📝 Homogenize the tissue sample using a homogenizer. Weigh 2.50 ± 0.05 g of the homogenized sample into a 15 mL centrifuge tube, add 4 mL of Reagent A and 150 µL of derivatization reagent, and shake the sample up and down to mix until it becomes a paste. Place the centrifuge tube in an 80°C water bath and incubate for 10 min. Remove the centrifuge tube, add 1 mL of Reagent B, close the centrifuge tube cap, mix well, and cool to room temperature (20–25°C); add 5 mL of extraction agent, close the centrifuge tube cap, and invert the tube up and down 20–25 times. Do not vortex. Centrifuge at 4000 r/min for 5 min (if the supernatant is less than 3 mL after centrifugation, shake the sample up and down 3–4 times and re-centrifuge). Transfer 3 mL of the upper layer into a 10 mL centrifuge tube and dry under a nitrogen or air stream in a 50–60°C water bath. Add 0.8 mL of cleanup agent, vortex for 1 min using a vortex mixer, then add 500 µL of sample reconstitution solution, and vortex for 5–6 s to mix thoroughly. Centrifuge at 4000 r/min for 5 min. It is strongly recommended to remove the upper organic phase. The remaining clear liquid is the test solution and should be analyzed immediately.
Homogenize the tissue sample using a homogenizer.
Weigh 2.50±0.05 g of the homogenized sample into a 15 mL centrifuge tube. Add 4 mL of Reagent A and 150 µL of derivatization reagent. Shake the sample up and down until it becomes a paste.
Place the centrifuge tube in an 80°C water bath and incubate for 10 min.
Remove the centrifuge tube, add 1 mL of Reagent B, close the cap, mix, and cool to room temperature (20–25°C). Add 5 mL of extraction solvent, close the cap, and invert the tube 20–25 times. Do not vortex.
Centrifuge at 4000 r/min for 5 min. (If the supernatant after centrifugation is less than 3 mL, shake the sample up and down 3–4 times and re-centrifuge.)
Transfer 3 mL of the upper layer to a 10 mL centrifuge tube and evaporate to dryness under a nitrogen or air stream in a 50–60°C water bath.
Add 0.8 mL of purification agent, vortex for 1 min, then add 500 µL of sample reconstitution solution, and vortex for 5–6 s to mix thoroughly.
Centrifuge at 4000 r/min for 5 min. It is strongly recommended to remove the upper organic phase. The remaining clear liquid is the test solution, which should be analyzed immediately.