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PHYTOCHEMICAL AND PHARMACOLOGICAL STUDIES OF ETHANOLIC EXTRACT FROM THE LEAF OF MANGROVE PLANT PHOENIX PALUDOSA ROXB

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The objective of this study was to evaluate the phytochemical constituents and pharmacological activity of the crude ethanolic extract of the leaves of Phoenix paludosa, collected from the sundarbans of Sathkhira range (specifically from Koromjol), Bangladesh. Phytochemical analyses were done by chemical group test. Antidiarrhoeal activities were assessed by castor oil-induced diarrhoea models in mice. Analgesic activities were studied using the model of acetic acid-induced writhing in mice, cytotoxic activity by brine shrimp lethality bioassay and antioxidant activity was measured by 1,1-diphenyl-2picrylhydrazyl (DPPH) methods. Steroids, glycosides, tannins, flavonoid and gums were found in the group tests. The extract exhibited moderate cytotoxicity with LC50 of 6.67 μg/mL and LC90 of 34.81 μg/mL and also showed statistically significant (p=0.009) analgesic effect. In antidiarrhoeal test, the extract increased the latent period significantly (p<0.05) and reduced the total number of passing liquid stool at all doses. When tested for in vitro antioxidant activity against stable DPPH, it was found to exhibit prominent free radical scavenging effect. The results obtained in this study suggested that P. paludosa has antidiarrhoeal and analgesic activities with moderate cytotoxic effect. It also has prominent antioxidant effect. These activity may be due to the presence of steroid, flavonoids and saponins in the extract.
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Malaysian Journal of Pharmaceutical Sciences
Vol. 8, No. 2, 5969 (2010)
© Penerbit Universiti Sains Malaysia, 2011
PHYTOCHEMICAL AND PHARMACOLOGICAL STUDIES OF
ETHANOLIC EXTRACT FROM THE LEAF OF MANGROVE PLANT
PHOENIX PALUDOSA ROXB.
ASHURA AKTER LIMA1, RAMENDU PARIAL2, MANISHA DAS3 AND
ASISH KUMAR DAS1
1Pharmacy Discipline, School of Life Science, Khulna University, Khulna-9208, Bangladesh
2Department of Biochemistry and Molecular Biology, University of Chittagong,
Chittagong-4331, Bangladesh
3Govt. Ayurvedic College and Hospital, Guwahati-14, Assam, India
The objective of this study was to evaluate the phytochemical constituents and pharmacological
activity of the crude ethanolic extract of the leaves of Phoenix paludosa, collected from the
sundarbans of Sathkhira range (specifically from Koromjol), Bangladesh. Phytochemical analyses
were done by chemical group test. Antidiarrhoeal activities were assessed by castor oil-induced
diarrhoea models in mice. Analgesic activities were studied using the model of acetic acid-induced
writhing in mice, cytotoxic activity by brine shrimp lethality bioassay and antioxidant activity was
measured by 1,1-diphenyl-2picrylhydrazyl (DPPH) methods. Steroids, glycosides, tannins,
flavonoid and gums were found in the group tests. The extract exhibited moderate cytotoxicity
with LC50 of 6.67 μg/mL and LC90 of 34.81 μg/mL and also showed statistically significant
(p=0.009) analgesic effect. In antidiarrhoeal test, the extract increased the latent period
significantly (p<0.05) and reduced the total number of passing liquid stool at all doses. When tested
for in vitro antioxidant activity against stable DPPH, it was found to exhibit prominent free radical
scavenging effect. The results obtained in this study suggested that P. paludosa has antidiarrhoeal
and analgesic activities with moderate cytotoxic effect. It also has prominent antioxidant effect.
These activity may be due to the presence of steroid, flavonoids and saponins in the extract.
Keywords: Phoenix paludosa, Analgesic, Antidiarrhoeal, Antioxidant, Ethanolic extract,
Cytotoxic
INTRODUCTION
The importance of medicinal plants in traditional health care practice and in providing
clues to new areas of drug research and biodiversity conservation is now well recognised.
About 80% of the world’s population relies on the use of traditional medicine, which is
predominantly based on plant material (WHO 1993). Scientific studies on a number of
medicinal plants indicated that promising phytochemical compounds can be developed
for many health problems (Gupta 1994). Herbal drugs have gained importance in recent
years because of their efficacy and cost effectiveness. These drugs are invariably single
plant extracts or fractions or mixtures of fractions/extracts from different plants, which
Corresponding author: parial.r@gmail.com
Ashura Akter Lima et al. 60
Malay J Pharm Sci, Vol. 8, No. 2 (2010): 5969
have been carefully standardised for their safety and efficacy (Subramoniam and
Pushpangadan 1999). Several plants of mangrove forest are used for medical purposes e.g.
Acanthus ebracteatus Vahl for chronic wound (Sujamnong 1979), the bark of Rhizophora
apiculata Blume for diarrhoea and wound (School of Thai Traditional Medicine 1981), and
the bark of Rhizophora mucronata for diarrhoea (Phongboonrod 1976). The plants in the
mangrove forest showed the potential as a source of antioxidant and cancer
chemoprevention agents (Bunyapraphatsara et al. 2003).
Phoenix paludosa (mangrove date palm) is a species of flowering plant in the palm
family, indigenous to coastal regions of India, Bangladesh, Thailand, Peninsular Malaysia
and Sumatra. Traditionally, P. paludosa is used as antipyretic and antiinflammatory agent
in the early days in various regions (Saronika 2002). P. paludosa fresh shoot has quinone
reductase induction activity but no antioxidant activity (Bunyapraphatsara et al. 2003).
Upon significant literature survey it was found that only a few research work has been
performed on P. paludosa Roxb. and no work has been performed on the leaf of this plant
to investigate their medicinal use in our locality. Hence, the present study was taken up to
investigate the antidiarrhoeal, antioxidant, analgesic and cytotoxic properties of the leaves
of P. paludosa Roxb. and to find out the chemical group present in the active plant parts.
METHODS
Animals
Young Swiss-albino mice aged 45 weeks with body weight (BW) of 2230 g were used for
the experiment. The mice were purchased from the animal house of Jahangirnagar
University, Bangladesh. The animals were housed in stainless steel cages at room
temperature (28±2°C) and 12/12 h light dark cycle. All animals were fed with standard
pellet diet (Vital Feed Ltd. Jessore, Bangladesh) and water ad libitum. The study was
approved by the University Teaching and Research Ethics Committee of the University of
Chittagong (Ref. 122009). All animal experiments were carried out in accordance with
Local Ethnical Committee Acts.
Plant Materials
The leaves of P. paludosa were collected from the sundarbans (Bangladesh part) of
Sathkhira range (specifically from Koromjol). The time of collection was in January 2009
during daytime. During collection, any type of adulteration was strictly prohibited. The
plants were mounted on paper and the sample was taxonomically identified by the expert
authority of Bangladesh National Herbarium, Mirpur and Dhaka (accession no. 34176).
Preparation of Plant Extract
The collected leaves were separated from undesirable materials or plants or plant parts.
The leaves were dried by shade drying for 20 days to ensure the active constituents were
free from decomposition and also to avoid any photochemical degradation. The leaves
were ground into a coarse powder with the help of a suitable grinder. The powder was
stored in an airtight container and kept in a cool, dark and dry place until analysis
commenced. 170 g of the powdered leaf was extracted with ethanol using soxhlet
61 Phytochemical and Pharmacological Studies of P. paludosa
Malay J Pharm Sci, Vol. 8, No. 2 (2010): 5969
apparatus. After extraction, ethanol was removed, firstly by means of a water bath and
then in an oven, yielding the extracted compound. The concentrate was designated as
crude ethanolic extract of P. paludosa leaves.
Phytochemical Screening
The aqueous extract of the plant was subjected to qualitative chemical screening for the
identification of the tannins alkaloids and flavonoids using standard procedures (Trease
and Evans 1996).
Test for tannins
The aqueous extract (1 mL) was mixed with 10 mL of distilled water and filtered. Ferric
chloride reagent (3 drops) was added to the filtrate. A blue-black or green precipitate
confirmed the presence of gallic tannins or catechol tannins, respectively.
Test for alkaloids
The aqueous extract (0.2 mL) was stirred and placed in 1% aqueous hydrochloric acid
(5 mL) on a steam bath. 1 mL of the filtrate was treated with Mayer’s reagent (3 drops)
while another portion was similarly treated with Dragendorff’s reagent. Turbidity or
precipitation with these reagents was considered as evidence for the presence of alkaloids.
Test for flavonoids
A portion of the aqueous extract (2 mL) was heated, and metallic magnesium and
concentrated hydrochloric acid (5 drops) were added. A red or orange colouration
indicated the presence of flavonoids.
Test for steroids
2 mL of acetic acid was added to 0.2 g of extract. The solution was cooled in ice, and then
concentrated sulphuric acid was carefully added. Colour development from violet to blue
or bluish-green indicated the presence of a steroidal ring, i.e., a glycone portion of cardiac
glycoside.
Test for saponins
1 g of extract was boiled with 5 mL of distilled water and filtered. 3 mL of distilled water
was added to the filtrate, and the mixture was shaken vigorously for about 5 minutes.
Frothing that persisted upon warming was taken as evidence of the presence of saponins.
Test for glycosides
A small amount of an alcoholic extract of the fresh or dried plant material was placed in
1 mL of water. A few drops of aqueous sodium hydroxide were added. The presence of
yellow colour was considered as an indication for the presence of glycosides.
Ashura Akter Lima et al. 62
Malay J Pharm Sci, Vol. 8, No. 2 (2010): 5969
Test for gums
5 mL solution of the extract was taken and then Molish reagent and sulphuric acid were
added. Red violet ring produced at the junction of two liquids indicated the presence of
gums.
Acetic Acid-induced Writhing in Mice
Analgesic activity of the ethanolic extract of P. paludosa leaves were tested using the model
of acetic acid-induced writhing in mice (Whittle 1964; Ahmed et al. 2004). Experimental
animals were randomly selected and divided into four groups. Each group consisted of
five mice. Group-I received distilled water only, groups-II (positive control) received
diclofenac intraperitoneally at a dose of 50 mg/kg BW, groups-III and IV received the
extract orally at 250 and 500 mg/kg BW, respectively. 45 min interval was allowed to
ensure proper absorption of the orally administered substances and 30 min for
intraperitoneally administered diclofenac. 0.25 mL acetic acid solution (0.7%, 10 mL/kg)
was administered intraperitoneally to every mouse. The number of abdominal
constrictions (writhing) and stretching with a jerk of the hind limb was counted after 15
min of acetic acid injection. The response of the extract and diclofenac treated groups were
compared with those of the control group (distilled water). Percentage protection against
writhing movement (% inhibition of writhing was taken as an index of analgesia) was
calculated as follows:
Percentage inhibition = Wt (control) Wt (test group)/ Wt (control);
Wt = mean number of writhing.
Tests for Antidiarrhoeal Activity
The experiment was performed according to the method described by Shoba and Thomas
(2001). Mice were fasted for 24 h and were randomly allocated to 4 groups of 5 animals
each. The animals were all screened initially by giving 0.5 mL of castor oil. Only those
showing diarrhoea were selected for the final experiment. Group I received distilled
water, groups III and IV received the extract orally (250 and 500 mg/kg respectively).
Group II was given Loperamide (50 mg/kg, orally) in suspension. After 60 min, each
animal was given 0.5 mL of castor oil and was placed in an individual cage, the floor of
which was lined with blotting paper which was changed every hour, observed for 6 h and
the characteristic diarrhoeal droppings were recorded. The latent period of each mouse
were also counted.
Assay for Cytotoxicity of P. paludosa Leaf Extract
Assay for brine shrimp lethality
It was used for probable cytotoxic action (Meyer et al. 1982; McLaughlin 1991). The eggs of
brine shrimp (Artemia salina Leach) were collected and hatched in a tank at a temperature
around 37°C with constant oxygen supply. Two days were allowed to hatch and mature
the nauplii. Stock solution of the sample was prepared by dissolving required amount of
63 Phytochemical and Pharmacological Studies of P. paludosa
Malay J Pharm Sci, Vol. 8, No. 2 (2010): 5969
extract in specific volume of pure dimethyl sulfoxide (DMSO). 4 mL of seawater was
given to each of the vials. Specific volumes of sample were transferred from the stock
solution to the vials to get final sample concentrations of 0.1, 0.5, 1, 10, 20, 40, 60, 80 and
100 μg/mL. In the control vials same volumes of DMSO (as in the sample vials) were
taken. With a pasteur pipette, 10 living nauplii were put into each of the vials. After 24 h,
the vials were observed and the number of nauplii that survived in each vial was counted.
From this, the percentage of lethality of brine shrimp nauplii was calculated for each
concentration of the extract.
Assay for Antioxidative Activity of P. paludosa Leaf Extract
The antioxidant activity of P. paludosa extract was assessed in comparison to standard
antioxidant ascorbic acid and vitamin E (Sigma, Germany) on the basis of scavenging
effect of the stable 2, 2-diphenyl-1-picrylhydrazyl (DPPH) free radical activity according to
established procedure (Brand-Williams, Cuvelier and Berset 1995). Five different
concentrations (1, 5, 10, 20 and 50 μg/mL) of plant extract (0.1 mL) was added to 3 mL of a
0.004% methanol solution of DPPH. Absorbance at 517 nm was determined after 30 min,
and the percentage inhibition activity was calculated from [(A0A1)/A0]x100, where A0 is
the absorbance of the control, and A1 is the absorbance of the extract/standard. The
inhibition curves were prepared and IC50 values were calculated.
Statistical Analysis
Statistical analysis for animal experiment was carried out using one-way ANOVA
followed by Dunnet’s multiple comparisons. The results obtained were compared with the
control group. p values <0.05 were considered to be statistically significant. The
concentration producing 50% of the maximum response (LC50 or IC50) was obtained by the
best visual fit from the plot of the individual experiments.
RESULTS
Phytochemical Screening
P. paludosa leaf revealed the presence of flavonoids, glycosides, tannins, gum and steroid
(Table 1).
Table 1: Phytochemical screening of P. paludosa leaf extract.
Test
Alkaloid
Flavonoids
Gum
Glycosides
Steroids
Inference
+
+
+
+
Note: + = present, = absent.
Analgesic Activity
In the acetic acid-induced writhing test, the ethanolic extract of leaves of P. paludosa
showed dose dependent decrease in the total number of writhings after 15 min of
administration of acetic acid intraperitoneally. The analgesic effect at the dose of 500
Ashura Akter Lima et al. 64
Malay J Pharm Sci, Vol. 8, No. 2 (2010): 5969
mg/kg body weight was significant (p=0.009) and the effect increased with the increase of
dose (Table 2).
Table 2: Effect of P. paludosa extract on acetic acid induced writhing of mice.
Group
(n=5)
Dose (mg/kg BW)
Writhing
(mean±SEM)
%
writhing
Percentage
writhing
inhibition
I (Control)
27.2±1.93
II (Diclofenac-Na)
50
6.8±2.13
25.00**
75
III (Et. extract)
250
24.8±1.46
91.18
8.82
IV (Et. extract)
500
18.6±1.60
68.38*
31.61
Notes: *p<0.05, **p<0.001, Et=ethanolic, n=number of rat, BW=body weight, SEM=standard error of mean.
Antidiarrhoeal Activity
In the castor oil-induced antidiarrhoeal model, the extract of leaves of P. paludosa
significantly increased the latent period in both 250 mg/kg body weight (p=0.020) and 500
mg/kg body weight (p=0.002) doses in the treated animals (Table 3). It was also observed
that there was a decreasing trend (p=0.052) in the number of liquid stools during the total
observation period at both doses of 250 and 500 mg/kg (Table 4).
Table 3: Effect of extract of leaves of P. paludosa on the latent period in castor oil-induced
diarrhoeal episode in mice.
Group (n=5)
Treatment
Dose (mg/kg BW)
Latent period (min;
mean±SEM)
I
Distilled water (solvent)
13.0±2.21
II
Loperamide
50
45.4±6.01*
III
Ethanol extract
250
25±3.52*
IV
Ethanol extract
500
36±4.48*
Notes: SEM=standard error of mean, n=number of rat, BW=body weight, *p<0.05.
Cytotoxic Activity
In brine shrimp lethality bioassay, test sample showed different mortality rate at different
concentrations (Table 5). The mortality rate of brine shrimp was found to increase with the
increase in concentration of the sample and the extract of leaves of P. paludosa showed
significant (p=0.001) toxicity to the brine shrimp nauplii. The concentrations of crude
extract for 50% mortality (LC50) and 90% mortality (LC90) were 16.67 µg/mL and 34.81
µg/mL, respectively.
65 Phytochemical and Pharmacological Studies of P. paludosa
Malay J Pharm Sci, Vol. 8, No. 2 (2010): 5969
Table 4: Effect of P. paludosa leaves extract on liquid defecation in the castor oil-induced
diarrhoeal episode in mice.
Type
(group)
1sth
2ndh
3rdh
4thh
5thh
6thh
Control (I)
3.00±1.26
3.80±0.37
2.40±0.68
2.40±0.68
1.00±0.45
0.40±0.40
Standard (II)
(p value)
0.00±0.00
(0.077)
0.00±0.00
(0.001)
0.20±0.20
(0.014)
0.20±0.20
(0.014)
0.00±0.00
(0.089)
0.00±0.00
(0.374)
250 mg (III)
(p value)
1.80±0.73
(0.436)
2.80±1.36
(0.598)
1.80±0.73
(0.684)
0.60±0.40
(0.052)
0.20±0.20
(0.141)
0.40±0.24
(1.000)
500 mg (IV)
(p value)
1.80±1.11
(0.497)
2.00±1.14
(0.128)
1.00±0.45
(0.532)
0.60±0.40
(0.052)
0.40±0.40
(0.347)
0.20±0.20
(0.667)
Note: Values are presented as mean±SEM, (n=5).
Table 5: Brine shrimp lethality bioassay of extract of P. paludosa leaves.
Extract concentration
(μg/mL)
Log
concentration
% of mortality
LC50 (μg/mL)
LC90 (μg/mL)
100
2
100
80
1.90309
100
60
1.778151
100
40
1.60206
100
20
1.30103
60
16.67
34.81
10
1
30
1
0
20
0.1
0.30103
0
0.5
1
0
Notes: LC50 and LC90 were determined from 24 h counts using probit analysis method described by
FINNEY computer program.
Antioxidant Activity
In the quantitative antioxidant assay using DPPH, the ethanolic extract of P. paludosa
leaves showed free radical scavenging properties, though the effect was more strong
(IC50=7.21 μg/mL) than one of the two standards, tocopherol/vitamin E (IC50>50 μg/mL)
but it showed weaker activity than the other standard, ascorbic acid (IC50=3 μg/mL)
(Table 6).
DISCUSSION
Phytochemical screening of the ethanol extract of P. paludosa leaf reveals the presence of
glycosides, saponins, gum, tannins and steroid. These compounds can show extensive
pharmacologic and other activities. In the acetic acid induced writhing test, the ethanolic
Ashura Akter Lima et al. 66
Malay J Pharm Sci, Vol. 8, No. 2 (2010): 5969
Table 6: DPPH free radical scavenging activity of ascorbic acid, vitamin E (tocoferol) and
P. paludosa leaf extract.
Test material
Concentration (μg/mL)
% scavenging activity
IC50 (μg/mL)
Ascorbic acid
1
21.86
5
86.16
3
10
96.77
20
97.41
50
97.28
Vitamin E
1
16.43
5
17.46
>50
10
23.29
20
23.67
50
27.17
P. paludosa extract
1
27.30
5
40.75
10
60.28
7.21
20
77.75
50
88.87
extract of leaves of P. paludosa showed dose dependent decrease in the total number of
writhing. From the observation it can be suggested that the ethanolic extract of the
P. paludosa leaves has mild analgesic activity. Analgesic activities are commonly exhibited
by the non-steroidal antiinflammatory drugs (NSAIDS). These NSAIDs exert
antiinflammatory effect principally by inhibiting the synthesis of prostaglandin (Vane
1971), an eicosanoid. Inhibition of prostaglandin synthesis may account for the analgesic
activity of the leaf extract as it contributes in large part to the analgesic activity of the
NSAIDs. Castor oil is made up of 90% ricinoleate (Mekeon, Lin and Stafford 1999), which
when metabolised is responsible for the observed effects of the oil. The active metabolite
ricinoleic acid is responsible for its diarrhoea inducing property, which diminishes Na+
and C1- permeability in the intestine (Gaginella and Phillips 1975); it is also associated
with endogenous stimulation of prostaglandins release (Zavala et al. 1998). Ethanol extract
of the leaf of P. paludosa was found to inhibit the severity of diarrhoea induced by castor
oil. It is possible that the extract was able to inhibit electrolyte permeability in the intestine
due to castor oil and or through the inhibition of prostaglandins release (Adzu et al. 2003).
We suggest that saponins and/or flavonoids present in the leaf extract might be
responsible for its antidiarrhoeal activity. The mortality rate of brine shrimp was found to
increase with the increase in concentration of the sample and the extract of leaves of
P. paludosa showed significant (p=0.001) toxicity to the brine shrimp nauplii. These results
tend to suggest possible antitumor, antibacterial or pesticidal activities. The free radical,
DPPH is widely used to test the ability of compounds to act as free radical scavengers or
hydrogen donors, and to evaluate antioxidant activity. It has also been used to quantify
antioxidants in complex biological systems in recent years. The DPPH method can be used
for solid or liquid samples and is not specific to any particular antioxidant component, but
applies to the overall antioxidant capacity of the sample. A measure of total antioxidant
67 Phytochemical and Pharmacological Studies of P. paludosa
Malay J Pharm Sci, Vol. 8, No. 2 (2010): 5969
capacity will help us understand the functional properties of the plant extract. In the
quantitative antioxidant assay using DPPH, the ethanolic extract of P. paludosa leaves
showed strong radical scavenging effect in comparison to the effect of the tocopherol
though it showed weak scavenging effect in comparison to the effect of standard ascorbic
acid. In a previous study on antioxidative activity of Phoenix dactylifera, antioxidant state
(plasma vitamin C, E and A and b-carotene) increased significantly on administration of
different extracts (Mohamed and Al-Okbi 2004). It has been recognised for some time that
several classes of flavonoids play significant roles in many physiological processes and
show antioxidant and fungicidal activity (Larson 1988) and are natural antihistamines.
Flavonoid and flavonol-lignan derivatives inhibit lipid peroxidation and are potent
quenchers of triplet oxygen.
CONCLUSION
Pharmacological evaluation of P. paludosa leaf extract reveals some interesting activities
like cytotoxicity, antidiarrhoeal, analgesic as well as the antioxidant activities of this plant.
From these we can assume that different active secondary metabolites are present in its
extracts and perhaps some of these compounds may function in a synergistic manner.
However, further studies are necessary to elucidate the mechanism lying with this effect.
This report may serve as a stepping stone for future research on the biological and
pharmacological activities of P. paludosa leaf extract.
ACKNOWLEDGEMENT
The authors would like to thank the Department of Pharmacy, Jahangirnagar University,
Bangladesh, for their cordial help in doing the study.
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diarrhoeal activity of Waltheria americana, Commelins cuelestris and Alternanthra repens,
Journal of Ethnopharmacology, 61: 4147.
... Their findings showed that fractions were completely insensitive to microbial growth, while the n-hexane, chloroform and methanol soluble fractions showed significant cytotoxicity having lethal concentration 50% (LC 50 ) values of 2.17 mg/mL, 2.77 mg/mL and 2.46 mg/mL, respectively. Similarly, Lima et al. (2010) prepared ethanolic extracts from leaves of P. paludosa and examined the cytotoxic activity using brine shrimp lethality bioassay. The results suggested that the ethanolic extract was moderately cytotoxic with LC 50 and LC 90 values of 6.67 mg/mL and 34.81 mg/mL, respectively. ...
... A sap mixture containing vitamin B complex with an iron tonic could provide more beneficial effects to the patients than either the sap or vitamin B complex or iron tonic alone. Lima et al. (2010) evaluated the antidiarrheal activity of the crude ethanolic extract of the leaves of P. paludosa using castor oilinduced diarrhea in mice. The extract showed dose dependant activity and reduce the frequency of liquid stools in rats. ...
... The onset of the diuretic effect was faster at a dose of 300 mg/ kg body weight than at 150 mg/kg body weight. In the same way, the analgesic activities of the ethanolic extract of leaves of P. padulsa were studied in acetic acid-induced writhing in mice by Lima et al. (2010). The authors found that leaves of P. padulsa had a significant (p ¼ 0.009) analgesic effect. ...
Article
Full-text available
Phoenix sylvestris is commonly known as Indian date and is native to India and southern portions of Pakistan. It is traditionally important and known for its nutritional values throughout the world. It is a rich source of carbohydrate, phenols, amino acids, flavonoids, tannins, alkaloids, terpenoids, dietary fibers, essential vitamins and minerals. Different parts of the plant exhibit diverse medicinal properties such as being antipyretic, cardiotonic, laxative, diuretic and antioxidant. In the present review, an attempt has been made to compile most of the information available regarding the distribution, cultivation, phytochemical characteristics, Ayurvedic properties, ethno-pharmacological, medicinal and non-medicinal uses of P. sylvestris in the last four decades.
... [15] In addition, studies have also shown that plants with alkaloids may have hypoglycemic effect via insulin-releasing mechanism and insulin-mimicking activity, thus improving postprandial hyperglycemia. [9] The presence of flavonoids in plant extract has also been reported to play an important role in plant biochemistry and physiology as enzyme inhibitor, antioxidants, [16] anti-diabetic, [17][18][19] anti-inflammatory, [20] antimicrobial, [21][22][23] antifungal, [21] and anticancer activities, thereby protecting the cells against all stages of carcinogenesis. [10] Furthermore, available report on phenolic compounds has also been demonstrated for their usefulness to prevent oxidative damage in cells, thereby acting as a free radical terminator. ...
... [15] In addition, studies have also shown that plants with alkaloids may have hypoglycemic effect via insulin-releasing mechanism and insulin-mimicking activity, thus improving postprandial hyperglycemia. [9] The presence of flavonoids in plant extract has also been reported to play an important role in plant biochemistry and physiology as enzyme inhibitor, antioxidants, [16] anti-diabetic, [17][18][19] anti-inflammatory, [20] antimicrobial, [21][22][23] antifungal, [21] and anticancer activities, thereby protecting the cells against all stages of carcinogenesis. [10] Furthermore, available report on phenolic compounds has also been demonstrated for their usefulness to prevent oxidative damage in cells, thereby acting as a free radical terminator. ...
Article
Background: Scabiosa columbaria L. is commonly known as the wild scabious and is used in South Africa to treat various ailments such as wound bruises, painful menstruation, heartburn, and colic. Despite its extensive traditional usage, the bioactive constituents of this plant are yet to be explored. Objective: The present study was carried out to identify the bioactive constituents of the ethanol extract of S. columbaria by phytochemical screening and gas chromatography-mass spectrometry (GC-MS). Materials and Methods: Sixty grams of the powdered leaves was sequentially extracted by ethanol and later tested for preliminary phytochemical screening and further subjected to GC-MS analysis. Results: The results showed the presence of alkaloids, flavonoids, phenolics, tannins, steroids, terpenoids, and saponins in the extract. The GC-MS analysis revealed the presence of 16 major compounds, with flavonoids (40.84%) being the most represented chemical class. Conclusion: The findings indicated that the plant possesses compounds with biological activities and therefore justifies its traditional usage in the treatment of skin and other diseases.
... [15] In addition, studies have also shown that plants with alkaloids may have hypoglycemic effect via insulin-releasing mechanism and insulin-mimicking activity, thus improving postprandial hyperglycemia. [9] The presence of flavonoids in plant extract has also been reported to play an important role in plant biochemistry and physiology as enzyme inhibitor, antioxidants, [16] anti-diabetic, [17][18][19] anti-inflammatory, [20] antimicrobial, [21][22][23] antifungal, [21] and anticancer activities, thereby protecting the cells against all stages of carcinogenesis. [10] Furthermore, available report on phenolic compounds has also been demonstrated for their usefulness to prevent oxidative damage in cells, thereby acting as a free radical terminator. ...
... [15] In addition, studies have also shown that plants with alkaloids may have hypoglycemic effect via insulin-releasing mechanism and insulin-mimicking activity, thus improving postprandial hyperglycemia. [9] The presence of flavonoids in plant extract has also been reported to play an important role in plant biochemistry and physiology as enzyme inhibitor, antioxidants, [16] anti-diabetic, [17][18][19] anti-inflammatory, [20] antimicrobial, [21][22][23] antifungal, [21] and anticancer activities, thereby protecting the cells against all stages of carcinogenesis. [10] Furthermore, available report on phenolic compounds has also been demonstrated for their usefulness to prevent oxidative damage in cells, thereby acting as a free radical terminator. ...
Article
Background: Scabiosa columbaria L. is commonly known as the wild scabious and is used in South Africa to treat various ailments such as wound bruises, painful menstruation, heartburn, and colic. Despite its extensive traditional usage, the bioactive constituents of this plant are yet to be explored. Objective: The present study was carried out to identify the bioactive constituents of the ethanol extract of S. columbaria by phytochemical screening and gas chromatography-mass spectrometry (GC-MS). Materials and Methods: Sixty grams of the powdered leaves was sequentially extracted by ethanol and later tested for preliminary phytochemical screening and further subjected to GC-MS analysis. Results: The results showed the presence of alkaloids, flavonoids, phenolics, tannins, steroids, terpenoids, and saponins in the extract. The GC-MS analysis revealed the presence of 16 major compounds, with flavonoids (40.84%) being the most represented chemical class. Conclusion: The findings indicated that the plant possesses compounds with biological activities and therefore justifies its traditional usage in the treatment of skin and other diseases.
... [15] In addition, studies have also shown that plants with alkaloids may have hypoglycemic effect via insulin-releasing mechanism and insulin-mimicking activity, thus improving postprandial hyperglycemia. [9] The presence of flavonoids in plant extract has also been reported to play an important role in plant biochemistry and physiology as enzyme inhibitor, antioxidants, [16] anti-diabetic, [17][18][19] anti-inflammatory, [20] antimicrobial, [21][22][23] antifungal, [21] and anticancer activities, thereby protecting the cells against all stages of carcinogenesis. [10] Furthermore, available report on phenolic compounds has also been demonstrated for their usefulness to prevent oxidative damage in cells, thereby acting as a free radical terminator. ...
... [15] In addition, studies have also shown that plants with alkaloids may have hypoglycemic effect via insulin-releasing mechanism and insulin-mimicking activity, thus improving postprandial hyperglycemia. [9] The presence of flavonoids in plant extract has also been reported to play an important role in plant biochemistry and physiology as enzyme inhibitor, antioxidants, [16] anti-diabetic, [17][18][19] anti-inflammatory, [20] antimicrobial, [21][22][23] antifungal, [21] and anticancer activities, thereby protecting the cells against all stages of carcinogenesis. [10] Furthermore, available report on phenolic compounds has also been demonstrated for their usefulness to prevent oxidative damage in cells, thereby acting as a free radical terminator. ...
... Mangroves are considered as one of the main productive coastal eco-systems. Mangroves have widely been used in traditional medicine to treat cancers and a number of natural compounds with different bio-activities have been reported from several mangrove plants [8]. S. hydrophyllacea (Family: Rubiaceae) is one of the common mangrove plants found mainly in South Asia including Sri Lanka [8,9]. ...
... Mangroves have widely been used in traditional medicine to treat cancers and a number of natural compounds with different bio-activities have been reported from several mangrove plants [8]. S. hydrophyllacea (Family: Rubiaceae) is one of the common mangrove plants found mainly in South Asia including Sri Lanka [8,9]. Several flavonoids, terpenoids and iridoids have been isolated from S. hydrophyllacea by previous researchers [9]. ...
Article
Purpose: To isolate active cytotoxic compounds from the hexane and chloroform extracts of the leaves of the mangrove plant, Scyphiphora hydrophyllacea C.F. Gaertn (Rubiaceae), grown in Sri Lanka. Methods: Dried pulverized leaves of S. hydrophyllacea were extracted with hexane and chloroform. Vacuum liquid chromatography (VLC), column chromatography (size exclusion chromatography, Sephadex LH-20) and reversed phase preparative recycling high performance liquid chromatography (HPLC) techniques were used to isolate three compounds (compounds 1, 2 and 3). The structures of the isolated compounds were established This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest
... The investigated plant Phoenix paludosa is abound with numerous phytochemicals like flavonoids, polyphenols, glycosides, gum, steroids, tannins (Lima et al., 2010;Saha et al., 2012;Samarakoon et al., 2016), phytosterols, triterpinoids (Alam et al., 2009) and so on. These ingredients are responsible for different pharmacological properties including antimicrobial and cytotoxic potency. ...
... The larvicidial toxicity of the above mentioned phytochemicals is responsible for the cytotoxic response of the plant and it may become clinically useful anticancer as well as pesticide agents as the brine shrimp assay is a considerable preliminary assessment of toxicity (Rahman and Islam, 2013). The exhibited cytotoxic property of the bark of Phoenix paludosa in this study supports the previous studies on cytotoxic potency of the leaf of the plant in different solvents (Alam et al., 2009;Lima et al., 2010;Samarakoon et al., 2016). Previous studies on the plant showed unreliable antimicrobial activity (Patra et al., 2014b). ...
Article
Full-text available
Plant origin is an important source of drugs. In this experiment, the ethanolic extract of the bark of Phoenix paludosa fractioned with three different solvents (Petroleum ether, Chloroform and Ethyl acetate), was investigated to evaluate the antibacterial and cytotoxic activity. Most popular disc diffusion method was applied to determine the antibacterial activity of 500 μg/disc crude extract of the plant against 12 pathogenic Gram positive and Gram negative bacteria. Cytotoxic test was performed by following the brine shrimp lethality bioassay at different concentrations (100, 50, 25, 12.5, 6.25 μg/ml) where Vincristine sulphate served as positive control. Though the antibacterial activity is not so promising compared to the standard Kanamycin, only the chloroform fraction of ethanolic extract exhibited activity against all experimental bacteria (zone of inhibition ranging from 7 to 15 mm), the plant has been proven to possess good cytotoxic activity, having LC50 value of 13.03, 9.24 and 7.47 μg/ml for petroleum ether, chloroform and ethyl acetate fraction of ethanolic extract of the bark of Phoenix paludosa, respectively.
... 13 It can be prescribed for gastroenteritis, cough, respiratory diseases, asthma, chest complaints, fevers, high blood pressure and fatigue. 13, 14 Several Phoenix species have a number of pharmacological activities like Anti-mutagenic, 15 Antioxidant, Anti-diarrheal, 16 Analgesic, 17 Anti-inflammatory. 18 Different classes of compounds were isolated and identified from genus Phoenix that contains lipids, phenolic acids, flavonoids, procyanidines, triterpenes, carbohydrates, vitamins, inorganic ions etc. 19 The literature available from all possible scientific sources showed very little research work on this species, whereas tribes claim that P. loureiroi was used in the treatment of various diseases and ailments, although there is no inbuilt scientific proof in support of the utility of this species as an analgesic, anti-inflammatory and antipyretics agent. ...
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The present research work was carried out the High Performance Thin Layer Chromatography (HPTLC) and Gas Chromatography-Mass Spectrometry (GC-MS) analysis and assessment of pharmacological activities of Phoenix loureiroi Kunth (Arecaceae) ethanolic leaves extract at doses of 200, 400 and 600 mg/kg, body weight, per os. Preliminary phytochemical screening, HPTLC and GC-MS studies were carried out according to the standard methods. The acute toxicity studies were conducted on Swiss albino mice as per Organization for Economic Cooperation and Development (OECD) guidelines 420. For the screening of analgesic activity, writhing test was conducted for peripheral analgesic activity whereas tail immersion test was conducted for central analgesic activity. Antipyretic activity was performed by using the yeast induced hyperpyrexia method and for the screening of anti-inflammatory activity carrageenan-induced rat paw edema method was used. Preliminary phytochemical screening of the ethanol extract of Phoenix loureiroi leaves (EEPLL) contains sterols, flavonoids, saponins, proteins, reducing sugar, tannins, and phenolic compounds. The HPTLC analysis method employed in this work resulted in good peak shape and enabled good resolution of quercetin present in the extract and GC-MS analysis showed a total of 25 peaks and led to the identification of 22 different phytoconstituents in the ethanolic extract. Lethal Dose 50 (LD 50) was above 2,000 mg/kg and no death was recorded. The prevailing study demonstrated that EEPLL possesses widespread analgesic, antipyretic and anti-inflammatory effects in dose dependent manner. It can be concluded that the ethanolic extract from Phoenix loureiroi leaves possesses promising analgesic, antipyretic and anti-inflammatory activities.
... This plant has been used as an antipyretic and an anti-inflammatory agent in Bangladesh [6]. A previous study has demonstrated that P. paludosa shoot has quinonereductase induction activity [7]. However, data available on possible cytotoxic activity or free radical scavenging activity of organic extracts of aerial parts of this mangrove plant are limited. ...
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Purpose: To investigate the anti-proliferative and antioxidant potentials of four different solvent extracts of Phoenix paludosa Roxb leaves. Methods: Four different solvent (hexane, chloroform, ethyl acetate and methanol) leaf extracts of the plant were tested for cytotoxicity against four cancer cells, viz, MCF-7 (oestrogen positive breast cancer cell line), MDA-MB-231 (triple negative breast cancer cell line), SK-BR-3 (breast adenocarcinoma) and ACHN (renal adenocarcinoma) as well as two normal cell lines, namely, HEK-293 (embryonic kidney cells) and MCF-10A (normal mammary epithelial cells)]. 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) and 2, 29-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) free radical scavenging assays were used to evaluate the antioxidant activity of the crude extracts. Results: The methanol extract showed the highest antioxidant activity (DPPH, half maximal inhibitory concentration (IC50) = 30.17 ± 6.21 μg/mL) and (ABTS, IC50 = 27.91 ± 3.21 μg/mL). Of the four extracts, methanol extract showed the strongest significant (p < 0.05) cytotoxicity to all four cancer cell lines at 24 and 48 h of incubation followed by the chloroform extract (IC50 of methanol extract (24 and 48 h): 36.71 ± 8.72 and 33.19 ± 5.53 μg/mL (MCF-7), 159.7 ± 32.09 and 141.9 ± 26.2 μg/mL (MDA-MB-231), 103.3 ± 18.9 and 75.39 ± 19.39 μg/mL (SKBR-3), 57.21 ± 3.72 and 43.16 ± 10.25 μg/mL (MCF-10A), 37.48 ± 5.75 and 26.99 ± 1.85 (ACHN) and 66.83 ± 14.26 and 60.34 ± 10.66 μg/mL (HEK-293)). Furthermore, the methanol extract was least cytotoxic to normal cell lines. Conclusion: The results obtained indicate that the methanol leaf extract of P. paludosa exhibit potent antioxidant and cytotoxic activities and has the potential of being developed into an anti-cancer agent. © Pharmacotherapy Group, Faculty of Pharmacy, University of Benin, Benin City, 300001 Nigeria. All rights reserved.
Article
The present study was designed to evaluate the effect of different solvents on the extraction of bioactive compounds, antioxidant and cytotoxic potentials of dwarf date palm (Phoenix loureiroi). The leaves were successively extracted with low to high polar solvents (petroleum ether, chloroform, ethyl acetate, methanol and water) using the Soxhlet method. Spectrophotometric determination of leaves extracts showed higher chlorophyll a and b contents, being 0.06 and 0.04 g/g extract, respectively while higher carotenoids content (0.94 g/g extract) was obtained in the chloroform extract. Moreover, higher total phenols (0.58 g GAE/100 g extract) and flavonoids (0.95 g RE/100 g extract) were found in the methanol extract. The HPLC-DAD analysis revealed that gallic acid (0.04 g/g extract) was higher in methanol extract in the method I and II. This extract also showed higher succinic acid (0.07 g/g extract) and citric acid (0.04 g/g extract) contents. The improved UPLC-PDA-MS results also confirmed that the presence of polyphenols (epicatechin, 0.09 g/g extract) was in the methanol extract of the leaves. Correspondingly, methanol extract showed higher antioxidant activity in DPPH• scavenging (IC50 = 6.45 μg/mL), NO (67.47%), lipid peroxidation inhibition (55.10%), ABTS•+ scavenging (16.60 mM TE/g extract), phosphomolybdenum (0.14 g AAE/g extract), FRAP (1899270 mM Fe (II)/g extract), ORAC (3388.80 mM TE/g extract) and O2-• scavenging (60.64%) assays. The cell viability test was studied on fibroblast L929 cell lines and showed no cytotoxicity effects in the methanol extract for up to 1000 µg/mL. Correlation study and principal component analysis demonstrated that antioxidant activity might be due the presence of polyphenols. Thus, it is suggested that the extraction of leaves polyphenols using polar solvents is a viable option in promising source of antioxidants.
Book
Marine plants such as algae (blue-green algae and seaweeds), seagrasses, mangrove plants, salt-tolerant or salt-loving plants (halophytes) and coastal sand dune plants are known to generate approximately 70% of oxygen on earth, and help regulate oxygen in the atmosphere. These plants are potential sources of nutrients and are also considered valuable for the development of new drugs owing to their unique bioactive compounds. This book provides the taxonomy, common name, global distribution, habitat, diagnostic features and pharmaceutical compounds (along with their activities) of 400 species of marine plants, accompanied by high quality illustrations. Biology and Ecology of Pharmaceutical Marine Plants is the first comprehensive book of its kind written by scientists from both the Marine Biology and Pharmacy disciplines to fill the long-felt need for a marine natural products book devoted exclusively to plants. It should be a standard reference for students, researchers and teachers of disciplines such as Pharmacy, Fisheries Science, Marine Biology, Life Sciences, Biotechnology and Biochemistry, as well as a valuable guide for pharmaceutical companies involved in the development of new drugs from marine plants.
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Mangrove areas are rich in medicinal and edible plants. Biological screening of the plants in this study may lead to drug and health product development. The biological tests includes antioxidation, antilipid peroxidation and cancer chemoprevention. Fifty seven samples of 32 species were tested. Calyces of Sonneratia caseolaris exhibited the strong antioxidant activity followed by stamens of Sonneratia caseolaris, calyces of S. alba, Cynometra ramiflora seeds, Xylocarpus rumphii fruit peel and branches. Some edible pods including Bruguiera parviflora, Ceriops decandra, C. tagal, Rhizophora mucronata etc., were also active. Some of these pods also exhibited antilipid peroxidation such as Bruguiera parviflora and Rhizophora mucronata. Besides, Cynometra ramiflora (seeds), Lumnitizera racemosa (leaves), Nypa fruticans (inflorescences) and Sonneratia caseolaris, exhib-ited strong antilipid peroxidation. Plants possessed cancer chemoprevention activities were Bruguiera gymnorrhiza (pods), Acanthus ebracteatus (leaves), Avicennia marina (leaves), Flagellaria indica (young shoot), Phoenix paludosa (young shoot) and Trianthema decandra (aerial part), of which Bruguiera gymnorrhiza pods exhibited strongest activity.
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This study was carried out to evaluate the antioxidant and anti-inflammatory activity of methanolic and water extracts of the edible portion of date fruits (Phoenix dactylifera of date fruits (Phoenix dactylifera of date fruits (L., Family Palmae) and methanolic extract of date seeds in adjuvant arthritis in rats, a model of chronic inflam-mation. Acute oral toxicity of the different extracts was carried out to determine their safety as functional foods. The results revealed that oral administration of the methanolic and aqueous extracts of edible portion of date fruits suppressed the swelling in the foot significantly by 67.8 and 61.3% respectively, while the methanolic extract of date seeds showed significant reduction by 35.5%. Antioxidant state (plasma vitamin C, E and A and β-carotene) increased significantly on administration of different extracts, while plasma level of MDA reduced significantly. Methanolic extract of edible portion of the fruit was the best in reducing foot swelling, ESR and plasma fibrinogen and in normalizing the plasma level of antioxidants. Adjuvant arthritic rats showed significant reduction in body weight gain and food efficiency ratio. Administration of any of the extracts pro-duced significant increase in body weight gain and food efficiency ratio. Acute lethal toxicity test showed that the safest extract was the methanolic of edible portion of the fruit followed by the water extract that showed complete safety up to 6 g/kg mice weight. Methanolic extract of the seed showed LD50 equal to 6.75 g/kg mice.
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Many enzymes and secondary compounds of higher plants have been demonstrated in in vitro experiments to protect against oxidative damage by inhibiting or quenching free radicals and reactive oxygen species. The roles of many other compounds as potential antioxidants can be inferred by their similarity to synthetic antioxidants of related structure. The evidence supports at least a partial antioxidant role in vivo for many classes of plant metabolite.
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A method, utilizing brine shrimp (Artemia salina Leach), is proposed as a simple bioassay for natural product research. The procedure determines LC (50) values in microg/ml of active compounds and extracts in the brine medium. Activities of a broad range of known active compounds are manifested as toxicity to the shrimp. Screening results with seed extracts of 41 species of Euphorbiaceae were compared with 9KB and 9PS cytotoxicities. The method is rapid, reliable, inexpensive, and convenient as an in-house general bioassay tool.
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A phytotherapeutic approach to modern drug development can provide many invaluable drugs from traditional medicinal plants. Search for pure phytochemicals as drugs is time consuming and expensive. Numerous plants and polyherbal formulations are used for the treatment of liver diseases. However, in most of the severe cases, the treatments are not satisfactory. Although experimental evaluations were carried out on a good number of these plants and formulations, the studies were mostly incomplete and insufficient. The therapeutic values were tested against a few chemicals-induced subclinical levels of liver damages in rodents. Even common dietary antioxidants can provide such protection from liver damage caused by oxidative mechanisms of toxic chemicals. However, experiments have clearly shown that plants such as Picrorrhiza kurroa, Andrographis paniculata, Eclipta alba, Silibum marianum, Phyllanthus maderaspatensis and Trichopus zeylanicus are sufficiently active against, at least, certain hepatotoxins. Screening plants for antihepatitis activities remains in its infancy. P. kurroa, E. alba, Glycyrrhiza glabra, A. paniculata and P. amarus are likely to be active against Hepatitis B virus. In the case of severe liver damage, most of the liver cells die or turn into fibrotic state. In this case, the treatment should include in addition to the therapeutic agents, agents which can stimulate liver cell proliferation. For developing satisfactory herbal combinations to treat severe liver diseases, plants have to be evaluated systematically for properties such as antiviral activity (Hepatitis B, Hepatitis C, etc), antihepatotoxicity (antioxidants and others), stimulation of liver regeneration and choleretic activity. The plants with remarkable activities for each of the above properties have to be identified. Single plant may not have all the desired activities. A combination of different herbal extracts/'fractions is likely to provide desired activities to cure severe liver diseases. Development of such medicines with standards of safety and efficacy can revitalise treatment of liver disorders. hepatoprotective activity.
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The antiradical activities of various antioxidants were determined using the free radical, 2,2-Diphenyl-1-picrylhydrazyl (DPPH*). In its radical form. DPPH* has an absorption band at 515 nm which dissappears upon reduction by an antiradical compound. Twenty compounds were reacted with the DPPH* and shown to follow one of three possible reaction kinetic types. Ascorbic acid, isoascorbic acid and isoeugenol reacted quickly with the DPPH* reaching a steady state immediately. Rosmarinic acid and δ-tocopherol reacted a little slower and reached a steady state within 30 min. The remaining compounds reacted more progressively with the DPPH* reaching a steady state from 1 to 6 h. Caffeic acid, gentisic acid and gallic acid showed the highest antiradical activities with a stoichiometry of 4 to 6 reduced DPPH* molecules per molecule of antioxidant. Vanillin, phenol, γ-resorcylic acid and vanillic acid were found to be poor antiradical compounds. The stoichiometry for the other 13 phenolic compounds varied from one to three reduced DPPH* molecules per molecule of antioxidant. Possible mechanisms are proposed to explain the experimental results.
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The first prescriptions for castor oil may well have been enscrolled on papyrus; certainly, the oil's cathartic properties have been recognized and employed therapeutically for centuries. Like some of its modern counterparts, castor oil has been prescribed at times without adequate pharmacological definition; indeed, its mechanisms of action are defined vaguely in classical pharmacological references, and only then in terms of an "irritant" or "stimulant" effect on the gut (1, 2). The past 15 years have seen a new interest develop in the drug and its actions on the gastrointestinal tract. A major incentive has been the insight given by this ancient remedy to the relationship between malabsorption of dietary fat and diarrhea. In 1890, Meyer (3) identified the active component of the oil as ricinoleic acid and this reference still constitutes a foundation for discussion of the drug's actions. Ricinoleic acid is a C~8, monounsaturated (at C9-1o), monohydroxylated