Biology Reference
In-Depth Information
Table 3.12 Reliability of Eqs. (
3.76
) and (
3.77
) in the determination of the amount of 2-MV Pch
(lide)
b
in the presence of 2-MV Pchl(ide)
a
Amount of 2-MV Pchl
(ide) added error
SD
Amount of 2-MVPchl
(ide)
b
calculated
(pmol/ml)
Percentage error between
amounts added and
calculated (%)
(pmol/ml)
Mean percentage
error
SD (%
%)
Pchl(ide)
a
Pchl(ide)
b
131.0
121.9
131.0
7.5
1.0
6.8
131.0
61.0
56.4
7.5
170.3
122.0
118.0
3.3
131.0
36.6
31.8
13.1
103.0
78.4
77.3
1.4
103.6
52.3
56.9
8.8
103.6
104.5
99.6
4.7
133.2
117.6
122.7
4.3
281.2
156.8
154.1
1.7
296.0
182.9
185.7
1.5
2-MV Pchl(ide)
a
/2-MV Pchl(ide)
b
molar ratio which was calculated as described
above. The reliability of calculating the amount of 2-MV Pchl(ide)
b
in the presence
of 2-MV Pchl(ide)
a
amounted to 1.0
6.8 % (Table
3.12
).
3.8.1.4 Sample Calculation of the Amount of 2-MV Pchl(ide) b
in a Mixture of 2-MV Pchl(ide) b and 2-MV Pchl(ide) a
1. An ether solution containing 199.0 pmol of 2-MV Pchl(ide)
a
and 402.4 pmol of
Pchl(ide)
b
per ml was prepared.
2. Two fluorescence emission spectra elicited by excitation at 435 and 463 nm were
recorded on the ether solution at 77
K.
3. The net fluorescence emission amplitudes at 635 and 643 nm respectively, for
the 2-MV Pchl(ide)
a
and 2-MV Pchl(ide)
b
components were calculated using
Eqs. (
3.76
) and (
3.77
). The apparent (E440 F635)/(E43 F643) fluorescence ratio
amounted to 0.0731/0.3397
0.2152.
4. The authentic molar ratio, of 2-MV Pchl(ide)
a/
2-MV Pchl(ide)
b
was calculated
from the apparent (E440 F635)/(E43 F643) fluorescence ratio with Eq. (
3.78
).
It amounted to 0.4829.
5. The calculated amount of 2MV-Pchl(ide)
b
in the mixture (412.0 pmol/ml) was
determined from the relation:
¼
2-MV Pchl
ð
ide
Þ
a
=
2-MV Pchl
ð
ide
Þ
b
¼
0
:
4829
where 2-MV Pchl(ide)
a
, as determined by 293
K fluorescence emission, amounted
to 402.4 pmol/ml.
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