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
none cited
Clinical
Clinical Research
Therapeutic / Clinical

Applications

Extraction of unconjugated Bile acids from Human Serum on Kinetex 2.6µm Polar C18 100x2.1 Column

Analytes

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Compound Name
Actigall
CID
465690
Molecular Formula
C24H40O4
Molecular Weight
392
No. Chiral Centers:
1
No. Hydrogen Bond Acceptors
4
No. Hydrogen Bond Donors
3

Conditions

LC Conditions (App ID: 24195)
Column:Kinetex 2.6 µm Polar C18 100 Å, LC Column 100 x 2.1 mm, Ea
Brand Name:Kinetex
Part No:00D-4759-AN
Elution Type:Gradient
Mobile Phase:A: 2mM Ammonium acetate (pH 6.9)
B: Methanol/Acetonitrile (50-50)
Gradient Profile:
Step No.Time(min)%A%B
105545
293070
39.53070
49.515545
5125545
Flow Rate:400 mL/min
Temperature:50°C
Detection:Mass Spectrometer (MS) amu @ 50°C

Sample Preparation Details
Description:Impact Protein Precipitation, 2mL Square Well Filter Plate, 2/Pk
Part Number:CE0-7565
Pre Treatment Note:Sample Prep Protocol Dispense: 400 uL methanol into the wells of the Impact plate Add: 100 ?L 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.
SPE Load:Load Sample
SPE Evaporation:Evaporate to dryness
SPE Reconstitute:0 µL initial mobile phase

Order Items Used in This Application

Similar Applications
No Data