Extraction of Amphetamines from Urine using Strata-X-C Microelution & Kinetex 2.6u Biphenyl, 50x2.1

Extraction of Amphetamines from Urine using Strata-X-C Microelution & Kinetex 2.6u Biphenyl, 50x2.1
LC Conditions
Column

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

Brand

Kinetex

Part No

00B-4622-AN

Phase Name

Biphenyl

Elution Type

Gradient

Mobile Phase

A: 0.1% formic acid in water

B: 0.1% formic acid in methanol

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

90

10

2

3

5

95

3

4

5

95

4

4.01

90

10

5

6

90

10

Flow Rate

0.50 mL/min

Temperature

25°C

Detection

Tandem Mass Spec (MS-MS) @ (22°C)

Detection Info
Sample Notes

Extraction of Amphetamines from Human Urine Using Strata-X-C u-Elution Plate Sorbent: Strata-X-C µ Elution 96-well Plate Condition: 200 uL methanol Equilibrate: 200 uL water Load: 400 uL diluted human urine (200 uL sample diluted 1:1 with water) Wash 1: 200 uL 2% formic acid/water Wash 2: 200 uL methanol Elute: 2x25uL 5% ammonium hydroxide/acetonitrile: methanol (60:40) Direct injection: injection of 2 uL sample directly HPLC: Kinetex Biphenyl, 2.6u, 50x2.1; A=0.1% FA/water, B=0.1% FA/methanol; Flow rate=0.5 mL/min; Inj. vol=2uL. Step Total Time(min) Flow Rate (µl/min) A (%) B (%) 0 0.00 500 90.0 10.0 1 3.00 500 5.0 95.0 2 4.0 500 5.0 95.0 3 4.01 500 90.0 10.0 4 6.0 500 90.0 10.0

Sample Preparation Details
Description

Novum SLE, MINI 96-Well Plate, 1/Pk

Part No

8E-S138-FGA

Pretreatment Note

Sample pre-treatment Dilute 150 uL of human plasma (spiked with 25 ng/mL and 125 ng/mL of cortisone and prednisolone respectively) with 150 uL of 50 mM sodium phosphate dibasic heptahydrate, pH unadjusted. Mix briefly (3-5 sec). Sample loading: Load the sample from pre-treatment step above onto the Novum plate and apply a short and gentle pulse of vacuum (~ 10" of Hg for 20 secs) until the sample has completely entered the media. Wait for 5 minutes. Elution: Dispense 1000 uL of ethyl acetate onto the SLE media and allow the solvent to elute by gravity (~ 5 min elution time) and collect the eluant. Apply vacuum at 5" of Hg for 45 secs to complete the extraction. NOTE: Prolonged application of vacuum will result in elution of plasma out of the SLE media and into the final extracted solvent. Dry down: Evaporate the final extract to complete dryness under slow stream of N2 at 40°C Reconstitute: Reconstitute the dry residue in 150 uL of initial mobile phase fortified with cortisol-D4.

Load

Load 150.00 µL Sample diluted in 150

Elution 1

1000 mL Ethyl acetate

Evaporation

Evaporate to dryness

Reconstitute

150 µL 10mM ammonium acetate in water:methanol(50:50)

Order Items Used in This Application

Kinetex 2.6 µm Biphenyl 100 Å, LC Column 50 x 2.1 mm, Ea
00B-4622-AN

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

View Product

Novum SLE, MINI 96-Well Plate, 1/Pk
8E-S138-FGA

Novum SLE, MINI 96-Well Plate, 1/Pk

View Product

  • 1. Amphetamine
  • 2. Methamphetamine
  • 3. MDMA
  • 4. MDA
  • 5. MDEA
-OEChem-06211003152D<br><br> 23 23  0     1  0  0  0  0  0999 V2000<br>    3.7320    2.0950    0.0000 N   0  0  0  0  0  0  0  0  0  0  0  0<br>    3.7320    1.0950    0.0000 C   0  0  3  0  0  0  0  0  0  0  0  0<br>    2.8660    0.5950    0.0000 C   0  0  0  0  0  0  0  0  0  0  0  0<br>    2.8660   -0.4050    0.0000 C   0  0  0  0  0  0  0  0  0  0  0  0<br>    4.5981    0.5950    0.0000 C   0  0  0  0  0  0  0  0  0  0  0  0<br>    3.7320   -0.9050    0.0000 C   0  0  0  0  0  0  0  0  0  0  0  0<br>    2.0000   -0.9050    0.0000 C   0  0  0  0  0  0  0  0  0  0  0  0<br>    3.7320   -1.9050    0.0000 C   0  0  0  0  0  0  0  0  0  0  0  0<br>    2.0000   -1.9050    0.0000 C   0  0  0  0  0  0  0  0  0  0  0  0<br>    2.8660   -2.4050    0.0000 C   0  0  0  0  0  0  0  0  0  0  0  0<br>    4.4682    1.5200    0.0000 H   0  0  0  0  0  0  0  0  0  0  0  0<br>    2.6540    1.1776    0.0000 H   0  0  0  0  0  0  0  0  0  0  0  0<br>    2.2554    0.4873    0.0000 H   0  0  0  0  0  0  0  0  0  0  0  0<br>    4.2881    0.0581    0.0000 H   0  0  0  0  0  0  0  0  0  0  0  0<br>    5.1350    0.2850    0.0000 H   0  0  0  0  0  0  0  0  0  0  0  0<br>    4.9081    1.1319    0.0000 H   0  0  0  0  0  0  0  0  0  0  0  0<br>    4.2690   -0.5950    0.0000 H   0  0  0  0  0  0  0  0  0  0  0  0<br>    1.4631   -0.5950    0.0000 H   0  0  0  0  0  0  0  0  0  0  0  0<br>    4.2690    2.4050    0.0000 H   0  0  0  0  0  0  0  0  0  0  0  0<br>    3.1951    2.4050    0.0000 H   0  0  0  0  0  0  0  0  0  0  0  0<br>    4.2690   -2.2150    0.0000 H   0  0  0  0  0  0  0  0  0  0  0  0<br>    1.4631   -2.2150    0.0000 H   0  0  0  0  0  0  0  0  0  0  0  0<br>    2.8660   -3.0250    0.0000 H   0  0  0  0  0  0  0  0  0  0  0  0<br>  1  2  1  0  0  0  0<br>  1 19  1  0  0  0  0<br>  1 20  1  0  0  0  0<br>  2  3  1  0  0  0  0<br>  2  5  1  0  0  0  0<br>  2 11  1  0  0  0  0<br>  3  4  1  0  0  0  0<br>  3 12  1  0  0  0  0<br>  3 13  1  0  0  0  0<br>  4  6  2  0  0  0  0<br>  4  7  1  0  0  0  0<br>  5 14  1  0  0  0  0<br>  5 15  1  0  0  0  0<br>  5 16  1  0  0  0  0<br>  6  8  1  0  0  0  0<br>  6 17  1  0  0  0  0<br>  7  9  2  0  0  0  0<br>  7 18  1  0  0  0  0<br>  8 10  2  0  0  0  0<br>  8 21  1  0  0  0  0<br>  9 10  1  0  0  0  0<br>  9 22  1  0  0  0  0<br> 10 23  1  0  0  0  0<br>M  END
Compound Name

Amphetamine

CID

3007

Molecular Formula

C9H13N

Molecular Weight

135

No. Hydrogen Bond Acceptors

1

No. Hydrogen Bond Donors

1

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