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Silliker
Chem, Res. Center Crete, IL – Report of A Validation of LC-MS/MS Method for Folate Analys
11
Calibration Curve using Dionex 3000 ultimate HPLC and API 5500 MS (Qtrap) detector:
The linearity evaluation of calibration were performed with standards prepared in the elution solvent,
i.e., 10:90:1(v,v,w) :: formic acid : methanol : ascorbic acid and neutralized with ammonium hydroxide.
The calibration curves were extended to 7pts. Concentrations and consisted of: 1, 2, 5, 20, 50, 200 and
500 ng/mL.
Figure 2
:
Calibration Curve of different folate compounds (Dionex 3000 Ultmate HPLC and
5500 API MS detector). The color scheme for the calibration curve of different folate
compounds is as the following
.
Folate (295) ______,
5-CH3-THF (313)_______,
5-CHO-THF
(327)_______,
THF (299) _____,
10-CHO-FA (295)_________,
10-CH3-FA (309) ______.
CH=THF (282) _______. Conc
Calibration Values:
The calibration values based on the above calibration curve for different folate compounds are listed in
the following Table 5.
Table 5: Evaluation of Response Linearity against concentration of six target folate compounds
the values of linear regression
Folate
Compound
Transition
Slope
Intercept
r2
weighing
Calibration
Line Color
Folic Acid
295
0.02807 +0.00269 0.9996
1/x
Navy Blue
5-CH3-THF
313
0.02453 +0.01463 0.9931
1/x
Purple
5-CHO-THF
327
0.02746 +0.01105 0.9922
1/x
Red
THF
299
0.01553 -0.01530
0.9992
1/x
Orange
CH=CH-THF
282
0.1050 +0.00211 0.9965
1/x
Sky Blue
10-CH3-FA
309
0.03783 -0.01387
0.9979
1/x
Light Green
10-CHO-FA
295
0.04435 +0.00650 0.9996
1/x
Dark Green
Folate Compound
Concentration (ppb)
2011.06/Fol-22 (February 2016)
FOR ERP USE ONLY
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