1. Sample Preparation
Adding 5mL water into 5g beaten eggs, votex 1min; then adding 10mL acetonitrile, vortex 1min; Adding salt bag (2.CA8115.B001), quickly vortex
2mins, (Heat release is obvious, operation needs rapidly), centrifuge 6min under 4000rpm (stratified, superstratum is 10mL acetonitrile).

2. SPE columns Operation
SPE column： CNW Poly-Sery HLB SPE column
(2.CA3179.0001, 60mg, 3mL)
Purification mode：
Adding 2mL upper acetonitrile solution into SPE column, collect solution (control flow rate to 1d/s), evaporating with Nitrogen nearly dry, adding
mobile phase (20% acetonitrile + 80% 0.1%Formic acid solution), constant volume to 1mL; use 2.CF2102.0001 to filter, waiting for machine
detection.
Enrichment mode：
Evaporating 2mL supernate with Nitrogen nearly dry, add 2mL water and mix well, waiting testing.
Activation： 3mL Formic acid
Balance： 3mL Water
Add the sample： Control flow rate 1d/s
Wash the packing： 3mL Water
Rinse the packing： 2*2.5mL Formic acid
Collect eluent, evaporating with Nitrogen nearly dry, adding mobile phase and constant volume to 1mL, use 2.CF2102.0001 filter, waiting for machine
detection.

QuEChERS method：
Add 1mL supernate to 2mL purification pipe(2.CA8747.H001), vortex 1min; Centrifuge 6min under 4000rpm, evaporating with Nitrogen nearly dry, 1mL
mobile phase redissolve, use SCAA-104 to filter, waiting for machine detection.

3. HPLC Conditions
Column： Athena uhplc C18，( 8.2105UA.0001,2.1*50mm，1.8um)
Mobile phase： Acetonitrile：0.1% Formic acid Water=60：40
Flow rate： 0.2mL/min
Column Temperature： 30℃
Injection volumn： 10uL

4. Mass Spectrometer Conditions
MRM mode
Flow rate: 7L/min
Desolvation temperature: 300℃
Atomizer pressure: 35psi
Cracking voltage: 135V
Ionization mode: -ESI
Collision Energy: 10eV