Development of Analytical and Preparatory Techniques - Chlorophyll Biosynthesis and Technological Applications - page 101

Table 3.7 Determination of the reliability of equations 3.57 and 3.58 used to calculate the
excitation amplitude of MV Pchl(ide) and DV Pchl(ide) at 437 and 451 nm, respectively

Percentage error
between amount of
Pchlide added and
calcd (%)

Amount of Pchlide
added (pmol/ml)

Amount of Pchlide
(pmol/ml) Mean percentage error SD

MV
Pchlide

DV
Pchlide

MV
Pchlide

DV
Pchlide

MV
Pchlide

DV
Pchlide MV Pchlide DV Pchlide

4 16 4.2 15.8 5 1.25 2.24% 0.87 6.9%

3.97%

10 10 9.6 10.4 4 4

12 8 11.1 8.9 7.5 11.25

14 6 14.1 5.9 0.7 1.6

16 10

18 2 18 2 0 0
Note: The excitation spectra were recorded at an emission of maximum of 625 nm. Other
experimental details are as in materials and methods

4 16.4 3.6 2.5

3.6.4 Calculation of the Amounts of Monovinyl and Divinyl
Protochlorophyll(ides) in the Presence of DV
Mg-Protos in a Mixture of the Three Tetrapyrroles

This situation can be resolved by appropriate adaptation of Eqs. ( 3.48 )] and ( 3.49 ).
In this case there is no reason to change the wavelength assignment of ( E a F b ) and
( E c F d ) which was adopted in Eqs. ( 3.53 ) and ( 3.54 ) for MV Pchl(ides) and DV Pchl
(ides), respectively. As may be recalled, in these equations, X(E a F b ) referred to the
deconvoluted net Soret excitation amplitude of the pentacoordinated MV Pchl(ides)
By (0-0) (E437F625) transition at 437 nm and Y(E c F d ) referred to that of the DV
Pchl(ides) Bx (0-0) (E451F625) transition at 451 nm. Therefore, in this context,
( E a F b ) refers to (E437F625) and represents the Soret excitation amplitude at
437 nm, when the excitation spectrum of the tetrapyrrole mixture containing
MV + DV Pchl(ides) and DV Mpe is recorded at an emission wavelength of
625 nm. Likewise ( E c F d ) represents ( E 451 F 625) and refers in the same F 625-nm
excitation spectrum to the Soret excitation amplitude of the mixture at 451 nm. On
the other hand the ( Eg Fh ) terms in Eqs. ( 3.48 ), ( 3.49 ), ( 3.50 ), ( 3.51 ) and ( 3.52 )
refer to ( E 424 F 625) and represent the Soret excitation amplitude at 424 nm in the
F 625-nm excitation spectrum (Table 3.7 ).
With the aforementioned assignments, Eqs. ( 3.48 ) and ( 3.49 ) transform into:

MV Pchl ð ides Þð E437F625 Þ¼½ð E437F625 Þ Q 1 ð E451F625 Þ

Q 2 ð E424F625 Þ= Q 3 (3.59)

DV Pchl ð ides Þð E451F625 Þ¼½ð E451F625 Þ Q 4 ð E437F625 Þ

Q 5 ð E424 F625 Þ= Q 6 (3.60)