International Journal of Research Publication and Reviews, Vol 3, no 3, pp 1779-1785, March 2022

International Journal of Research Publication and Reviews

Journal homepage: [Link] ISSN 2582-7421

Phyllanthus emblica (Amla) : Extraction and Determination by

laboratory content

Chaitanya Joshia,Harshal Borseb

Postgraduate scholar, Postgraduate scholar, K.T.H.M College Nashik

ABSTRACT:

Emblica officinalis Gaertn. or Phyllanthus emblica Linn, usually called Indian gooseberry or amla, is arguably the foremost necessary healthful plant within the
Indian ancient system of medication, the piece of writing. numerous elements of the plant are accustomed to treat a spread of diseases, however the foremost
necessary is that the fruit. The fruit is employed either alone or together with different plants to treat several ailments like respiratory disease and fever; as a drug,
laxative, liver tonic, refrigerant, stomachic, restorative, alterative, antipyretic, medication, hair tonic; to forestall ulceration and upset stomach, and as a biological
process. diagnosis studies have shown that amla possesses antipyretic, analgesic, medicament, antiatherogenic, adaptogenic, cardioprotective, gastroprotective,
antianemia, anti hypercholesterolemia, wound healing, medication, antiatherosclerotic, hepatoprotective, nephroprotective, and neuroprotective properties.
additionally, experimental studies have shown that amla and a few of its phytochemicals like acid, ellagic acid, pyrogallol, some nor sesquiterpenoids, corilagin,
geraniin, elaeocarpus, and prodelphinidins B1 and B2 additionally possess antineoplastic effects. Amla is additionally reportable to possess neuromodulatory,
chemomodulatory, chemopreventive effects, atom scavenging, inhibitor, medication, antimutagenic and immunomodulatory activities, properties that are
efficacious within the treatment and hindrance of cancer. This review for the primary time summarizes the results associated with these properties and
additionally emphasizes the aspects that warrant future analysis to ascertain its activity and utility as a cancer preventive and therapeutic drug in humans.

Keywords: Antioxidant; Polyphenol; Flavonoid; Amla; Lemon Peel; Cucumber Peel

Introduction:

The biological antioxidants are compounds that protect biological systems against the probably harmful effects of processes or reactions that cause
excessive oxidation. Nowadays aerobic stress is one of the extraordinary problems with fashionable society. utterly totally different air pollution,
smoking, ultraviolet radiation etc. Ends up in aerobic stress of cells, causes various diseases like skin disease, melanomas or pic aging of the skin,
cancer, heart disease, inflammation, arthritis, immune systems decline, brain pathology and cataracts etc. The consumption of fruits And vegetables
has been found to be associated with lowering of these diseases as they contain an outsized amount of polymer compounds, antioxidants and
flavonoids. In various studies it has been found that Associate in Nursing Antioxidants can inhibit or delay the oxidation of an oxidisable substrate in
an extremely chain reaction so antioxidants seem to be necessary in bar of these diseases (Ali et al. 2010; Ames 1983; Ames et al. 1993; Feskanich et
al. 2000; Halliwell 1996). The extraction of polymer compounds is also printed as a result of the separation of medicinally active portions from plant
tissues pattern selective solvents through traditional extraction procedures. The foremost criteria of extraction technique is separating the soluble and
insoluble components and jilting entirely insoluble Cellular mass. The extraction products of plants have relatively advanced mixtures covering sort of
groups of plant metabolites either in liquid kind or semi-solid state (Imran et al. 2011; Ramamoorthy et al. 2007). the general techniques of extraction
of healthful plants embrace maceration, infusion, percolation, digestion, decoction, hot continuous extraction (soxhlet), aqueous-alcoholic extraction by
fermentation, counter current extraction, microwave assisted extraction, ultrasound extraction (sonication), essential fluid extraction (SFE), phytonic
extraction (with hydro-fluoro-carbon solvents), etc. For the aromatic plants, three sorts of hydro-distillation techniques (water distillation, steam
distillation, steam and water distillation), hydrolytic maceration followed by distillation technique, expression methodology and enfleurage
methodology (cold fat extraction) is also used. variety of the foremost recent ways that of extraction for aromatic plants embrace head house saddlery
technique, solid half micro-extraction, body part extraction technique, micro-distillation, thermo-micro-distillation, and molecular distillation
techniques (Ics-Unido 2006).

Materials:

Three different types of fruits & vegetables were purchased from native market of Nashik, Maharashtra (India). These embrace Amla (Emblica
officinalis), Lemon (Citrus limon) and Cucumber (Cucumis sativus). For the current study the components of fruit taken are area units, edible a part of
amla, part lemon and part cucumber. Folin-Ciocalteu chemical agent, soda, acid, Quercetin, Methanol, chloride, metallic element acetate, Butylated
radical dissolver (BHT), vitriol, inorganic phosphate, ammonium ion molybdate of AR grade were purchased from native provider of Nashik.
 International Journal of Research Publication and Reviews, Vol 3, no 3, pp 1779-1785, March 2022 1780

Methods:

All the fruits, vegetables taken were washed with water and desired components were dried in a kitchen appliance at 70 0c. At the moment the
components were grinded and sieved with mesh aperture size of two metric linear units. The portion of fruits (1gm) was homogenized with totally
different concentrations of binary compound wood spirit answer (0-70% methanol). The stuff was stirred with a magnetic stirrer at 900 revolutions per
minute at temperature for half-hour. The extract was centrifuged at 3000 revolutions per minute for twenty minutes. The supernatant was removed and
filtered with whatman paper four. Finally the supernatant was held on within the dark. equally another extraction was done keeping the methanol: water
(50:50 v/v) magnitude relation constant however varied the quantity of sample dose (1gm, 1.5 gm, 2.0gm, 2.5gm).

Determination of total phenolic content

The total phenolic resin content of the fruit and vegetable extracts were calculable by the strategy represented by Singleton and Rossi (1965) with
slight modification and reported as acid equivalent (GAE) per gram of fruit/vegetable. acid in varied concentrations (20-140)

Determination of antioxidant activity

The inhibitor activity of fruit and vegetable extracts were calculable by phosphomolybdenum assay and reported as Butylated chemical group
dissolving agent equivalent (BHTe) per gram of fruit/vegetable. completely different concentrations of Butylated chemical group dissolving agent
(25-125 μg/ml) in water were ready as customary. 0.3 milliliter of fruit extracts were taken to take a look at tubes. The chemical agent answer was
ready by combining ten milliliter of zero.6 M sulfuric acid, ten milliliter of twenty eight metric linear unit inorganic phosphate and ten milliliter of
four metric linear unit ammonia molybdate into a beaker and three milliliter chemical agent answer was further to all or any the tubes. 0.3 milliliter
of fuel served as blank. All the tubes were incubated at ninety five oC for ninety min. The tubes were cooled to temperature and also the optical
density was measured at 695 nm mistreatment ultraviolet illumination photometer (UV-1800,Shimadzu).

Determination of total flavonoid content

The total flavonoid content of the fruit and vegetable extracts were calculable by mistreatment chloride quantitative chemical analysis technique
and reported as Quercetin equivalents (Qe) per gram of fruit/vegetable. Quercetin in varied concentrations (12.5-100 μg/ml) was ready in fuel as
customary. 0.5 milliliter of every fuel extracts (1:10) were taken in take a look at tubes and one.5 milliliter fuel, 0.1ml of 100% chloride, 0.1
milliliter of one M metal acetate and a pair of.8 milliliter water were further individually to every tubes. All the tubes were incubated at
temperature for thirty min. Optical density was measured at 415 nm by a mistreatment ultraviolet illumination photometer (UV-1800,Shimadzu).

Results and Discussion

The total polyphenol content, antioxidant activity and flavonoid content was determined by folin-ciocalteu method, phosphomolybdenum assay
and aluminum chloride colorimetric technique respectively as stated above.

Total Polyphenol content

The standard curve (y = 0.012 x, r2 = 0.986) for determination of total polyphenol content was obtained by measuring OD of standard solution of
Gallic acid and is shown in Figure 1. The Total phenolic content of all the three samples/extracts were then estimated using this standard curve and
results were shown in Figure 2.

Effect of solvent Concentration

It is seen from the Figure a pair of.a that polyphenol content of Emblica officinalis extract will increase with increase in concentration of wood
alcohol within the solvent. The utmost quantity of polyphenol extracted was 112.52 mg GAE/g at seventieth wood alcohol in water nearly adequate
to the quantity (111.72 mg GAE/g) obtained at five hundredth wood alcohol in water. The lemon tree (peel) and Cucumis sativus (peel)
additionally show a rise in total polyphenol content with an increase in concentration of wood alcohol, however the rise isn't terribly high when
five hundredth wood alcohol concentration. The utmost quantity of polyphenol extracted was nine.7 and 7.4 mg GAE/g for peel and cucumber peel
severally at seventieth wood alcohol in water. Therefore, five hundredth wood alcohol in water was taken as optimum solvent concentration for
any study.

Effect of amount of dry sample powder

The 50% wood alcohol resolution was to extract total polyphenol from all the 3 samples by variable the quantity (1-2.5 g) of dry powder of
fruit/vegetable, the results obtained were shown in Figure a pair of.b. It is seen from the figure that total phenolic resin content of extract will
increase with increase in quantity of dry sample. For 2.5 g of sample it's 194, 16.072, 10.0548 mg GAE for amla, lemon and cucumber peel
severally. It is seen that Emblica officinalis shows most phenolic resin content as actual fruit half is taken for study. Each lemon tree and Cucumis
sativus shows a comparable quantity of phenolic resin content as waste half was taken for the study (peel). tho' amla contains an excellent quantity
of phenolic resin compounds, the quantity of phenolic resin compounds in lemon and cucumber peels are ok to extract. so these materials thrown
away everyday from households as waste is used for extraction of valuable phenolic resin compounds
 International Journal of Research Publication and Reviews, Vol 3, no 3, pp 1779-1785, March 2022 1781

Figure 1 Standard curve for determination of Total phenolic content

Figure 2: (a) Effect of solvent concentration (b) Effect of amount of sample on extraction of total polyphenols compounds

Total Flavonoid content

The standard curve (y = zero.005 x, r2 = 0.994) for determination of total flavonoid content was obtained by chloride quantitative chemical analysis
technique as Quercetin equivalents and is shown in Figure three. The whole flavonoid content of all the 3 samples/extracts were then calculable
victimization of this commonplace curve and results were shown in Figure four.

Effect of solvent Concentration

It is seen from Figure four.a that total flavonoid content of all the 3 sample extracts will increase with increase in concentration of wood alcohol within
the solvent however the rise isn't terribly sharp. the utmost quantity of flavonoid extracted was nineteen.2,18.1,17.1 mg /g of amla, lemon peel, &
cucumber peel severally, at seventieth wood alcohol in water nearly adequate to quantity (111.72 mg GAE/g) obtained at five hundredth wood alcohol
in water. Therefore, five hundredth wood alcohol in water was taken as optimum solvent concentration for any study. Diankov et al. (2011) additionally
found that solvent concentration doesn't have an effect on considerably the extraction rate. It also can be seen from the figure that a similar trend was
obtained in all the 3 samples with relevant solvent concentration and also the quantity of flavonoid content is additionally comparable.
 International Journal of Research Publication and Reviews, Vol 3, no 3, pp 1779-1785, March 2022 1782

Effect of amount of dry sample powder

The 50% wood alcohol resolution was to extract total flavonoid from all the 3 samples by variable the quantity (1-2.5 g) of dry powder of
fruit/vegetable, the results obtained were shown in Figure four.b. It is seen from the figure that total flavonoid content of extract will increase with
increase in quantity of dry sample. For 2.5 g of sample it's twenty two.41, 21.34, 20.28 mg Qe for amla, lemon and cucumber peel severally. It is seen
that in terms of total flavonoid content all the 3 samples are comparable. tho' amla contains a higher quantity of flavonoids, the quantity gift in lemon
and cucumber peels is almost adequate to it. Therefore, these waste materials are used for extraction of valuable flavonoids rather than expensive amla
fruit

Figure 3: Standard curve for determination of Total flavonoid content

Figure 4: (a) Effect of solvent concentration (b) Effect of amount of sample on extraction of total flavonoids

Antioxidant activity
For determination of total inhibitor activity Phospho atomic number 42 assay and metal chelating activity are reportable as Butylated radical dissolver
(BHT) equivalents. The quality curve (y = zero.013 x, r2 = 0.992) obtained is shown in Figure five. The inhibitor activity of all the 3 samples/extracts
was a calculable victimization commonplace curve and results were shown in Figure vi.

Effect of solvent Concentration

It is seen from the Figure vi.a that inhibitor activity of all the 3 sample extracts will increase with increase in concentration of wood alcohol within the
solvent however the rise isn't terribly sharp. The utmost inhibitor activity shown was eight.54, 2.91& 1.75 mg BHTe/g of amla, lemon peel, &
cucumber peels severally, at seventieth wood alcohol in water. It also can be seen from the figure that most inhibitor activity was shown by amla.
Lemon and cucumber pees additionally show the inhibitor activity so these are used as a low cost supply of antioxidants.
 International Journal of Research Publication and Reviews, Vol 3, no 3, pp 1779-1785, March 2022 1783

Effect of amount of dry sample powder

All the 3 samples were analyzed for his or her inhibitor activity by variable the number (1-2.5 g) of dry powder of fruit/vegetable; the results obtained
were shown in Figure half-dozen.b. It is seen from the figure that total inhibitor activity of extract will increase with increase in quantity of dry sample.
For 2.5 g of sample it's thirteen.132, 3.332, 2.352 mg of BHT/ two.5 g for amla, lemom and cucumber peel severally. It is seen that in terms of inhibitor
activity amla is superior to lemon and cucumber peel. although amla shows higher inhibitor activity, smart activity was additionally shown by lemon
and cucumber peels
.

Figure 5: Standard curve for determination of Antioxidant activity

Figure 6: (a) Effect of solvent concentration (b) Effect of amount of sample on antioxidant activity

Conclusions

In this study, five hundredth binary compound solvent extracts from 3 fruit and vegetable samples gave higher amounts of total polyphenol and
inhibitor activity. It is all over that polyphenol content is proportional to inhibitor property of sample as amla having higher polyphenol content shows
higher inhibitor property compared to lemon and cucumber peels. It also can be understood that there's no direct relationship between total phenolic
 International Journal of Research Publication and Reviews, Vol 3, no 3, pp 1779-1785, March 2022 1784

resin compounds present within the sample and also the total flavonoid content. Amla having a terribly high quantity of total phenolic resin content
shows similar flavonoid content with lemon and cucumber peels. So lemon and cucumber peels are thought-about as a low cost supply of flavonoid,
and may be used expeditiously for its extraction. These materials, otherwise waste, are used as potential supply of antioxidants for industrial application
additionally.

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