ABBOTTNUTRITIONDIVISION
TITLE
PAGENO.
Determination of TransVitaminK
1
byHPLC andFluorescenceDetection
11 of 14
C
I
=W x 40,000=0.05527x 40,000=2210.8µg/L
3) Concentration of Intermediate II Standard
C
II
=W x 8,000=0.05527 x 8,000=442.16µg/L
4) Concentration ofWorkingStandards
C
1
=Wx 240=0.05527x 240=13.26µg/L
C
2
=Wx 480=0.05527x 480=26.53µg/L
C
3
=Wx 800=0.05527x 800=44.26µg/L
b) Measurement of PeakArea
All peak areaswere determinedwith an integratorwhichmeasured the relative intensities of each
peak.
c) Calculation of TransVitaminK
1
StandardConcentration.
Assume the followingdatawere collected
Concentration
PeakArea (beginning)
PeakArea (end)
Total K
1
(µg/L)
(cis)
(trans)
(cis)
(trans)
44.22
71419
602905
73296
635102
26.53
43414
369164
55876
384373
13.26
21716
185545
26581
190061
1) Calculation of TransVitaminK
1
Fraction in theStandard
For all of the standards in the table, the average trans vitaminK
1
fraction is
T
a
=0.8885
2) TransVitaminK
1
StandardConcentration
C
1t
=C
1
xT
a
=13.26x 0.8885=11.78
g/L
C
2t
=C
2
xT
a
=26.53x 0.8885=23.57
g/L
C
3t
=C
3
xT
a
=44.22x 0.8885=39.29
g/L
d) Preparation of theStandardCurve
The trans vitaminK
1
peak areas at the beginning and the end of the analysiswere averaged.
TransK
1
Concentration (µg/L)
Average peak area
39.29
619004
23.57
376769
11.78
187803
Regression of concentration onpeak area yields
T=
A
2
A
1
+A
2
=
602905
71419+602905
=0.8941
VitK-02
FORWORKINGGROUP/ERPUSEONLY
DONOTDISTRIBUTE
