Have you ever tried the extraction of the compound by using the bligh&dyer protocol from a homogenized tissue sample?

Extraction

1. Homogenize 100-200 mg frozen zebrafish in 1 mL ice-cold methanol.

2. Add 2 mL ethyl acetate, vortex, and centrifuge (10,000 g, 10 min, 4°C).

3. Collect supernatant, evaporate solvent under nitrogen/vacuum.

4. Reconstitute in 100-200 μL HPLC-compatible solvent (50% methanol).

HPLC ConditionsHPLC Conditions

1. C18/C8 column

2. Gradient: water-acetonitrile (0.1% formic acid)

3. Flow rate: 0.5-1 mL/min

4. Detection: UV (245 nm) or mass spectrometry

References

Alderman et al. (2012) and Takahashi et al. (2014)

Notes

- Validate protocol for your specific conditions

- Consult local regulations for animal handling

- Ensure laboratory safety with organic solvents

Optimize and adjust as needed.