Data Supplement

• Supplemental Data -

  Supplemental Figure S1. Mass spectrometric fragmentation pathways of cantharidin-lysine amide conjugate and cantharidin-lysine imide conjugate in UPLC-Q/Orbitrap MS.

  Supplemental Figure S2. Representative extracted ion chromatograms of the conjugates of cantharidin with amino acids in PBS incubation.

  Supplemental Figure S3. Typical MS/MS spectra of the conjugates of cantharidin with amino acids in PBS incubation.

  Supplemental Figure S4. The stability of cantharidin in PBS and plasma incubation.

  Supplemental Figure S5. ¹H NMR spectra of chemically synthesized cantharidin-aniline imide conjugate.

  Supplemental Figure S6. ¹³C NMR spectra of chemically synthesized cantharidin-aniline imide conjugate.

  Supplemental Figure S7. Representative extracted ion chromatograms and MS/MS spectra of cantharidin-aniline imide conjugate from PBS incubation and chemical synthesis.

  Supplemental Figure S8. Representative extracted ion chromatograms of the conjugates of cantharidin with amino acids and N-acetyl-lysine found in the liver sample.

  Supplemental Figure S9. The relative content of hepatic cantharidin-N-acetyl-lysine amide conjugate in cantharidin-treated mice.

  Supplemental Figure S10. Representative extracted ion chromatograms and product ion spectra of cantharidin and its metabolites.

  Supplemental Figure S11. Effects of P450 inhibitor 1-aminobenzotriazole (1-ABT) on levels of the conjugates and its metabolites in the urine sample of cantharidin-treated mice.

  Supplemental Figure S12. Covalent binding and metabolism of cantharidin in mouse liver microsomal incubation.

  Supplemental Figure S13. The typical extracted ion chromatogram and product ion spectrum of amide conjugate of cantharidin with protein lysine residues obtained from liver samples of cantharidin-treated mice after proteolytic digestion.

  Supplemental Figure S14. Relative content of amide and imide conjugates of cantharidin with lysine or arginine in the PBS incubation for 3 h at different pH values.

  Supplemental Table S1. Conjugates of cantharidin with amino acid and N-acetyl-lysine in PBS incubation.