A retrospective study on capsicum

(C. Annum) to elicit the findings of

capsaicinoid and electrolyte content.

Bell peppers - Solanaceae

BACKGROUND

Bell pepper - is a cultivator group of species Capsicum annum belonging to family

Solanaceae originating from south and Central America. Fruit can widely vary in colour,
shape, size. Cultivators of the plant produce fruits of difference colours which includes red,
yellow, orange, green, chocolate/brown. The main aim of the project is to derive the
proximate principles of bell pepper in relation in order to determine it approximately in terms
of sensory attributes. In order to study biochemical and nutritional profile of bell pepper with
various proximities of principles, physiochemical properties of the bell pepper samples were
recorded. In a project to identify as well as study relating to large issues & use a persuasive
rationale to justify the reasons for this study.

SCIENTIFIC CLASSIFICATION:

Kingdom : Division - Magnoliophya

Class : Mangnoliopsida
Subclass : Asteridae
Order : Solanales
Family : Solanaceae
Genus : Capsicum
Species : C. Annuum
BINOMIAL NOMENCLATURE: C. Annuum

INTRODUCTION

Ideal temperature for growing capsicum includes warm soil, ideally 21-29 degree must be
placed in a moist environment but not waterlogged.
On estimation the carbohydrate content is high in the green bell pepper than red bell pepper
hence it gives an estimated determination of higher nutritional value. On sample processing
the bell pepper had potassium in adequate ratio which is beneficial when consumed as well
creates more space for anti-oxidants to fight against free radicals circulating in the blood
stream.

With adequate trace elements such as sodium, calcium, dietary fibres and phosphorus among
which the bell pepper sample comprises of more dietary fibres considered as more nutritive
content. Normal bacterial intestinal flora such as Escherichia coli were included in the
investigation with the aqueous extract of green and red bell pepper with chilled distilled water
as a measure of susceptibility of the microbe to the formulation was determined using “ Broth
tube dilution method “.

The conclusive study with the results suggested that the above microbiological techniques
were evaluated and confirmed using a highly sensitive (RT-PCR).Pungency of the pepper
fruit is attributed mainly to capsaicinoid concentration it adds flavour to food when used as
species. According to the reports, it’s primarily used as an recognised therapeutic effects for
treating gastric ulcers and rheumatoid arthritis. This group of capsicum fruits are lamides of
branched fatty acids with 9-11 carbon which has higher concentration of capsaicin
concentration (CAP) with few morphotypes as exception such as C.pubescens and does
suggest difference between C. Annum and C.pubescens.

HYPOTHESIS:

The present study is to derive the result on bell pepper used as in traditional Indian medicine
for many chronic diseases as a expectant dietary management. On determining many
properties of the fruit, various nutritive and phytochemical characters were further subjected
to biological & pharmacological study for therapeutic purpose. In order to achieve the
bactericidal property RT-PCT was ordered to exhibit the arrangement on a agarose gel was a
qualitatively analysed.

METHODS & MATERIALS

A sample of regional morphotypes of pepper were collected from backyard were used to
determine an analysis report on carbohydrates, proteins and ions in it. Each sample consists
of red and green bell pepper along with aqueous solution to extract the pulp of capsicum.
With the available sample solution, few experiments were carried to find the calculative
variables of sodium, potassium and carbohydrate content.

With the corresponding damply to lot of immature pods harvested from one plant.
Color in fresh fruit were measured based on mendelian characters of pods, size and shape of
the fruit. Vitamin C content rich fresh fruits were brought into healthy life to create a better
immunity as well to eliminate free radicals from the bloodstream. Samples obtained were
grounded with oxalic acid of 0.4% in a ration of 1:10. The absorbed sample was send for
centrifugation 660 rpm for 25mins. Later 1 mL of the supernatant was mixed with buffer
solution of sodium acetate and along with 2,6-dichlorophenol indophenol solution. With the
absorbance of solution will be measured with spectrophotometer with an wavelength of
520nm and on the basis of L-ascorbic acid (Vitamin C) calculations were carried out. The
estimated value of concentrated Vitamin C were reported as ascorbic acid accounting for
100g fresh weight.

Total phenolic contents were determined to find the fresh fruit sample without seeds stored at
-20 degree calcium for the next 16hours of time, before grounding them it should be
transferred to a stainless steel blended for 30seconds after which the demonised water is in a
ration of 1:10, with an ice bath contenting for next 15mins in a stirring mechanism and
centrifuging them for 3640 rpm. Mainly carried out to find the phenolic content with added
0.1 mL of supernatant which is now mixed with 2.8mL of deionized with 2% of sodium
carbonate.
On further calculation of phenol content and flavonoid were determined according to the
aluminium chloride colorimetric method.
Beta-carotene analysis was carried out only on the samples of ripened capsicum that had
reached maturity. The suspended material was centrifuged at 2500rpm continued for 15mins
followed by preserving at 20 degree Celsius until further analysis of the flavonoids from the
fruit species.

BACKGROUND

The two major components obtained from C.annum are capsaicin (C) and dihydrocapsaicin
(DHC). The viable difference between them is the presence of carbon-carbon double bonds.
Capsaicin (8-Methyl-N-vanillyl-6-nonenamide) with a molecular formula: C18H27NO3 and
molecular weight : 305.42.

2D – DIMENSIONAL VIEW

This is mainly acting as a neuropeptide releasing agent that are selective for primary sensory
peripheral neurons. Also utilized as topical creams, this capsaicin aids in controlling
peripheral nerve pain and are also useful in controlling chemotherapy and radiotherapy
induced mucositis. Capsaicin subjected as an alkaloid with notable thermal stability with
retaining tendency even boiling and are slightly water soluble, whereas it’s soluble in ethanol
and vegetable oils.

There are multiple possibilities to separate capsaicin and DHC but herein this review we
shall follow the chromatographic technique with liquid mobile conducted at a very high
pressure which offers the possibility of automating and computer processing the methods.
All suitable methods are drawn to list out the analysis assessing the distribution of
capsaicinoids in foods.

METHODS

PLANT MATERIAL

The fruits of C. annuum were collected from the local laboratory cultures for thesis purpose.
These samples used for assays were possesses, labelled and numbered as well stored in the
Biochemistry plant refrigerator. In the preparation of the eluent mixture, anhydrous
methanol is used as standard solution along with solutions of capsaicin and DHC of the same
brand were used up.

METHODOLOGY

Prepared with prior grinding, stored in bag containers with pod samples from each of the 10
different samples taken out of the freezer and were allowed to cool down to reach room
temperature prior to the preparation. Special progression were carried out in order to
prevent the loss of moisture condensation on the pod sample. The represented fruit sample
is well grounded with a GM 200 (Grindomix) mill to pass a 20 screen mesh. The powered
were accurately measured and weighed into 2 vials along with 2ml of N-
dimethylformamide (DMF) were added to the vial and tightly sealed with Teflon-lined
screw caps.

Extraction was carried out at 80 Degree C in a dry block heater continued for 1hr of time.
After which all samples were churned every 15min during the 1 hr period in order to assure
to proper mixing of samples with the standard solution. Post-churning all the samples were
removed from the heat block with proper use of hand-glove, following centrifuged for 5mins
in a Hettich Rotofix 32A Rotor and supernatants were eventually decanted into a 10mL
volumetric flask. This same procedure was repeated three more times for a total of 4
extractions and the content of flask were brought to volume of DMF.

ANALYSIS OF CAPSAICINOID FOR HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Samples of the extract of C. Annuum were placed into an autosampler measured in vials and
used directly for HPLC analysis for short run procedure of Morales. The chromatograms
were performed unit wise and equipped with Jasco PU-1580 Intelligent HPLC Pump and UV-
970 type optical absorption detector having spontaneous adjustable wavelength accounting
between 190-590mm.

Separations were accomplished with a C18 NUCLEOSIL 100 column which is filled with
particles accounting for 5mm diameter. The samples used in the testing phase were all the
fragments of C. Annuum obtained randomly from different batches for relative findings. In
order to establish a proper eluent composition in separating 2 main components from C.
annuum and once isolated from other component it will be preserved separately to look
after the structural biogenetics. In the process of chromatographic migration of capsaicin
and dihydrocapsaicin are at progressive rise in the polarity state of mobility. In all
determinations, the eluent used are methanol-water mixtures with different compositions.
The components involved in the separation were carefully monitored at 220nm.

H20 %
3.5

2.5

1.5

0.5

0
0.5 1 1.5 2 2.5 3

FIG: Above graphic form representing the chromatographic C. Annuum extract

ANALYSIS OF PROTEINS IN C. ANNUUM

The main aim is to detect the presence essential nutritive protein from capsaicin from the
extracted solution. Thermal stability is retained thus the hydrogen bonds of secondary and
tertiary protein structure while the protein increase in size due to denaturation of their
quaternary structural arrangement. This involves 5% of acetic acid solution, another 5% of
acetic acid which is now diluted with 100ml of distilled water. All the above reagents are
used to elicit the levels of proteins mixture composition.

PROCEDURE

1. Take 5ml of given extract solution in a new test tube

2. Heat the upper part of the test tube over the open burned flame.
3. Monitor the changes carefully as the turbidity or haziness will appear if the presence
of protein is present.
4. Accordingly, add few drops of 5% of acetic acid are slowly added from one end of the
test tube allowed for coagulation.
5. Add 1ml of test solution, add 1 or 2 drops of the indicator of pH (4.6-6.7)
6. Allow it to settle and add 1% of acetic acid drop by drop until the precipitation is
obtained.

RESULT
 Presence of protein casein and it’s precipitation proves the analysis.

Species Moisture (%) Protein (%) Ascorbic acid

(mg/100g)
Capsicum Annuum L 85.7% 1.35 92.8

FEHLING’S TEST

This test is a chemical test used mainly to differentiate between reducing and non-
reducing sugars. Objective of the experiment is to assess the presence of carbohydrate in
the extracted c. annuum solution.

PRINCIPLE

The carbohydrates possess potentially free carbonyl groups such as ketones or aldehydes
can eventually act as an reducing agent when mixed with appropriate reagent. The Fehling’s
solution appears deep blue in colour and comprises of copper sulphate mixed with
potassium sodium tartrate which is a strong alkali.

Further on heating the extracted solution with Fehling’s solution, thus once added it will
oxidizes the aldoses to corresponding aldonic acids. Copper ion in the solution reduces to
form insoluble yellow or reddish pink colour precipitate. On the other hand, the ketones are
oxidized to break down short chain fatty acids on exposure to the solution.

REQUIREMENTS

Reagents used :

1. Fehling’s solution A : Dissolve 7g of CuSo4.7H20 in 100ml of water

2. Fehling’s solution B : Dissolve 24g of KOH and 34.6g of potassium sodium tartrate in
100ml of NS water
3. Fehling’s solution : Mix equal proportions of both the solution A and B just before
using it in the experiment.
4. Sample (5% Glucose, 5% Sucrose, 5% Fructose and starch)

Materials Required :

1. Pipettes
2. Test tubes – 3
3. Test tube stand
4. Test tube holder

Equipment :
 1. Water bath

RESULT

The extracted solution appearance turns yellowish in colour on adding the reagent A
which does indicate an positive sign for presence of reducing sugars. Aromatic aldehydes
cannot be detected via this test.

REAGENT USED SAMPLE COLOUR OBSERVATION RESULT

SOLUTION

FEHLING’S C. Annuum Copper ion The extracted C. Annuum is

SOLUTION A extract solution oxidises to contents of the soluble and are
insoluble yellow c. annuum oxidised in
colour appear on the Fehling’s
top of the solution A thus
solution it also
possessing
reducing sugar
in it.

FEHLING’S C. Annuum Copper ions are The solution Thus, C.

SOLUTION B extract solution Oxidised to give shows dark red Annuum
Soluble reddish in colour along soluble in
pink colour side adding the oxidizing the
appearance on reagent Fehling’s
adding the reagent
solution B

PAGE REFERENCE

1. https://www.researchgate.net/figure/Nutritional-composition-of-green-chilli-
fruits_tbl2_263713896
2. https://microbenotes.com/fehlings-test/
3. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4348314/
4. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8224274/