Simplifying Natural Product Analysis with Data- Independent Acquisition Workflow

Significance of the Topic

Natural products are a rich source of unique bioactive molecules but present complex analytical challenges. High-resolution mass spectrometry with data-independent acquisition expands the detectable compound range and enhances confidence in identification, crucial for natural product research and quality control.

Objectives and Study Overview

This article demonstrates a user-friendly DIA acquisition and processing workflow on the Shimadzu LCMS-9030 Q-TOF platform. Using an extract of Aconitum carmichaeli, it details the integration of hardware and software tools to achieve comprehensive compound coverage and streamlined identification.

Methodology and Instrumentation

Sample Preparation and Chromatography

• Extraction of 500 mg Aconitum carmichaeli in 70 percent methanol with agitation for thirty minutes
• Filtration through a 0.2 micrometer syringe filter and direct injection of 10 microliter
• Separation on a C18 column maintained at forty degrees Celsius using a gradient of water formic acid and acetonitrile at 0.4 milliliter per minute

Mass Spectrometry and Data Acquisition

• Shimadzu LCMS-9030 Q-TOF with electrospray ionization source
• DIA parameters including precursor range one hundred to one thousand mass to charge, twenty mass isolation window, collision energy spread fifteen to fifty five electronvolts
• High speed acquisition up to one hundred hertz and heated gas flows for optimal desolvation

Data Processing

• LabSolutions LCMS software for method setup
• LabSolutions Insight Explore for deconvolution, formula prediction, library searches and in silico fragmentation

Key Results and Discussion

From the Aconitum carmichaeli extract, over five thousand components were detected within a twelve minute window, yielding two hundred sixty unique precursor ions. A deconvoluted feature at m z four fifty four point two seven nine seven was annotated as fuziline. This compound matched the NIST twenty twenty library with a score of eighty nine and received an Assign score of eighty eight in public database searching. In silico fragmentation supported structural assignments of key fragment ions.

Benefits and Practical Applications

• Comprehensive compound coverage enables thorough profiling of complex natural matrices
• User friendly software reduces setup complexity and accelerates DIA adoption
• Automated deconvolution and formula prediction enhance identification confidence
• Integration of local and public spectral libraries streamlines unknown discovery

Future Trends and Potential Applications

• Advanced machine learning models for automated compound annotation and scoring
• Expansion of public and in house spectral libraries for greater coverage
• Integration of multi omics data to correlate metabolite profiles with biological activity
• High throughput screening workflows for rapid natural product dereplication

Conclusion

The combination of the Shimadzu LCMS-9030 Q-TOF with LabSolutions software provides a robust, streamlined DIA workflow for natural product analysis. This approach delivers enhanced coverage, confidence in identification, and efficient data processing, facilitating discoveries in pharmacognosy, quality control, and industrial applications.

References

1. Chua Chun Kiang Udi Jumhawan Chia Shao Hua Doriane Toinon Simplifying Natural Product Analysis with Data Independent Acquisition Workflow Shimadzu Application News First Edition March 2021
2. NIST Mass Spectral Library Version 2020 National Institute of Standards and Technology
3. Shimadzu Corporation LabSolutions LCMS and LabSolutions Insight Explore Version 599 User Manuals 2021