Quantitation of 5-Methyltetrahydrofolate in Cerebrospinal Fluid Using Liquid Chromatography-Electrospray Tandem Mass Spectrometry



Fig. 1
HPLC -ESI-MS/MS ion chromatogram of 5-MTHF 100 nM standard. [5-MTHF 1 (m/z 460.3 > 313.3), 5-MTHF 2 (m/z 460.3 > 194.5), and 5-MTHF-13C5 (m/z 465.3 > 313.3)]



A327622_1_En_19_Fig2_HTML.gif


Fig. 2
HPLC -ESI-MS/MS ion chromatogram of 5-MTHF abnormal QC (30 nM). [5-MTHF 1 (m/z 460.3 > 313.3), 5-MTHF 2 (m/z 460.3 > 194.5), and 5-MTHF-13C5 (m/z 465.3 > 313.3)]



 






3.2 Data Analysis




1.

Instrumental operating parameters are given in Table 1.


Table 1
HPLC -ESI-MS/MS operating conditions































































(a) HPLC (5-MTHF)a

Column temp
40 °C
  

Flow rate
0.375 mL/min
  

Gradient
Time (min)
Mobile phase A (%)
 
0
100
 
1.5
0
 
2
0
 
2.1
100

(b) MS/MS tune settingsb

Entrance potential (V)
10

Curtain gas (psi)
20

CAD gas
Medium

Ion spray (V)
5500

Temp (°C)
500

GS 1 (psi)
50

GS 2 (psi)
50

Resolution Q1 and Q3
Unit


aOptimized for Shimadzu prominence liquid chromatography system equipped with Phenomenex Synergi-Hydro, 4 μm, 150 × 3 mm analytical column; Mobile phase A: 0.1 % formic acid in water; Mobile phase B: 0.1 % formic acid in methanol

bOptimized for ABSciex 4000QTRAP®. Tune settings may vary slightly between instruments

 

2.

Data are analyzed using Analyst software version 1.6.2 (AB Sciex).

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Oct 21, 2016 | Posted by in BIOCHEMISTRY | Comments Off on Quantitation of 5-Methyltetrahydrofolate in Cerebrospinal Fluid Using Liquid Chromatography-Electrospray Tandem Mass Spectrometry

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