Extraction of Opiates-NIDA5 Using Strata X-C on Kinetex 2.6µm C18 50 x 2.1mm ID by LCMS

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: 0.1% Formic acid in H2O

B: 0.1% Formic acid in ACN

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

95

5

2

0.24

95

5

3

1.25

70

30

4

2.5

70

30

5

2.51

5

95

6

2.8

5

95

7

2.81

95

5

8

4.6

95

5

Flow Rate

450.00 mL/min

Temperature

0°C

Detection

Mass Spectrometer (MS) @ (22°C)

Detection Info

Sample Preparation Details

Description

Strata-X-C 33 µm Polymeric Strong Cation, 30 mg / 3 mL, Tubes, 50/Pk

Part No

8B-S029-TBJ

Pretreatment Note

Blown down samples (after evaporation) were reconstituted in 1mLof initial mobile phase (A/B=95/5).

Condition 1

1 mL Methanol at 2 mL/min

Condition 2

1 mL Water at 2 mL/min

Load

Load 1.00 mL Sample diluted in 100 mM KH2PO4 (pH 6)

Wash 1

1 mL 0.1N HCl at 2 mL/min

Wash 2

1 mL Methanol at 2 mL/min

Dry

2 mins @ 10 of Hg pressure

Elution 1

0.5 mL 5% ammonium hydroxide in methanol at 1 mL/min

Elution 2

0.5 mL 5% ammonium hydroxide in methanol at 1 mL/min

Evaporation

Evaporate to dryness

Reconstitute

1000 µL Initial mobile phase (A/B=95/5)