PHARCHM 2 LAB
NOCEJA, Alyssa Marie P.
ONG, Jan Winifred C.
PAMINTUAN, Anne Katherine C.
PANGANIBAN, Perla Andrea L.
PEJANA, Ina Christine B.
Mr. Jay Jazul
Ms. Grace Alunan
August 7, 2010
The word saponin was derived from the Latin word sapo which means soap. Saponins are
generally known as non-volatile, surface-active compounds that are widely distributed in nature,
occurring primarily in the plant kingdom. They are high-molecular-weight glycosides consisting of a
sugar moiety (component) linked to a aglycone. This combination of polar (sugar moiety) and non polar
(aglycone) explains their soap-like behavior in aqueous solutions. The classical definition of saponins is
based on their surface activity. The amphiphilic nature of saponins dominates their physical properties in
solution. They are strongly surface active, form stable foams; act as emulsifying agents and form micelles
in much the same way as detergents.
II. Chemical Structure
Saponins are high-molecular-weight glycosides which consists of a non polar aglycone coupled
with one or more sugar moieties. The general formula for glycosides is:
The aglycone or non-saccharide portion of the saponin molecule is called the genin or sapogenin.
Depending on the type of genin present, the saponins can be divided into three major classes:
1. Triterpene glycosides
2. Steroid glycosides
3. Steroid Alkaloid glycosides
The aglycones are normally hydroxylated at C-3 and certain methyl groups are frequently
oxidized to hydroxymethyl, aldehyde, or carboxyl functionalities. All saponins have in common the
attachment of one or more sugar chains to the aglycone. Monodesmosidic saponins have a single sugar
chain normally attached at C-3. Bidesmosidic saponins have two sugar chains, often with one attached
through an ether linkage at C-3 and one attached through an ester linkage at C-28 (for triterpene saponins)
or an ether linkage at C-26 (steroid and steroid alkaloid saponins). Tridesmosidic saponins have three
sugar chains and are seldom found.
Saponins are often subdivided into three main classes, the tritepenoid saponins, steroid saponins,
and the steroid alkaloid saponins.
Monocotyledons are the main source of steroid saponins with the Liliaceae,
Dioscoreaceae and Agavaceae providing many representatives. The most important saponin-bearing
genera are: in the Liliaceae family Trillium, Chlorogalum, Smilax, Nolina, Agapanthus; in the Agavaceae
the Agave, Yucca, and Manfreda; in the Dioscoreaceae the Dioscorea. Of the dicotyledons, the genera
Digitalis (Scrophulariaceae) and Trigonella (Leguminosae) have been found to contain steroid saponins.
Steroid alkaloid saponins (glycoalkaloids, azasteroids)
The Solanaceae family constitutes the largest source. Glycoalkaloids are usually found in
all organs of the plant, with the highest concentrations in regions of high metabolic activity, such as
flowers, unripe berries, youg leaves and sprouts.
In the Solanaceae family, although steroid alkaloids have been isolated from all organs,
the complex glycoalkaloid mixtures are generally found in larger amounts in the roots than in the aerial
IV. Extraction Procedures
Extraction procedures in extracting saponins have to be mild as possible because certain
types of saponins can undergo different transformations. Some of those transformations are
enzymatic hydrolysis during water extraction, esterification of acidic saponins during alcohol
treatment, hydrolysis of labile ester groups, and transacylation. Choosing the best extraction
procedure and materials must be considered in the first place, to avoid the said transformations.
As in any extraction process, the extraction solvent, extraction conditions, such as
temperature, time, pH, solvent to feed ratio, and the properties of the feed material, such as
composition and particle size are the main factors that determine process efficiency and the
properties of the end product.
There are numerous and variety of methods in extracting saponin from a saponin-
containing plant. First method is by maceration by using methanol, ethanol, water or aqueous
alcohol as extracting solvent. Dried part of the plant is macerated in 180ml of the said solvents
with strength of 95% for three days at room temperature and the resulting extract is filtered
through filter paper. The residue from the filtration is extracted again twice using the same
procedure. The filtrates combined and then evaporated to dryness under reduced pressure. For
this method, it is said that methanol is the best solvent to use since it gave a double yield of the
crude extract when compared with the rest. Methanol is primarily used as an extracting solvent
because it is easily obtain and gives an efficient and the best result of extracts. Water is a less
efficient extraction solvent for saponins, unless, specifically water-soluble glycosides are desired.
Water, as an extracting material has the advantage of being easily lyophilized and gives a clear
extract. Depending on the proportion of water used for extraction, either monodesmosidic or
bidesmosidic saponins may be obtained. All in all, methanol is mostly used in many methods of
extracting saponin from a saponin-containing plant part or plant.
Some of many methods are the following: Method number two is by decoction. Dried
plant material was decocted in 300-ml of water at 60 degrees Celsius for 3 hours then, the extract
was filtrated. The filtrate was dried using freeze-dried technique. Method number three; the dried
plant material was soaked in 300-ml water for 24 hours prior to maceration with 95% ethanol
(180 ml). The rest of the extraction procedure was performed as described in the Method 1.
Method number four; the dried plant material was defatted with hexane (180 ml) for 24 hrs prior
to maceration with 95% ethanol (180 ml). The rest of the extraction procedure was performed as
described in the Method 1. Method number five, the dried plant material was extracted with 95%
ethanol (400-ml) using a Soxhlet apparatus set at 50 oC for 3 hours. The extract was filtrated and
dried under reduced pressure. Method number six, the dried plant material was extracted using
the same procedure as described in Method 3 except that after the plant material had been soaked
with water for 24 hrs; the water was squeezed out of the plant material prior to maceration with
95% ethanol. Method number seven, the dried plant material was soaked in 300-ml water. After
24 hrs, the water was squeezed out of the plant material. The plant material was extracted with
400-ml 95% ethanol using a Soxhlet apparatus set at 50 oC for 4 hrs. The extract was filtrated
and dried under reduced pressure. Method number eight, the dried plant material was soaked in
300-ml water. After 24 hrs, the water was squeezed out of the plant material. The plant material
was percolated with circulating 95% ethanol (200 ml) for three rounds. The residue was
extracted again twice using the same procedure. The combined extract was filtrated and dried
under reduced pressure. Method number nine is a defatting step, generally with petroleum ether,
applied before the extraction step or on the extract itself; or with N-butanol saturated, by
dissolving or suspending the extractives in water on the said mixture. Lastly, an optional
procedure like precipitation of saponins with diethyl ether or acetone can also be used.
%yield of the extract was calculated using the following equation.
%yield = Wcrude extract/Wdried plant x 100
Wcrude extract = weight of crude extract
Wdried plant = weight of dried plant material
V. Medicinal Uses
Triterpene saponins possess antimicrobial properties. It has strong fungicidal activity. Numerous
early examples of these saponins exist in the literature. The strongest activity is exhibited by
monodesmosidic saponins. Shorter carbohydrate chains lead to lower water solubility and weaker
antifungal activity.One example is Maesasaponin mixture isolated from Maesa lancelota which inhibits
growth of fungi at 50mcg/ml, Candida albicans at 100mcg/ml and Microsporum langeroni at 250mcg/ml.
Fungicidal activity to Candida albican strains can be influenced by the variation of the etherglycosidically
bonded carbohydrate units and the aglycosidically bonded oligosaccharide at C28 of the aglycone.
Furostanol saponins generally have antifungal MIC values 125mcg/ml to 1000mcg/ml and antiyeast MIC
values from 12.5 to 10 mcg/ml. Saponins also possess antiviral properties. The oleanane-type inhibits the
DNA synthesis of herpes simplex virus type 1, while the ursane-type saponin inhibits its viral caspid
protein synthesis. Herpes simplex virus is inactivated at 250µg/ml. Many other types of saponins
inactivate different kinds of viruses. These include human type A and B influenza viruses, polio
virus(250µg/ml), vaccinia, vesicular stomatitis virus, adenovirus type 6, rubella virus, rhinovirus, human
immunodeficiency virus (HIV)(0.5 mg/ml). The problem however is that most saponins had low
therapeutic indexes thus, small increase in concentration gives toxic effects. Though it elicits
antimicrobial activity on both prokaryotic and eukaryotic organisms, saponins show weak antibacterial
activity. It is also found that tetraglycoside saponins have stronger activity than triglycoside saponins.
Some examples of saponins which show antibacterial activities are 5β-spirostan-3β-ol saponins and
jujubogenin saponins. It has an MIC (Minimum Inhibitory Concentration) of 10 µg/ml.
Large numbers of saponins exhibit an anthelmintic activity but many find no practical application
because of the parallel irritation of mucous membranes. Some of the common helmiths killed by saponins
are liver flukes Fasciola hepatica and Dicrocoelium spp. at concentrations of around 0.005mg/ml. Some
saponins also elicit an antiprotozoal activity against Plasmodium falciparum. Saponin fractions
(31.8µg/ml) had slightly better antiplasmodial activity thsn pure glnoside A (42.3µg/ml).
Saponins are known for their high cytotoxic activities, this property enables some saponins to
also have antitumor properties. Many saponins have been proven to have antitumor activities but are not
widely used because of its small therapeutic window. One exception to this is a saponin from Acer
negundo which has active yet high therapeutic index, giving good survival rates at doses of around
1.5mg/kg. Antineoplastic (P-388 leukemia) activity has also been reported. Some saponins also elicit
antimutagenic activity at 200µg.
Saponins also have spermicidal activities. The mechanism of action involves disruption of
spermatozoid plasma cell membrane. It gives 100% immobilization at 0.5% to 0.25% levels. Strong
inhibition of spermatozoid motility was associated with a high hemolytic index.
Saponins produce a general and non-specific ability to produce local irritation, especially of
mucous membranes. This takes place in nasal cavity, throat, the bronchi, the lungs and in kidney epithelia.
One effect is sneezing provoked by saponins in powder form. The irritation of the throat and respiratory
tract increases respiratory fluid volume by drawing more water into the bronchial secretions, hence
diluting the mucus and reducing its viscosity. The surface activity of saponins also renders the sputum
less viscid, making it more mobile and easier to eject. Another is its amphiphilic nature that causes them
to spread out as monomolecular film at the back of the throat and thus aid the elimination of mucus.
The diuretic action of many saponins can be traced to a local irritation of kidney epithelia.
Diuresis caused by certain saponin-containing plants such as Ononis, Herniana, Betula, and Solidago
species is relatively mild, and the effect may just as well originate from the accompanying flavonoids and
essential oils. An alternative theory is that the potassium content of these plants is in effect the diuretic
agent. The flowering tips of the branches of Solidago virgaurea, S. serotina, and S. canadensis are all
used in the treatment of inflammatory complaints of the urinary tract and urinary caculi.
Saponins are also involved in cholesterol metabolism. Oleanolic acid saponins from Aralia
mandshurica (Asteraceae), Calendula officinalis (Asteraceae) and Beta vulgaris (Chenopodiaceae) fed by
mouth reduce total lipid content, triglyceride content and cholesterol content of rats by up to 37%, 30%,
and 25%, respectively. Alfalfa saponins are reported to depress concentrations of lipids and cholesterol in
the livers of mice. Soybean saponins reduce intestinal uptake of cholesterol in the rat.
Crdiovascular activity in saponins have also been recorded. Antiarrhythmic activity in rats and
rabbits has been demonstrated by panaxatriol saponins from Panax notoginseng. Seed saponins of
Ziziphus jujuba are hypotensive in normal rats and cats. Vasodilatory effects have been shown by crude
saponins of Panax notoginseng.
Many saponins are capable of inducing the release of adrenocorticotropic hormone (ACTH) and
corticosterone in rats but this does not appear, all the same, to be a general phenomenon of triterpene
saponins. Certain saponin-containing plants have capillaro-protective properties. It has been known that
seed extracts of horse chestnut (Aesculus hippocastanum, Hippocastanaceae) have beneficial effects on
The best known of the saponin antiulcerogenic drugs is glycyrrhiza. Protective effects in various
experimentally induced gastric ulcer models in rats have been shown by a saponin-containing ethanol
extract of Pyrecantha staudtii (Icacinaceae) leaves. Ginseng flower saponins were effective in treating
The root extract of Platycodon grandiflorum (Campanulaceae) has a stronger analgesic
effect than a 100-200 mg dose of acetylsalicylic acid (aspirin). Barbatosides A and B from Dianthus
barbatus are both analgesic and anti-inflammatory. Analgesic effects are also shown by saponins from
Panax notoginseng, from Crossopteryx febrifuga, from Dolichos falcatus and from the bark of Zizyphus
rugosa. Root saponins of Bupleurum chinese (Umberlliferae) produce antipyretic effects on rabbits.
Immunomodulators boost the immune system of an organism, and immunosuppressive
factors, which block the immune system. Immunomodulators includes immunostimulants which are
employed during chemotherapy, in the treatment of infectious diseases, and to stimulate the immune
response after vaccination. Saponins from Quillaja saponaria (Rosaceae), when administered orally,
stimulate both Th1 immune response and the production of cytotoxic T lymphocytes against exogenous
antigens which makes it ideal for use in subunit vaccines and vaccines directed against intacellular
pathogens as well as for therapeutic cancer vaccines. Some are restricted for human vaccination use due
to its high toxicity and undesirable haemolytic effect. Plants important for their tonic and so-called
“adaptogenic” properties are Panax ginseng, Eleutherococcus senticosus, and Aralia mandshurica, all
from the family Araliaceae.
Certain chinese herbal medicines are reputed to have sedative activity: Polygala
tenuifolia roots (Polygalaceae), Ziziphus jujube seeds (Rhamnaceae), and Panax japonicum rhizomes
Many saponins isolated from plant sources produce an inhibition of inflammation.
Fruticesaponin B, a bidesmodic saponin with unbranched saccharide was shown to have the highest
inflammatory activity. Aecsin also show anti inflammatory action at oral dose of 50mg/ml.
Saponins from Ficus platyphylla are believed to have profound central nervous system activities.
It protects from seizures at 25 to 100 mg/kg.
Ilexosides A-F and G-I from Ilex crenata exhibit antiallergic activity. They are approximately 10
times more potent as glcyrrhizin in inhibiting histamine release from mast cells.
Cytotoxic / Antitumor
Plant ( type of saponin Microorganism killed/
Pharmacologic activity Dose
and part used) Action
Maesa lancelota 50µg/ml Epidemophyton
100µg/ml Candida albicans
Panax notoginseng Aphanomyces
Colubrina retusa 50 µg/ml Candida albicans
Solanum chrysotrichum 100 µg/ml Aspergillus niger
200 µg/ml Candida albicans
Antiyeast Capsicum annuum 12.5 to 10 µg/ml
Colubrina retusa 10 µg/ml Mycobacterium
Capsicum annuum > 1000 µg/ml Gram + and -
Maesa lancelota 250 µg/ml Herpes simplex virus
Camellia sinensis Human influenza a and
Aesculus chinensis 100 µg/ml HIV
Furcraea foetida 4 µg/ml Mutant p53
Aralia dasyphylla 1.2 µg/ml KB cells
0.02 µg/ml HeLa-S3 cells
Acanthophyllum 10 µg/ml Lymphocyte
Ononis, Herniana, - -
Flowering tips of the - Treatment of
branches of Solidago inflammatory
virgaurea, S. serotina, complaints of the
S. Canadensis urinary tract and
Aralia mandshurica - Reduce total lipid
Calendula officianalis - content, triglyceride
Beta vulgaris - content and cholesterol
content of rats
Alfalfa saponins - Depress concentrations
of lipids and cholesterol
in the livers of mice
Panax notoginseng - Antiarrhythmic action
Ziziphus jujuba -
Panax notoginseng - Vasodilatory action
Effects on capillary fragility
and venous stasis/ venous
Aesculus - Contains capillo-
hippocastanum (horse protective properties
Glycyrrhiza (best - -
known of the saponin
Dianthus barbatus Both analgesic and anti-
Ziziphus rugosa (bark)
Chinese herbal medicines Polygala tenuifolia
Ziziphus jujube (seeds)
Ficus platyphylla 25 to 100 mg/kg
a. Antiallergy Ilex crenata
Glycoalkaloid α- 1.0 – 0.0001 M Inhibits the growth of
tomatine (tomato) Fusarium oxysporum,
Glycoalkaloid α- Active against
Solanine (potato Trichoderma viride,
and other fungal stain
Glucoside and Active against the
galactoside fungus Trichophyton
Deltonin and deltoside Active against Fusarium
from Dioscorea solani and
deltoids Phytophthora infestans
Pennogenin glycoside 6.25 µg/ml Antifungal activity
from Dracaena mannii against all strains –
Sarsasapogenin 0.5 µg/ml to 8 µg/ml Active against Candida,
triglycoside As-1 from Cryptococcus,
Asparagus officinalis Trichophyton,
(Liliaceae) Microsporum and
Asterosaponins from Inhibit influenza virus
Asterias forbesi, multiplication
Acanthaster planci and
Costus speciosus 5-500 µg/100g body Abortifacient
Afromontoside from 100 µg/ml
Diosgenin and 10 µg/ml