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Extraction of Timolol from Serum using Strata-X-C 10 mg Plate & Kinetex 2.6u Biphenyl 50x2.1 Column

Sample Preparation Details

Condition 1 :0.5 mL Methanol
Condition 2 :0.5 mL Water
Load :Load 375.00 µL Sample diluted in 375
Wash 1 :0.5 mL 2% Formic acid in water
Wash 2 :0.5 mL Methanol
Elution 1 :0.125 mL 5% Ammonium hydroxide in Acetonitrile/Methanol (60:40)
Elution 2 :0.125 mL 5% Ammonium hydroxide in Acetonitrile/Methanol (60:40)
Elution 3 :0.125 mL 5% Ammonium hydroxide in Acetonitrile/Methanol (60:40)
Evaporate: :Evaporate to dryness
Notes :No dry down step was performed. 1µL of final eluent was injected.

Application Detail (App ID: 22982)

Part No:00B-4622-AN
Dimensions:50 x 2.1 mm ID
Elution Type:Gradient
Elution A:0.1% formic acid in water
Elution B:0.1% formic acid in acetonitrile
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 95 5
2 0.5 95 5
3 3 5 95
4 3.5 5 95
5 3.51 95 5
6 5.5 95 5

Flow Rate:0.5 mL/min
Col. Temp: 25 °C
Detection: Tandem Mass Spec (MS-MS) @ (22°C)
Note:Strata X-C 10 mg Plate Protocol

Condition
500 µL methanol
Equilibrate
500uL water
Load
750 µL diluted serum
(375 µL plasma diluted 1:1 with 4% phosphoric acid in water)
Wash1
500 µL 2% formic acid in water
Wash 2
500 µL methanol
Elute
375 µL(3x125 µL) 5% ammonium hydroxide in acetonitrile:methanol(60:40)
Inject
1 µL

Note: Spiked conc. of analyte was 1 ng/mL serum

Analyte Details

Analyte Detail not available

1
Timolol
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