recovery rate of the sample is between 92.80% and 104.20%. This proves that the method has good repeatability, high precision, accuracy and reliability, and fully meets the analysis requirements.

Core findings

This study conducted a systematic analysis of black truffles from Anshan, Liaoning, Kunming, Yunnan, and Gongshan, Yunnan, and found that there were significant regional differences in the content of their core active ingredients.

Comparison of polysaccharide content in black truffles

Polysaccharide is one of the important active ingredients of black truffle and has good antioxidant activity and hypoglycemic activity. The study used the phenol-sulfuric acid method for determination.

• Polysaccharide content of water extract: After direct water extraction, the polysaccharide content of black truffles from Anshan, Liaoning Province was the highest, followed by Kunming, Yunnan, and finally Gongshan, Yunnan.

• Polysaccharide content of alcohol precipitate: After ethanol precipitation and purification, the polysaccharide content of black truffles in various regions increased, and the content ranking changed: the polysaccharide content of black truffles in Gongshan, Yunnan was the highest, followed by Kunming, Yunnan, and finally Anshan, Liaoning.

L-ergothioneineineineineineineineine content comparison

L-ergothioneineineineineineineineine, as a safe, non-toxic, multi-functional thiohistidine derivative, has multiple effects such as anti-inflammatory, prolonging cell survival cycle, and promoting the formation of nerve cells. This study used liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determination.

• The content measurement results show that the black truffles from Kunming, Yunnan, have the highest L-ergothioneineineineineineineineine content, which is significantly higher than the L-ergothioneineineineineineineineine content in black truffles from Gongshan, Yunnan and Anshan, Liaoning. This clearly indicates that black truffles originating from Kunming, Yunnan, have greater potential in antioxidant and anti-aging applications.

Application prospects

The data results of this study provide solid theoretical support and data support for the precise development and application of black truffle in the fields of cosmetics, medicine and food.

1. Directional screening of raw materials: To develop products focusing on moisturizing and repairing functions, raw materials from Yunnan Gongshan with extremely high alcohol-precipitated polysaccharide content can be preferred; if the core appeal is antioxidant, raw materials from Yunnan Kunming with the highest L-ergothioneineineineineineineineine content should be preferred.
2. Quality control standardization: The LS-MS/MS detection method established in this study is accurate and reliable, and can be used as a key technical means for quality control and standardization of black truffle raw materials and its derivative products, ensuring the stability and controllability of product efficacy.
3. Product efficacy empowerment: Research has confirmed that China’s native black truffles are rich in high-value active ingredients, providing strong scientific endorsement for its innovative applications in high-end skin care products, functional foods, and medical dressings.

Research summary

This study systematically analyzed the polysaccharide and L-ergothioneineineineineineineineine content in black truffles from different origins, and revealed that black truffles from Yunnan performed well in terms of L-ergothioneineineineineineineineine and alcohol-precipitated polysaccharides. Among them, the Kunming origin had the highest L-ergothioneineineineineineineineine content, and the Gongshan origin had the most prominent alcohol-precipitated polysaccharide content.

Compared with traditional raw materials with a single active ingredient, black truffle extract has both the repairing and antioxidant capabilities of polysaccharides and L-ergothioneineineineineineineineine, achieving a natural synergy of multiple physiological functions. This study provides a reliable basis for the precise selection and efficacy of black truffle in high-end skin care products and functional foods, and also lays a scientific foundation for the in-depth development and quality control of natural raw materials.