Stewart Postharvest Review

An international journal for reviews in postharvest biology and technology

Degradation of chlorophyll during processing of green vegetables: a review

Shalini Gaur,1* US Shivhare2 and Jasim Ahmed3

1
Department of Biotechnology and Environmental Sciences, Thapar Institute of Engineering and Technology, Patiala,
Punjab, India.
2
Department of Chemical Engineering and Technology, Panjab University, Chandigarh, India.
3
Department of Food Science and Agricultural Chemistry, Macdonald Campus of McGill University, Ste-Anne-de-Bellevue,
Montreal, Canada.

Abstract
Purpose of review: Retention of greenness, which is attributed by the pigment chlorophyll, is desired by consumers and processors of
thermally processed green vegetables. However, the retention of chlorophyll during the cooking/processing of green vegetables has
been recognised as a problem. To overcome this problem, researchers have explored various techniques including alkaline salt solu-
tions and high-temperature short-time treatments. The kinetics of chlorophyll degradation have also been studied with respect to water
activity, pH, temperature, pressure, etc. This review is an effort to put together the findings of various researchers on chlorophyll degra-
dation.
Recent findings: The traditional approach to model only the retained chlorophyll has been criticised. With newer and more precise
techniques to detect and quantify the degradative intermediates and products of chlorophyll, attempts are being made to model further
degradation of pheophytins to pheophorbides. Newer information is also being generated to decipher the mechanism of chlorophyll
degradation in senescent tissues and during processing, and multi-response models are being introduced to model the chlorophyll de-
gradative reactions in foods.
Directions for future research: More precise modelling approaches need to be introduced to model chlorophyll degradation in view
of newer analytical instrumentation and processing methods becoming available, and considering the role of associated enzymes and
other process parameters.

Keywords: Chlorophyll; green vegetables; colour; reaction kinetics; activation energy

Abbreviations Introduction
FDA Food and Drug Administration The appearance of food is one of the main parameters that
defines its quality and corresponds to the things we perceive
HTST High-temperature Short-time by the sense of sight, mainly the colour [1]. According to
psychologists, we capture about the 83% of the surround-
*Correspondence to: Shalini Gaur, Department of Biotech- ings with sight and only 17% with the rest of our senses [2].
nology and Environmental Sciences, Thapar Institute of Since colour is a major sensory attribute in the determina-
Engineering and Technology, Patiala 147004, Punjab, India. tion of product acceptability, it is of critical importance for
email: gaurshalini@gmail.com the food industry to minimise the loss of green colour,
which most vegetables contain. Chlorophyll, the pigment
Stewart Postharvest Review 2006, 5:14 responsible for green colour is degraded during thermal
Published online 01 October 2006 processing.
doi: 10.2212/spr.2006.5.14
Degree of greenness is important in determining the final
quality of thermally-processed vegetables. During thermal
processing, the natural cellular structures disintegrate result-
ing in amenability of the pigment to various enzymatic and
non-enzymatic reactions, which finally leads to brownish
degradation products [3]. Gupte and Francis [4] reported
© 2006 Stewart Postharvest Solutions (UK) Ltd.
Gaur et al. / Stewart Postharvest Review 2006, 5:14
Figure 1. Reactions of chlorophyll.
Chlorophyll
Heat, acid Mg2+ Phytol Chlorophyllase
Mg2+, phytol
Pheophytins Methyl chlorophyllide
Heat, acid Heat, acid
Pheophorbide
that the colour changes (from bright green to olive brown)
occurring during processing can be attributed to the conver-
sion of chlorophyll a and b to their corresponding pheo-
phytins, which is ultimately perceived by the consumer as a
loss of quality. Minimising chlorophyll degradation during
the processing of vegetables is still an industrial challenge.
Chlorophyll: structure and reactivity
Chlorophyll a and b are typically found in higher plants, occur-
ring in an approximate ratio of 3:1. Both chlorophyll a and b are
magnesium-containing derivatives of the tetrapyrrole phorbin
[5]. Chlorophyll a has a methyl group at the C-3 carbon atom,
while a formyl group is bonded to the same carbon atom in chlo-
rophyll b. Chlorophyll a appears blue-green while chlorophyll b
appears yellow-green [8]. In addition to structural differences
between chlorophyll a and b, their thermal stabilities are also
different. Chlorophyll a was reported to be thermally less stable
than chlorophyll b [6–7]. The higher thermal stability of chloro-
phyll b has been attributed to the electron-withdrawing effect of
its C-3 formyl group [5].
In green cellular tissues subjected to various processing condi-
tions, chlorophyll undergoes distinct types of degradation reac-
tions. The changes in the chlorophyll molecule during heating
have been discussed by Jaques et al [3]. De-metalation and
epimerisation were observed during heat treatment, and pro-
longed heating leads to additional de-methoxycarbonylation of
the molecule. De-methoxycarbonylation also occurs during the
canning of vegetables. De-phytylation of chlorophyll is achieved
enzymatically during fermentation and storage and is often
observed together with de-metalation.
In food products, chlorophyll degradation studies revealed that
chlorophyll degrades to pheophorbide via pheophytins or
chlorophyllide [6] (Figure 1). Schwartz and Lorenzo [8] demon-
strated that chlorophyll degradation terminates at pheophorbide.
However, studies by Matile et al. [9] and Ginsburg et al. [10]
have shown that chlorophyll degradation continues beyond
pheophorbide to colourless compounds. Heaton et al. [11] ob-
served that chlorophyll degradation of cabbage heads in cold
storage did not lead to pheophorbide accumulation, leaving the
degradation of pheophorbide to colourless by-products as the
only explanation. The accepted mechanism of chlorophyll deg-
radation in fruits and vegetables and strategies for the control of
chlorophyll degradation in food products have been reviewed by
Heaton and Marangoni [12].
Various researchers have modelled the whole pathway of chlo-
rophyll degradation to pheophytin and subsequently from pheo-
phytins to pheophorbide. Weemaes et al. [13] studied the com-
bined effects of pressure and temperature on changes in broccoli
juice greenness. The authors found that there was further degra-
dation of pheophytins when a higher temperature (70oC) was
applied in combination with elevated pressure. The loss of
greenness under these conditions was modelled by two consecu-
tive first-order degradation steps. The temperature dependence
of the rate constants for both degradation steps could be de-
scribed by the Arrhenius equation, with activation energies in-
creasing with pressure. In another study, Weemaes et al. [14]
determined the kinetic parameters for the two-step chlorophyll
degradation. They concluded that the kinetic parameters for the
first degradation step were in accordance with the kinetic pa-
rameters for pheophytinisation of the total chlorophyll content,
with an activation energy of 69 kJ/mol. The second degradation
step for subsequent degradation of pheophytins was character-
ised by an activation energy of 105.49 kJ/mol.
Kinetics of chlorophyll degradation
Kinetic modelling has created interest in food processing, as it
provides the possibility of controlling changes in food quality
during processing and storage. Most of the kinetic research in
food science is limited to microbial and enzymatic kinetics;
however, it could be applied to other quality changes with proc-
ess time and storage. At pre-defined conditions, the rate of a
reaction and the temperature dependence of a food system can
be controlled and predicted in advance. This approach is known
as kinetic modelling. One of the limitations in kinetic modelling
is selecting the appropriate model for a reaction (or a series of
reactions). In food science, most researchers have opted for a
simple model, either a first-order or zero-order model, to fit their
experimental data. Most of the reported data on chlorophyll or
colour degradation were fit using simple first-order or fraction-
conversion kinetic models (equations 1–4).
Some conventional models to describe
chlorophyll degradation
The kinetic parameters, namely rate constant and activation
energy, provide useful information on thermal degradation of
chlorophyll. The first-order reaction is represented by the
following equation:
ln(C C o ) = −kt …(1)
where, C is measured pigment concentration or tristimulus
colour value at time t; Co denotes measured pigment concen-
tration or colour value(s) at zero time; k is the rate constant
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Gaur et al. / Stewart Postharvest Review 2006, 5:14
Figure 2. Kinetic model for chlorophyll degradation using multi-response
approach.
Chlorophyll
k3
k1
Chlorophyllide
Pheophytin
k4
k2
Pheophorbide
during heating (1/min) and t is heating time (min). The Ar-
rhenius equation to relate the dependence of the rate constant
with temperature is represented as:
k = Ao exp(− E a RT ) …(2)
where Ao is the frequency factor (1/min), Ea is the activation
energy (kJ/mol), R is the universal gas constant
[8.314 J/ (mol.K)] and T is absolute temperature (K).
Fractional conversion is a convenient variable often used in
place of concentration and, for irreversible first-order reac-
tion kinetics, the rate constant at constant temperature can be
determined through fractional conversion, f:
f = (C o − C ) (C o − C ∞ ) …(3)
where C∞ is the concentration at infinite time signifying that
the reaction is completed.
Fractional conversion technique using first-order reaction
kinetics changes to the following form:
ln(1 − f ) = ln (C − C ∞ ) (C o − C ∞ ) = −kt …(4)
where –Co is chlorophyll content at zero time, to; C is the
chlorophyll content at any time t and C∞ is chlorophyll con-
tent at infinite time such that all the chlorophyll has been
converted to pheophytins.
Kinetic modelling: multi-response approach to
chlorophyll degradation
Reactions in foods are complex in nature, sometimes consist-
ing of a series (cascade) of reactions. Therefore, multi-
response modelling can be useful for describing such a cas-
cade. van Boekel [15] has described chlorophyll degradation
by multi-response modelling (Figure 2). It implies simply
that more than one response is taken into account, where the
measured responses have one or more parameter in common.
Uni-response modelling consists of measuring only one re-
sponse (eg, chlorophyll concentration) or that of only one
product (eg, chlorophillide). However, it is possible to meas-
ure the concentration of more than one reactant/product at the
same time using multi-response modelling. Multi-response
modelling has advantages over uni-response modelling due to
the fact that more precise estimates of the parameters can be
obtained and the proposed kinetic model can be tested more
rigorously.
Appropriate modelling of a complex reaction network takes
into account identification of the main reactants and products,
identification of primary and secondary reaction routes, stud-
ies of the effect of temperature, effect of reactant concentra-
tion, and environmental conditions such as pH, ionic strength
and specific ions.
The general approach is to predict a mechanism for the reac-
tion network, and the mechanism should then be translated
into a mathematical model. This can be done by setting up
differential equations, one for each component involved. In
the case of only two or three reaction steps, it may still be
possible to find an analytical solution for the differential
equations, but in most cases one will have to resign to nu-
merical integration. Nowadays this is possible with the help
of computers and softwares. The parameters in the model are
the kinetic parameters (rate constants, activation enthalpy/
energy and entropy) and they need to be estimated by fitting
the model to the experimental data. Van Boekel [15] has de-
scribed the handling of chlorophyll degradation in pickles
and coleslaw using a multi-response approach.
Schwartz and von Elbe [16], in a study of chlorophyll degra-
dation in processed spinach puree, suggested an equimolar
stoichiometric first-order relationship for the degradation of
pheophytins to pyropheophytins as a function of time
(equation 5).
dy
= k1 x − k 2 y …(5)
dt
where x is chlorophyll concentration, y is pheophytin concen-
tration, and k1 and k2 are the degradation rate constants for
chlorophylls and pheophytins, respectively. The authors con-
cluded that pheophytin degradation follows the first-order
reaction kinetics. In vitro studies on chlorophyllase, the en-
zyme responsible for conversion of chlorophyll to chloro-
phyllide and pheophytins to pheophorbide, have also been
performed [17].
Chlorophyll degradation has been shown to follow different
pathways depending on the commodity [11]. However, the
mechanisms and kinetics of those reactions have only been
partially characterised. Heaton and Marangoni [12] summa-
rised the accepted mechanism of chlorophyll degradation in
fruits and vegetables and extended it to include the degrada-
tion of pheophorbides into colourless compounds. Takamiya
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Table 1. Chlorophyll degradation kinetic parameters of selected green to study the rate of chlorophyll degradation under various
environmental conditions. Tijskens [20] stated that the ulti-
Product Activation energy Reference
(kcal/mol)
mate goal of modelling is to predict the future behaviour in
any circumstance while generating more knowledge about
Spinach [55] the process under study. Another article in this issue covers
143 tristimulus colour measurement techniques and colour kinet-
Chlorophyll a 35 ics and, therefore, this review has focused on only the kinet-
Chlorophyll b
ics of chlorophyll degradation.
Chlorophyll a aw 0.75 19 [56]
Chlorophyll a aw 0.62 21 Thermal degradation of chlorophylls (both a and b) in broc-
Chlorophyll b aw 0.75 20 coli juice was studied spectrophotometrically by Weemaes et
Chlorophyll b aw 0.62 20
al. [13]. Degradation of chlorophyll occurred at temperatures
Peas 18.08 [57] above 60oC. Chlorophyll degradation followed first-
order kinetics and chlorophyll a was found to be more
Asparagus 15.20 heat sensitive than chlorophyll b. Temperature dependencies
Spinach 25.2 [16] of rate constants were described by the Arrhenius
Chlorophyll a 22.5 equation with activation energies of 71.04 ± 4.89 kJ/mol and
Chlorophyll b 67.11 ± 6.82 kJ/mol for chlorophyll a and b, respectively.
20.7
Pheophytin a 15.7
Pheophytin b Ahmed et al. [21] studied the effect of particle size on chlo-
rophyll content and total colour of green chili puree. Both
Spinach [41] chlorophyll content and green colour varied significantly
with processing temperature and mesh size. Chlorophyll con-
Chlorophyll a 15.0 ± 0.9
Chlorophyll b 22.0 ± 0.3 tent increased with increased mesh fineness, but decreased
linearly with processing temperature. Up to a 60% loss in
Chlorophyllide a 19.7 ± 0.1 chlorophyll was noted after heating at 100oC for 15 min.
Chlorophyllide b 22.8 ± 0 .4

Broccoli juice [13]
Chlorophyll a 16.99 ± 1.17
Chlorophyll b 16.06 ± 1.63
Total chlorophyll
16.52 ± 1.44
et al. [18], in a study on mechanism of chlorophyll degrada-
tion using gene cloning, determined that after successful re-
moval of phytol and Mg2+ from the chlorophyll molecule by
the use of chlorophyllase and magnesium dechetalase, pheo-
phorbide is cleaved and reduced to yield a colourless, open
tetrapyrrole intermediate.
Several researchers have studied the rate and extent of chlo-
rophyll degradation and conversion [13]. The use of a col-
ourimeter (tristimulus colour L, a and b values) to measure
the colour of green vegetables has been advocated by various
investigators [13, 19] based on its superiority over conven-
tional spectrophotometric methods of chlorophyll measure-
ment. Analytical methods of solvent (acetone and ether) ex-
tracted chlorophyll determination are found to be time con-
suming compared with rapid colour measurement techniques.
Colour measurement studies, though useful in measuring the
ultimate criteria of appearance of foods, are not helpful in
deducing the mechanism of chlorophyll degradation in green
tissues during thermal processing. Therefore, there is a need
Schwartz and Lorenzo [22], in a study on continuous aseptic
processing and storage, concluded that oxidation during stor-
age was not a dominant factor in chlorophyll degradation and
colour loss. A major problem in thermal processing of green
vegetables is the continuous decrease in pH values due to
acid formation. To avoid this problem, Ryan-Stoneham and
Tong [23] conducted chlorophyll degradation studies in a
specially designed reactor with an online pH control
capability. The authors concluded that the activation energy
was independent of pH, with a magnitude varying between
17 and 17.5 kcal/mol for both chlorophyll a and b. The deg-
radation rate was found to decrease log-linearly as the pH
increased. The rate constant and activation energy of chloro-
phyll degradation computed by various researchers is given
in Table 1.
Effect of pH on chlorophyll degradation
A number of researchers [4, 24] have demonstrated higher
stability of chlorophylls at alkaline pH conditions, and nu-
merous patents have also been granted for addition of alka-
line compounds to conventionally heat-processed green vege-
tables. Gupte and Francis [4] reported that the use of magne-
sium carbonate in combination with high-temperature short-
time (HTST) processing (at 150oC) initially improved chloro-
phyll retention in pureed spinach. However, the effect was
not stable during storage. Magnesium compounds resulted in
the formation of hard white crystals of magnesium-
ammonium-phosphate [25]. Ahmed et al. [19] studied the
colour degradation kinetics of coriander leaf puree however,
they worked on puree at unmodified pH.
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Gaur et al. / Stewart Postharvest Review 2006, 5:14
The extent of chlorophyll degradation depends on the thermal
severity of the process. The possibility of reducing the sever-
ity of the process from a chemical point of view, while main-
taining a microbiologically equivalent heat intensity, has
been investigated [6] and was accomplished by increasing the
temperature and reducing the processing time [4]. It is well
known that acidic foods (pH ≤4.5) do not require processing
above 100oC for their preservation, as heat-resistant food
spoilage microorganisms are unable to thrive at acidic pH. It
is, therefore, relevant to study the effect of combining pH
with a less drastic thermal process, in order to obtain maxi-
mum retention of chlorophyll in thermally-processed food.
The effect of pH (3–8) and microbial growth on green colour
degradation of blanched broccoli during storage was investi-
gated by Gunawan and Barringer [26]. Colour degradation
accelerated when broccoli was submerged in Mcllvaine’s
buffer at decreasing pH. Pheophytinisation followed first-
order reaction kinetics. The logarithmic values of reaction
rate constants were linearly correlated to the ambient pH up
to pH 7. Acids containing a benzene ring were found to cause
a faster colour change than acids with simple chains due to
their hydrophobicity.
Role of enzymes in chlorophyll degradation
Enzymatic degradation of chlorophyll in senescent plant tis-
sues has been extensively studied. Recent reviews [27, 28]
describe the role of chlorophyllase, magnesium dechetalase
and pheophorbide a oxygenase in chlorophyll degradation.
Among these enzymes, chlorophyllase (chlorophyll chloro-
phyllido-hydrolase, EC 3.1.1.14) catalyses the first step in
chlorophyll catabolism and removes the phytol chain from
the porphyrin ring to form chlorophyllide.
Lipoxygenase is widely distributed in vegetables and is in-
volved in off-flavour development and colour loss. The latter
is due to hydroperoxide and radical formation by oxidation of
lipids, which can destroy chlorophyll and carotenoids during
frozen storage [29, 30]. Lopez-Ayerra et al. [31] concluded
that lipid oxidation was related to chlorophyll degradation in
spinach. Peroxidases are assumed to play an important part in
chlorophyll degradation, a process accompanying ripening
and senescence in most fruits and vegetables. Peroxidase is
responsible for off-flavour development during storage of
canned products, especially in non-acidic vegetables which
often exhibit high levels of activity. They impair not only the
sensory properties and, hence, the marketability of a product,
but its nutritive value as well [29]. Martinez et al. [32] re-
ported that chlorophyll degrading peroxidase activity in
strawberry and canola seeds decreased with chlorophyll con-
tent in their ripening stage, while the reverse occurred in
mustard.
Kato and Shimizu [33] reported that a phenolic peroxidase
H2O2 system was involved in in vitro bleaching of chloro-
phyll. The phenolics active in the system were derivatives of
monophenols having an electron-attracting group in the para
position. In another study on chlorophyll degradation in to-
bacco leaves, Kato and Shimizu [34] suggested that not only
solubilised chlorophyll but also membrane (thylakoid) and
protein (light harvesting chlorophyll- protein complex) bound
chlorophyll were bleached by a phenolic-peroxidase–H2O2
system. The authors concluded that deterioration of the sub-
cellular environment induced by senescence accelerates chlo-
rophyll bleaching by a peroxidase-catalysed reaction. In a
recent study, Yamamuchi et al. [35] concluded that in peroxi-
dase mediated chlorophyll degradation, peroxidase oxidises
the phenolic compounds with hydrogen peroxide and forms
the phenoxy radical. The phenoxy radical then oxidises chlo-
rophyll to colourless low molecular weight compounds
through the formation of C-13-hydroxychlorophyll-a and a
bilirubin-like compound as an intermediate.
Chlorophyll complexation with zinc salts
One approach for retention of green colour in canned green
vegetables is incorporation of a chemical additive to stabilise
chlorophyll. These additives may be classified into two
groups: (1) alkalising or alkalising-buffer agents such as hy-
droxides and carbonates [4] and (2) metallic salts such as the
chlorides and acetates of zinc and copper [36]. The drawback
in using alkalising agents is that the intensity of the green
colour decreases gradually when the canned green vegetable
is stored at room temperature [4]. It is well known that addi-
tion of alkaline salts leads to textural loss due to dissolution
of cellulose in cell walls and vitamin loss. In contrast, replac-
ing the magnesium in the chlorophyll molecule with a zinc
ion results in a stable green compound [37].
Metals such as zinc and copper have long been known to be
introduced into the chlorophyll-porphyrin ring. Formation of
green metallocomplexes of chlorophyll derivatives during
thermal processing is known as “regreening” and is consid-
ered a promising method to preserve the colour of canned
green vegetables [37]. The spectral curves of zinc pheo-
phytins a and b have been published and the similarity be-
tween these curves and those for the chlorophylls a and b,
respectively, have been indicated [36].
Humphery [38] concluded that the metal chlorophyll deriva-
tives form bonds that are more resistant to acid and heat than
the naturally occurring magnesium chlorophyll complexes.
The metallo-chlorophyll complexes have been characterised
to some extent. It has been found that the formation of copper
complexes occurred more readily than zinc complexes [37].
Copper salts were shown to be 30 times more reactive than
zinc salts in heated spinach puree [36]. Bobbio and Guedes
[39] compared the stability of copper chlorophyll and magne-
sium chlorophyll against light, O2, temperature (40 and
60oC), Na2S2O5 (sodium metabisulphite), benzoyl peroxide
and acid. The use of copper chlorophyll was found to be
much more stable than magnesium chlorophyll. Use of cop-
per chlorophyll complexes are however, not permitted in the
USA [40]. The differential stability of the zinc complexes
and the chlorophylls in acidified solutions provided the basis
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for spectrophotometric estimation of zinc pheophytins,
chlorophylls, and pheophytins in mixtures in diethyl ether or
80% acetone [36].
Applications and patents
Metallo-chlorophyll complexes have a potential use in the
food industry for preserving the desirable fresh green colour
of vegetables. Colour stabilisation can be implemented in the
traditional retorting of chlorophyll-containing vegetables. A
patent has been issued for the preservation of the green col-
our of canned vegetables [42], and further investigations us-
ing alkaline conditions and HTST processing have been pat-
ented [43]. The Veri-green process is a patented procedure in
which blanching of green vegetables is performed in the pres-
ence of zinc salts [3]. Leake and Kirk [44] have also patented
a process that includes blanching of vegetables in an aqueous
ZnCl2 (zinc chloride) solution followed by thermal treat-
ments. Another patent leading to colour improvement of
green vegetables through the aqueous packing solution con-
taining zinc or copper ions has been obtained by La Borde
and von Elbe [45]. Donato et al. [46] patented a process for
improving the colour and texture of vegetables by blanching
in aqueous solution containing zinc ions at a concentration of
500 ppm. The United States Food Drug Administration
(FDA) has responded by limiting the amount of added zinc in
canned vegetables to 75 ppm and has given temporary per-
mits to a number of processors to test market canned green
beans [40].
Reactivity of chlorophylls with zinc
Schanderl et al. [47] achieved a ratio of 1:1 between com-
plexed metal and chlorophyll in a pure solution. LaBorde and
von Elbe [48] proposed that metal ions reacted only with
chlorophyll derivatives and not with native chlorophyll.
Pheophytin a, pyropheophytin a and pheophorbide a react
readily with zinc, however the b form of chlorophyll deriva-
tives have been known to be less reactive [37, 40]. Tonucci
and von Elbe [49] further reported that pheophytin a did not
react to form a complex as rapidly as pyropheophytin a did.
La Borde and von Elbe [48] stated that zinc-pyropheophytin
a could be formed via two reactions, either from further reac-
tion of zinc-pheophytin a to form zinc-pyropheophytin or
through zinc complexation with pyropheophytin a. However,
Canjura et al. [41] concluded that, of the two pathways, con-
version to zinc-pyropheophytin a from the metallo-
pheophytin derivative would be the more favoured reaction.
Canjura et al. [41] blanched peas in a zinc solution of
50–500 mg/L concentration followed by processing in a par-
ticle cell reactor, simulating the continuous flow aseptic proc-
essing system. They reported that the complex formation
increased during heat processing and was dependent on zinc
absorbed by peas during blanching. Processing between
121 and 125oC promoted formation of zinc-pheophytin a
while treatment at 130–145oC led to formation of zinc-
pyropheophytin a.
Factors involved in zinc-pheophytinisation
Studies have indicated that the formation of complexes de-
pend on zinc concentration, pigment concentration and pH
[40]. In addition, other constituents such as proteins, phytic
acid, etc, capable of binding zinc, also affect complex forma-
tion. According to Lin and Schyvens [50], blanching
temperature and zinc concentration were the main factors
affecting the colour of sterilised green bean, whereas the
interaction between blanching temperature and blanching
time was a secondary factor. The optimum blanching
condition to improve the colour of sterilised endive was
found to be 50oC in a blanching solution containing 200 ppm
zinc ions.
Canjura et al. [41], in a study on zinc absorption into pea
tissue, concluded that the complex formation was dependent
on the concentration of available zinc ion, duration of contact
between the zinc solution and pea tissue, and temperature.
The authors suggested the use of metal salts coupled with
continuous flow aseptic processing technology for preserva-
tion of the green colour in vegetables.
Metallo-chlorophyll complex formation in heated vegetables
has been shown to be pH dependent [51]. Jones et al. [36]
reported a nearly 40% increase in zinc complex formation
during heating of fresh spinach in zinc chloride solution be-
tween pH 6.8 and 8.5. However, the rate of copper complex
formation was unaffected between pH 3.8 and 8.5. The
amount of zinc complex formed increased 11-fold as the pH
of spinach puree increased from pH 4.0–8.5 but decreased at
pH 10.0 [40]. A decrease in the reaction rate at higher pH
values was attributed to the lower solubility of zinc chloride
at higher pH and the effective removal of zinc ions from the
reaction medium. LaBorde and von Elbe [40] further sug-
gested that the formation of zinc complexes during heat treat-
ment would be most rapid in vegetables containing the great-
est amount of chlorophyll and processed with the greatest
amounts of zinc salts. Also, given the FDA’s limit of 75 ppm
of zinc ion in canned vegetables, the success of this process
may be a function of the species and quality of the fresh
vegetable. Since high pH values also decrease the formation
of pheophytin from chlorophylls, optimum regreening can be
achieved by processing and storage in buffered process solu-
tions at a pH that allows the reactions to occur at the maxi-
mum rate with minimal salt precipitation.
Tonucci and von Elbe [49] monitored the reaction of chloro-
phyll a derivatives with zinc ions by high pressure liquid
chromatography and studied the kinetics of metal-complex
formation. They stated that the true order of the complex
formation reaction was second order; the reaction was first
order with respect to concentration of the chlorophyll a de-
rivative and also first order with respect to the concentration
of zinc ion. Steric hindrance from peripheral groups and
charge distribution on the tetrapyrrole ring were found to
have an effect on the rate of reaction of chlorophyll deriva-
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Gaur et al. / Stewart Postharvest Review 2006, 5:14
tives with zinc ions. The displacement mechanism of type
SN2 has been indicated for incorporation of the metal ions
into porphyrins.
Influence of light on chlorophyll degradation
Foods are available in various packaging materials and glass
is commonly used for beverages, edible oils, dressings and
milk products. During sale, these packages are displayed on
shelves and are exposed to light which may, depending on
the sensitivity of the respective product, impair food quality.
In addition to oxidative deterioration of lipids, vitamins and
proteins there are losses in natural pigments and colour of
packaged food products. The sensitivity of many foods to
light is due to their pigment content, which is stimulated by
quantas of the visible spectral region, acting as photo-
sensitisers. The energy is transferred directly from the photo-
sensitisers to the food components or oxygen. The latter is
converted into the reactive singlet form and induces oxida-
tion [52].
While the effects of light on foods have been studied in depth
[53], the effects of optical filters in packaging materials, spe-
cifically with regard to the effects of wavelengths, are still
unclear. Thron et al. [54] studied the influence of light on a
model system (sunflower oil spiked with chlorophylls) based
on the wavelengths of the visible spectral range. The sensitiv-
ity to oxidation was determined by the oxygen consumption
during exposure time. The resulting effects of oxidation, such
as the degradation of the chlorophyll derivatives and the for-
mation of pentane as a secondary oxidation product, were
found to be at a maximum after exposure. The greatest quan-
tum-dependent sensitivity was detected at around 650 nm and
the lowest at around 500 nm. It was shown that the quanta
absorbed by the absorption maximum of chlorophyll at
650 nm are most effective on photo-oxidation. This study
suggested that while selecting packaging for chlorophyll-
containing foods, the critical spectral ranges around 400 nm
(yellow-green) and 650 nm (blue-green) need special consid-
eration. Foods mainly irradiated by fluorescent light need
more protection in the lower wavelength range, and foods
irradiated by natural daylight in the longer wavelength.
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