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

Sample Preparation Details

Condition 1 :1 mL Acetonitrile
Condition 2 :1 mL Acetonitrile/Water (65:35)
Load :Load 1.60 mL Sample diluted in Acetonitrile/Water (50:50)
Wash 1 :0.5 mL Acetonitrile/Water (65:35) at 1 mL/min
Wash 2 :0.5 mL Acetonitrile/Water (65:35) at 1 mL/min
Elution 1 :0.5 mL Acetonitrile at 1 mL/min
Elution 2 :0.5 mL Acetonitrile at 1 mL/min
Evaporate: :Evaporate to dryness
Reconstitute :100 µL ethyl acetate/acetonitrile (5:95)
Notes :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

Application Detail (App ID: 19313)

Part No:00B-4462-E0
Dimensions:50 x 4.6 mm ID
Elution Type:Gradient
Elution A:0.05 % Formic acid
Elution B:5 mM Ammonium acetate + 0.1 % Formic acid in methanol
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 92 8
2 5 92 8
3 205 0 100
4 290 0 100
5 360 92 8

Flow Rate:1 mL/min
Col. Temp: 35 °C
Detection: Mass Spectrometer (MS) @ amu (450°C)
Detector Info: AB Sciex API 4000™
Note:SecurityGuard™ ULTRA Guard Cartridge System extends column lifetime.
- SecurityGuard ULTRA Cartridges, UHPLC C18 for 4.6mm ID Columns, 3/Pk Part No.: AJ0-8768
- Holder Part No.: AJ0-9000

Analyte Details

Analyte Detail not available

1
25-Hydroxy vitamin D3 (25-OH D3)
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2
25-Hydroxy vitamin D3-d6 (25-OH D3-d6)

3
25-Hydroxy vitamin D2 (25-OH D2)
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