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16CONTENT OF CAROTINOIDS IN THE LEAVES OF SOME SPICES
ZARUHI VARDANYAN Doctor of Biological Sciences, Professor, Chair of Chemistry and Biology Vanadzor State University named after Hovhannes Tumanyan, Vanadzor, Armenia
ANUSH GEVORGYAN
PhD in Biology, Associate Professor, Chair of Chemistry and Biology Vanadzor State University named after Hovhannes Tumanyan, Vanadzor, Armenia
GOHAR AVETISYAN
Master's student, Specialization: Biology, Vanadzor State University named after Hovhannes Tumanyan, Vanadzor, Armenia
Annotation- The article presents the results of research the carotene content in the leaves of certain spicy plants: Dill (Anethum graveolens L.), Basil (Ocimum basilicum L. .), Coriander (Coriandrum sativum L. .), Parsley (Petroselinum crispum Fuss.). The quantity of carotenoids in parsley and dill leaves is equal and exceeds the quantity of carotenoids in coriander leaves by 1.325 times and in basil leaves by 3.759 times. The quantity of carotenoids in dill and parsley leaves is closer to the quantity of chlorophylls. Stemming from our research, based on the quantitative predominance of carotenoids, the population should be recommended to use dill and parsley as spices, as immune system stimulants and antioxidants.
Key words - optical density, leaf extract, wet weight, dry weight, pigment volume concentration
Introduction
With the emergence of pigments, including carotenoids in plant tissues, the history of the origin of organic nature has faced a greatest aromorphosis, the ultimate result of chemical and subsequently, biological evolution as well. Carotenoids are yellow, orange, or red pigments synthesized in the cells of bacteria, fungi, and higher plants. They represent the most widespread group of plant pigments. Their number exceeds 600. The most physiologically active components of that large group of chloroplast pigments in the leaves of green plants are a- and P-carotenes, as well as their oxygen-containing derivatives, xanthophylls, equally called carotenoids. As additional pigments, they absorb the energy of light and transfer it to chlorophyll a. The synthesis of carotenoids in the plant tissue is carried out more actively in the dark and is suppressed under the light. [15]
According to the findings of a number of studies, carotenoids carry out a certain role in the flowering period of plants; their quantity is relatively high in the stigma and pollen grains. During this period, the quantity of carotenoids partially decreases in the leaves and increases in stamens and petals. [10]
Carotenoids carry a grave importance in terms of maintaining the water balance of a human body. Our bodies obtain vitamin A as a result of carotenoids transformation. The daily requirement of carotenoids is less than 6 mg. [16]
Studies have shown that carotenoids contribute to the prevention of the development of a number of diseases in the human body. Carotenoids have anti-carcinogenic, anti-toxic, immunomodulatory effects and their use as immunomodulators in the combined treatment of malignant tumors is clinically recommended. [12]
Research carried out by Frederick Khachik has demonstrated that the addition of carotenoids in food is justified from the point of view of disease prevention. [2]
According to in vitro and in vivo laboratory studies presented by A. Bendich, beta-carotene can protect phagocytic cells from auto-oxidative effects, activate the proliferation of T and B lymphocytes, stimulate the tumoricidal capacities of effector T cells, macrophages, cytotoxic T cells,
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and natural killer cells, as well as increase the synthesis of certain interleukins. [1] Carotenoids also affect the regulation of cells' cholesterol metabolism. [11]
Carotenoids have antioxidant activity and affect cell membranes. [7]. They participate in the modification of the structure of proteins in the cell membrane and in signal receiving. [3;4] An important factor is the stimulating effect of carotenoids on the immune system, particularly their tumor suppressive effect. Carotenoids inhibit the development of fibrosarcoma by activating immune reactions [6]; the findings of in vitro studies particularly emphasize the suppressing effect of lycopene and P-carotene on the growth of breast tumor cells. [5]
The Material and Method
Data from the literature demonstrate that out of up to 600 types of carotenoids contained in the human daily diet, the body assimilates only six: a-carotene, P-carotene, lycopene, cryptoxanthin, neoxanthin, and lutein [16].
Taking into consideration the fact that spicy plants are included in humans' daily diet, we studied the carotene content in the leaves of certain spicy plants: Dill (Anethum graveolens L.), Basil (Ocimum basilicum L. .), Coriander (Coriandrum sativum L..), Parsley (Petroselinum crispum Fuss.).
The test samples were taken from a Gugark Region greenhouse farm in Lori Region, Armenia.
We determined the content of pigments - chlorophylls and carotenoids - by determining the optical density. [9]: The optical density of pigments was determined using a spectrophotometer (UV-Vis SPECTROPHOTOMETER SL150) with a wavelength of 665 nm and 649 nm for chlorophylls, and 440.5 nm for carotenoids. Spectrophotometric analysis is a relatively more accurate quantitative method for determining pigments in leaves. The pigment concentration is determined by means of optical density. The density of the extract is determined on a spectrophotometer by the absorption maximum of chlorophyll a and b in the red part of the spectrum, and in the case of carotenoids under the maximum absorption wave. The experiments were carried out in the Natural Science Complex Laboratory of Vanadzor State University.
The pigment content is expressed in mg of wet and dry weight (calculated per 1 g) in % of wet and dry mass per unit surface of the leaf.
During the experiment, we took 0.2 g of fresh leaves of each investigated plant. We prepared a 96% alcohol solution of chlorophyll from the leaves [14].
The concentration of pigments according to optical density indicators was determined by Vernon's formula in 96% ethyl alcohol solution. [13]
Ca = 11,63 • D665 - 2,39 • D649 Cb = 20,11 • D649 - 5,18 • D665 Ccar=4,695 • d440,5 - 0 268 • (Ca+b)
The quantity of chlorophyll a and chlorophyll b was determined by Vernon's formula:
Ca + Cb = 6,10 x D665 + 20,04 x D649
The carotenoid concentration was determined with Wettstein's formula. [8] Ckar = 4.695 x D440.5 - 0.268 (Cxl a +C xl b) ) [13]
According to the content of pigments in the leaf extract, we determine the content of pigments in the researched samples.
By weight of sample extract volume "
V • С
А =_
P-1000
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Where
C is the pigment concentration mg/l, V is the volume of the extract in ml (4 ml), P is the mass of the sample (0,2 g),
A is the pigment content in plant sample mg/g wet weight [13] Findings
As a result of the research, the following data were obtained:
Table 1. Concentration of pigments indifferent plant species
Plant species Optical density under different wavelengths Ca, mg/l Cb, mg/l Ccar, mg/l
d440,5 nm d649 nm d665 nm
Dill (Anethum graveolens L.) 1,690 0,301 0,700 7,422 2,427 5,295
Basil (Ocimum basilicum L.) 1,691 0,808 1,557 16,177 8,18 1,411
Coriander (Coriandrum sativum L.) 1,683 0,639 0,500 4,288 10,26 4,003
Parsley (Petroselinum crispum Fuss.) 1,689 0,311 0,664 6,979 2,815 5,305
The obtained findings reveal that the highest content of chlorophyll a is observed in the leaves of the Ocimum basilicum L. species.
According to the quantity of chlorophyll a, the chain of the plants studied is as follows:
Basil (Ocimum basilicumL) > Dill (Anethumgraveolens L.), > Parsley (Petroselinum crispum Fuss.) > Coriander Coriandrum sativum L• •)•'
The concentration of chlorophyll a in basil leaves exceeds the concentration of chlorophyll a in dill by 2.18 times, parsley by 2.32 times, and coriander by 3.77 times.
The concentration of chlorophyll b is highest in coriander leaves.
According to the quantity of chlorophyll b, the chain of the plants studied is as follows:
Coriander Coriandrum sativum L. •) > Basil (Ocimum basilicum L. •) > Parsley (Petroselinum crispum Fuss.) > Dill (Anethum graveolens
The concentration of chlorophyll b in coriander leaves exceeds that of chlorophyll b in basil leaves by 1.25 times, parsley leaves by 3.64 times, and dill leaves by 4.23 times.
The concentration of carotenoids is highest in parsley leaves.
And according to the concentration of carotenoids, the chain of researched plants is as follows:
Parsley (Petroselinum crispum Fuss.)> Dill (Anethum graveolens L.)> Coriander (Coriandrum sativum L. •) > Basil (Ocimum basilicum L. •):
The quantity of carotenoids in parsley and dill leaves is equal and exceeds the quantity of
carotenoids in coriander leaves by 1,325 times, and in basil leaves by 3,759 times. _Table 2. The quantity of pigments in sample plants_
Plant species Quantity of pigments in plant samples mg/g, wet weight Chlorophyll a:b
Chlorophyll a Chlorophyll b Carotenoid
Dill (Anethum graveolens L.) 0,148 0,0485 0,106 3.05
Basil (Ocimum basilicum L.) 0,323 0,1636 0,028 1.97
Coriander (Coriandrum sativum L.) 0,0856 0,205 0,080 0.42
Parsley (Petroselinum crispum Fuss.) 0,139 0,0563 0,106 2
The comparison of chlorophyll a+b , chlorophyll a/b, and carotenoid quantities is presented in Graph 1:
1.
lj2. _3. 4,
3,5 3 2,5 2 1,5 1 0,5 0
Anethum graveolens L. Ocimum basilicum L. Coriandrum sativum L. Petroselinum crispum Fuss
a+b 0,196 0,486 0,29 0,19
a/b 3,05 1,97 0,42 2
car 0,106 0,028 0,08 0,106
According to the literature data, the normal chlorophyll/carotenoid ratio is 3, i.e., the quantity of chlorophylls exceeds that of carotenoids by three times. [10]:
The findings of our studies are as follows:
• In Anethum graveolens L species, the quantity of chlorophylls a and b exceeds that of carotenoids by 1.85 times;
• In the plant Ocimum basilicum L. it exceeds by 17.37;
• In the plant Coriandrum sativum L. it exceeds by 3.63,
• In the plant Petroselinum crispum it exceeds by 1.84.
As a result, it turns out that the quantity of chlorophylls is the highest in basil leaves. The quantity of carotenoids in dill and parsley leaves is closer to that of chlorophylls, and the ratio of chlorophylls to carotenoids in coriander leaves is close to normal.
Summary
The study of the pigment concentration in the leaves of some spicy plants - Dill (Anethum graveolens L.), Basil (Ocimum basilicum L..), Coriander (Coriandrum sativum L.), Parsley (Petroselinum crispum Fuss.) revealed that the quantity of carotenoids in parsley and dill leaves is equal to 0.106mg/g weight wet. The quantity of carotenoids in parsley and dill leaves is equal and exceeds the quantity of carotenoids in coriander leaves by 1.325 times and in basil leaves by 3.759 times. As a result, it turns out that the quantity of chlorophyll is the highest in basil leaves. The quantity of carotenoids in dill and parsley leaves is closer to the quantity of chlorophylls, and the ratio of chlorophylls to carotenoids in coriander leaves is close to normal. Stemming from our research,
based on the quantitative predominance of carotenoids, the population should be recommended to use dill and parsley as spices, as immune system stimulants and antioxidants.
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