Applications
25-OH Vitamin D2 and D3 Control from Serum on Kinetex 2.6µm C18 after PPT
19314
Separation Mode: Reversed Phase
Reversed Phase
C18
HPLC
Vitamins
Clinical
Therapeutic / Clinical
Therapeutic / Clinical

Applications

25-OH Vitamin D2 and D3 Control from Serum on Kinetex 2.6µm C18 after PPT
Analytes
1
2
3
Compound Name
Calcifediol
Details
LC Conditions (App ID: 19314)
Column:
Brand Name:
Part No:
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
20.08928
33.40100
44.80100
56928
Flow Rate:
1 mL/min
Temperature:
35°C
Detection:
Mass Spectrometer (MS) amu @ 22°C
Sample Preparation Details
Description:
Strata-X 33 µm Polymeric Reversed Phase, 30 mg / 3 mL, Tubes, 50/Pk
Part Number:
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)

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