For developers of health supplement products, the Alliance has three professional experts in the field of natural chemistry, who are experienced in the preparation, refinement, separation and identification of extracts . In addition to the extract library mentioned above, the plant resources they can provide also include plant material sources, crude extract preparation, refined extract/fractionation preparation, process optimization, rapid prediction of active ingredients, separation and identification of pure compounds. However, the Alliance only plans to provide services for the separation and identification of pure compounds in the form of industry-university cooperation.

New drug development researchers (users) still have high concerns about the development risks after crude extract library screening. They believe that the time from extract to active ingredient is too long, the re-prepared extract cannot reproduce the biological activity, or the active ingredient cannot be confirmed because the activity is lost during the purification and separation process. To solve this problem, we will use the strategy of activity-guided separation and purification - mass spectrometry/molecular network analysis - targeted separation and purification to provide users with services to quickly identify and separate active ingredients.

The explanation is as follows: Activity-guided separation and purification is a traditional strategy used in natural product development. It requires close cooperation between the natural product chemistry team and the basic medical research team. After each separation, activity tests are performed to find effective stratification, and then the stratification is separated and tested for activity again. This process is repeated until a pure compound with activity is found. However, due to the above-mentioned problems, the effectiveness of this strategy is quite limited, so a better natural product development strategy is needed.

Since the late 1960s, mass spectrometry has been widely used to detect metabolites in biological samples. Non-standard tandem mass spectrometry is currently one of the main strategies used to discover and annotate new metabolites in metabolomics research. Non-standard tandem mass spectrometry can detect the chemical characteristics of all ionized compounds in a mixed sample. The so-called "chemical characteristics" refers to the molecular ion peak spectrum of all fragments produced after each compound is impacted. Generally speaking, compounds with similar structures will produce the same fragments after impact, and the same molecular ion peak spectrum will appear in mass spectrometry analysis. Therefore, by performing similarity comparison based on the "chemical features" of these mass spectrometry results, the compounds in the mixed sample can be classified according to structure, and the correlation between each compound can be analyzed, and then presented in a network-connected visual manner to generate a "molecular network". This network can be combined with the results of sample activity analysis to further identify the compound group that is the source of activity; at the same time, it can also be compared with existing public mass spectrometry databases to identify known compounds and thus identify new active compounds. This is also the strategy for the development of emerging natural products in the past few years.

Currently, molecular network analysis is mainly performed using the GNPS database (Global Natural Product Social Molecular Networking, GNPS-MN). For example, as shown in the figure below, high-throughput screening is first used to identify candidate extracts, followed by activity-guided separation and purification (no more than 2 cycles), and mass spectrometry/molecular network analysis is performed simultaneously on the active and adjacent inactive layers, and annotation is performed using the correlation between the content of the compounds in the active and inactive layers, so that active compound groups can be found. At the same time, during the annotation process, comparisons are made with the existing public mass spectrometry database to mark known compounds, which means that active compounds that cannot be annotated may be novel active compounds.
For potential new active compounds, we will conduct targeted purification separation: after each separation, we will use tandem mass spectrometry to track the separation of the molecular ion peak spectrum containing the new target compound until the pure compound is separated. In other words, this strategy can not only quickly find active ingredients, but also discover new active ingredients .
In addition, compared with the traditional method of using activity-guided separation and purification to try to find active ingredients, this new strategy saves the time and cost required for activity verification in the process, and can accelerate the development and application of new natural product drugs .