Extraction of Analgesics from Plasma Using Strata X-A on Kinetex 2.6u, C18, 50x2.1mm by LC-UV

LC Conditions

Column

Kinetex 2.6 µm C18 100 Å, LC Column 50 x 2.1 mm, Ea

Brand

Kinetex

Part No

00B-4462-AN

Phase Name

C18

Elution Type

Gradient

Mobile Phase

A: 20 mM KH2PO4(pH 7.0)

B: Methanol

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

90

10

2

1

90

10

3

1.01

30

70

4

5.5

30

70

Flow Rate

0.40 mL/min

Temperature

0°C

Detection

UV-Vis Abs.-Variable Wave.(UV) @ (22°C)

Detection Info

Sample Preparation Details

Description

Strata-X-A 33 µm Polymeric Strong Anion, 30 mg / 1 mL, Tubes, 100/Pk

Part No

8B-S123-TAK

Pretreatment Note

Elution solvent was applied in 2 aliquots of 500uL followed by evaporation to dryness at 45 deg C under N2 and reconstitute with 500 uL initial mobile phase (20mm KH2PO4(PH 7.0)/ MeOH=90/10) spiked with 5ug/mL Fenoprofen (IS).

Condition 1

1 mL Methanol at 1 mL/min

Condition 2

1 mL Water at 1 mL/min

Load

Load 0.50 mL Sample diluted in Water

Wash 1

1 mL 50 mm Na acetate, PH 7.0 at 1 mL/min

Wash 2

1 mL Methanol at 1 mL/min

Dry

2 mins at 10 of Hg

Elution 1

1 mL 5% FA/MeOH at 1 mL/min

Evaporation

Evaporate to dryness

Reconstitute

500 µL 20mm KH2PO4(PH 7.0)/ MeOH=90/10