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Collaborative Study Report: Method 2011.20 Nucleotides by HPLC-UV
Page 27
internal standard quantifier
566.2
298.0
21
18
120
internal standard qualifier
566.2
280.0
37
18
80
a
Analyte = vitamin D
3
-PTAD adduct, Internal standard ion =
d6
-vitamin D3-PTAD adduct
b
DP = declustering potential
c
EP = entrance potential
d
CE = collision energy
e
CXP = collision cell exit potential
J. Calculations
(a) Concentration of d6-vitamin D
2
in Vitamin D
2
Stable Isotope Labelled Stock Standard Solution:
10000
x
1%
1cm
max)
abs(λ
2
D2conc
2
E
SS SILD
SS SILD
where: SILD
2
SS
D2conc
= concentration of SIL
d6
-vitamin D
2
in stock standard (units: µg/mL)
SILD
2
SS
abs(λ max)
= UV absorbance of stock standard at 265 nm (units: none)
= extinction coefficient for vitamin D
2
in ethanol [461] (units: dL
/g.cm)
10000 = concentration conversion factor (units: g/dL to µg/mL
(b) Concentration of
d6
-vitamin D
3
in Vitamin D
3
Stable Isotope Labelled Stock Standard Solution:
10000
x
1%
1cm
max)
abs(λ
3
D2conc
3
E
SS SILD
SS SILD
where: SILD
3
SS
D3conc
= concentration of SIL
d6
-vitamin D
3
in stock standard (units: µg/mL)
SILD
3
SS
abs(λ max)
= UV absorbance of stock standard at 265 nm (units: none)
= extinction coefficient for vitamin D
3
in ethanol [485] (units: dL/
g.cm)
10000 = concentration conversion factor (units: g/dL to µg/mL
(c) Concentration of
d6
-vitamin D
2
in Internal Standard:
1000
10
1.0
x
SS SILD
SILIS
D2conc
2
D2conc
where: SILIS
D2conc
= concentration of SIL
d6
-vitamin D
2
in internal standard (units: ng/mL)
SILD
2
SS
D3conc
= concentration of SIL
d6
-vitamin D
2
in stock standard (units: µg/mL)
1000 = concentration conversion factor (units: µg/mL to ng/mL)
(d) Concentration of
d6
-vitamin D
3
in Internal Standard:
1000
10
1.0
x
SS SILD
SILIS
D3conc
3
D3conc
where: SILIS
D3conc
= concentration of SIL
d6
-vitamin D
3
in internal standard (units: ng/mL)
SILD
3
SS
D3conc
= concentration of SIL
d6
-vitamin D
3
in stock standard (units: µg/mL)
1000 = concentration conversion factor (units: µg/mL to ng/mL)
(e) Concentration of non-labelled vitamin D
2
in Vitamin D
2
Non-Labelled Purity Standard Solution:
10000
x
1%
1cm
max)
abs(λ
2
D2conc
2
E
PS NLD
PS NLD
where: NLD
2
PS
D2conc
= concentration of vitamin D
2
in purity standard (units: µg/mL)
NLD
2
PS
abs(λ max)
= UV absorbance of purity standard at 265 nm (units: none)
= extinction coefficient for vitamin D
2
in ethanol [461] (units: dL/
g.cm)
10000 = concentration conversion factor (units: g/dL to µg/mL)
(f)
Concentration of non-labelled vitamin D
3
in Vitamin D
3
Non-Labelled Purity Standard Solution:
10000
x
1%
1cm
max)
abs(λ
3
D3conc
3
E
PS NLD
PS NLD
where: NLD
3
PS
D3conc
= concentration of vitamin D
3
in purity standard (units: µg/mL)
NLD
3
PS
abs(λ max)
= UV absorbance of purity standard at 265 nm (units: none)
= extinction coefficient for vitamin D
3
in ethanol [485] (units: dL/
g.cm)
10000 = concentration conversion factor (units: g/dL to µg/mL)
(g) Concentration of non-labelled vitamin D
2
in Non-Labelled Working Standard Solution:
1%
1cm
E
1%
1cm
E
1%
1cm
E
1%
1cm
E
2016.05 (FEBRUARY 2017) VITD-18
MLT REPORT
FOR ERP USE ONLY
DO NOT DISTRIBUTE