Applications
25-OH Vitamin D2 and D3 Standards from Serum on Kinetex 2.6µm C18 after PPT
19313
Separation Mode: Reversed Phase
Reversed Phase
C18
HPLC
Vitamins
Clinical
Therapeutic / Clinical
Therapeutic / Clinical

Applications

25-OH Vitamin D2 and D3 Standards from Serum on Kinetex 2.6µm C18 after PPT

Analytes

1
2
3
Compound Name
Calcifediol
CID
5283731
Molecular Formula
C27H44O2
Molecular Weight
400
No. Hydrogen Bond Acceptors
2
No. Hydrogen Bond Donors
2

Conditions

LC Conditions (App ID: 19313)
Column:Kinetex 2.6 µm C18 100 Å, LC Column 50 x 4.6 mm, Ea
Brand Name:Kinetex
Part No:00B-4462-E0
Elution Type:Gradient
Mobile Phase:A: 0.05 % Formic acid
B: 5 mM Ammonium acetate + 0.1 % Formic acid in methanol
Gradient Profile:
Step No.Time(min)%A%B
10928
25928
32050100
42900100
5360928
Flow Rate:1 mL/min
Temperature:35°C
Detection:Mass Spectrometer (MS) amu @ 450°C

Sample Preparation Details
Description:Strata-X 33 µm Polymeric Reversed Phase, 30 mg / 3 mL, Tubes, 50/Pk
Part Number:8B-S100-TBJ
Pre Treatment Note:Note: Avoid over drying eluates to avoid loss of analytes and/or internal standard, which will impact both precision and accuracy of data Reconstitute with 100 µL ethyl acetate/acetonitrile (5:95), and vortex mix for 5 seconds Centrifuge for 10 min at 2000g to settle any precipitate Transfer clear liquid to amber-colored autosampler vial and cap
SPE Condition 1:1 mL Acetonitrile
SPE Condition 2:1 mL Acetonitrile/Water (65:35)
SPE Load:Load 1.60 mL Sample diluted in Acetonitrile/Water (50:50)
SPE Wash 1:0.5 mL Acetonitrile/Water (65:35) at 1 mL/min
SPE Wash 2:0.5 mL Acetonitrile/Water (65:35) at 1 mL/min
SPE Elution 1:0.5 mL Acetonitrile at 1 mL/min
SPE Elution 2:0.5 mL Acetonitrile at 1 mL/min
SPE Evaporation:Evaporate to dryness
SPE Reconstitute:100 µL ethyl acetate/acetonitrile (5:95)

Order Items Used in This Application

Similar Applications
Kinetex 2.6 µm C18 100 Å, LC Column 50 x 4.6 mm, Ea
Reversed Phase