| 1UDCA |
|---|
| 2UDCA-D4 |
| 3GCDCA |
| 4GCDCA-D4 |
| 5CA |
| 6CA-D4 |
| 7GDCA |
| 8GDCA-D4 |
| 9TDCA |
| 10TDCA-D4 |
| 11CDCA |
| 12CDCA-D4 |
| 13DCA |
| 14DCA-D4 |
| 15LCA |
| 16LCA-D4 |
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Applications
Extraction of unconjugated Bile acids from Human Serum on Kinetex 2.6µm Polar C18 100x2.1 Column
24195
Separation Mode: Reversed Phase
Reversed Phase
Polar C18
HPLC
Gastrointestinal Agents
Clinical
Clinical Research
Therapeutic / Clinical
Extraction of unconjugated Bile acids from Human Serum on Kinetex 2.6µm Polar C18 100x2.1 Column
Analytes
Details
LC Conditions (App ID: 24195)
Column: |
Brand Name: Kinetex |
Part No: |
Phase Name: Polar C18 |
Sample Note: Sample Prep Protocol
Dispense: 400 uL methanol into the wells of the Impact plate
Add: 100 uL of doubly stripped Serum sample (spiked with analytes at 200ng/mL) directly into the organic solvent in each well of the plate.
Vortex: 2 minutes at maximum possible speed.
Wait: Allow 5 minutes for completion of protein precipitation.
Vacuum: Place the Impact plate onto a suitable 96-well SPE manifold followed by positioning a 96-well collection plate inside, under the Impact plate. Vacuum at 5" of Hg until filtrate is collected completely.
Dilute & inject: Dispense 300 uL of mobile phase A (or water) into the collection plate, vortex for 30 secs at a high speed and inject on LC-MS-MS
Note: A doubly stripped serum sample was employed for extraction purposes to eliminate the potential bias due to presence of any endogenous bile acids, leading to erroneous analyte quantitation.
Table 1. % Absolute Recovery for Bile acids from Human Serum Extraction on an Impact Protein Precipitation Plate
Analyte % Recovery % CV (N=5)
UDCA 91% 1.1
GCDCA 90% 3.7
CA 88% 4.8
GDCA 90% 4.4
TDCA 94% 3.5
CDCA 90% 4.5
DCA 88 4.6
LCA 90% 6.9 |
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Phenomenex is a technology leader committed to developing novel analytical chemistry solutions that solve the separation and purification challenges of researchers worldwide