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Extraction of Mono Hydroxy Vitamin D2 and D3 from Phree on Kinetex C18 2.6u 30x3 mm

Sample Preparation Details

Part Number :8E-S133-TGB
Description :Phree Phospholipid Removal, 30 mg / well, 96-Well Plates, 2/Pk
Load :Load Sample
Evaporate: :Evaporate to dryness
Reconstitute :0 µL initial mobile phase

Application Detail (App ID: 21868)

Part No:00A-4462-Y0
Dimensions:30 x 3 mm ID
Elution Type:Gradient
Elution A:0.1% formic acid/water
Elution B:0.1% Formic acid/methanol
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 40 60
2 0.5 5 95
3 2 5 95
4 2.01 40 60
5 3.5 40 60

Flow Rate:0.6 mL/min
Col. Temp: ambient
Detection: Mass Spectrometer (MS) @ amu (22°C)
Detector Info: SCIEX API 5000™
Note:Instrument: API 5000 (AB SCIEX)
Ionization Source: APCI, Positive Polarity
Ionization Source Parameters
Curtain Gas: 25.0 psi
Gas 1 (Neb Gas): 40.00 psi
ihe: ON
CAD (Collision Gas): 10.0
NC: 5.00 µA
TEM: 360 °C
Note: Gas 2 is not available in APCI source
MRM Transitions and Mass Dependent Parameters:
Compound Q1, Da Q3, Da Dwell, msec DP, V EP, V CE, V CXP, V
Vit D2 (1) 395.3 209.3 45 69 10 25 10
Vit D2 (2) 395.3 269.2 45 69 10 25 10
IS (D3-3H2) 386.2 257.2 45 69 10 25 10
Vit D3 (1) 383.2 257.2 45 69 10 23 10
Vit D3 (2) 383.2 211.1 45 69 10 34 10


Analyte Details

Analyte Detail not available

1
25 Hydroxy vitamin D3-2H3
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2
25 Hydroxy vitamin D3
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3
25 Hydroxy vitamin D2
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