Nicotine and Metabolites (Ion Suppression) by SPE using Gemini NX-C18, LC-MSMS (Fig.5)
LC Conditions
Column
Gemini 3 µm NX-C18 110 Å, LC Column 50 x 2 mm, Ea
Brand
Gemini
Part No
00B-4453-B0
Phase Name
NX-C18
Elution Type
Gradient
Mobile Phase
A: 20mM Ammonium Bicarbonate
B: 100% Acetonitrile
Gradient Profile
Step No.
Time(min)
%A
%B
1
0
90
10
2
3
25
75
3
3.1
90
10
4
5
90
10
Flow Rate
500.00 mL/min
Temperature
25°C
Detection
Tandem Mass Spec (MS-MS) @ (22°C)
Detection Info
Sample Preparation Details
Description
Strata-X-C 33 µm Polymeric Strong Cation, 60 mg / 3 mL, Tubes, 50/Pk
Part No
8B-S029-UBJ
Pretreatment Note
Reconstitution Solvent: 500uL Acetonitrile/20mM Ammonium bicarbonate (10:90)
Condition 1
2 mL Methanol at 1 mL/min
Condition 2
2 mL Ammonium acetate buffer at 1 mL/min
Load
Load 2.00 mL Sample diluted in 0.5 diluted urine sample
Wash 1
2 mL Ammonium acetate buffer at 1 mL/min
Wash 2
2 mL 30% Methanol at 1 mL/min
Dry
> 10 Hg for 5 min to remove residual water
Elution 1
2 mL 1.5% Ammonium hydroxide methanol solution at 1 mL/min
Elution 2
2 mL 1.5% Ammonium hydroxide methanol solution at 1 mL/min
Evaporation
Evaporate to dryness
Reconstitute
500 µL 500uL ACN/20mM Ammonium bicarbonate (10:90)
Order Items Used in This Application
- 1. Nornicotine
- 2. 3-OH-Cotinine
- 3. Anabasine
- 4. Cotinine
- 5. Nicotine
-OEChem-06211003142D<br><br> 23 24 0 1 0 0 0 0 0999 V2000<br> 2.5896 1.3184 0.0000 N 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.2647 -1.7694 0.0000 N 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.3987 0.7306 0.0000 C 0 0 3 0 0 0 0 0 0 0 0 0<br> 4.2077 1.3184 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.8987 2.2694 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.8987 2.2694 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.3987 -0.2694 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.5326 -0.7694 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.2647 -0.7694 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.5326 -1.7694 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.3987 -2.2694 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.9511 0.4491 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.5177 0.7814 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.7741 1.5705 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.5051 2.3983 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.8339 2.8860 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.9635 2.8860 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.2922 2.3983 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.0000 1.1268 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 1.9957 -0.4594 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.8016 -0.4594 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 1.9957 -2.0794 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.3987 -2.8894 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 1 3 1 0 0 0 0<br> 1 6 1 0 0 0 0<br> 1 19 1 0 0 0 0<br> 2 9 1 0 0 0 0<br> 2 11 2 0 0 0 0<br> 3 4 1 0 0 0 0<br> 3 7 1 0 0 0 0<br> 3 12 1 0 0 0 0<br> 4 5 1 0 0 0 0<br> 4 13 1 0 0 0 0<br> 4 14 1 0 0 0 0<br> 5 6 1 0 0 0 0<br> 5 15 1 0 0 0 0<br> 5 16 1 0 0 0 0<br> 6 17 1 0 0 0 0<br> 6 18 1 0 0 0 0<br> 7 8 1 0 0 0 0<br> 7 9 2 0 0 0 0<br> 8 10 2 0 0 0 0<br> 8 20 1 0 0 0 0<br> 9 21 1 0 0 0 0<br> 10 11 1 0 0 0 0<br> 10 22 1 0 0 0 0<br> 11 23 1 0 0 0 0<br>M END
Compound Name
Nornicotine
CID
412
Molecular Formula
C9H12N2
Molecular Weight
148
No. Hydrogen Bond Acceptors
2
No. Hydrogen Bond Donors
1
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