133 / 363
© 2015 AOAC INTERNATIONAL
F. Calculations
(
a
) The amount of ΔDi-0S in μg/g, representing unsulfated CS in
the sample, is calculated as follows:
where P
0
= peak area of ΔDi-0S in sample chromatogram; b
0
=
y-intercept of calibration curve for disaccharide ΔDi-0S; m
0
= slope
of calibration curve for disaccharide ΔDi-0S; V = volume of Test
Solution 1 = 100 mL; W = sample weight, in g; and D = dilution
factor = 50.
(
b
) The amount of ΔDi-4S in μg/g, representing CSA in the
sample, is calculated as follows:
where P
4
= peak area of ΔDi-4S in sample chromatogram; b
4
=
y-intercept of calibration curve for disaccharide ΔDi-4S; m
4
= slope
of calibration curve for disaccharide ΔDi-4S; V = volume of Test
Solution 1 = 100 mL; W = sample weight, in g; and D = dilution
factor = 50.
(
c
) The amount of ΔDi-6S in μg/g, representing CSC in the
sample, is calculated as follows:
where P
6
= peak area of ΔDi-6S in sample chromatogram; b
6
=
y-intercept of calibration curve for disaccharide ΔDi-6S; m
6
= slope
of calibration curve for disaccharide ΔDi-6S; V = volume of Test
Solution 1 = 100 mL; W = sample weight, in g; and D = dilution
factor = 50.
(
d
) The amount of ΔDi-di(2,6)S in μg/g in the sample is
calculated as follows:
where P
2,6
= peak area of ΔDi-di(2,6)S in sample chromatogram;
b
6
= y-intercept of calibration curve for disaccharide ΔDi-6S; m
6
=
slope of calibration curve for disaccharide ΔDi-6S; V = volume of
Test Solution 1 = 100 mL; W = sample weight, in g; D = dilution
factor = 50; and F = molecular weight conversion between ΔDi-6S
and ΔDi-di(2,6)S = 1.190.
(
e
) The amount of ΔDi-di(4,6)S in μg/g in the sample is
calculated as follows:
where P
2,6
= peak area of ΔDi-di(4,6)S in sample chromatogram;
b
6
= y-intercept of calibration curve for disaccharide ΔDi-6S; m
6
=
slope of calibration curve for disaccharide ΔDi-6S; V = volume of
Test Solution 1 = 100 mL; W = sample weight, in g; D = dilution
factor = 50; and F = molecular weight conversion between ΔDi-6S
and ΔDi-di(4,6)S = 1.190.
(
f
) The amount of ΔDi-tri(2,4,6)S in μg/g in the sample is
calculated as follows:
Figure 2015.11A. Stock standard solution chromatogram.
Peak assignments and approximate retention times: (
1
)
Δ
Di-
0S (2.5 min), (
2
)
Δ
Di-6S (5.9 min), (
3
)
Δ
Di-4S (6.3). Not shown:
Δ
Di-di(2,4)S (9.6 min),
Δ
Di-di(2.6)S (9.9 min), and
Δ
Di-tri(2,4,6)
S (12.0 min).
Figure 2015.11B. CS rawmaterial from bovine trachea
sample chromatogram. Peak assignments: (
1
)
Δ
Di-0S,
(
2
)
Δ
Di-6S, and (
3
)
Δ
Di-4S.
Figure 2015.11C. CS rawmaterial from shark cartilage
sample chromatogram. Peak assignments: (
1
)
Δ
Di-0S,
(
2
)
Δ
Di-6S, (
3
)
Δ
Di-4S, (
4
)
Δ
di-di(2,6)S, and (
5
)
Δ
di-di(4,6)S.
Candidates for 2016 Method of the Year
132