Parameter Set Anti-HIV Drugs - LC-MS/MS

• Reconstitute the Internal Standard Mix.
• Add 800 μl Internal Standard Mix to 12 ml Precipitation Reagent to form mixture A.
• Pipette 50 μl research sample/calibrator/MassCheck® control into a 1.5 ml reaction vial.
• Add 25 μl Extraction Buffer.
• Mix and incubate 2 min at ambient temperature (do not centrifuge).
• Add 250 μl of mixture A and mix 30 s minimum (vortex).
• Centrifuge 5 min at 15000 x g.
• Dilute the supernatant with Dilution Buffer prior to injection depending on the instrument sensitivity.

Group 1
0.00–0.20 min       0 % Mobile Phase 2
0.21–2.00 min     70 % Mobile Phase 2
2.01–2.90 min   100 % Mobile Phase 2
2.91–3.50 min       0 % Mobile Phase 2
Group 2
0.00–0.20 min       0 % Mobile Phase 2
0.21–0.60 min     50 % Mobile Phase 2
0.61–2.40 min     90 % Mobile Phase 2
2.41–3.00 min       0 % Mobile Phase 2
Group 3
0.00–0.20 min       0 % Mobile Phase 2
0.21–2.30 min     90 % Mobile Phase 2
2.31–3.10 min       0 % Mobile Phase 2