Development of Analytical and Preparatory Techniques - Chlorophyll Biosynthesis and Technological Applications - page 111

6. Since the sample contained a total amount of 86.2 pmol/ml of MV + DV Chl(ide)
a, the concentration of MV Chl(ide) a amounted to (86.24/100) 90.29 ¼ 77.87

pmol/ml. That of DV Chl(ide) a amounted to 86.2 77.87 ¼ 8.37 pmol/ml.

3.7.5 Calculation of the Net Fluorescence Amplitudes
at 475 and 498 nm of MV and DV Chlorophyll(ide)
b Respectively in Mixtures of the Two Compounds

Calculation of the net Soret excitation amplitudes of MV and DV Chl(ide) b in
mixtures of these two tetrapyrroles was also achieved by adaptations of Eqs. ( 3.37 )
and ( 3.38 ).
As was mentioned earlier the optimum wavelengths for this analysis were
determined to be in the Soret region, at 475 and 498 nm for MV and DV Chl(ide)
b, respectively. Adaptation of Eqs. ( 3.37 ) and ( 3.38 ) was achieved via the following
assignments:
X

deconvoluted net Soret excitation amplitude of MV Chl(ide) b (E475 F660)
at 475 nm;
Y

¼

deconvoluted net Soret excitation amplitude of DV Chl(ide) b
(E498 F666) at 498 nm;
( E a F b )

¼

¼ (E475 F660) ¼ Soret excitation amplitude of the MV + DV mixture

at 475 nm,
when the excitation spectrum is recorded at an emission wavelength of 660 nm;
( E C F D ) Soret excitation amplitude of the MV + DV mix-

ture at 498 nm, when the excitation spectrum is recorded at an emission wavelength
of 666 nm.
By substitution of the above values for X, Y, ( E a F b ) , and ( E c F d ) in Eqs. ( 3.37 )
and ( 3.38 ) the latter transform into:

¼ ( E498 F666 ) ¼

DV Chl ð ide Þ a ð E498 F666 Þ¼ ð E475 F660 Þð E475 F660 Þ= ½ k 6 ð 1 = K 3 Þ

(3.69)

MV Chl ð ide Þ b ð E475 F660 Þ¼ ð ½ E475 F660 Þð E498 F666 Þ= k 8 ð 1 = K 4 Þ

(3.70)

where K 3 and K 4 are as defined by Eq. ( 3.39 ) and Where:
k 5 ¼ DV Chl ð ide Þ b ð E475 F660 Þ= DV Chl ð ide Þ b ð E498 F666 Þ

k 6 ¼ MV Chl ð ide Þ b ð E475 F660 Þ= MV Chl ð ide Þ b ð E498 F666 Þ

(3.71)

k 7 ¼ MV Chl ð ide Þ b ð E498 F666 Þ= MV Chl ð ide Þ b ð E475 F660 Þ

k 8 ¼ DV Chl ð ide Þ b ð E498 F666 Þ= DV Chl ð ide Þ b ð E475 F660 Þ