Development of Analytical and Preparatory Techniques - Chlorophyll Biosynthesis and Technological Applications - page 70

k 0 1 ¼ Pchlide a ð E400 F633 Þ= Pchlide a ð E440 F640 Þ

k 0 2 ¼

ð Þ= ð Þ

Proto E400 F633 Proto E440 F640

k 0 3 ¼

ð Þ= ð Þ

Copro E400 F633 Copro E440 F640

(3.16)

k 0 4 ¼

Pchlide a ð E400 F622 Þ= Pchlide a ð E440 F640 Þ

k 0 5 ¼ Proto ð E400 F622 Þ= Proto ð E440 F640 Þ
k 0 6 ¼

Copro ð E400 F622 Þ= Copro ð E440 F640 Þ

fluorescence emis-
sion amplitude of Pchlide at 640 nm which is elicited by a 440-nm excitation of the
hexane-extracted acetone fraction. Proto (E440 F640)

According to the above terminology:Pchlide (E440 F640) ¼

fluorescence emission
amplitude of Proto at 640 nm which is elicited by a 440-nm excitation of the
hexane-extracted acetone fraction; etc. By solving Eqs. ( 3.13 ), ( 3.14 ) and ( 3.15 )
for Pchlide (E440, F640) the following equation is obtained:

¼

C 0 1 C 0 2 ð C 0 3

(3.17)

Pchlide ð E440 F640 Þ¼½ð E440 F640 Þ ð E400 F633 Þ E400 F622 Þ=

Where (E440 F640) is calculated from Eqs. ( 3.11 )or( 3.12 ) and were

C 0 1 ¼ k 0 2 K 0 2 : K 4 = K 0 3 1 =

C 0 2 ¼ K 0 2 = K 0 3 (3.18)

C 0 3 ¼ K 0 1 þ K 0 2 : K 0 5 = K 0 3

and

K 0 1 ¼ k 0 1 = k 0 2 1

K 0 2 ¼ k 0 3 = k 0 2 1

K 0 3 ¼ k 0 6 k 0 5 : k 0 3 = k 0 2 (3.19)

K 0 4 ¼ k 0 5 = k 0 2

K 0 5 ¼ k 0 5 : k 0 1 = k 0 2 k 0 4

The values of the k 0 constants were calculated according to Eq. ( 3.16 ). The
fluorescence amplitudes used in Eq. ( 3.16 ) were derived from the emission spectra
of standard Pchlide, Proto and Copro dissolved separately in acetone/H 2 O/
0.1NNH4O(9:2:1, v/v) that was previously extracted with hexane (Rebeiz
et al. 1975a ). The emission spectra of the standards were elicited by two successive
excitations at 440 and 400 nm. The k 0 values used for the calculations of the K 0 and
C constants were the mean of at least five determinations performed on five
different concentrations of standard Pchlide, Proto, and Copro. The K 0 constants
were calculated from Eq. ( 3.19 ) and the C 0 constants from Eq. ( 3.18 ). Under our
instrumental conditions the values obtained were 0.03 for C 1 , 0.0 for C 2 0 , and 0.99
for C 3 0 .