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Antioxidant and Antimicrobial Activity of Alpinia officinarum

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Alpinia officinarum is a rhizome belonging to the family zingeberaeceae. Hydro alcoholic extract by hot and cold maceration and methanol extract by percolation process Qualitative phytochemical analysis of extract of Alpinia officinarum rhizome showed a majority of the compound including tannins, alkaloids, flavonoids and saponins. Hydroalcoholic extract prepared by hot maceration process was found to contain more phenol and flavonol and it was measured as 50.1 mg/g and 54.02 mg/g, respectively. All the three extracts showed moderate to potent antimicrobial activity against the Bacillus cereus, Staphylococcus aureas, Pseudomonas auroginosa, Escherichia coli. None of the extracts showed antifungal activity against Aspergillus niger and Candida albicans. All the three extracts showed a concentration dependent radical scavenging activity by inhibiting diphenylpicrylhydrazyl free radical at the same time hydroalcoholic extract prepared by hot maceration process showed better reducing and total antioxidant activity.
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Indian Journal of
Scientific Publication of Indian Pharmaceutical Association
Pharmaceutical Sciences
January-February 2010, 72(1):1-148
ISSN 0250-474X
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Indexed with PubMed
Indian Journal of Pharmaceutical Sciences Volume 72 • Issue 1January - February 2010 • pp 1-120
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Indian Journal of Pharmaceutical Sciences 145January - February 2010
*Address for correspondence
E-mail: Pharmarsrividya@yahoo.com
Antioxidant and Antimicrobial Activity of Alpinia
officinarum
A. R. SRIVIDYA*, S. P. DHANABAL, V. K. MISRA AND G. SUJA
Department of Pharmaceutical Biotechnology, J. S. S. College of Pharmacy, Rocklands, Ooty-643 001, India.
Srividya et al.: Antioxidant and antimicrobial activity of Alpinia offi cinarum
Alpinia offi cinarum is a rhizome belonging to the family zingeberaeceae. Hydro alcoholic extract by hot and cold
maceration and methanol extract by percolation process Qualitative phytochemical analysis of extract of Alpinia
offi cinarum rhizome showed a majority of the compound including tannins, alkaloids, fl avonoids and saponins.
Hydroalcoholic extract prepared by hot maceration process was found to contain more phenol and fl avonol and it was
measured as 50.1 mg/g and 54.02 mg/g, respectively. All the three extracts showed moderate to potent antimicrobial
activity against the Bacillus cereus, Staphylococcus aureas, Pseudomonas auroginosa, Escherichia coli. None of the
extracts showed antifungal activity against Aspergillus niger and Candida albicans. All the three extracts showed a
concentration dependent radical scavenging activity by inhibiting diphenylpicrylhydrazyl free radical at the same
time hydroalcoholic extract prepared by hot maceration process showed better reducing and total antioxidant activity.
Key words: Alpinia offi cinarum, antimicrobial, antioxidant, cold and hot maceration, free radical scavenging,
hydroalcoholic extract
This paper deals with the antioxidant and
antimicrobial activity of Alpinia officinarum which
is not scientifically proven. Alpinia officinarum is
a rhizome belonging to the family, zingeberaeceae,
cultivated in South East Asia. This rhizome is
characterized by dark reddish brown colour which
has a strong aromatic odour. Ethnomedical uses for
this rhizome is found to be against rheumatism,
bronchial catarrh, bad breath, ulcers, whooping
cough in children, throat infections to control
incontinence[1-3]. Plant material was collected in
the month of May 2007 from the local market in
Ooty and authenticated at the Medicinal Survey
and Collection Unit, Government Arts College,
Ootacamund, Tamil Nadu, India. The dried rhizomes
of Alpinia of cinarum were powdered and extracted
with 50 % ethanol by hot and cold maceration. The
extract was ltered and the ltrate was evaporated to
dryness in a rotary evaporator to yield dark brown
semi-solids. The powdered rhizome was extracted
with methanol by percolation method. The extract
yielded a dark brown semi solid. All the extracts were
stored in the refrigerator till use[4].
The extracts obtained were tested for the qualitative
chemical tests for the identification of various
phytoconstituents such as alkaloids, carbohydrates,
protein, amino acids, steroids and sterols, glycosides,
flavonoids, tannins, triterpinoids, fixed oil[5,6]. Total
phenol content of the extract was determined by using
Folin-Ciocalteu method[7]. This test is based on the
oxidation of phenolic groups with phosphomolybdic
and phosphotungstic acids. After oxidation a green–
blue complex formed is measured at 750 nm. In a
series of test tubs, 0-4 ml of the extract (1 mg/ml
to 0.1 mg/ml) in methanol was taken, mixed with 2
ml of Folin-Ciocalteu reagent in distilled water (1:
10) and 1.6 ml of sodium carbonate. After shaking,
it was kept for 2 h for the reaction to take place.
Then absorbance was measured at 750 nm. Using
gallic acid monohydrate, standard curve was prepared
and the linearity was obtained in the range of 1-10
µg/ml. Using the standard curve, the total phenol
content was obtained. The total phenol content was
expressed as gallic acid equivalent in mg/g of the
extract[7]. The extract 0.5 ml was separately mixed
with 1.5 ml of methanol, 0.1 ml of 10% AlCl3, 0.1
ml of 1 M potassium acetate and 2.8 ml of distilled
water. After incubation at 30º, the absorbance of the
reaction mixture was measured at 415 nm. Using
rutin, standard curve was prepared and linearity was
obtained in the range of 1-10 µg/ml. Using the
standard curve total avonol content was expressed
as rutin equivalent, in mg/g or percentage w/w of the
extract[8].
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Indian Journal of Pharmaceutical Sciences146 January - February 2010
Evaluation of antimicrobial activity was performed
by Cup plate method. Sterile nutrient agar/Sabouraud
dextrose agar plates were prepared by pouring the
sterilized media in sterile Petri plates under aseptic
conditions. The test organism 0.1 ml was spread on
agar plates. Cups were made at the size of 5 mm
diameter, in the agar plates using the sterile borer.
Drug as well as the standard and DMSO solvent
control were added into the pores separately. The
plates were maintained at +4º for 1 h to allow the
diffusion of solution into the medium. All the plates
containing bacteria were incubated at 37° for 24 h
and that of fungi at 28° for 48 h[9].
In vitro antioxidant activity was evaluated by
diphenyl picryl hydrazyl (DPPH) radical scavenging
method. The principle of this assay is based on
the measurement of the scavenging ability of the
antioxidant towards the stable radical. The free radical
DPPH is reduced to the corresponding hydrazine
when it reacts with hydrogen donors, this stability is
evaluated by the decolouration assay which evaluates
the decrease in absorbance at 518-528 nm produced
by the addition of the antioxidant to a DPPH solution
in ethanol or methanol. The assay was carried out
in a 96 well microtitre plate. To 200 µl of DPPH
solution, 10 µl of each of the test sample or the
standard solution were added separately in the wells
of the microtitre plate. The final concentration of
the test and the standard solution used were 1000
µl to 0.9765 µl. The plates were incubated at 37º
for 20 min and the absorbance of each solution was
measured at 490 nm using ELISA reader against the
corresponding test and standard and the remaining
DPPH was calculated. IC50 is the concentration of
the sample required to scavenge 50% of DPPH free
radical[10]. Percentage of inhibition is calculated by
subtracting the absorbance of the sample from the
absorbance of the control divided by absorbance of
the control.
The reducing power of the extract was determined
according to the method of Oyaizu[11]. Different
concentrations of the extract (100-1000 µl) in 1
ml of distilled water were mixed with phosphate
buffer and potassium ferricyanide. The mixture was
incubated at 50º for 20 min. A portion (2.5 ml) of
trichloroacetic acid (10%) was added to the mixture,
which was then centrifuged at 3,000 rpm for 10 min.
The supernatant solution (2.5 ml) was mixed with
distilled water (2.5 ml) and ferric chloride (0.5 ml of
0.1%) and the absorbance was measured at 700 nm.
Increased absorbance of the reaction mixture indicated
increased reducing power. Ascorbic acid was used as
a reference standard; phosphate buffer (PH 6.6) was
used as blank solution[11]. The absorbance of the nal
reaction mixture of two parallel experiments were
taken was expressed as mean±standard deviation.
Total antioxidant activity of the extract was evaluated
by the phosphomolybdenum method. The assay is
based on the reduction of Mo (V1) to Mo (V) by
the extract and the subsequent formation of a green
phosphate/Mo(V) complex at acidic pH.
The extract 0.1 ml was combined with 3 ml of
the reagent solution (0.6 M sulphuric acid, 28 mM
sodium phosphate and 4 mM ammonium molbdate).
The tubes containing the reaction solution were
incubated at 95º for 90 min. The absorbance
of the solution was measured at 695 nm using
spectrophotometer against blank after cooling to room
temperature. Methanol (0.3 ml) in the place of extract
was used as the blank. The antioxidant activity is
expressed as the number of equivalent of ascorbic
acid[11,12].
TABLE 1: QUALITATIVE PHYTOCHEMICAL ANALYSIS
OF ALPINIA OFFICINARUM RHIZOME
Constituents HA(HM) HA(CM) ME(PER)
Alkaloids + + +
Glycosides - - -
Terpenoids + + -
Saponins + + +
Tannins + + +
Protein and amino
acids
+++
Flavonoids + + +
Steroids + + -
HA(HM) is hydroalcoholic extract prepared by hot maceration process,
HA(CM) is hydroalcoholic extract prepared by cold maceration process
and ME(PER) represents methanol extract prepared by percolation process.
+indicates the presence and - indicates the absence.
TABLE 2: QUANTITATIVE PHYTOCHEMICAL
CONSTITUENTS ANALYSIS OF ALPINIA
OFFICINARUM
Name of the extract Total phenol
content
(mg/g)
Total avonol
content (mg/g)
HA(HM) 50.1 54.02
HA(CM) 41.35 36.36
ME(PER) 30.6 27.64
HA(HM) is hydroalcoholic extract prepared by hot maceration process,
HA(CM) is hydroalcoholic extract prepared by cold maceration process
and ME(PER) represents methanol extract prepared by percolation process.
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Indian Journal of Pharmaceutical Sciences 147January - February 2010
Qualitative phytochemical analysis of extract of
Alpinia of cinarum rhizome showed a majority of the
compound including tannins, alkaloids, avonoids and
saponins and the results are tabulated in the Table 1.
The total phenol content of the extracts of Alpinia
officinarum rhizome is expressed as gallic acid
equivalent and the total avonol content of the extract
is expressed as rutin equivalent and hydroalcoholic
extract prepared by hot maceration process was
found to contain more phenol and avonol and it was
measured as 50.1 and 54.02 mg/g, respectively. The
results for the total phenol and favonol were tabulated
in the Table 2. All the extracts at 1000 µg/ml showed
significant anti bacterial activity in comparison
with the standard but none of the extracts showed
antifungal activity even at higher concentrations. The
results for the antimicrobial activity are tabulated in
the Table 3. All the three extracts showed moderate
to potent antibacterial activity against the B. cereus,
Staph. aureas, P. aeruginosa and E. coli. None of the
extracts showed antifungal activity against Aspergillus
niger and Candida albicans. The hydroalcoholic
extract prepared by hot maceration showed minimum
inhibitory concentration in the range of 31.25 to
500 µg/ml. The results for the minimum inhibitory
concentration were tabulated in Table 4. The
hydroalcoholic extract prepared by hot maceration
process showed a concentration-dependent radical
scavenging activity by inhibiting DPPH free radical.
All the three extracts showed moderate reducing
power when compared to ascorbic acid which has
taken as the standard antioxidant. Hydroalcoholic
extract prepared by hot maceration showed better
antioxidant activity than the other extracts. The results
TABLE 3: ANTIMICROBIAL ACTIVITY OF ALPINIA OFFICINARUM RHIZOME
Microorganism Zone of Inhibition (mm)
HA(HM) HA(CM) ME(PER) Standard
1000 µg 500µg 250µg 1000µg 500µg 250 µg 1000µg 500 µg 250 µg
Gram Positive Tetracycline 25
Bacillus cereus 27 23 18 22 17 13 19 17 12 25
Staphylococcus
aureus
38 29 24 29 27 26 27 26 18 35
Gram negative
Pseudomonas
aeruginosa
17 15 14 16 14 13 12 12 09 20
Escherichia coli 23 18 13 17 16 11 09 07 04 19
Fungi Amphotericin B
Candida albicans 08 06 05 06 00 00 00 00 00 11
Aspergillus niger 06 03 00 04 00 00 00 00 00 08
HA(HM) is hydroalcoholic extract prepared by hot maceration process, HA(CM) is hydroalcoholic extract prepared by cold maceration process and ME(PER)
represents methanol extract prepared by percolation process. Vehicle control containing DMSO did not produce any inhibition
TABLE 4: MINIMUM INHIBITORY CONCENTRATION
OF ALPINIA OFFICINARUM RHIZOME
Microorganisms MIC of Rhizome extract (µg/ml)
HA (HM) HA(CM) ME(PER)
Gram Positive
Bacillus cereus 250 500 250
Staphylococcus aureus 31.25 125 250
Gram negative
Pseudomonas auroginosa 250 500 >1000
Escherichia coli 250 500 >1000
Fungi
Candida albicans 500 1000 >1000
Aspergillus niger 500 500 >1000
HA(HM) is hydroalcoholic extract prepared by hot maceration process,
HA(CM) is hydroalcoholic extract prepared by cold maceration process and
ME(PER) represents methanol extract prepared by percolation process. MIC
is Minimum inhibitory concentration expressed in µg/ml
TABLE 5: ANTIOXIDANT ACTIVITY OF ALPINIA OFFICINARUM RHIZOME
Name of the Extract IC 50 Value of DPPH (µg/ml) Reducing power Total antioxidant activity nM
equivalent of ascorbic acid
HA (HM) 95.41±1.10 60.71±0.36 2.868±0.044
HA(CM) 123.43±0.78 59.8±0.05 2.528±0.025
ME(PER) 137.33±1.10 59.70±0.05 2.542 ±0.116
Ascorbic acid 2.75±0.09 3.50±0.05 -
HA(HM) is hydroalcoholic extract prepared by hot maceration process, HA(CM) is hydroalcoholic extract prepared by cold maceration process and ME(PER)
represents methanol extract prepared by percolation process.
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Indian Journal of Pharmaceutical Sciences148 January - February 2010
for the antioxidant activity were tabulated in the Table
5.
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Accepted 20 January 2010
Revised 26 October 2009
Received 19 June 2009
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... It possesses antibacterial activity due to change in the internal pH of the bacterial cell along with protein denaturation thereby damaging the cytoplasmic members of bacteria.18,19 It also shows excellent antimycobacterial,[20][21][22][23][24][25][26] anticancer,20,21,[23][24][25][26][27] ...
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Despite the threat of coronavirus infection, the Siddha system of medicine, India's traditional medicine, plays an important role in southern India, particularly in Tamilnadu. It contributed considerably not only in the first wave of Covid-19, but also in the second wave. The Government of Tamilnadu developed Siddha COVID-19 treatment centers for asymptomatic, mild, and moderate COVID-19 positive patients in 2020. The TPEC COVID Care Centre initiated at Vellore also one of the Centers that can be managed by Siddha medicines and Siddhar's Yogam. As of July 14, 2021, about 4525 COVID positive patients had been treated with Siddha integrated treatment at Vellore alone in the first and second waves. Kaba Sura Kudineer, Thalisathy Review Article Thillaivanan et al.; JPRI, 33(44B): 504-516, 2021; Article no.JPRI.73602 505 Vadagam, Amukkara Chooranam Mathirai, Bramanandha Bairavam Mathirai, and Adathodai Manapagu are indeed the five Siddha classical preparations used to manage the symptoms of COVID-19 positive patients at TPEC COVID Care Centre in Vellore. This Siddha medical practice is effective in conditions of symptoms and helps in the reduction of clinical outcomes. A pilot study at the same site confirmed the Siddha classical preparation's safety and effectiveness. A feedback analysis study performed at the same center also revealed that the above-mentioned Siddha classical preparations are beneficial in symptomatic treatment without causing any side effects. The medicines utilized in this study are typically proposed in other COVID care centers also in Tamilnadu. This review attempted to analyze the preclinical and clinical efficacy of Siddha Classical medicines used at that Centre for the management of COVID-19.
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Despite the threat of coronavirus infection, the Siddha system of medicine, India's traditional medicine, plays an important role in southern India, particularly in Tamilnadu. It contributed considerably not only in the first wave of Covid-19, but also in the second wave. The Government of Tamilnadu developed Siddha COVID-19 treatment centers for asymptomatic, mild, and moderate COVID-19 positive patients in 2020. The TPEC COVID Care Centre initiated at Vellore also one of the Centers that can be managed by Siddha medicines and Siddhar’s Yogam. As of July 14, 2021, about 4525 COVID positive patients had been treated with Siddha integrated treatment at Vellore alone in the first and second waves. Kaba Sura Kudineer, Thalisathy Vadagam, Amukkara Chooranam Mathirai, Bramanandha Bairavam Mathirai, and Adathodai Manapagu are indeed the five Siddha classical preparations used to manage the symptoms of COVID-19 positive patients at TPEC COVID Care Centre in Vellore. This Siddha medical practice is effective in conditions of symptoms and helps in the reduction of clinical outcomes. A pilot study at the same site confirmed the Siddha classical preparation's safety and effectiveness. A feedback analysis study performed at the same center also revealed that the above-mentioned Siddha classical preparations are beneficial in symptomatic treatment without causing any side effects. The medicines utilized in this study are typically proposed in other COVID care centers also in Tamilnadu. This review attempted to analyze the preclinical and clinical efficacy of Siddha Classical medicines used at that Centre for the management of COVID-19.
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