Herbal Liposomes: Natural Network for Targeted Drug Delivery System

Abstract

Herbal medicines have tremendous therapeutic potential that can explored across various effective drug delivery system. Decoctions, herbal teas, tinctures, glyceritum, oxymel, and use much soap, herbal tablets, herbal capsules, and herbal cream, herbal books, and prepared the confection of the most commonly available forms of dosage. The less use of herbal formulations in recent decades due to their lack of standardization. It is possible to use plant extract and isolated constituents to overcome this problem. But these phytoconstituents are suffering from drawbacks, mostly due to problems with stability and low lipid solubility. Novel drug delivery such as liposomes plays an important role in problem solving. Infact, compliance with the patient also improves. The review article discusses the recent status of new herbal liposomal formulations and describes the different ways in which these formulations are prepared.

Full text

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*Corresponding author: E-mail: vrsalunkhe@rediffmail.com;
Journal of Pharmaceutical Research International
33(29B): 31-41, 2021; Article no.JPRI.67442
ISSN: 2456-9119
(Past name: British Journal of Pharmaceutical Research, Past ISSN: 2231-2919,
NLM ID: 101631759)
Herbal Liposomes: Natural Network for Targeted
Drug Delivery System
Vijay R. Salunkhe
1*
, Prasanna S. Patil
1
, Ganesh H. Wadkar
1
and Somnath D. Bhinge
1
1
Rajarambapu College of Pharmacy, Kasegaon, Dist. Sangli, Maharashtra, India.
Authors’ contributions
This work was carried out in collaboration among all authors. All authors read and approved the final
manuscript.
Article Information
DOI: 10.9734/JPRI/2021/v33i29B31586
Editor(s):
(1) Dr. Sawadogo Wamtinga Richard, Scientific Research and Innovation, Burkina Faso.
Reviewers:
(1) Viviane Fonseca Caetano, Universidade Federal de Pernambuco, Brazil.
(2) María Cristina Sifuentes Valenzuela, Universidad Nacional Autónoma de México, México.
Complete Peer review History:
http://www.sdiarticle4.com/review-history/67442
Received 01 February 2021
Accepted 05 April 2021
Published 24 May 2021
ABSTRACT
Herbal medicines have tremendous therapeutic potential that can explored across various effective
drug delivery system. Decoctions, herbal teas, tinctures, glyceritum, oxymel, and use much soap,
herbal tablets, herbal capsules, and herbal cream, herbal books, and prepared the confection of the
most commonly available forms of dosage. The less use of herbal formulations in recent decades
due to their lack of standardization. It is possible to use plant extract and isolated constituents to
overcome this problem. But these phytoconstituents are suffering from drawbacks, mostly due to
problems with stability and low lipid solubility. Novel drug delivery such as liposomes plays an
important role in problem solving. Infact, compliance with the patient also improves. The review
article discusses the recent status of new herbal liposomal formulations and describes the different
ways in which these formulations are prepared.
Keywords: Herbal liposomes; targeted drug delivery; preparation.
Review Article

Salunkhe et al.; JPRI, 33(29B): 31-41, 2021; Article no.JPRI.67442
32
1. INTRODUCTION
India officially recognised alternative
health programs have a long, safe, and
reliable use of many herbal medicines
[1]. Millions of Indians often use herbal
medicines as herbs, home remedies,
natural foods and medicines as self-medicinal
products or as non-allopathic products
[2,3]. Herbal medicines are thought to
be as ancient as human beings. Plant or
plant preparations have been commonly
used in medicine since ancient times. Up to now,
phytomedicines have been commonly used in
most of the world's population [4].
Herbal medicines are believed to be as
old as human beings. Since ancient times
plants materials have also been widely
used in medicine. Phytomedicines have also
been widely used in most of the world's
population thus far [5]. Moreover, Advancement
in the Ayurvedic herbal medicine has
been revolutionized from the showing of
phytochemicals and pharmacological
activities to elucidating their mechanisms of
action and sites of action [6,7]. Some
drawbacks of herbal medicines and
phytochemicals, such as highly acidic pH
instability, problems with solubility and
absorption, can lead to medication levels
below plasma therapy concentration,
resulting in little or no therapeutic effects [8].
Incorporating novel drug delivery new
technologies into active plant substances
minimizes presystemic metabolism, drug
degradation in the gastrointestinal tract,
distribution / accumulation of drugs in
nontargeted tissues and organs, thereby
reducing side effects and increasing
therapeutic effectiveness and eventually patient
compliance [9]. In this situation we need a
capable drug delivery system by using
various biomaterials such as biodegradable
nanomaterial [10]. Therefore liposomes
considered as a novel targeted drug delivery
system tool. Liposomes can be prepared
according to different methods. They may vary in
their dimensions, composition (different
phospholipids and cholesterol contents),
charge (resulting from the charges of the
composing phospholipids), and structure
(multilamellar liposomes consisting of several
concentric bilayers, separated by aqueous
compartments or unilamellar liposomes,
consisting of only one phospholipid
bilayer surrounding one aqueous compartment)
[11].
1.1 Novel Drug Delivery System
1.1.1 Liposomes
The integration of the drug into the carrier
network takes place in novel drug delivery
technology or alters the drug structure at the
molecular level to achieve distribution efficiency.
New ideas for monitoring pharmacodynamics,
pharmacokinetics, immunogenicity, non-specific
toxicity, drug biorecognition and efficacy have
been developed. Such new approaches, also
known as targeted drug delivery are based on an
interdisciplinary approach integrating science
of polymers, bioconjugate, chemistry
pharmaceutical and molecular biology. The novel
drug delivery system is built to distribute drugs
continuously over a longer duration of circulation,
at a consistent and reproducible rate. Potential
benefits of this concept include prevention of
drug-related side effects due to controlled blood
levels rather than fluctuating blood levels,
improved patient compliance due to decreased
dosage frequency and reduced total medication
dosage [12]. Bangham first described liposomes
during the study of cell membranes in 1965 [13].
He found the liposomes to be vesicles structures
composed of hydrated bilalyers that develop
spontaneously as water disperses phospholipids
[14,15]. More studies have been carried out on
liposomes and their use in various fields, such as
medicine and science [16]. The term liposome
derives from two Greek words: "Lipos" means fat
and "Soma" means body [17]. A liposome is a
vesicle made out of the same material of a cell
membrane. Usually these consist of
phospholipids, which are molecules that consist
of a tail and a head portion [18]. The head is
hydrophilic, while the tail consisting of a long
hydrocarbon chain is hydrophobic. Phospholipids
are typically found in bilayer form.The main
reason it has for advancing work into liposomes
is largely because liposomes can simulate
biological cells.This also ensures that liposomes
are extremely biocompatible, making them an
ideal candidate for a drug delivery system, with
uses ranging from enzymes, antibacterials,
antiviral drugs, antiparasite drugs, fungicides,
transdermal carriers, vaccine diagnostic tools
and adjuvants. Antitumor and antifungal agents
in liposome form are commercially available
today [19]. Its amphipathic nature is a typical
feature of bilayer-forming lipids. A collection of
polar heads attached covalently to one or two
hydrophobic hydrocarbon tails.Once such lipids,
e.g. phosphatidyl glycerol, phosphatidyline or
phosphatidyl ethanolamine, are exposed to an

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33
aqueous environment, interactions among
themselves (hydrophilic interaction between the
polar head groups and van der Waals
interactions between hydrocarbon chains and
hydrogen bonding with to water molecules) lead
to condition formation of closed bilayers.The size
of the liposomes can differ, ranging from smallest
vesicles (20 nm in diameter) to visible liposomes
under an optical microscope (diameter equal to
or greater than 1 µm or greater than the size of
living cells) [20]. A drug encapsulated by
liposomes achieves a long-lasting therapeutic
standard as medication must first be released
from liposomes before metabolism and excretion
[21]. Liposomal products have also been
extensively employed mostly in past decade to
improve drug delivery efficiency across multiple
routes of administration. The major benefits of
topical liposomal drug formulations are accrued
by their demonstrated ability: (i) Reduced
adverse effects and incompatibilities arising from
exceptionally high systemic absorption of
medications (ii) Boost substantially the
accumulation of drugs also at site of
administration as a result of the high substantive
rates of biological membranes liposomes (iii) The
number of hydrophobic and hydrophilic drugs
could be readily integrated. Liposomes are often
biodegradable, non-toxic and are designed for
large-scale preparations. In addition, they also
provide selective passive targeting of tumor
tissues, increased efficacy and therapeutic index,
increased stability through encapsulation,
reduced toxicity of encapsulants, improved
pharmacokinetic effects, and site specificity
Ligand coupling to achieve the flexibility of active
targeting.Structural liposomes consist primarily of
two components: Phospholipids and cholesterol
[22]. Liposomes can also be divided into two
categories: (1) Multilamellar vesicles (MLV) and
(2) unilamellar vesicles. Unilamellar vesicles can
also be classified into two categories: (1) Large
unilamellar vesicles and (2) small unilamellar
vesicles. The vesicle has one single phospholipid
bilayer sphere in unilamellar liposomes that
surrounds the aqueous solution. Vesicles have
an onion-structure in multilamellar liposomes.
Classically, several monolayer vesicles of
smaller size can develop within other vesicles to
form a multilayer structure of concentrated
phospholipid spheres separated by a layer of
water [23].
1.1.2 Methods of liposome preparation
For liposome preparation, the following methods
are used:
1. Passive loading techniques
2. Active loading technique.
Passive loading techniques include three
different methods:
A) Mechanical dispersion method
B) Solvent dispersion method
C) Detergent removal method
A) Mechanical dispersion methods are the
following types:
 Lipid film hydration by hand shaking, non-
hand shaking or freeze drying
 Micro-emulsification
 Sonication
 French pressure cell
 Membrane extrusion
 Dried reconstituted vesicles
 Freeze thawed liposomes
B) Solvent dispersion methods are following
types:
 Ethanol injection
 Ether injection
 Double emulsion vesicles
 Reverse phase vesicles
 Stable plurilamellar vesicles
C) Detergent removal methods are following
types:
 Detergent(cholate, alkylglycoside, Triton X-
100) removal from mixed micelles by
 Dialysis
 Column chromatography
 Dilution
 Reconstituted Sendai virus enveloped
vesicles
1.1.2.1 Quercetin liposomes
Quercetin is well known as an effective
antioxidant which protects against damage from
free radicals or reactive oxidative species (ROS)
associated with oxidative stress.In particular, the
antioxidant quercetin activity may be attributable
to the ability of catalase to protect cells from
oxidative stress and damage by scavenging
H
2
O
2
. The capacity of quercetine as an
anticancer agent. When used in combination,
quercetin appears to increase the cytotoxicity of
cisplatin to ovarian cancer in the various murine
models of cancer.Low dissolution in
gastrointestinal fluids leads to slow

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34
gastrointestinal absorption of quercetin due to its
poor aqueous solubility, which results in low
bioavailability. Its solubility needs to be enhanced
and different methods have been used to
achieve this, including the development of more
water-soluble quercetin compounds, the use of
liposomes in formulating approaches. Quercetin
liposomes were developed taking advantage of
the ability of quercetin to bind copper.Quercetin
powder was applied directly to preformed
liposomes cholesterol (CHOL) and (2-distearoyl-
sn-glycero-3-phosphocholine (DSPC) containing
copper [24]. Another approach used to prepare
quercetin-loaded liposomes is the thin film
hydration process. Phospholipids, cholesterol,
and quercetin were dissolved in 25 ml of
chloroform-methanol mixture (4:1) at a constant
molar ratio.In the rotary evaporator, the mixture
is evaporated to remove traces of solvent and
form thin film. At room temperature, hydrate the
film with phosphate buffer (pH 7.4) for 1 hour.
The dispersion of the vesicles was then
homogenized by means of a sonicator probe.
Optimized liposomal formulations were explored
using surface-response methodology (RSM) [25].
1.1.2.2 Curcumin liposomes
Curcumin is one of the most widely studied
bioflavonoids today and many studies have
confirmed its antioxidant, anti-inflammatory, anti-
cancer, chemoprotective and gastroprotective
properties [26]. Curcumin is a natural compound
with many antitumor properties and specificity of
the tumor cells, but it is poor in bioavailability and
water solubility. Liposomes increase the
therapeutic index of curcumin by shielding the
drug from enzymatic degradation, and surface
modulation helps polyethylene glycol (PEG) to
circulate long-term. Curcumin has such a range
of anti-cancer effects including tumor blocking,
tumor growth avoidance, and invasion and
metastasis inhibition [27]. Curcumin liposomes
prepared using hydration method of thin film and
using dilution method of polyol. In thin film, lipid
hydration method was dissolved in chloroform,
placed in round bottom flask then dried under
reduced pressure in rotary evaporator to form
thin lipid film on the flask's inner surface. The
phospholipid film was kept in PBS buffer (pH
7.4), sonicated for 30 minutes at 4°C and
rehydrated afterwards. Liposomes under UV
lamp is sterilized for 2h [28]. Method for diluting
polyol without residual organic solvents.
Curcumin liposomes have a molar ratio of 9:1
and consist of phosphatidylcholine (PC)
hydrogenated and cholesterol (CH). The polyol
solvents used are propylene glycol (PG),
polyethylene glycol 400 and glycerine (PEG-
400). Extrusion was applied following suspension
[29].
1.1.2.3 Paclitaxel liposomes
Some of the most promising paclitaxel anticancer
drugs particularly effective in treating breast and
ovarian cancer suffer from problems such as low
water solubility and low bioavailability. Products
currently available in non-aqueous vehicles
containing Cremophor EL
®
(polyethoxylated
castor oil) can produce allergic reactions as well
as precipitation when administered intravenously
in an aqueous diluent.The lack of any effective
vehicle delivery restricts and delays the
widespread clinical use of this drug. It is
therefore strongly recommended to develop
alternative formulation of paclitaxel with strong
aqueous solubility and reduced side effects [30].
Paclitaxel loaded liposomes have been
developed with the objective of enhancing the
cancer treatment effects. The encapsulating
Paclitaxel liposomes were prepared in 20 percent
ethanol using a thin film hydration process using
PTX-saturated 20 percent ethanol, bovine serum
albumin (BSA) solution and bovine serum
albumin (BSA) solution, and the amount of PTX
loading in the liposome [31].
1.1.2.4Colchicine liposomes
Colchicine, an alkaloid present in the plants
Colchicum autumnale and Gloriosa
extractsSuperb, is effective in treating acute gout
and dermatological disorders such as
leukocytoclasticvasculitis, psoriasis, and the
syndrome of sweets. Oral colchicine
administration also has dose-dependent side
effects, suggesting the need to develop
alternative dosage form which topically delivers
colchicine to the affected joints. The vesicular
systems were widely used for the delivery of
transdermal and dermal drugs.The vesicular
system is heavily dependent on its physico-
chemical properties. Deformable vesicles such
as elastic liposomes have become more effective
in improving the transport of drugs. The elastic
liposomal formulation of Colchicine was prepared
using traditional rotary method of sonication by
evaporation40±1.0°C. At room temperature, the
obtained vesicles were swollen for 2 hours to
obtain large multilamellar vesicles (LMLV).
LMLVs have been sonicated at 4°C and 40 W for
20 minutes to prepare smaller vesicles. The

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sonicated vesicles is extruded by a sandwich of
polycarbonate membranes 100 and 200 nm [32].
1.1.2.5 Silymarin liposomes
A hepatoprotective agent, Silymarin has poor
bioavailability for oral use. The new form of
dosage drug doesn't affect the liver and
inflammatory cells directly. To develop a lecithin-
based silymarin carrier system by incorporating a
phytosome-liposome approach to enhance oral
bioavailability and target specificity to improve
liver hepatoprotection. The liposome for silymarin
was prepared using film hydration method. It is
focused on the fact that phytosomal silymarin is
more stable in the gastric environment to
improve silymarin bioavailability while liposomal
silymarin has the highest capacity to be captured
and modulated by macrophages, Kupffer cells,
and infiltrated WBC viz., neutrophils, monocytes,
etc.This process makes it possible to target
silymarin-targeted inflammation in the
formulation. Liposomes are prepared by the film
hydration method. Put 10 mg of silymarin (S),
different amounts of soybean
phosphatidylcholine and cholesterol into round
bottom flask and dissolve them in a chloroform
methanol mixture (1:9). At 40°C the solvent was
then evaporated to form a thin film on a rotary
evaporator under vacuum. Dry in a vacuum
desiccator overnight, eliminated traces of solvent
from the film. The film was prepared in
phosphate buffered saline (PBS, pH 7.4)
containing different quantities of cryoprotectants
(mannitol and sucrose) at 100 RPM for 1 hour at
50°C hydrated to prepare the liposomal
suspension. After 5 cycles the liposome vesicle
size decreased at 20,000 psi under high
pressure homogenization. The liposomes had
been placed in a deep refrigerator at 80°C
overnight. The frozen liposomes were lyophilised
at low pressure and placed in the airtight
container at 4°C [33].
1.1.3 Liposomal neem gel
In order to improve its effectiveness, different
synthetic drugs and herbal drugs are
incorporated into liposomes. The incorporation of
herbal extracts into liposomes can eliminate the
side effects associated with synthetic lipids.
Azadirachta indica leaves have strong
antibacterial activity which confirms the great
potential of bioactive neem compounds.
Azadirachta indica aqueous extract and alcoholic
extract, alcoholic leaf extract, have been found to
be more aggressive against the bacterial
species. This extract has been integrated into the
liposomes to boost its skin delivery function.
Methanolic Neem Extract (MeNE) was
introduced by hydration of thin film into the
liposomes. A lipid layer was prepared by
dissolving in the chloroform-methanol mixture
(2:1 v / v) accurately weighed amounts of
Methanolic Neem Extract (MeNE), soya lecithin
and cholesterol in the round bottom flask
containing glass beads.Rotary evaporation at 45-
50°C, under reduced pressure, eliminated the
solvent mixture from the lipid process to produce
the thin layer of lipids on the flask wall and bead
surface. The dried lipid film was hydrated in
phosphate buffer pH (6.5) at a temperature of 60
± 2°C. The dispersion was left to stand at the
room temperature for 2-3 hours to completely
swell the lipid membrane and obtain a vesicle
suspension [34].
1.1.3.1 Capsaicin liposomes
Capsaicin is natural compound and has poor
bioavailability for oral use. For enhanced oral
bioavailability, capsaicin, an important
medication for treatment of the neuropathic pain,
can be encapsulated into liposome.Another
promising method in development of liposome
formulations may be the excellent in vitro-in vivo
correlation (IVIVC) of capsaicin-loaded
liposomes, which has the additional benefit of
reducing animal testing. Thin film hydration
process used to formulate liposomes. Capsaicin,
soybean lecithin with a phosphatidylcholine was
dissolved in a single-neck flask in 20 ml of
absolute ethanol and subjected to ultrasound
until the solution was clear and transparent. The
solution was evaporated using rotary evaporator
to eliminate ethanol before adding cholesterol
(0.2 g), sodium cholate (0.8 g) and isopropyl
myristate (0.8 g), and dissolving further in 20 ml
ethanol. The evaporation process was repeated
again to remove residual solvent, leaving a film-
like complex at the bottom of the flask. Double
distilled water hydrated the dried lipid film to
create a final solution with different
concentrations (2, 8, 15 mg-mL-1, called FI, F2
and F3 formulation, respectively).
The different preparations were stored at a 4°C
before further investigations into the efficiency of
encapsulation, the stability test and the effect of
different capsaicin concentrations on liposomal
systems [35].
1.1.3.2 Baicalein liposomes
Baicalein exhibited suppression against nitric
oxide / prostaglandin E2 production and cancer

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cell proliferation [36]. Because of its low oral
bioavailability it is inconvenient and troublesome
to use BAI. To enhance its bioavailability grow
nano liposomes containing flavonoids (BAI-LP).
Baicalein-loaded nanoliposomes (BAI-LP) were
prepared by hydration of thin films.The lipid
phase was prepared in a 25ml recovery flask by
dissolving correctly weighted amounts of
soybean phosphatidylcholine (SPC), cholesterol
and baicalein in 6ml dichloromethane-methanol
(2:1, V / V) mixture. The mixture was withdrawn
at 40°C by rotary evaporation, creating a thin
layer of lipids on the eggplant-shaped bottle wall.
The thin film was then purged for 5 minutes with
nitrogen.The lipids film was hydrated at 60°C in
an eggplant-shaped bottle with 10ml of ultrapure
water, accompanied by 15–30 minutes of
sonication.Samples were filtered using 450 nm
and 220 nm membrane filters to obtain yellowish
Baicalein-loaded nanoliposomes (BAI-LP) [37].
1.1.3.3 Brucine liposomes
Brucine itself is known for relieving arthritic and
chronic pain as an analgesic and anti-
inflammatory treatment. The major
pharmacodynamic activities include pain relief,
swelling reduction, and circulation
enhancement.Because of its high occurrence of
side effects, including violent seizures and even
lethal poisoning, the possible use of brucine is
extremely limited. Therefore, it is essential to find
a suitable formulation also for therapeutic
application of this molecule to minimize its
harmful side effects while preserving or possibly
improving its efficacy.Develop brucine liposome
to reduce side effects. Modified ethanol-dripping
method was used in brucine liposome
preparation. A lipid ethanol solution (lecithin:
cholesterol = 6:1, w / w), sodium deoxycholate,
tween-80 and brucine (16:4:4:1, w / w) was
discharged into mannitol solution (5.3 mg / ml).In
the final suspension the ethanolic-lipid liquid ratio
to the aqueous phase was 1:9 (v / v).Then the
suspension was sonicated for 20 min with an ice
water-bath and 72 h freeze-dried. The dry
powder may be rehydrated 3 min before
application [38].
1.1.3.4 Asparagus racemosus liposomes
Antiulcer, antioxidant, immunomodulatory,
antidiabetic, antidiarrheal, phytoestrogenic,
antiaging and adaptive properties have been
recorded for the pharmacological activity of
Asparagus racemosus root extract
(AR).Asparagus root has moisturizing, cooling,
anxiety, constipation, galactose regulation,
aphrodisiac, diuretic, rejuvenating, digestive,
stomachic and antiseptic properties, so it can be
used as a digestive asparagus root. The
Asparagus racemosus root has many beneficial
effects recommended for treating nervous
disorders, dyspepsia, vomiting, tumors and
inflammation.Preparation of thin film hydration
(TF) for Asparagus racemosus root extract
liposomes, reverse phase evaporation (REV),
and polyol dilution (PD) methods with different
lipid extract ratios.In a 250 ml round bottom flask,
thin film hydration (TF)-total lipid mixture (100
mg) with a PC to CHOL molar ratio of 7:3 has
been dissolved in 20 ml chloroform.The solvent
was eliminated from the lipid process under
reduced pressure, by rotating at 35°C until a dry
film was deposited on the flask wall. The dry thin
film was hydrated with an aqueous solution
containing various weight ratios of Asparagus
racemosus to total lipid at 0:5, 1:5, 2:5 and 3:5 in
a 20 ml isotonic phosphate buffer pH 7.0 above
phase transition temperature to obtain a
homogeneous white liposome suspension. The
suspension was left for further 2 hours at the
room temperature to achieve full swelling of a
lipid membrane. Use ultracentrifugation at 4°C
for 1 h, the unentrapped Asparagus racemosus
was isolated.Liposomes with isotonic phosphate
buffer pH 7.0 were washed several times until
the concentration of Asparagus racemosus in the
supernatant was < 1.0 percent. Before usage,
the filtered liposomes were moved to a container
and placed at 4ºC.Reverse-phase evaporation
(REV)method- The total lipid mixture (100 mg)
with a PC-to-CHOL molar ratio of 7:3 was
dissolved in 60 ml dichlotromethane in a round
bottom flask, after which the aqueous phase
containing different weight ratios of Asparagus
racemosus to total lipid (0:5, 1:5, 2:5 and 3:5 in
20 ml isotonic phosphate buffer , pH 7.0) was
injected into the lipid solution by means of a 22
gauge hypodermic needle with a 5 ml syringing
needle. With a glass stopper, the flask was
immediately sealed and placed in an ultrasonic
bath. At 7° C the mixture was sonicated. For 10
minutes the water-in - oil emulsion (w/o) is
formed. The emulsion was then transferred to a
rotary evaporator and, under reduced pressure at
35°C, the solvent was slowly evaporated until a
viscous gel was formed. Finally the initiation of
the evaporation culminated in a homogenous
aqueous dispersion. Liposomal dispersions were
subsequently subjected to complete removal of
the remaining organic solvent traces in a rotary
evaporator at 35 to 37ºC for approximately 30
minutes. Finally, the obtained liposome
dispersion was purified by ultracentrifugation in

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Table 1. Liposomal formulations
Formulations
Active ingredients
Biological activity
Method of preparation
Reference No
Quercetin loaded liposomes Quercetin Wound healing Thin film hydration [25]
Curcumin liposomes
Curcumin
Anticancer
Thin film hydration
[27]
Curcumin liposomes Curcumin Anti-inflammatory Thin film hydration [28]
Curcumin loaded thiolated polymer
coated liposomes
Curcumin Aphthous ulcer Thin film hydration [29]
Paclitaxel liposomes Paclitaxel Anticancer Thin film hydration [30]
Colchicine liposomes Colchicine Anti-gout Rotaryevaporation sonication method [32]
Silymarin liposomes Silymarin Hepatoprtective activity Reverse
evaporation technique
[33]
Liposomal neem gel Methanolic neem extract Antimicrobial
Thin film hydration [34]
Capsaicin liposomes Capsaicin Anti-inflammatory Thin film hydration [35]
Baicalein liposomes Baicalein Antitumor Thin film hydration [37]
Brucine liposomes Brucine Anti-inflammatory Modified ethanol dripping method [38]
Rutin liposomes Rutin Antioxidant activity Film dispersion
Method
[40]
Guggul liposomes Guggul Anti-inflammatory Sonication method [41]
Rutin trihydrate
Liposomes
Rutin Topical administration
as antioxidant
Thin film hydration method [42]
Doxorubicin PEG liposomes Doxorubicin Anti-cancer Evaporation method [43]
Herbal liposome Ketoconazole Seborrheic Dermatitis Thin film hydration [44]
QUE/RES-loaded elastic liposomes Quercetin Anti-cancer Thin lipid film method [45]
EGCG Quercetin hybrid liposomes Quercetin Antioxidant activity Thin film hydration [46]
Paclitaxel liposomes
Paclitaxel
Anti-proliferative
Thin film hydration
[47]
Paclitaxel-loaded stealth liposomes Paclitaxel Anti-cancer thin film hydration technique [48]
Asparagus racemosus liposomes Asparagus racemosus Anti-inflammatory Thin-film hydration, reverse-phase
evaporation and polyol dilution
[49]
Cosmeceutical liposomes avobenzone and arbutin Cosmetic Purpose Thin film hydration and reverse-phase
evaporation
[50]
PEGylated Liposomes Meloxicam Anti-cancer Thin film hydration [51]
Arbutin Liposomes Arbutin Skin-whitening activity Film dispersion method [52]
Vitamin loaded topical liposomal Vitamine E Anti-oxidant Film dispersion method [53]

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the same way as the TF method, and stored at
4°C until further use. Polyol dilution (PD)
method-Concentrate the total lipid mixture (100
mg) with PC to 50°C, and dissolve CHOL with a
molar ratio of 7:3 in 4 ml of propylene glycol.
Preheat aqueous phase to 50°C with various
weight ratios of AR to total lipids (0:5, 1:5, 2:5
and 3:5) in 16 ml of isotonic phosphate buffer
(pH 7.0) Then slowly infused. Lipid solution, and
mix for 45 minutes. The resulting liposome
dispersion was purified by ultracentrifugation in a
manner similar to that previously described by
the TF method and stored at 4°C until needed
[39].
1.1.3.5 Rutin liposomes
Rutin (quercetin-3-O-rutinoside) is a natural
glycoside to the flavonoid. This exhibits essential
scavenging properties both in vitro and in vivo on
oxygen radicals. It also exhibits several other
significant pharmacological properties, including
antiviral, vasoprotective, and anti-inflammatory.
Rutin shows low water solubility, which leads to
poor efficacy or bioavailability in oral absorption.
Therefore, the development of novel rutin
delivery systems to improve the therapeutic
effectiveness of rutin is of considerable interest.
Developing rutin liposome to improve therapeutic
efficacy of the rutin. Nanoparticles of rutin-loaded
liposome were prepared using film dispersion
method.The delivery systems were prepared in
ethanol media (10 ml), containing 0.15:5:1 rutin-
PC-CH under 40° C rotary evaporator and 1 h 60
r / min. In a rotary evaporator at 40°C for 30 min,
thin lipid film products were low hydrated using
phosphate buffer (15 mL, pH 6.8, and containing
Tween 80) at low speed. The mixtures were
sonicated for 10 min and then filtered through a
microporous filter membrane of 0.22μm and
matured at 4°C overnight [40].
1.1.3.6 Guggul liposomes
The antiinflammatory effects were identified
using a gum resin, i.e. guggul, obtained from
Commiphora Mukul. The preparation of guggul-
liposomes was accomplished by the use of
different cholesterol and guggul lipid
concentrations with phenyl butazone by
sonication method. Guggle lipid is accurately
weighed and dissolves on agitator at 700 rpm in
10 milliliters of distilled water until full dissolution,
another mixture of phenylbutazone and
cholesterol in ethanol was prepared until thin
layer formation. Both the mixtures were
diversified by 5 percent PVA solution up to 20 ml
and attuned volume. The mixture was sonicated
(3 cycles for 5 min) to form fine guggul-liposomal
vesicles [41].
Herbal liposomes are mostly studies as
compared synthetic drugs the details are
depicted in Table 1 with their method of
preparation, active ingredients and biological
activity.
2. CONCLUSION
As per the example discussed we have
concluded an application of new drug delivery
systems to phytoconstituents that result in
increased bioavailability, increased solubility and
permeability, dose reduction and consequently
side effects. With the development of
standardization, extraction, identification
techniques, scientists can now focus their
research on the creation of herbal medicines that
can suit the targeted delivery, substantially
reduced doses and side effect properties of the
conventional medicine method.
CONSENT
It is not applicable.
ETHICAL APPROVAL
It is not applicable.
COMPETING INTERESTS
Authors have declared that no competing
interests exist.
REFERENCES
1. Bhinge SD, Bhutkar MA, Randive DS,
Wadkar GH, Todkar SS, Development and
evaluation of antimicrobial polyherbal gel,
Ann. Pharm. Fr. 2017;75(05):349-358.
Available:https://doi.org/10.1016/j.pharma.
2017.04.006
2. Vaidya A, Devasagayam T, Current status
of herbal drugs in India: An Overview, J.
Clin. Biochem. Nutr. 2006;1-11.
3. Chavan RR, Bhinge SD, Bhutkar MA,
Randive DS, Wadkar GH, Todkar SS.
Green synthesis of silver and iron
nanoparticles using fresh plant extracts of
blumea eriantha dc: evaluation of its hair
growth promoting activity and partial
characterization, J Cosmet. Dermatol.
2020;20(04):1283-1297.
DOI: https://doi.org/10.1111/jocd.13713

Salunkhe et al.; JPRI, 33(29B): 31-41, 2021; Article no.JPRI.67442
39
4. Shejawal KP, Randive DS, Bhinge SD,
Bhutkar MA, Todkar SS, Mulla AS, Jadhav
NR. Green synthesis of silver, iron and
gold nanoparticles of Lycopene extracted
from tomato: Their characterization and
cytotoxicity against COLO320DM, HT29
and Hella cell, J. Mater. Sci.: Mater. Med.
2021;32:19.
DOI: 10.1007/s10856-021-06489-8
5. Chavan RR, Bhinge SD, Bhutkar MA,
Randive DS, Wadkar GH, Todkar SS.
Urade MN, Characterization, antioxidant,
antimicrobial and cytotoxic activities of
green synthesized silver and iron
nanoparticles using alcoholic Blumea
eriantha DC plant extract, Mater. Today
Commun. 2020;24:101320.
Avialble:https://doi.org/10.1016/j.mtcomm.
2020.101320
6. Randive DS, Shejawal KP, Bhinge SD,
Bhutkar MA, Wadkar GH, Jadhav NR.
Green synthesis of Silver and iron
nanoparticles of isolated
proanthrocynidine: Its Characterization,
antioxidant, antimicrobial and cytotoxic
activities against COLO320DM and HT29,
J. Genet. Eng. Biotechnol. 2020;18(43):1-
11.
Avialble:https://doi.org/10.1186/s43141-
020-00058-2
7. Randive DS, Bhinge SD, Sayyad SF,
Wadkar GH. Comparative standardization
of marketed formulations of fermented
biomedicine – Arjunaristha, Indonesian J.
Pharm. 2016;27(04):220-225.
8. Kulkarni G. Herbal drug delivery systems:
an emerging area in herbal drug research,
J. Chronother. Drug Deliver. 2011;2:113-
119.
9. Uhumwangho MU, Okor RS, Current
trends in the production and biomedical
applications of liposomes: a review. J.
Med. Biomed. Res. 2005;4:9-21.
10. Randive DS, Shejawal KP, Bhinge SD,
Bhutkar MA, Wadkar GH, Jadhav NR.
Green synthesis of gold nanoparticles of
isolated citrus bioflavonoid from orange:
Characterization and in vitro cytotoxicity
against colon cancer cellines COLO
320DM and HT29, Indian Drugs.
2020;57(08):61-69.
11. Rooijen NV. Liposomes, encyclopedia of
immunology (Second Edition). 1998;1588-
1592.
Avialble:https://doi.org/10.1006/rwei.1999.
0407
12. Sharma P, Verma S, Misri P, Global need
for novel herbal drug formulations, Int. J.
Pharmacog. Phytochem. Res.
2016;8:1535-1544.
13. Bangham AD. Surrogate cells or trojan
horses. The discovery of liposomes. Bio
Essays News Rev. Mol. Cell. Dev. Biol.
1995;17:1081–1088.
14. Rai M, Ingle Ap, Bansod S, Kon K./
Nanotechnology in diagnosis, treatment
and prophylaxis of infectious diseases.
2015;133-149.
Avialble:https://doi.org/10.1016/B978-0-12-
801317-5.00009-8
15. Cheepsattayakorn A, Cheepsattayakorn R.
Roles of nanotechnology in diagnosis and
treatment of tuberculosis. J. Nanotechnol.
Diagn. Treat. 2013;1:19 25.
16. Verma N, Roshan A, liposomes: A targeted
drug delivery system, AMS. 2015;2:65-70.
17. Samad A, Sultana Y, Aqil M. Liposomal
drug delivery systems: An update review,
Curr. drug deliv. 2007;4:297-305.
18. Dwivedi C, Verma S. Review on
preparation and characterization of
liposomes with application, J. Scientific &
Innov. Res. 2013;2:486-508.
19. Sanarova E, Lantsova A, Oborotova N,
Orlova O, Polozkova A, Dmitrieva M,
Nikolaeva N. Liposome drug delivery. J.
Pharm. Sci. Res. 2019;11:1148-1155.
20. Pathak S, Mishra R, Kumar S, Prakash G,
Parthasarthy R. Effect of cholesterol
concentration on size of liposome, IOSR J.
Pharm. Biol. Sci. 2012;1:50-53.
21. Bozzuto G, Molinari A, Liposomes as
nanomedical devices, Int. J. Nanomed.
2015;10:975–999.
DOI: 10.2147/IJN.S68861
22. Srinivas Murthy BR, Vishakha K, Prakash
P, Jamal Basha D, Govardhan T, Prathap
M, Formulation and in vitroevaluation of
liposomes loaded with mupirocin. Int. J.
Res. Pharm. Nano Sci. 2015;4(3):162-174.
23. Princely S, Dhanaraju MD. Design,
formulation, and characterization of
liposomal-encapsulated gel for transdermal
delivery of fluconazole. Asian J. Pharm.
Clin. Res. 2018;11:417-424.
24. Chen KTJ, Anantha M, Leung AWY,
Kulkarni JA, Militao GGC, Wehbe M,
Sutherland B, Cullis PR. Characterization
of a liposomal copper(II)-quercetin
formulation suitable for parenteral use,
Drug Deliv. Transl. Res. NP. 2019;1:202-
215.

Salunkhe et al.; JPRI, 33(29B): 31-41, 2021; Article no.JPRI.67442
40
25. Jangde R, Singh D. Preparation and
optimization of quercetin-loaded liposomes
for wound healing, using response surface
methodology, Nanomed Biotechnol.
2016;44(2):635-641.
26. Jovičić D, Jozinović A, Grčević M,
Aleksovska ES, Šubari D. Nutritional and
health benefits of curcumin, Food in Health
and Disease, scientific-professional journal
of nutrition and dietetics. 2017;6(1):22-27.
27. Jadia1 R, Kydd J, Piel B, Rai P. Liposomes
aid curcumin’s combat with cancer in a
breast tumor model, Oncomedicine.
2018;3:94-109.
28. Sinjari B, Pizzicannell J, D’Auror M,
Zappacost R, Gatt V, Fontana A, Trubiani
O, Diomede F. Curcumin/liposome
nanotechnology as delivery platform for
anti-inflammatory activities via
NFkB/ERK/pERK pathway in human dental
pulp treated with 2-hydroxyethyl meth
acrylate (HEMA), Frontiers in Physiology.
2019;10:1-11.
29. Kaneramit LK, Aiemsumang P, Kewsuwan
P. Development of curcumin liposome
formulations using polyol dilution method,
Songklanakarin J. Sci. Technol.
2016;38(6):605-610.
30. Sharma N, Kumar V. Liposomal paclitaxel:
Recent trends and future perspectives. Int.
J. Pharm. Sci. Rev. Res. 2015;31(1):205-
211.
31. Taguchi YOK, Sakuragi M, Imoto S,
Yamasaki K, Otagiri M. Preparation,
characterization, and in vitro/in vivo
evaluation of paclitaxel- bound albumin-
encapsulated liposomes for the treatment
of pancreatic cancer, ACS Omega.
2019;4(5):8693-8700.
32. Pal SH, Puneet U, Kumar TA, Jain S.
Elastic liposomal formulation for sustained
delivery of colchicine: in vitro
characterization and in vivo evaluation of
anti-gout activity, The AAPS Journal.
2009;11:54-64.
33. Kumar N, Rai A, Reddy ND, Raj PV, Jain
P, Deshpande P, et al. Silymarin
liposomes improves oral bioavailability of
silybin besides targeting hepatocytes, and
immune cells, Pharmacological Reports.
2014;66:788–798.
34. Singh A, Vengurlekar P, Rathod S. Design,
development and characterization of
liposomal neem gel. Int. J. Pharma Sci
Res. 2014;5:140-148.
35. Zhu Y, Wang M, Zhang J, Peng W,
Firempong CK, Deng W, et al. Improved
oral bioavailability of capsaicin via
liposomal nano formulation: preparation, In
vitro drug release and pharmacokinetics in
rats, Arch. Pharm. Res. 2015;38:512–521.
36. Ohtsuki T, Himeji M, Fukazawa H, Tanaka
M, Mimura HYA. High-yield production of
scutellaria radix flavonoids (Baicalein,
Baicalin and Wogonin) by liquid-culture of
Scutellaria baicalensis root-derived cells,
Braz. Arch. Biol. Technol. 2009;52:291-
298.
37. Li K, Zhang H, Gao L, Zhai Y, Shi M, Li J,
et al. Preparation and characterization of
baicalein-loaded nanoliposomes for
antitumor therapy, Journal of
Nanomaterials. 2016;1-9.
38. Xiang-qi QIN, Yuan YUAN, Chang-sheng
LIU , Qiu-yun WANG, Xi SHE, Bai-can
YANG. Preparation of liposomal brucine
and its pharmaceutical/pharmacodynamics
Characterization, Acta Pharmacol Sin.
2007;28(11):1851–1858.
39. Nathsiree P, Kanin R, Lalana K, Neti W,
Narong S. Experimental and therapeutic
medicine. 2016;2790-2796.
40. Guo Y, Shen LX, Lu YF, Li H-Y, Min K, Li
LF, Yu CY, Zheng X. Preparation of rutin-
liposome drug delivery systems
andevaluation on their In vitro antioxidant
activity, Chin. Herb. Med. 2016;8:
371-375.
41. Paliwal S, Sharma J, Tilak A, Sohgaura A,
Gupta C, Dave V. Synergistic effect of
guggul liposomes lipid nano-vesicles for
the treatment of inflammation, Int. J. Lif.
Sci. Bioengg. 1-13.
42. Aparajita V, Padmini R. Formulation and
characterization of rutintrihydrate
liposomes for topical delivery, Int. J.
Pharm. Res. 2015;7(3):29-37.
43. Alyane M, Barratt G, Lahouel M. Remote
loading of doxorubicin into liposomes by
transmembrane pH gradient to reduce
toxicity toward H9c2 cells, Saudi Pharm. J.
2016;24(2):166-175.
44. Dave V, Sharma S, Yadav RB, Agarwal U.
Herbal liposome for the topical delivery of
ketoconazole for the effective treatment of
seborrheic dermatitis, Appl. Nanosci.
2017;7:973–987.
45. Cadena PG, Pereira MA, Cordeiro RBS,
Cavalcanti IMF, Neto BB, Pimentel MCCB,
Filho JLL, Silva VL, Santos-Magalhães NS.
Nanoencapsulation of quercetin and
resveratrol into elastic liposomes,
Biochimica et Biophysica Acta.
2013;1828:309–316.

Salunkhe et al.; JPRI, 33(29B): 31-41, 2021; Article no.JPRI.67442
41
46. Chen W, Zou M, Ma X, Lv R, Ding T, Liu
D. Coencapsulation of EGCG and
quercetin in liposomes for optimum
antioxidant activity. J. Food Sci.
2019;84(1):111-120.
47. Wang H, Xue Y, Di W, Sun L, Chen L.
Preparation process of paclitaxel
liposomes and its anti-proliferative effect
on human gastric cancer MGC803 cells,
Biomed. Res. 2016;27(2):360-363.
48. Xu Y, Meng H. Paclitaxel-loaded stealth
liposomes: Development, characterization,
pharmacokinetics, and biodistribution, Artif.
Cells Nanomed. Biotechnol.
2016;44(1):350-355.
DOI: 10.3109/21691401.2014.951722
49. Plangsombat N, Rungsardthong K,
Kongkaneramit L, Waranuch N, Sarisuta
N. Anti-inflammatory activity of liposomes
of Asparagus racemosus root extracts
prepared by various methods, Exp. Ther.
Med. 2016;12(4):2790-2796.
DOI: 10.3892/etm.2016.3661
50. Liu JJ, Nazzal S, Chang TS, Tsai T.
Preparation and characterization of
cosmeceutical liposomes loaded with
avobenzone and arbutin. J. Cosmet. Sci.
2013;64(1):9-17.
51. Shaji J, Menon I. PE gylated liposomes of
meloxicam: Optimization by quality by
design, In vitro characterization and
cytotoxicity evaluation, Pharm.
Nanotechnol. 2017;5(2):119-137.
DOI:10.2174/22117385056661704281521
29
52. Wen AH, Choi MK, Kim DD. Formulation of
liposome for topical delivery of arbutin,
Arch. Pharm. Res. 2006;29(12):1187-
1192.
DOI: 10.1007/BF02969312
53. Padamwar MN, Pokharkar VB.
Development of vitamin loaded topical
liposomal formulation using factorial design
approach: Drug deposition and stability,
Int. J. Pharm. 2006;31;320(1-2):37-44.
DOI: 10.1016/j.ijpharm.2006.04.001
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