2026-10-14T09:49:50Z https://api.figshare.com/v2/oai

oai:figshare.com:article/33886145 2026-09-17T11:04:21Z category_1 category_4 category_146 category_7 category_21 category_873 category_39 category_45 category_734 category_133 portal_63 item_type_2 month_year_09_2026

Fully Automated Purification of Small Molecules Using Online MicroSPE-UPLC-SPE Venkata Ganesh Jarubula (25014947) Gerd Grewing (25014950) Pauline Stark (25014953) Maximilian Frey (5243549) Dario Esposto (25014956) Simon Isfort (25014959) Marvin Hempel (25014962) Guillaume Patoine (9038313) Henriette Uthe (12163740) Alain Tissier (746524) Gerd U. Balcke (4901056) Biophysics Biochemistry Space Science Medicine Biotechnology Chemical Sciences not elsewhere classified Ecology Sociology Biological Sciences not elsewhere classified Plant Biology validated using plant throughput peak fractionation spe structural annotation preserving volatile compounds enables fully automated eliminating drying steps based structure elucidation approach enables milligram fully automated purification collecting narrow fractions successful purification consuming purification well format system allows single day several sesquiterpenoids scale isolation pump loading postcolumn microspe online microspe metabolites remains metabolite enrichment low sensitivity key challenge fungal extracts directly eluted deuterated solvents 1 ml Structural annotation of metabolites remains a key challenge in metabolomics, as NMR-based structure elucidation for every unknown UPLC peak is limited by low sensitivity and time-consuming purification. To overcome these constraints, we developed an online microSPE-UPLC platform that enables fully automated, high-throughput peak fractionation. The system allows the injection of up to 1 mL of organic extract onto microSPE cartridges in a 96-well format for metabolite enrichment and cleanup. A dual-pump loading and refocusing strategy ensures quantitative transfer to UPLC, maintaining high chromatographic resolution while collecting narrow fractions on postcolumn microSPE. Fractions can be directly eluted with deuterated solvents for NMR analysis, eliminating drying steps and preserving volatile compounds. This approach enables milligram-scale isolation of natural products per compound within a single day. The workflow was validated using plant and fungal extracts, enabling the successful purification and characterization of several sesquiterpenoids and a diterpenoid. 2026-09-17T00:00:00Z Dataset Media 10.1021/acs.analchem.6c00842.s002 https://figshare.com/articles/media/Fully_Automated_Purification_of_Small_Molecules_Using_Online_MicroSPE-UPLC-SPE/33886145 CC BY-NC 4.0