PHENOLIC ACIDS AND THEIR VITR0 EFFECTS

INTRODUCTION
The term "phenolic acids'' refers to aromatic acids that have a phenolic ring and a carboxyl functional
group. One of the most common classes of plant phenolics is made up of flavonoids and phenolic acids.
A greater comprehension of flavonoid concentration and biological activities may reveal that they have
therapeutic potential due to their importance to plants and human health. It would also be helpful to
anticipate and monitor the quality of medicinal herbs.
The hydroxyl, carboxyl, and at least one aromatic ring are all present in these molecules. One of the
simplest phenolic acids is salicylic acid. The salicylic acid molecule's structure is shown below.

One of the main types of phenolic substances found in plants are phenolic or phenol carboxylic acids,
which are a form of phytochemical commonly known as a polyphenol. They can be found in a wide
range of plant-based meals, with the largest concentrations being in seeds, fruit and vegetable skins, and
leafy greens. Rarely do they exist in free form; often, they are present in bound forms like amides,
esters, or glycosides. The phenolic acids are mainly divided into two sub-groups:
● Hydroxycinnamic acid
● Hydroxybenzoic acid
In Vitro Antioxidant Activity of Phenolic Acids
In vitro antioxidant efficacy of phenolic acids is substantially higher than that of renowned antioxidant
vitamins. Foods frequently include hydroxycinnamic acids, which are quinic acid or glucose-based
simple esters generated from cinnamic acid. Chlorogenic acid is the most prevalent soluble bound
hydroxycinnamic acid found. (a combined form of caffeic and quinic acids). The four most prevalent
hydroxycinnamic acids are;
● Ferulic
● Caffeic
● P-coumaric
● Sinapic acids

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On the other hand, hydroxybenzoic acids, which are generated from benzoic acid, share a C6-C1
structure. They are present as lignin-bound cell wall fractions and are hydrophilic (conjugated with
sugars or organic acids). In comparison to hydroxycinnamic acids, hydroxybenzoic acids typically occur
in low concentrations in foods like red fruits, onions, black radish, etc. The four hydroxybenzoic acids
that are most frequently seen are;
● P-hydroxybenzoic
● Protocatechuic
● Vanillic
● Syringic acids

BIOSYNTHESIS OF PHENOLIC ACIDS

The plants generate a vast array of low-molecular-weight substances that are referred to as "natural
products'' or secondary metabolites, which are typically derived from the phenylpropanoid,
isopropanoid, alkaloid, and fatty acid pathways and confer the metabolic flexibility required for biotic
and abiotic responses. Phenolic acids are a varied family of plant polyphenols that have received the
most attention in research. They are created through the phenylpropanoid route's shikimic acid, during
the monolignol pathway as by-products, during the breakdown of cell wall polymers like lignin, and in
some cases by microbes as well. The majority of phenolics in bacteria, fungi, and plants are produced
via the shikimate or shikimic acid pathway, which works by converting simple carbohydrate molecules
(produced by the pentose phosphate cycle and glycolysis) into phenylalanine and tryptophan. In a multi

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step metabolic process, larger molecules of carbohydrates decompose into glucose, which subsequently
transform to dehydroshikimic and shikimic acids. While shikimic acid changed into phenylalanine,
dehydroshikimic acid changed into gallic acid, a basic hydroxybenzoic acid. While some plants and
grasses convert 4-hydroxycinnamic with the aid of tyrosine ammonia lyase, phenylalanine ammonia
lyase (PAL) converts phenylalanine into trans-cinnamic acid and releases ammonia, which serves as the
key point for the synthesis of phenolic acids as well as in primary and secondary metabolism. (TAL).
The generation of phenolics is significantly influenced by the PAL gene expression and its protein
function through metabolite channeling, feedback regulation, and post transcriptional modifications.
Phenylpropanoids and flavonoids are lacking in bacteria due to the lack of PAL and chalcone synthase,
however some marine bacteria, including Streptomyces maritimes, Streptomyces verticillatus, and
Sorangium cellulosum, are known to have PAL. A PAL protein was found in cyanobacteria in 2007 by
Moffitt and colleagues. It shares similarities with yeast and plant PALs in terms of its tertiary and
quaternary structure, and it associates with gene clusters involved in the biosynthesis of secondary
metabolites. Enzyme substitution treatment for phenylketonuria patients has been recommended as an
alternative to prokaryotic PAL.
In recent years, knowledge of the phenylpropanoid pathway's enzymes has increased, providing useful
information for metabolite and enzyme engineering. Esterified phenolic acids are incorporated into the
plant cell wall and are released during acid/alkali hydrolysis. Cinnamic acid is transformed into
p-coumaric acid by the activity of monooxygenase following the addition of a hydroxyl group at the
pera position. (use cytochrome P450 as oxygen binding site). Caffeic and ferulic acids are created
through the hydroxylation and oxymethylation of p-coumaric acid, respectively. These phenolic acids
have a C6-C3 structure and are the precursors to lignin and other phenolic compounds. Cinnamic acid
changed into benzoic acid and its derivatives by losing two carbon atoms.
Biotechnological methods have been employed for phenolic acid synthesis on a big scale. The control of
several biosynthetic pathways for numerous secondary metabolites of chemical-pharmaceutical
relevance is revealed using cell and tissue culture models, which are also employed to increase their
synthesis. The benefit of using plant tissue culture in flasks and bioreactors over the conventional
technique (growing the entire plant in a natural setting or on an agricultural farm) is that it offers
improved control over the limits of the environment, such as seasonal change and geographic locations.
A potential method for phenolic acid mass synthesis has been provided through biotechnology.

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METABOLISM OF PHENOLIC ACIDS

Studies of intestinal, hepatic, and microbial metabolism conducted in vitro and in vivo have validated
the relationship between parent phenolic acids and their metabolites. In addition to dietary sources, the
bacteria in the colon can also make phenolic acids by breaking down other types of polyphenols. Dietary
phenolic acids, which are present in all plants and are quite protective in oxidative stress situations.
After being absorbed from the GI tract, they are heavily metabolized in humans (particularly caffeic and
ferulic acids), where they undergo methylation, glucuronidation, and sulfation derivatives that modify
their structural makeup and have biological consequences. The hydroxyl group of phenolic acids is
reduced by sulfation and glucuronidation, which lowers their antioxidant activity. With the exception of
the metabolites of caffeic and ferulic acids, there is a dearth of information available about the biological
activity of phenolic acids. The metabolic fates of ferulic, caffeic, and sinapic acids have also been well
investigated. These acids are metabolized mostly by intestinal bacteria. According to reports, 11-25% of
the entire quantity consumed is excreted as free or bound ferulic acid. The excretory system
predominantly eliminates the phenolic acid metabolism's excretory products in the form of feces and
urine.

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BIOAVAILABILITY OF PHENOLIC ACIDS
The degree to which phenolic acids exhibit biological activity is principally influenced by their
bioavailability, which is the percentage of their absorption, digestion, and metabolism after entering the
circulatory system. Numerous epidemiological and experimental studies have shown that phenolic acids
have a preventive effect against degenerative illnesses such as cancer, diabetes, inflammation, and many
others. Due to their potent antioxidant nature, their health impacts are especially well-known. However,
while phenolic acids got less attention than flavonoids, we must expand our database on their
bioavailability in order to better understand their impacts on health.
The table below, which has been adjusted from published data, summarizes the bioavailability of several
phenolic acids. The bulk of studies on hydroxycinnamic acids have focused on caffeic, ferulic, and
chlorogenic acids. When hydroxycinnamic acids are consumed in their free form, they are quickly
absorbed from the stomach and small intestine and then conjugated by detoxifying enzymes. We only
have a little amount of information for investigations on the bioavailability of hydroxybenzoic acids
since their dietary content is substantially lower than that of hydroxycinnamic acids. The most often
researched hydroxybenzoic acid for metabolism and absorption is gallic acid, which supports its
excellent absorption capacity.

Phenolic Dose Intake Metabolites and aglycone

acid (duration)
Tsample (h) Determined 24 h urinary
{after last dose} (μM) Excretion (dose
%)

1.94 mg/day 21 days 12 ND 42.5

Ferulic acid 9.7 mg/day 21 days 12 1.1 51.8

48.55 mg/day 21 days 12 7.6 38.6

100 μmol/kg 1(GI) 0.33 30.39 --

Caffeic acid
10 mg/kg 1(GI) 0.5 6.0 --

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 45 mg/day 8 days 12 108.7 40.9

110 mg/day 1(GI) 0.5 ≈550 13.7

125 mg/kg 1(GI) 2.0 60 --

500 mg

p-Coumaric 100 μmol/kg 1(GI) 0.16 139 --

acid

Gallic acid 100 μmol/kg 1(GI) 1 3.51 --

50 mg/kg 1(GI) -- -- --

Chlorogenic
248 mg/kg 1(GI) 0.5-1.0 ≈8 --
acid

88.6 mg/day 8 days 12 113.4 58.8

36.3 mg/kg 1(GI) 0.5 0.96 --

Rosmarinic acid 50 mg/kg 1(GI) 0.5 4.63 5.47

200 mg/kg 1(GI) -- -- 31.8

EXTRACTION AND QUANTIFICATION OF PHENOLIC ACIDS

Extraction of phenolic compound(s) from plant materials depends on the nature of sample matrix and
chemical properties of desired phenolic(s) such as number of aromatic rings and hydroxyl groups.
Different solvents such as water, acetone, meth/eth/prop-anol, ethyl acetate are used to extract the
phenolic compounds. After extraction, analytes are separated by chromatographic techniques (TLC,
column, HPLC) and structure elucidation by elemental analysis, FT-IR, NMR, UV–vis.,

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GC—MS/HRMS and X-ray crystallography. Chromatographic techniques viz. HPLC and GC alone or
in combination with mass spectrometry provide the best results.
Folin-Ciocalteu and Folin-Denis assay are the commonly used spectrophotometric methods for
measurement of total phenolic content. Phenolic acids, flavonoids and tannins are the most important
phenolic compounds for GC analysis. High Performance Liquid Chromatography (HPLC) is the most
favored method used to separate and quantify the phenolic compound(s). Selection of mobile phase,
column, column temperature, types of detectors, and sample purification are the major factors affecting
the HPLC. Methanol, acetonitrile and their aqueous forms are leading mobile phases for phenolics
quantification through HPLC in gradient elution.
C18 or reverse phase column are used predominantly with 10-30 cm long, 3.9-4.6 mm internal diameter
and 3–10 μm particle size, but development of new column types with 3–25 cm length, 1–4.6 internal
diameter and 1.7-10’μm superficial porous particles are used in advanced HPLC methods. Identification
of phenolics in UV–vis and photodiode array detectors is carried out at 190-380 nm, colorimetric,
fluorimetric, photodiode coupled with fluorescence are used. Schematic representation of different
extraction and structure determination methods used for plant phenolics are given below;

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APPLICATIONS OF PHENOLIC ACIDS
Although their exact function in plants is still unclear, phenolic acids are known to serve a variety of
functions, including protein synthesis, allelopathy, nutrient intake, structural components, enzyme
activity, and photosynthesis. The primary polyphenols generated by plants are phenolic acids, which are
renowned for their wide range of biological uses. Phenolic acids serve as the progenitor of bioactive
compounds frequently employed in the food, cosmetics, and pharmaceutical sectors. The main benefit of
employing phenolic acids is that natural bacteria can metabolise them, making them a vital replacement
for man-made chemicals that are equally bad for the environment. Following is a summary of the many
uses of phenolic acids in pictures that draws inspiration from literature.

PHENOLIC ACIDS AS SIGNALING MOLECULES

Plant phenols, especially phenolic acids, function as signalling molecules during the beginning of the
development of arbuscular mycorrhizal and legumerhizobia symbioses and can also be involved in plant
defence. In addition to serving as precursors for phenolic lipids, these compounds present themselves as
an alternate source of carbon for some diazotrophs under unfavourable environmental circumstances.
When a legume plant's seed germinates and develops into a seedling, phenolic acids are promptly
produced by the plant's ascending roots. Recently, certain phenolic acids have been discovered in the
nodules of Vigna mungo. These phenolic acids induce the generation of indole acetic acid and also
control Rhizobium sp. morphogenesis. A number of phenolic acids were found to either promote or
repress the expression of the nod gene in Rhizobium trifolii. This work's potential mechanism of action,

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which claimed that the impact of phenolic acids depended on their chemical makeup, concentration, and
strain-specificity, was also explored.

PHENOLIC ACIDS IN FOOD

The average daily intake of phenolic acids is 211 and 11 mg/day for hydroxycinnamic and
hydroxybenzoic acids, respectively, according to a 1998 German research, while the population under
observation exhibited variation from 6 to 987 mg of phenolic acids. Numerous studies have shown that a
diet high in fruits and vegetables is healthier because they act primarily as antioxidants and prevent
illnesses like cancer, coronary heart disease, and stroke that are brought on by oxidative damage. The
examination of phenolic acids' potential function in plant life is simply one of many inquiries; its
significance for food safety is another large topic of interest. They have been employed to improve food
items' organoleptic (flavor, astringency, and hardness), chromatic, sensory, nutritional, and antioxidant
attributes. Additionally, researchers have looked at how phenolic acids affect fruit maturation, inhibit
enzymatic browning, and serve as food preservatives. The most common phenolic acids present in the
majority of fruits and cereal grains, respectively, are caffeic and ferulic acids (free/esterified). About
two-thirds of the total dietary phenolics in plant-based foods are accounted for by flavonoids alone, but
phenolic acids also play a significant role and make up the remaining one-third. Because of these simple
phenolic acids' antioxidant behaviour and potential health advantages, the scientific community is
becoming increasingly interested in them. People get phenolic acids every day from foods including
vegetables, fruits, cereals, tea, coffee, and spices since they are so common. According to a diet chart,
this amounts to roughly 25 mg per day. Even though there are several in vitro studies that support the
antioxidant and health preventive benefits of all phenolic acids, the precise mechanism behind these
actions is still poorly understood.

PHENOLIC ACIDS AS ANTIOXIDANTS

UV radiation exposure can lead to skin cancer and accelerated ageing of the skin by causing sunburn,
DNA damage, and connective tissue degeneration. According to epidemiological research, sunscreens
and sunblockers cannot entirely prevent UV-induced skin cancer. Therefore, in order to cure the
disorders caused by UV irradiation, we must look for novel focused chemopreventive strategies.
Because reactive oxygen species (ROS) like superoxide (•O2 ), hydroxyl (•OH), and peroxyl (•RO2 )
can have harmful effects on the skin, numerous studies have looked at antioxidant compounds as a way
to strengthen the body's natural defences against oxidative stress-related illnesses. Researchers
concentrated on phenolic acids, flavonoids, and other phenolics as well as natural antioxidants and their
derivatives for this. Although phenolic compounds are recognised as direct antioxidants, they have
shown indirect antioxidant action by activating endogenous defence mechanisms and having favourable
regulatory effects on signalling pathways. Antioxidants are phenolic acids. (due to the reactivity of
phenol moiety; hydroxyl substituent on aromatic ring). Although there are a number of mechanisms for
the antioxidant activity of phenolic acids (because of the reactivity of the phenol moiety), radical

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scavenging via hydrogen atom donation is thought to be the primary mechanism. In phenolic acids,
substitutes on the aromatic ring alter the stabilisation of the structure, which in turn affects the capacity
to quench radicals. The antioxidant properties of free, esterified, glycosylated, and nonglycosylated
phenolics have been described and shown to differ, indicating that distinct phenolic acids have varied
antioxidant activity.

PHENOLIC ACIDS AS ANTIDIABETIC AGENTS

Due to an imbalance between the production of free radicals and a person's capacity to oxidise them,
diabetes is recognised as an oxidative stress condition. Because ROS, which are weakly neutralised by
antioxidants, cause organ damage, oxidative stress is strongly linked to inflammation and a wide range
of metabolic illnesses. The activity of free radicals is inhibited by antioxidants in a variety of ways, and
phenolic substances, especially phenolic acids (which have strong antioxidant and free radical
scavenging capacity), combat oxidative stress and its drawbacks by blocking the enzymes that produce
ROS. The phenolic acids have an impact on how insulin and glucose receptors work. (have a crucial role
in diabetes). They stimulate the translocation of GLUT4 through the PI3K/Akt and AMP activated
protein kinase pathways and increase the expression of the glucose transporter GLUT2 in the
insulin-producing pancreatic beta-cells. Both chlorogenic and ferulic acids have been shown to stimulate
transporters similarly and to be effective antidiabetic drugs. The finest of the phenolic acids' abilities is
their ability to block -glucosidase and -amylase, two crucial enzymes involved in the breakdown of
dietary carbohydrates into glucose.

PHENOLIC ACIDS AS ANTIMICROBIAL AGENTS

Among all the phenolics found in faecal water, phenolic acids are the ones that are most prevalent. They
function as food preservatives and display antibacterial action. The chemical structure of phenolic acids,
specifically the saturated chain length, location, and number of substitutions in the core benzene ring,
has been used to identify their antibacterial potentials. The antibacterial activity of phenolic acids is
lower than that of their methyl and butyl esters. Alkyl chain length increases significantly with activity;
for example, phenolic acid oligomers have more activity than their monomers. The quantity of hydroxyl
(OH) and methoxy (OCH3) functional groups in a given chemical determines the antibacterial activity
of the hydroxybenzoic and hydroxycinnamic acids, and their structure-function connections do not
adequately account for the variety of these compounds. Hydroxybenzoic acids, like other weak organic
acids, followed the pattern of un-dissociated acid diffusion through the membrane for antibacterial
action through the acidification of cytoplasm, which causes cell death. Therefore, the main determinants
of phenolic acids' solubility in microbial membrane and related antibacterial capability are their pKa and
lipophilicity. The pH also has a significant impact since it determines the charge on the carboxyl
(COOH) group, ring replacements (hydroxyl and methoxy groups), and side chain saturation. Phenolic
acid antibacterial activity is negatively correlated with pH variation. Hydroxycinnamic acids lose a
considerable amount of their antibacterial action when their double linkages are reduced.

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PHENOLIC ACIDS IN CANCER CURE AND TREATMENT
According to a report by the globe Health Organization (WHO), more people die from cancer each year
than from AIDS, malaria, and TB combined. By 2030, it is predicted that this figure would increase by
80%. Cancer is one of the greatest health concerns in the globe. According to a research from the Indian
Council of Medical Research (ICMR), the number of cancer patients in India alone will rise significantly
by 25%. Evidence from epidemiology shows that eating a diet high in antioxidant-rich fruits and
vegetables considerably lowers the chance of developing various cancers, indicating that certain dietary
antioxidants may be useful in reducing the incidence and mortality of cancer. The prevention and
treatment of cancer are greatly aided by phenolic acids and their derivatives, such as hydroxybenzoic
and hydroxycinnamic acids. Plant phenolics may present a potential in this area, and many clinical trials
are still ongoing. Between the 1940s and 2006, more than half of all anticancer prescription medications
licenced globally were natural compounds or their derivatives. They limit the creation of genotoxic
compounds, inhibit the activity of mutagen-transforming enzymes, control heme-containing phase I
enzymes, carcinogen-detoxifying phase II enzymes, and also prohibit the development of DNA adducts,
all of which contribute to the reduction of tumour start. The majority of phenolic compounds work in
various ways to prevent or treat certain cancers.

CONCLUSION
Based on their phenolics content, cereals, vegetables, and fruits can minimise the risk of disease and
improve human health. The scientific community is focused on improving the extraction and
purification processes for these naturally given molecules because of the vast range of health advantages
and industrial applications of phenolic compound(s). Non-flavonoid polyphenols, such as phenolic
acids, are an important family of polyphenols that are often employed in human diets. Depending on
how many vegetables, fruits, and whole grains one consumes each day such as apples, mangos, berries,
plums, cherries, kiwis, citrus fruits, onions, tea, coffee, and flour produced from whole wheat, rice,
maize, or oats one should consume 200 mg of phenolic acids everyday or more. The nutritional
supplements grape seed extract and green tea extract both contain phenolic acids. The supplements are
normally sold as antioxidants, but recent research has shown that whole grains, vegetables, and fruits
offer more health advantages than antioxidant supplements, leaving a huge market opportunity for their
extraction. Phenolic acids are easily absorbed through the walls of the gastrointestinal system and are
good for human health because they may function as antioxidants and prevent cell damage brought on
by free-radical oxidation processes. Phenolic acids boost human beings' ability to fight inflammation
when consumed regularly. Because phenolic acids serve as precursors to other important bioactive
compounds that are frequently required by the pharmaceutical, cosmetic, and food sectors, their demand
is particularly high in the industrial sector. The rising demand for phenolic acids is also related to their
efforts to protect the environment as they are broken down by natural microbes.

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