Region

Applications

Pain Panel using Strata-X-Drug B and Kinetex Phenyl-Hexyl 2.6u 50x4.6mm

Sample Preparation Details

Condition 1 :1 at 1.0
Condition 2 :1 at 1.0
Load :Load 0.20 mL Sample diluted in 0.1% Formic acid/water
Wash 1 :1 mL 0.1% Formic acid/water at 1.0 mL/min
Wash 2 :1 mL 30% methanol at 1.0 mL/min
Dry :3 to 4 mins at high vacuum (10" of Hg)
Elution 1 :0.5 mL 2% ammonium hydroxide/methanol: acetonitrile (1:1) at 1.0 mL/min
Elution 2 :0.5 mL 2% ammonium hydroxide/methanol: acetonitrile (1:1) at 1.0 mL/min
Evaporate: :Evaporate to dryness
Reconstitute :200 µL HPLC mobile phase A/B=85/15
Notes :Load: Samples from hydrolysis Using b-Glucoronidase as below To 200 µL Urine (calibrator, control) in a 1.5 mL micro centrifuge tube, add 20 µL combined IS (Internal Standards) spiking solution. Vortex 10-15 seconds followed by adding 100 µL 0.1 M Ammonium acetate buffer (pH 4.0) to each tube, mix/vortex another 15 seconds. Add 40 µL ß-Glucoronidase solution (100,000 units/mL). Vortex 10-15 seconds. Incubate for 2 hours in a shaker at 55 °C to complete hydrolysis of the glucuronides. Add 400 µl of 0.1% Formic acid and vortex for another 60 seconds. Centrifuge the hydrolyzed samples at 21,000 g for 10 minutes (to separate any proteins). Collect supernatant for further cleanup via Solid Phase Extraction (SPE). Wash 1: 1 mL 0.1% FA Wash2: 1 mL 30% methanol Dry: 3 to 4 mins at high vacuum (~10” of Hg) Elute: 2x500 uL 2% ammonium hydroxide/methanol: acetonitrile (1:1) Blow Down: To dryness under nitrogen at 40-45o C Reconstitute: With 200 uL of 85/15 (A/B) of LC mobile phase (Note: 1. No conditioning or equilibration needed in extraction) 2.Total of 15 internal standard used for the quantitation of 41 analytes.

Application Detail (App ID: 20847)

Part No:00B-4495-E0
Dimensions:50 x 4.6 mm ID
Elution Type:Gradient
Elution A:10mM ammonium formate
Elution B:0.1% Formic acid/methanol
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 95 5
2 4 0 100
3 5 0 100
4 5.1 95 5
5 7 95 5

Flow Rate:0.6 mL/min
Col. Temp: ambient
Detection: Mass Spectrometer (MS) @ amu (22°C)
Detector Info: AB Sciex API 5000™
Note:MS/MS Parameters
Detection: API 5000 MS/MS, TurboIonspray, positive polarity
CAD Gas: 7; GS1, GS2 = 50; curtain gas: 20, ESI spray voltage: 5500; Temp: 650 deg C

Analyte Details

Analyte Detail not available

1
Morphine
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2
Morphine-d6
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3
Hydromorphone
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4
Hydromorphone-d6
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5
Naloxone
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6
Codeine
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7
Oxymorphone
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8
Naltrexone
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9
Amphetamine
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10
Amphetamine-d5

11
6-MAM
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12
Oxycodone
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13
Hydrocodone
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14
Hydrocodone-d6
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15
Methamphetamine
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16
MDA
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17
MDMA
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18
MDEA
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19
Norfentanyl
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20
Tramadol
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21
Benzoylecgonine
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22
Benzoylecgonine-d3
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23
Meperidine
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24
Meperidine-d4
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25
Normeperidine
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26
Normeperidine-d4
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27
Meprobamate
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28
Norbuprenorphine
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29
PCP
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30
PCP-D5
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31
Fentanyl
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32
Fentanyl-d5
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33
EDDP
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34
Flurazepam
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35
Norpropoxyphene
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36
Norpropoxyphene-d5
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37
Sufentanil
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38
Buprenorphine
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39
Propoxyphene
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40
Carisoprodol
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41
Methadone
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42
Methadone-d3
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43
Midazolam
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44
Lorazepam
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45
Clonazepam
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46
Hydroxyalprazolam
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47
Oxazepam
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48
Flunitrazepam
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49
Alprazolam
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50
Temazepam
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51
Nordiazepam
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52
Nordiazepam-d5
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53
Diazepam
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54
6-MAM-d3
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55
Methamphetamine-d5
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