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A LC-MS/MS Method for Quantifying the Schisandrin B and Exploring Its Intracellular Exposure Correlating Antitumor
Bosu Meng1,2, Shouhong Gao1,2, Jihui Chen3
1College of Traditional Chinese Medicine, Yunnan University of Traditional Chinese Medicine, Kunming 650500, Yunnan, China.
Schisandrin B (Sch.B) effectively inhibits colorectal cancer cell growth. This study developed a method to track Sch.B distribution within cancer cells, revealing higher concentrations in mitochondria than the nucleus, aiding in understanding its antitumor mechanism.
Area of Science:
- Pharmacology
- Analytical Chemistry
- Cancer Biology
Background:
- Schisandrin B (Sch.B) exhibits antineoplastic properties against colorectal cancer.
- The precise mechanism of Sch.B's action remains unclear.
- Intracellular drug distribution patterns can elucidate drug mechanisms.
Purpose of the Study:
- To establish a sensitive UHPLC-MS/MS method for quantifying Sch.B in colorectal cancer cells.
- To investigate the intracellular spatial distribution of Sch.B within cancer cells.
- To correlate drug distribution with its antineoplastic effects.
Main Methods:
- Development and validation of an ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) assay.
- Sample preparation via methanol protein precipitation.
- Analysis using an Atlantis T3-C18 column with gradient elution.
- Cell viability and apoptosis assays (IC50 determination).
Main Results:
- A validated UHPLC-MS/MS method with high sensitivity and accuracy (R > 0.99, recovery 85-91%).
- Sch.B demonstrated dose-dependent inhibition of HCT116 cell proliferation, with significant suppression at 75 μM (IC50).
- Intracellular Sch.B levels peaked at 36 hours in nucleus and mitochondria, with higher accumulation in mitochondria.
Conclusions:
- The developed UHPLC-MS/MS method is suitable for determining Sch.B in colorectal cancer cells.
- Sch.B exhibits significant antitumor activity against HCT116 cells.
- Mitochondrial accumulation of Sch.B may play a crucial role in its observed antineoplastic effects.
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