Testosterone from Female Human Plasma by LC/MS/MS using Strata-X-A and Kinetex 1.7 µm C18
LC Conditions
Column
Kinetex 1.7 µm C18 100 Å, LC Column 30 x 2.1 mm, Ea
Brand
Kinetex
Part No
00A-4475-AN
Phase Name
C18
Elution Type
Gradient
Mobile Phase
A: 0.1% Formic Acid +1 mM Amm Formate in Water
B: 0.1% Formic Acid +1 mM Amm Formate in ACN
Gradient Profile
Step No.
Time(min)
%A
%B
1
0
90
10
2
2.5
10
90
3
3.5
10
90
4
3.6
90
10
Flow Rate
0.40 mL/min
Temperature
0°C
Detection
Mass Spectrometer (MS) @ (22°C)
Detection Info
Sample Preparation Details
Description
Strata-X-A 33 µm Polymeric Strong Anion, 30 mg / 3 mL, Tubes, 50/Pk
Part No
8B-S123-TBJ
Pretreatment Note
Following evaporation of elution solvent @ 50-55 C under gentle nitrogen stream; Add 50 uL 25% hydroxylamine solution and heat at 60-65 C for 5-10 min, then add 200 uL 5% aqueous formic acid and vortex the tubes Transfer the solution to autosampler vials and inject 25 uL on column
Condition 1
2 mL Methanol
Condition 2
2 mL Water
Load
Load 1.25 mL Sample diluted in Water
Wash 1
0.6 mL 50% Methanol in Water
Dry
Dry for 5 min under high vacuum
Elution 1
0.3 mL Methanol
Elution 2
0.3 mL Methanol
Evaporation
Evaporate to dryness
Reconstitute
250 µL 200 uL 5% aqueous formic acid
Order Items Used in This Application
- 1. Testosterone
- 2. Testosterone-d3
-OEChem-06211003152D<br><br> 49 52 0 1 0 0 0 0 0999 V2000<br> 8.6500 2.1128 0.0000 O 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.0000 -2.1948 0.0000 O 0 0 0 0 0 0 0 0 0 0 0 0<br> 7.3931 -0.1425 0.0000 C 0 0 2 0 0 0 0 0 0 0 0 0<br> 7.3931 0.8575 0.0000 C 0 0 2 0 0 0 0 0 0 0 0 0<br> 6.5271 -0.6425 0.0000 C 0 0 1 0 0 0 0 0 0 0 0 0<br> 5.6610 -0.1425 0.0000 C 0 0 1 0 0 0 0 0 0 0 0 0<br> 4.7510 -0.6493 0.0000 C 0 0 2 0 0 0 0 0 0 0 0 0<br> 6.5271 1.3575 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.6610 0.8575 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 8.3393 1.1623 0.0000 C 0 0 2 0 0 0 0 0 0 0 0 0<br> 8.3393 -0.4472 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.5431 -1.6840 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 8.9229 0.3575 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.7430 -1.6909 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.6451 -2.2117 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 7.3931 1.8575 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.8242 -0.0852 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.7587 0.3507 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.8763 -0.6132 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.8076 -2.2406 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.8680 -1.6981 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 7.4830 -0.9877 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 7.2664 -1.0618 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.3972 0.2825 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.9256 1.8325 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.1285 1.8325 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.4490 1.4401 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.0504 0.7499 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 7.9018 1.6016 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 8.0883 -1.0141 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 8.8767 -0.7564 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.7612 -2.2644 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 7.1523 -1.5685 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 9.3838 -0.0572 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 9.3838 0.7723 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.2478 -2.6877 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.0460 -2.6846 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 8.0131 1.8575 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 7.3931 2.4775 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.7731 1.8575 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.2324 0.3814 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.4343 0.3968 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.3787 0.3459 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.7635 0.9706 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.1388 0.3554 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.6718 -0.0279 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.2647 -0.7146 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.8100 -2.8606 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 9.2566 2.2406 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 10 1 1 1 0 0 0<br> 1 49 1 0 0 0 0<br> 2 21 2 0 0 0 0<br> 3 4 1 0 0 0 0<br> 3 5 1 0 0 0 0<br> 3 11 1 0 0 0 0<br> 3 22 1 6 0 0 0<br> 4 8 1 0 0 0 0<br> 4 10 1 0 0 0 0<br> 4 16 1 1 0 0 0<br> 5 6 1 0 0 0 0<br> 5 12 1 0 0 0 0<br> 5 23 1 1 0 0 0<br> 6 7 1 0 0 0 0<br> 6 9 1 0 0 0 0<br> 6 24 1 6 0 0 0<br> 7 14 1 0 0 0 0<br> 7 17 1 0 0 0 0<br> 7 18 1 1 0 0 0<br> 8 9 1 0 0 0 0<br> 8 25 1 0 0 0 0<br> 8 26 1 0 0 0 0<br> 9 27 1 0 0 0 0<br> 9 28 1 0 0 0 0<br> 10 13 1 0 0 0 0<br> 10 29 1 0 0 0 0<br> 11 13 1 0 0 0 0<br> 11 30 1 0 0 0 0<br> 11 31 1 0 0 0 0<br> 12 15 1 0 0 0 0<br> 12 32 1 0 0 0 0<br> 12 33 1 0 0 0 0<br> 13 34 1 0 0 0 0<br> 13 35 1 0 0 0 0<br> 14 15 1 0 0 0 0<br> 14 20 2 0 0 0 0<br> 15 36 1 0 0 0 0<br> 15 37 1 0 0 0 0<br> 16 38 1 0 0 0 0<br> 16 39 1 0 0 0 0<br> 16 40 1 0 0 0 0<br> 17 19 1 0 0 0 0<br> 17 41 1 0 0 0 0<br> 17 42 1 0 0 0 0<br> 18 43 1 0 0 0 0<br> 18 44 1 0 0 0 0<br> 18 45 1 0 0 0 0<br> 19 21 1 0 0 0 0<br> 19 46 1 0 0 0 0<br> 19 47 1 0 0 0 0<br> 20 21 1 0 0 0 0<br> 20 48 1 0 0 0 0<br>M END
Compound Name
Testosterone
CID
6013
Molecular Formula
C19H28O2
Molecular Weight
288
No. Hydrogen Bond Acceptors
2
No. Hydrogen Bond Donors
1
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