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Journal of Ethnopharmacology 104 (2006) 113–118
Vitamin C content and antioxidant activity of the fruit and of the Ayurvedic
preparation of Emblica officinalis Gaertn.
P. Scartezzini a,∗, F. Antognonia, M.A. Raggib, F. Polia, C. Sabbioni b
aFaculty of Pharmacy, Department of Biology, University of Bologna, Via Irnerio 42, 40126 Bologna, Italy
bFaculty of Pharmacy, Department of Pharmaceutical Sciences, University of Bologna, Via Belmeloro 6, 40126 Bologna, Italy
Received 13 October 2004; received in revised form 27 July 2005; accepted 26 August 2005
Available online 13 October 2005
Abstract
Emblica officinalis Gaertn. is one of the most important plants of ¯
Ayurved, the traditional Indian medicine. In this ancient medicine, the fruit of
Emblica officinalis is processed according to a method named “Svaras Bhavana”, whereby the therapeutic potential of the plant is enhanced by
treating the main herb with its own juice. For many years, the activity of the fruits was attributed to the high content of ascorbic acid; however,
this has recently been questioned. The aim of the paper is to clarify this matter. A reliable and feasible HPLC method with diode array detection
has been developed for the determination of ascorbic acid in Emblica fruit and particularly in Emblica fruit processed according to the Ayurvedic
method. The antioxidant effects have also been evaluated in comparison to the real levels of Vitamin C by different antioxidant tests. The data
obtained show that the Emblica fruit contains ascorbic acid (0.40%, w/w), and that the Ayurvedic method of processing increases the healthy
characteristics of the fruit thanks to a higher antioxidant activity and a higher content of ascorbic acid (1.28%, w/w). It has also been found that
Vitamin C accounts for approximately 45–70% of the antioxidant activity.
© 2005 Elsevier Ireland Ltd. All rights reserved.
Keywords: Emblica officinalis;¯
Ayurveda; Ascorbic acid; Liquid chromatography; Antioxidant activity
1. Introduction
Emblica officinalis Gaertn. (Phyllanthus emblica L.;
Dichelactina nodicaulis Hance; Emblica arborea Raf.; Phyllan-
thus glomeratus Wall.; Cicca emblica Kurz.; Diasperus emblica
Kuntze) is a medium to large deciduous tree belonging to a
small subgenus of trees of the Euphorbiaceae growing in India,
Sri Lanka, Pakistan, Uzbekistan, S.E. Asia, and China. Emblica
(Amla in Hindi) grows wild and is cultivated up to 1400m a.s.l.;
in India, the most common cultivars are “Chakaiya”, “Banarsi”,
and “Francis” (Scartezzini and Speroni, 2000). The previous
name, Phyllanthus emblica L., was attributed by Linneaus with
reference to a peculiar characteristic of this plant. The branches
of this tree are oddly flattened in the manner of a leaf; the
flowers bloom from the edges of these leaf-like branches, thus
the name Phyllanthus, from the Greek words “phyllon” (leaf)
and “anth`os” (flower). The name Emblica derives almost cer-
∗Corresponding author.
E-mail address: scartezzini@biocfarm.unibo.it (P. Scartezzini).
tainly from the “corruption” of the Sanskrit name “Amlika”,
although some authors believe that it could derive from the
“corruption” of the Arabic word “Embelgi” used by Arabic
physicians to name its fruit. In Sanskrit, Emblica has many
synonyms: Amalaki (pure, clean), Dhatriphala (nurse fruit),
Amritaphala (fruit of immortality), and others. All of these syn-
onyms show how important this plant is in traditional Indian
medicine. In Malaysia, this plant is so renowned that a city and
a river bear its name: Malacca. The fruits of Emblica are widely
consumed raw, cooked, or pickled, but they are also principal
constituents of many Ayurvedic preparations (Scartezzini and
Speroni, 2000). Indeed, Emblica is one of the most important
plants of ¯
Ayurved, the Indian Traditional Medicine. According
to the two main classic texts on ¯
Ayurved, Charak Samhita and
Sushrut Samhita, Amalaki is regarded as “the best among reju-
venative herbs”, “useful in relieving cough and skin disease”,
and “the best among the sour fruits”. There are two historically
ascertained events in which this plant was used for medical pur-
poses: the famine of 1939–1940 and the cases of scurry in the
Indian army in Nassirdab, today known as Rajasthan, in 1837
(Srinivasan, 1944). For many years, the therapeutic potential
0378-8741/$ – see front matter © 2005 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.jep.2005.08.065
114 P. Scartezzini et al. / Journal of Ethnopharmacology 104 (2006) 113–118
of the fruits was attributed to their high ascorbic acid con-
tent, about 1g Vitamin C per 100mL of fresh juice (Kapoor,
1990). Because of the presence of tannins, the ascorbic acid does
not oxidise even in dried fruit, thus maintaining its antiscurvy
property unchanged. Many studies have been published on the
comparison between the effects of ascorbic acid and Emblica.
The fruit extract has many pharmacological activities: it inhibits
micronucleiformation, sister chromatid exchanges,clastogenic-
ity and mutagenicity induced by metals such as lead, aluminum,
cadmium, nickel, and caesium; it preserves against radiations
(Scartezzini and Speroni, 2000), possesses antidiabetic activity
(Sabu and Kuttan, 2002), inhibits clastogenicity of benzopyrene
and cyclophosphamide (Sharma et al., 2000; Haque et al., 2001),
is gastroprotective (Al-Rehaily et al., 2002), cytoprotective, and
immunomodulating (Sai Ram et al., 2002). In recent years,
new pharmacological activities have been found for Emblica:
it has cytoprotective activity against chromium (Sai Ram et al.,
2003), protects against oxidative stress in ischemic-reperfusion
injury (Rajak et al., 2004), shows antivenom capacity (Alam
and Gomes, 2003), ameliorates hyperthyroidism and hepatic
lipid peroxidation (Panda and Kar, 2003), displays antipro-
liferative activity on MCF7 and MDA-MB-231 breast cancer
cell lines (Lambertini et al., 2003), shows antitussive activ-
ity (Nosal’ova et al., 2003), and induces apoptosis in Dalton’s
Lymphoma Ascites and CeHa cell lines (Rajeshkumar et al.,
2003).
In contrast with previous authors, Ghosal et al. (1996)
asserted that Emblica fruits do not contain ascorbic acid,
either in the free or in the conjugated form, but contain two
new hydrolysable tannins of low molecular weight, namely
emblicanin A (2,3-di-O-galloyl-4,6-(S)-hexahydroxydiphe-
noyl-2-keto-glucono-␦-lactone) and emblicanin B (2,3,4,6-
bis-(S)-hexahydroxydiphenoyl-2-keto-glucono-␦-lactone), and
other tannins, such as punigluconin (2,3-di-O-galloyl-4,6-
(S)-hexahydroxydiphenoylgluconic acid) and pedunclagin
(2,3,4,6-bis-(S)-hexahydroxydiphenoyl-d-glucose). The two
emblicanins exhibited a very strong antioxidant action; more-
over, they improved the efficacy of Vitamin C in reducing
dehydroascorbic acid to ascorbic acid (Ghosal et al., 1996;
Bhattacharya et al., 2000a). Some studies published later
examined the pharmacological activities of these tannins
and reported that they exhibit antioxidant activity in vitro
and in vivo. In particular, they preserve erythrocytes against
oxidative stress induced by asbestos (Ghosal et al., 1996), show
antioxidant effects in rat brain (Bhattacharya et al., 2000a),
exert a prophylactive effect against neuroleptic-induced tardive
dyskinesia (Bhattacharya et al., 2000b), show antioxidant
activity against ischemia reperfusion (Bhattacharya et al.,
2002), antiulcerogenic effects (Sairam et al., 2002), and
anticataractogenic effects in vitro (Suryanarayana et al., 2004).
The goal of the present paper was to evaluate the extent to
which Vitamin C content determines the antioxidant activity
of the fruit, and to what extent the traditional Ayurvedic pro-
cessing method (called Svaras Bhavana) might influence the
biological activity of the Ayurvedic preparation. In Ayurvedic
terminology, Svaras Bhavana means “to increase the potency
by treating the main herb with its own juice”. For this purpose,
a simple and rapid liquid chromatographic (HPLC) method
was developed to determine Vitamin C content in dried and
frozen Emblica fruit, in Emblica fruit processed according to the
Ayurvedic method, and in an industrial extract (Merck). Their
respectiveantioxidantactivitieswere evaluatedby meansof2,2-
azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium
salt (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) tests in
extracts having equal ascorbic acid levels.
2. Materials and methods
2.1. Materials
The wild fruit of Emblica were authenticated by Dr. M.R.
Uniyal, Maharishi ¯
Ayurveda Product Ltd., Noida, India. The
commercial Ayurvedic samples of Emblica analysed in this
study (Emblimap—MA-390), batch no. April 02 and April
01, were bought from Maharishi ¯
Ayurveda Product Italy (VR,
Italy). The dry extract of Emblica was supplied by Dr. Phyllis
Carter, External Corporate Communications of Merck KGaA
(Darmstadt, Germany), batch no. K4908265 (titre in total small
hydrolysable tannoids 69%). Waters C18 Spherisorb 5 m
ODS2 HPLC columns (Ireland) was used. Solvents (water and
methanol) were of HPLC grade and purchased from Carlo Erba
Reagenti S.p.a. (Milan, Italy). The Folin–Ciocalteu reagent and
the DPPH radical and ABTS were bought from Sigma (Milan,
Italy). Ascorbic acid (85%, m/m), phosphoric acid, dipotas-
sium peroxidisulfate (K2S2O8) and NaOH were obtained from
CarloErba ReagentiS.p.a.Ultrapure water(18.2Mcm−1)was
obtained by means of a Millipore (Milford, MA, USA) MilliQ
apparatus.
2.2. Apparatus
The HPLC apparatus consisted of an Agilent (Waldbronn,
Germany) 1100 series isocratic pump and an Agilent 1100 series
diode array (DAD) detector. The data system consisted of an HP
CORE Chemstation LC 3D (Waldbronn, Germany).
2.3. Standard solution and sample preparation
The stock solution of ascorbic acid was 10 mg mL−1in water.
All dilutions subsequent were made in water.
Five hundred milligrams of the finely powdered fruit mate-
rial or Ayurvedic preparation was extracted once with 50mL of
water for 30min. All samples were filtered through a 0.22 m
membrane filter from Millipore (Malsheim, France).
2.4. HPLC method
2.4.1. Chromatographic conditions
TheDADdetector wasset in the200–350 nmrange andquan-
titative analysis was performed at 243nm with reference on at
308nm. Separationwas achievedusinga Waters C18 Spherisorb
5m ODS2 Column. The mobile phase was constituted of a
mixture (97.5:2.5, v/v) of phosphate buffer 100mM, pH 2.5,
P. Scartezzini et al. / Journal of Ethnopharmacology 104 (2006) 113–118 115
and methanol. The flow rate was 0.4mL min−1and the injec-
tion loop was 20L.
2.4.2. Calibration curves
Standard solutions of ascorbic acid, in the 1–50gmL−1
concentration range, were prepared and injected in the HPLC
system.
The analyte peak area values were plotted against the corre-
sponding concentrations of the analyte (expressed as gmL−1),
and the calibration curve was constructed by means of the least-
square method.
2.4.3. Sample analysis
An aliquot of the sample, after suitable dilution with water,
was injected into the HPLC.
Every sample solution was injected in triplicate. The area
value of ascorbic acid peaks obtained injecting the extract of
Emblica into the HPLC was interpolated on the calibration
curve.
2.4.4. Precision assays
A single sample of Emblica was analysed six times. Percent-
age standard deviations (R.S.D.%) were obtained for repeata-
bility (intra-day precision) and intermediate precision (inter-day
precision).
2.4.5. Accuracy assays
Known amounts of ascorbic acid standard solution were
added to known amounts of Emblica sample. The method accu-
racy was evaluated by calculating the difference between the
spiked sample peak areas and the original sample peak areas,
then comparing these differences with the peak area obtained
by injecting standard solutions having the same concentration
as the sample spiking.
2.5. Total polyphenol content
Total polyphenol content of Emblica was determined by the
Folin–Ciocalteu colorimetric method (Ragazzini and Veronesi,
1973). The methanolic Emblica extract was mixed with 0.5mL
of the Folin–Ciocalteu reagent and 0.5 mL of 10% Na2CO3, and
the absorbance was measured at 760nm after 1 h incubation at
room temperature. Total polyphenols were expressed as gallic
acid equivalents.
2.6. Scavenging activity on the DPPH radical
The antioxidant activity of the different Emblica extracts was
assayed by their scavenging effect on the DPPH radical (Wang
et al., 1998). Emblica extracts were first analysed by HPLC for
the quantitative determination of ascorbic acid content and then
diluted in order to have exact concentrations of ascorbic acid in
the range 0.01–0.5mg mL−1(corresponding to 0.001–0.05%).
Dilutionwas also performedon pureascorbicacid solution.Both
Emblica samples and pure Vitamin C solution were added to a
methanol solution of DPPH (1×10−4M), shaken vigorously,
and kept in the dark for 30min. The absorbance of samples
was measured with a spectrophotometer (Perkin-Elmer 554) at
517 nm against a blank of methanol. Positive controls consisted
of ascorbic acid. The antioxidant activity of each extract, con-
taining same concentration of ascorbic acid, was determined as
follows, according to the percentage of DPPH decolouration:
Decolouration(%) =(AbsBlank −Abssample)×100
AbsBlank
All tests were run five times and averaged.
2.7. ABTS radical cation decolourization assay
The antioxidant capacity of each extract of Emblica was
evaluated by studying its ability to bleach the 2,2-azinobis-(3-
ethylbenzothiazoline-6-sulfonic acid) radical (ABTS•+)(Miller
et al., 1993). Concentrations of ascorbic acid and dilutions for
the assay were performed as described above. ABTS•+was pro-
duced by the reaction of ABTS solution (2mM) with 70 mM
potassium persulfate (K2S2O8) (both solutions were prepared
in bidistilled water) for 12–16h, in the dark and at room tem-
perature. The stock solution was diluted in phosphate buffered
saline (PBS) so as to achieve an absorbance of 0.70±0.02 at
734nm. Nine hundred and ninety microliters of diluted ABTS
solution was mixed with 10L of sample. The absorbance at
734nm was taken at 30 ◦C exactly 1min after initial mixing.
The percentage decrease of absorbance was calculated as fol-
lows:
Inibition(%) =(AbsBlank −Abssample)×100
AbsBlank
All test were run five times and averaged.
3. Results and discussion
3.1.1. HPLC method
Attempts were made to evaluate Vitamin C content in
Emblica extracts using previously published methods (Finley
and Duang, 1981; Kim et al., 2002; Valls et al., 2002). In our
experimental conditions, separation of the ascorbic acid peak
from interference of the matrix was unsatisfactory; therefore,
a new HPLC procedure was developed. The analysis was per-
formed using a C18 reverse phase column, and the composition
(% methanol), the pH, and the flow rate of the mobile phase were
investigated. The best conditions were as follows: the mobile
phaseconsisted ofamixture of100 mMphosphatebuffer,pH2.5
(97.5%),and methanol(2.5%), witha flow rateof 0.4mL min−1.
The DAD detector was set in the 200–350nm range, and the
quantitative analyses were carried at 243nm. Under these con-
ditions, a neat peak of ascorbic acid was detected at 9.9 min. The
spectrum was recorded and maximum absorbance was found at
243nm (data not shown).
A good linearity was found from 1 to 50gmL−1ascor-
bic acid, and the linear regression equation was y=145.44x−
68.938 (rc=0.9996), where yis the peak height, expressed
as arbitrary units, and xis the ascorbic acid concentration,
116 P. Scartezzini et al. / Journal of Ethnopharmacology 104 (2006) 113–118
Table 1
Analytical parameters of the HPLC procedure for the ascorbic acid quantitation
Parameter Ascorbic acid (gmL−1)
Linearity range 1–50
Regression equationay=145.44x−68.938
Correlation coefficient (rc) 0.9996
Analyte concentration 10 25 50
Repeatability (R.S.D.%)b2.36 0.98 1.1
Intermediate precision (R.S.D.%)b3.20 1.90 1.48
Limit of detection (LOD) 0.01
Limit of quantification (LOQ) 0.04
ay: peak area and x: concentration (gmL−1).
bn=6.
expressed as gmL−1. Limit of quantitation (LOQ) and limit of
detection (LOD) values, calculated according to the USP XXV
Edition Guidelines (United States Pharmacopoeia, 2003), were
0.04 and 0.01gmL
−1, respectively.
Repeatability and intermediate precision values, obtained at
concentration values minimum, medium, and high, were in the
range 0.98–3.2%. The validation parameters are reported in
Table 1.
Thevolume ofwater andthe timerequired foroptimal extrac-
tion (50–200mL and 0.5–24 h, respectively) were also investi-
gated. Results indicate that the best quantitative extraction of
VitaminCwas obtained bytreating 500mg ofmatrixwith 50mL
of water for 30 min. The extract was injected into the HPLC after
suitable dilutions with water.
To confirm that the extraction procedure under these con-
ditions was quantitative, two successive extracts of the same
sample of Emblica fruit were injected separately into the HPLC
apparatus. In the second extraction step, the analyte peak area
was 1% of that obtained in the first extraction step. These results
indicate that the extraction procedure is easy, rapid, and reliable.
The chromatogram of an extract of processed Emblica
recorded at 243nm with reference off showed two peaks very
close to each other (Fig. 1a). A comparison between the spectra
of these peaks with that of pure ascorbic acid confirmed that
the retention time of the analyte was 9.9min. Moreover, it is
possible to eliminate the interference peak by setting the ref-
erence of the detector (which is automatically subtracted from
the signal at 243nm) at 308 nm (Fig. 1b, reference on). In fact,
the absorbance values of the interference spectrum at 243 and
308 nm were the same, while the absorbance of ascorbic acid at
308nm is practically equal to zero.
3.2. Application to Emblica extracts
The above-described procedure was used to analyse differ-
ent products of Emblica, and in particular frozen fruit, dried
Fig. 1. Chromatograms of an extract of processed fruit recorded at 243nm with
detector reference: (a) off and (b) at 308nm.
fruit, processed (Ayurvedic preparation) fruit, and a commercial
Merck extract. As shown in Table 2, ascorbic acid is present in
all the examined products, ranging from 0.37% in dried fruit to
2.00% in the Merck extract. The precision was calculated for
processed Emblica, and gave good repeatability and inter-day
precision values, lower than 3%.
The accuracy of the method, obtained by means of recovery
assays, gave satisfactory results, always between 102 and 104%.
The dried and frozen fruits contain about the same amount
of Vitamin C (0.4%), while a higher level of Vitamin C was
found in the processed fruit (Ayurvedic preparation). In fact,
the Ayurvedic preparation (called “Svaras Bhavana”) consists
in mixing a specific quantity of the dried fruit powder with its
own fresh juice for a few hours; then, it is dried and powdered
again (fruit processed once). This powder is again mixed with
the juice of fresh Emblica, dried, and powdered again; this is
the second processing. The procedure is repeated for up to 21
times. The Ayurvedic procedure aims to increase the health effi-
ciency of the product (The Ayurvedic Formulary of India, 2003).
Indeed, processed fruit contained up to three times more ascor-
bic acid than the dried unprocessed ones; also, the total content
of polyphenolic compounds was higher (Table 2), suggesting
that the Ayurvedic method of processing the fruit increased its
beneficial properties. The difference between dried and frozen
fruit is also worth noting. Both of them have a very similar ascor-
bic acid content, and this suggests that the tannins present in the
fruit may prevent the oxidation of Vitamin C during the drying
Table 2
Ascorbic acid and total polyphenol content in different products of Emblica officinalis
Frozen fruit Dried fruit Processed fruit Merck extract
Ascorbic acid (%) 0.40 ±0.01 0.37 ±0.01 1.28 ±0.01 2.00 ±0.01
Total polyphenols content (GAEa) 201.8 ±0.8 206.9 ±0.9 255.2 ±1.2 200.1 ±1.3
aExpressed as gallic acid equivalents.
P. Scartezzini et al. / Journal of Ethnopharmacology 104 (2006) 113–118 117
process, thereby maintaining the nutritional quality of the fruit
unaltered.
The amount of total polyphenols in the matrices analysed was
performed with the aim of verifying if the increase in ascorbic
acid was accompanied by a similar increase in the content of
these compounds, which are also involved in antioxidant activ-
ity. Results showed that processed Emblica fruits had a higher
level of total polyphenols (about 25%) than the other matrices
(Table 2).
3.3. Antioxidant activity
The antioxidant activity of all extracts was evaluated with
the ABTS and DPPH tests. These methods are rapid, sensitive,
reproducible, and require simple conventional laboratory equip-
ment. They were selected for their different characteristics. In
fact, the DPPH test is particularly suitable for the evaluation of
antioxidant activity of crude extracts (Poli et al., 2003), while
the ABTS one is usually applicable also to biological tissues (Re
et al., 1999).
The antioxidant assays were performed on aqueous extracts
preparedas describedabove with fivereplicates persample. S.D.
ranged from 0.2 to 1.5%.
DPPH test results (Fig. 2) indicate that, when the con-
centration of ascorbic acid in the samples was higher than
0.25mg mL−1(corresponding to 0.025%), the bleaching power
of the solutions was too high; for this reason, it is not possible
to estimate the antioxidant role of Vitamin C versus the other
compounds.
The results of the ABTS test are plotted in Fig. 3. Also, this
test cannot be applied to samples with an ascorbic acid concen-
tration higher than 0.25 mgmL−1for the same reason described
above.
In any case, all extracts exhibited an antioxidant activity even
at the lowest concentration tested, i.e. 0.01mg mL−1.
A comparison between antioxidant activity of ascorbic acid
standard solution and those of the three different Emblica prod-
ucts, measured at the same concentration of ascorbic acid, shows
that the processed fruit has the highest antioxidant activity. The
resultsof bothtests alsoshow thatthe antioxidant activityof pro-
cessed and dried Emblica does not depend only upon Vitamin
Fig. 2. Antioxidant activity assayed by DPPH test (expressed as % bleaching)
of extracts from different matrices of Emblica, containing known amount of
ascorbic acid (() pure ascorbic acid; () processed fruit; () dried fruit; (䊉)
Merck extract).
Fig. 3. Antioxidant activity assayed by ABTS test (expressed as % bleaching)
of extracts from different matrices of Emblica, containing known amount of
ascorbic acid (() pure ascorbic acid; () processed fruit; () dried fruit; (䊉)
Merck extract).
C content. In fact, in aqueous extracts of processed and dried
fruit, Vitamin C accounts for 45–70% of the antioxidant activ-
ity. In particular, the antioxidant activity of processed fruit is due
to ascorbic acid for only 60% or less, while in the other prod-
ucts this percentage increases. These results are in agreement
with those indicating that the presence of emblicanins and rutin
favours the conversion of dehydroascorbic acid to ascorbic acid
(Ghosal et al., 1996), and confirm that the Ayurvedic method
of processing increases the amount of antioxidant compounds
present in the final product.
4. Conclusions
Until the 1990s, it was believed that Emblica officinalis con-
tained a large amount of Vitamin C. Ghosal et al. (1996) pub-
lished a paper where he asserted that the fruit of this plant does
not contain any ascorbic acid, but contains emblicanins which
are responsible for the biological activity of the fruit. Our results
indicate that these fruits do indeed contain ascorbic acid, albeit
in a smaller quantity than previously reported (Kapoor, 1990).
The evaluation of antioxidant activity shows that it is due in a
large percentage to presence of ascorbic acid. It is however pos-
sible that the tannins identified by Ghosal et al. (1996) favour
the recycling of ascorbic acid, thereby increasing the antioxidant
activity of the Emblica fruit.
TheHPLC methoddescribed hererepresents areliable proce-
dure for analysis of ascorbic acid in Emblica officinalis: indeed,
the selectivity was increased in order to obtain a good separation
between ascorbic acid and interference peaks, allowing a rapid
and accurate determination, which could be helpful for scientific
as well as commercial applications.
Theresults ofthis paper alsoindicate thatthe Ayurvedic tradi-
tionalmethod ofprocessing thefruit (Svaras Bhavana) increases
its beneficial characteristics. In fact, the amount of ascorbic acid
and polyphenols found in processed fruit as well as the antioxi-
dant activity was higher than in dried fruit.
References
Alam, M.I., Gomes, A., 2003. Snake venom neutralization by Indian medic-
inal plants (Vitex negundo and Emblica officinalis) root extract. Journal
of Ethnopharmacology 86, 75–80.
118 P. Scartezzini et al. / Journal of Ethnopharmacology 104 (2006) 113–118
Al-Rehaily, A.J., Al-Howiriny, T.A., Al-Sohaibani, M.O., Rafatullah, S.,
2002. Gastroprotective effects of “amla” Emblica officinalis on in vivo
test models in rats. Phytomedicine 9, 515–522.
Bhattacharya, S.K., Bhattacharya, D., Muruganandam, A.V., 2000b. Effect of
Emblica officinalis tannoids on a rat model of tardive dyskinesia. Indian
Journal Experimental Biology 38, 945–947.
Bhattacharya, S.K., Bhattacharya, D., Sairam, K., Ghosal, S., 2002. Effect
of bioactive tannoid principles of Emblica officinalis on ischemia-
reperfusion-induced oxidative stress in rat heart. Phytomedicine 9,
171–174.
Bhattacharya, A., Ghosal, S., Bhattacharya, S.K., 2000a. Antioxidant activ-
ity of tannoid principles of Emblica officinalis (amla) in chronic stress
induced changes in rat brain. Indian Journal Experimental Biology 38,
877–880.
Finley, J.W., Duang, E., 1981. Resolution of ascorbic, dehydroascorbic and
diketogulonic acids by paired-ion reversed-phase chromatography. Journal
of Chromatography 207, 449–453.
Ghosal, S., Tripathi, V.K, Chauhan, S., 1996. Active constituents of
Emblica officinalis: Part 1—the chemistry and antioxidant effects of two
hydrolysable tannins, emblicanin A and B. Indian Journal of Chemistry
35B, 941–948.
Haque, R., Bin-Hafeez, B., Ahmad, I., Parvez, S., Pandey, S., Raisud-
din, S., 2001. Protective effects of Emblica officinalis Gaertn. in
cyclophosphamide-treated mice. Human & Experimental Toxicology 20,
643–650.
Kapoor, L.D., 1990. Handbook of Ayurvedic Medicinal Plants. CRC Press,
Boca Raton.
Kim, H.S., Jang, D.K., Woo, D.K., Woo, K.L., 2002. Simultaneous determi-
nation of the water soluble vitamins in multi-nutrients tablets by reversed-
phase high-performance liquid chromatography. Journal Food Science and
Nutrition 7, 12–17.
Lambertini, E., Piva, R., Khan, M.T., Lampronti, I., Bianchi, N., Borgatti, M.,
Gambari, R., 2003. Effects of extracts from Bangladeshi medicinal plants
on in vitro proliferation of human breast cancer cell lines and expression
of estrogen receptor alpha gene. International Journal of Oncology 24,
419–423.
Miller, N.J., Rice-Evans, C., Davies, M.J., Gopinathan, V., Milner, A., 1993.
A novel method for measuring antioxidant capacity and its application to
monitoring the antioxidant status in premature neonates. Clinical Sciences
84, 407–412.
Nosal’ova, G., Mokry, J., Hassan, K.M., 2003. Antitussive activity of the fruit
extract of Emblica officinalis Gaertn. (Euphorbiaceae). Phytomedicine 10,
583–589.
Panda, S., Kar, A., 2003. Fruit extract of Emblica officinalis ameliorates
hyperthyroidism and hepatic lipid peroxidation in mice. Pharmazie 58,
753–755.
Poli, F., Muzzoli, M., Sacchetti, G., Tassinato, G., Lazzarin, R., Bruni, A.,
2003. Antioxidant activity of supercritical CO2extracts of Helichrysum
italicum. Pharmaceutical Biology 41, 379–383.
Ragazzini, E., Veronesi, G., 1973. Quantitative analysis of phenolic com-
pounds after thin-layer chromatographic separation. Journal of Chro-
matography 77, 369–375.
Rajak, S., Banerjee, S.K., Sood, S., Dinda, K.A., Gupta, Y.K., Maulik,
S.K., 2004. Emblica officinalis causes myocardial adaptation and protects
against oxidative stress in ischemic-reperfusion injury in rats. Phytother-
apy Research 18, 54–60.
Rajeshkumar, N.V., Pillai, M.R., Kuttan, R., 2003. Induction of apoptosis in
mouse and human carcinoma cell lines by Emblica officinalis polyphenols
and its effect on chemical carcinogenesis. Journal of Experimental and
Clinical Cancer Research 22, 201–212.
Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., Rice-Evans,
C., 1999. Antioxidant activity applying an improved ABTS radical
cation decolorization assay. Free Radical Biology and Medicine 26,
1231–1237.
Sabu, M.C., Kuttan, Ramadasan, 2002. Anti-diabetic activity of medicinal
plants and its relationship with their antioxidant property. Journal of
Ethnopharmacology 81, 155–160.
Sai Ram, M., Neetu, D., Deepti, P., Vandana, M., Ilavazhagan, G., Kumar,
D., Selvamurthy, W., 2003. Cytoprotective activity of amla (Emblica
officinalis) against chromium (VI) induced oxidative injury in murine
macrophages. Phytotherapy Research 17, 430–433.
Sai Ram, M., Neetu, D., Yogesh, B., Anju, B., Dipti, P., Pauline, T., Sharma,
S.K., Sarada, S.K.S., Ilavazhagan, G., Kumar, D., Selvamurthy, W., 2002.
Cyto-protective and immunomodulating properties of amla (Emblica offic-
inalis) on lymphocytes: an in-vitro study. Journal of Ethnopharmacology
81, 5–10.
Sairam, K., Rao, Ch.V., Dora Babu, M., Vijay Kumar, K., Agrawal, V.K.,
Goel, R.K., 2002. Antiulcerogenic effect of methanolic extract of Emblica
officinalis: an experimental study. Journal of Ethnopharmacology 82,
1–9.
Scartezzini, P., Speroni, E., 2000. Review on some plants of Indian traditional
medicine with antioxidant activity. Journal of Ethnopharmacology 71,
23–43.
Sharma, N., Trikha, M., Raisuddin, S., 2000. Inhibitory effect of
Emblica officinalis on the in vivo clastogenicity of benzo(a)pyrene and
cyclophosphamide in mice. Human & Experimental Toxicology 19,
377–384.
Srinivasan, M., 1944. Indian goosberry (Phyllanthus emblica). Nature 153,
684.
Suryanarayana, P., Kumar, P.A., Saraswat, M., Petrash, J.M., Reddy, G.B.,
2004. Inhibition of aldose reductase by tannoid principles of Emblica
officinalis: implications for the prevention of sugar cataract. Molecular
Vision 10, 148–154.
The Ayurvedic Formulary of India, Part I, 2003. Second revised English ed.,
Government of India, pp. 350–354.
United States Pharmacopoeia, 25th ed., 2003. United States Pharmacopoeial
Convention, Rockville, MD, pp. 2439–2442.
Valls, F., Sancho, M.T., Fernandez-Muino, M.O., Alonso-Torre, S., Checa,
M.A., 2002. High-pressure liquid chromatographic determination of
ascorbic acid in cooked sausages. Journal of Food Protection 65,
1771–1774.
Wang, M., Li, J., Rangarajan, M., Shao, Y., La Voie, E.J., Huang, T.C., Ho,
C.T., 1998. Antioxidant phenolic compounds from sage (Salvia offici-
nalis). Journal Agricultural and Food Chemistry 46, 4869–4873.
