AOAC SPIFAN ERP Meeting Book-March 16, 2016

2011.06 (Fol-22) w/SLV FOR ERP USE ONLY DO NOT DISTRIBUTE

Silliker Chem, Res. Center Crete, IL – Report of A Validation of LC-MS/MS Method for Folate Analysis

Table 14: Evaluation of Response Linearity against concentration of six target folate compounds

FA

5-CH3-THF

5-CHO-THF

Std Level

Conc. (ng/mL)

Cal. Conc (ng/mL)

Cal. Conc (ng/mL)

Cal. Conc (ng/mL)

Accuracy

Accuracy

Accuracy

Std D Std C Std B Std A

2.5 5.0

2.04 5.89 101 499

81.6% 117.8% 101.0% 99.8%

2.69 4.75 97.0 503

107.6% 95.0% 97.0% 100.6%

2.50 4.85 104 496

100.0% 97.0% 104.0% 99.2%

100 500

79.2 357

-1.18E+03 1.50E+03

-7.85E+01 1.21E+03

Intercept

Slope

0.9998

0.9999

0.9998

R^2

THF

10-CH3-FA

10-CHO-FA

Std Level

Conc. (ng/mL)

Cal. Conc (ng/mL)

Cal. Conc (ng/mL)

Cal. Conc (ng/mL)

Accuracy

Accuracy

Accuracy

Std D Std C Std B Std A

2.5 5.0

2.41 4.87 108 493

96.4% 97.4% 108.0% 98.6%

2.75 4.30 105 496

110.0% 86.0% 105.0% 99.2%

2.55 4.86 101 499

102.0% 97.2% 101.0% 99.8%

100 500

-5.69E+02

-213

72.4

Intercept

597

6.19E+02

1.22E+03

Slope

0.9994

0.9997

0.9999

R^2

Injection Solvent = 1% acetic acid /methanol/1% ascorbic acid (v/v/w 80:20:1) Calibration Curve using Dionex 300 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.

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