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Файл:Ординатура / Хирургия / Библиотека им академика М.И. Перельмана / Книга_5958_Библиотеки_им_академика_М_И_Перельмана
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have been developed since then. Furthermore, the extraction time was greatly
reduced compared to the traditional extraction method. To summarize,
microwave-assisted extraction methods have evolved enough to be used on an
industrial scale in place of traditional eugenol extraction methods.
72
MAE has
a number of advantages, including a quick extraction time, low energy usage,
high extraction efficiency, and minimum target component degradation.
While MAE provides the benefits listed above, it also has a few disadvantages.
MAE is difficult to perform on fully dry and extremely moist materials when a
nonpolar solvent such as hexane is used as the extracting solvent. Further-
more, when the target compounds and/or solvents are non-polar, microwave
efficiency can be lowered and rendered useless when the extraction fluid has a
high viscosity.
73
6.8.3 Supercritical Carbon Dioxide Extraction
Supercritical carbon dioxide (SC-CO
2
) extraction is another successful
method for obtaining eugenol from clove buds. Extensive research has
proven that SC-CO
2
extraction is highly effective. An example of the result of
the study conducted by Gopalakrishnan et al. is that SC-CO
2
extraction ex-
tracts more clove oil than liquid extraction does. This study examined two
different experiments, in which they discovered that increasing the flow rate
of CO
2
during supercritical extraction yields higher eugenol yields. The
aforementioned two tests by Wenqiang et al. examined the clove bud oil
composition and found that it was quite similar whether extracted using SC-
CO
2
or other traditional extraction procedures (solvent extraction, steam
distillation, and molecular distillation).
74
Though numerous scientific
studies have proven the ability of SC-CO
2
extraction to efficiently harvest
clove oil, less research has been done to maximize the SC-CO
2
extracted
eugenol yield. To prove the efficacy of the SC-CO
2
extraction procedure for
the recovery of eugenol from clove bud oil, further research is required. SC-
CO
2
is cheap and comes in large quantities, ensuring that no changes are
made to bioactive molecules and that their curative or functional charac-
teristics are not compromised. It’s also capable of solubilizing lipophilic
compounds and is easily removed from products after the process is com-
plete and is a generally accepted environmentally beneficial technique.
75
A simplified process-scale SFE system consists of two extraction tanks: one
equipped with tempera ture controllers or pressure valves to maintain the
appropriate extraction conditions, and the other, which is fitted on both
ends with temperature controllers as well as pressure valves to keep the
extraction conditions consistent. In order to extract the fluid from the ex-
traction tank, pumps are utilized.
76
Pumps utilized for circulation of fluid
are found in the system s, too. Wettable powders can be stored for long
periods of time due to their fast hydration process. A valve is placed in
the separator’s bottom part to collect the product that flows from the sep-
arator. Supercritical fluid extraction is now seen as a feasible alternative to
traditional extraction methods. Solvents in the supercritical state have
122 Chapter 6
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physical-chemical properties that are halfway between liquid and gas, in-
creasing the solvent’s extractive capacity. Due to their density, these fluids
have a high solvation power, while their high diffusion and low viscosity
values offer the necessary penetration power into the solid matrix. The
solvents used in constructing an SFE process are crucial, and a wide range of
compounds can be used as solvents in this approach.
77
Super critical ex-
traction is now widely recognized as a more ecologically friendly alternative
to traditional phase separation that avoids the majority of the disadvantages.
Separating moderately polar compounds can also be carried out with organic
modifiers such as methanol, isopropanol, and acetonitrile. On the other
hand, despite the fact that its use is still moderate, it has risen steadily over
the last few years. This pattern exemplifies both contemporary technical
advances and the general advantages of this separation technique. Poly-
phenols, biogenic amines, and polyacetylenes were among the natural
compounds studied in vegetable-derived diets in addition to lipids. In
addition, multiple lipid classes were investigated simultaneously in complex
vegetal extracts and bilberry using the supercritical approach.
78
As the food
component industry catches up to its materials engineering and pharma-
ceutical counterparts and grows beyond extraction, SC-CO
2
extraction tech-
nology is gaining appeal as a high-value, environmentally safe, and effective
extraction method. SC-CO
2
addresses issues, including product purity, pro-
cess efficiency, wellness, and environmental impact that aren’t addressed by
traditional methods.
79
6.8.4 Ionic-liquid Extraction Method
Recently, researchers have concentrated their efforts on ionic liquid appli-
cations (ILs). Numerous studies have focused on extracting active com-
ponents from traditional herbs using ILs as alternative solvents to
circumvent the difficulties associated with standard methods of extracting
natural products (NPs) using organic chemical reagents. In recent years, the
use of ionic liquids (ILs) for analytical applications has evolved, resulting in
improvements in the quality and efficacy of extraction. Ionic liquids are a
type of liquid organic salt that interacts selectively with specific polar and
non-polar compounds via ion exchanges, stacking interactions, hydrophobic
interactions, or hydrogen bonds, thereby enhancing the selectivity of the
extraction process. For example, when paired with hydrophilic amino acids,
1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM][PF
6
]) extraction
efficiency is enhanced in contrast to standard organic solvent extraction.
Because ILs are employed in a number of extractive processes, notably MAE
and UAE, they must be stable enough to resist the high temperatures asso-
ciated with these techniques. Tetraalkylammonium cations and imidazo-
lium ions, such as tetrafluoroborate [BF
4
], bis(trifluoromethylsulfonyl)imide
[NTf
2
], trifluoromethanesulfonate [CF
3
SO
3
], and hexafluorophosphate [PF
6
],
exhibit excellent heat stability in this regard; additionally, organic anions
have been shown to be more stable.
80
Scientific Evaluation of the Role of Ocimum sanctum in Ayurvedic 123
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The IL extraction method is a viable alternative for recovering organic,
inorganic, and metal ions in plants and herbs with a high recovery rate.
Extensive research has been undertaken over the last two decades on IL-based
extraction, isolation, and purification strategies for bioactive compounds from
biomass. ILs and their mixes with water or organic solvents can be used
directly in solid–liquid extraction (SLE) of biomass-derived value-added
chemicals. Because ILs are ionic, they can interact with electromagnetic
waves; thus, in addition to even more conventional SLE techniques, IL-based
microwave- or ultrasonic-assisted extraction (MAE) as well as ultrasonic-
assisted extraction (UAE) may also be preferred due to their shorter
extraction times and higher extraction efficiencies. When the true matrix is a
liquid, such as a water-rich biomass extract, it is possible to perform IL-based
liquid–liquid extractions (LLE). Additionally, ILs may be integrated into a solid
matrix to minimize the process’s overall IL consumption, to increase the
extraction efficiency, and to simplify solvent/material recycling (IL modified
materials for solid-phase extractions, SPE). Indeed, this family of ILs has been
shown to be effective for extracting and purifying bioactive chemicals from
biomass, amino acids, and proteins in water and raw materials, as well as
stabilizing proteins, enzymes, and nucleic acids. It was concluded that the
predominant solvent for hydrophilic ILs should be IL aqueous solutions. This
is an especially interesting approach since it overcomes the high viscosity of
some ILs while increasing the solvent’s biocompatibility and lowering the
overall solvent cost. Additionally, ILs have been shown to be effective at
extracting small extractable compounds by increasing their solubility in
aqueous solutions through a hydrotropic or micelle-mediated effect, whilst
also avoiding the dissolution and the extraction of the lignocellulosic fraction,
as well as extracting proteins, that have a low solubility in water, according to
the works reviewed here. The development of the more cost-effective and
sustainable extraction as well as separation processes is important for the
recovery and commercialization of novel and low-cost bioactive products for
such nutraceutical, skincare products, and pharmaceutical industries, with
the goal of improving modern society’s quality in the near future, and in
which ILs could play a significant role as a catalyst.
81
6.8.5 Enzyme-assisted Extraction
A unique and convenient alternative to standard solvent extraction processes
is enzyme-assisted extraction (EAE): the method is novel and convenient
since the enzymes catalyze specific reactions without subjecting the targeted
products to severe conditions that could result in degradation. Additionally,
proteins such as cellulases, hemicellulases, and pectinases hydrolyze the cell
wall, increasing permeability and enabling metabolite release from plants.
By utilizing enzy mes such as lipases, proteases, and phospholipases, the
amount of solvent utilized in the extraction can be reduced. The most often
used enzymes in the extraction of oil from plants are cellulase, a-amylase,
and pectinase. These proteins can be obtained from fungi, bacteria, animals,
124 Chapter 6
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or plants or engineered, and their selective catalysis enables them to be used
to recover a specific bioactive chemical in high yields and with minimal en-
ergy waste. However, some limitations exist due to the high cost of the en-
zymatic strategy, the incomplete disruption of the cell wall, and also the
difficulty of commercial application owing to the enzymes’ variable actions in
response to environmental changes such as oxygen concentration, nutrient
variety, as well as operating temperature. Natural product extraction is ham-
pered by the structure of the cell membrane and cell wall, the micelles formed
by macromolecules like polysaccharides and proteins, and the aggregation
and denaturation of proteins at high temperatures during extraction.
82
EAE improves the extraction efficiency by hydrolyzing the cell membrane
and membrane components, and also macromolecules within the cell,
allowing the natural product to be released more easily. EAE is mostly
composed of cellulose, a-amylase, and pectinase. Polysaccharide is a
bioactive component of Traditional Chinese Medicine (TCM) Astragali
Radix. Zhang et al. reviewed and noted that the EAE of polysaccharide
isolated from Astragalus membranaceus radix using a variety of enzymes and
discovered that glucose oxidase degraded polysaccharide more effectively
than the other seven enzymes examined (amyloglucosidase hemicellulase,
bacterial amylase, fungal amylase, pectinase, cellulose, and vinozyme). By
adjusting the EAE setting with glucose oxidase, polysaccharide synthesis
increased by more than 250 percent as compared to the non-enzyme treated
approach. When chlorogenic acid was extracted from Eucommia ulmoides
leaves using cellulase and ionic solutions, the yield of chlorogenic acid was
significantly increased. Zhang et al. described in their review that using
pectinase and cellulase enzymes enhanced the yield of carotenoid and
lycopene extraction from tomato waste. In comparison to untreated solvent
extraction, cellulase and pectinase-treated samples produced sixfold and
tenfold more of the two target chemicals, respective ly.
83
6.8.6 Pressurized Liquid Extraction
Numerous research organizations have referred to pressurized liquid ex-
traction (PLE) as accelerated solvent extraction, improved solvent extraction,
pressurized fluid extraction, rapid fluid extraction, and high-pressure solv-
ent extraction. PLE extracts use high pressure. Due to the fact that high
pressure maintains solvent liquids above their boiling point, they have a
high solubility and diffusion rate of lipid solutes, as well as high solvent
penetration into the matrix.
84
When compared to other extraction methods,
PLE significantly reduced the extraction time and solvent consumption and
showed increased repeatability. Using pressurized liquid extraction, re-
searchers from the University of Macau and other institutions successfully
isolated a variety of natural components from the TCM, including saponins,
flavonoids, and essential oil. Some researchers believed PLE could not be
used to extract thermolabile chemicals due to the high extraction tempera-
ture, while others believed it could be employed due to the shorter extraction
Scientific Evaluation of the Role of Ocimum sanctum in Ayurvedic 125
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time in PLE. When PLE was employed at 200 1C to extract antioxidants from
grape pomace, Maillard reactions occurred. Gizir et al. successfully used PLE
to obtain the anthocyanin-rich extract from black carrots because anthocyanin
degradation is time-dependent and the high temperature and short duration
of the PLE extraction conditions could offset the disadvantage of the high
temperature used in the extraction.
85
This approach frequently uses water as
the solvent, eliminating the environmental and health hazards associated
with organic solvents. By operating at elevated temperatures (up to 200 1C)
and pressures (35–200 bar), PLE improves the quality of the extraction, which
can be performed dynamically, with the solvent being pushed continuously
through the vessel, or statically, with the solvent being refilled more often. At
elevated temperatures, the solvent’s viscosity decreases, allowing it to pene-
trate the matrix more effectively and extract the desired analytes; however, this
method cannot be used for thermolabile compounds and may result in the co-
extraction of other compounds due to the reduced selectivity of extraction at
higher temperatures. The increased pressure maintains the solvent in the li-
quid phase and breaks the cell walls of the plants, putting strain on the
matrix. Non-ionic surfactants, antioxidants, including ascorbic acid, CO
2
to
lower the aqueous pH, and drying agents are all alternatives to organic solv-
ents. Additionally, the PLE apparatus prevents light- and oxygen-sensitive
compounds from degradation and can be combined with other extraction
techniques like UAE to maximize its effectiveness.
86
6.8.7 Pulsed Electric Field Extraction
By altering membrane structures during extraction, pulsed electric field ex-
traction (PEF) significantly increases the extraction yield and minimizes the
extraction time. The success of PEF treatment is reliant on a number of par-
ameters, including the field intensity, the specific energy input, the pulse
count, and the treatment temperature. PEF extraction is a non-thermal
method that reduces the breakdown of thermolabile chemicals. Hou et al.
obtained the highest yield of ginsenosides (12.69 mgg
1
) utilizing PEF at a 20
kV cm
1
electric field intensity, a 6000 Hz frequency, a 70% ethanol–water
solution, and a 150 l h
1
velocity. The PEF extraction method yields more
ginsenosides than the MAE, heat reflux, UAE, or PLE methods. The entire
process of PEF extraction took less than a second and was much faster than
the other methods evaluated. Bouras et al. discovered that the PEF treatment
improved the phenolic content and antioxidant activity of Norway spruce bark
by eight-fold and thirty-fold, respectively, as compared to untreated samples.
87
6.9 Pharmacological Profile
6.9.1 Antimicrobial Properties
In 1952, Joshi CG and Magar examined O. sanctum’s antibiotic activity
against Escherichia coli, Staphylococcus aureus, Bacillus anthracis, Salmonella
126 Chapter 6
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spp., Vibrio spp., and Mycobacterium tuberculosis.
88
Jabeen et al. discovered
antimicrobial activity against Pasturella multocida, E. coli, S. aureus, B. sub-
tilis, and Salmonella typhimurium, as well as E. coli, Klebsiella spp., B. subtilis,
and S. aureus.
89
Kumar et al. conducted in vitro trials against specific in-
fections under laboratory conditions and examined the pathogen’s devel-
opment or suppression, as well as observed effects over a set time period.
90
Tulsi essential oil inhibits E. coli, Bacillus subtilis, Bacillus anthracis,
Staphylococcus aureus, Pseudomonas vulgaris, and Pseudomonas aeruginosa
significantly. These essential oils are high in caryophyllene, eugenol, and
methyl eugenol, which are all efficient against Anthrobacterglobiformis,
Bacillus megatherium, E. coli, and Pseudomonas sp. Eugenol is the most
therapeutic component of Tulsi, according to Grover and Rao in 1977.
91
Leaf
extracts, both aqueou s and alcoholic, have been shown to have antimicrobial
properties. The extract is effective against a number of enteric illnesses,
including E. coli, Klebsiella aerogens, Shigella dysenteriae, Pseudomonas aer-
uginosa, and Staphylococcus aureus. Additionally, the ant ibacterial activity of
seeds was examined, and it was observed that crude, supernatant, residual,
and dialyzed seed samples inhibited the growth of P. multocida, E. coli, B.
subtilis, and S. aureus. Figure 6.6. represents the pharmacological activities
of O. sanctum. Table 6.2 describes the clinical studies of O. sanctum extracts.
Figure 6.6 Pharmacological activities of O. sanctum.
Scientific Evaluation of the Role of Ocimum sanctum in Ayurvedic 127
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Table 6.2 Clinical studies involving Ocimum sanctum extract.
a
Tulsi extract Participants Duration Dosage Outcomes Ref.
Tulsi leaves caps 40 male adults
T2DM
(45–55 year)
6.5 weeks 3 g per day before meal Significantk postprandial
glucose & fasting blood glucose
92
Tulasi leaves caps 30 adults obesity
(17–30 year)
8 weeks 250 mg per day 2 daily
before meal
Improved BMI, lipid profile
(except TC), TG, and IR
93
Tulsi powder leaves 30 adults T2DM 2 weeks 2 g per day Significantk post-prandial
glucose & fasting blood
94
Fresh tulsi leaves 3 adults T2DM 5 weeks Fresh leaves 3 daily Considerable decrease in blood
glucose reaching near normal
levels
95
Tincture from tulsi 200 adults Gouty
Arthritis
12 weeks 10 drops 3 times per day Significant reduction in serum
uric acid
96
Aqueous tulsi leaves 100 adults MetS
(Z40 year)
12 weeks 5 ml/ 2 day before
meals
Improved lipid profile, fasting
blood glucose, and BP
97
Tulsi
leaves þglibenclamide
drug
60 adults T2DM
(30–65 year)
13 weeks 300 mg per day
Tulsi þ5mg
glibenclamide
Significant kfasting blood &
postprandial glucose reduced
HBA1c
98
Aqueous tulsi leaves 40 adults T2DM
(45–55 year)
8 weeks 500 g per day Significant improvement in lipid
profile
99
Powder tulsi leaves 90 male adults
T2DM/MetS
(40–60 year)
12 weeks 2 gper day Improved T2DM symptoms:
kpolydipsia, kpolyphagia, & BP
100
Whole plant decoction 10 adults T2DM 12 weeks 14 g per day Reduced blood glucose in 9
T2DM adults
101
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Powder whole plant tulsi 5 adults
psychosomatic
(60–80 year)
12 weeks 3 g 2 per day Significant improvement in lipid
profile
102
Fresh juice tulsi leaves 50 female adults,
hypotensive
(18–30 year)
4 weeks Juice 2 per day Significant changes in blood
pressure
103
Ethanolic tulsi 22 healthy adults
(22–37 year)
4 weeks 300 mg per day before
food
Reduction in lipid profile, in 6
participants
104
Fresh juice 75% tulsi 20 adults,
hypertension
(45–64 year)
10 days 30 ml per day Significant kblood pressure 105
Tulsi powder leaves 27 adults T2DM/
MeS (45–65 year)
4 weeks 1 g per day in morning
before meal
Improved lipid profile, blood
glucose, glycated proteins
(HbA1c) & UA.
105
a
BMI ¼ body mass index measured by weight (kg)/height (m
2
); BP ¼blood pressure; HbA1C¼glycosylated hemoglobin; IR¼insulin resistance;
MetS ¼ metabolic syndrome; T2DM ¼type 2 diabetes mellitus; TC¼total cholesterol, TG¼triglycerides; UA ¼ uric acid.
Scientific Evaluation of the Role of Ocimum sanctum in Ayurvedic 129
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Tulsi extract was found to be highly effective against an antibiotic-
resistant strain of Staphylococcus aureus. Its leaf extract showed consider-
able antibacterial activity against drug-resistant Salmonella enterica serovar
Typhi when coupled with chloramphenicol (C) and trimethoprim (Tm)
(S. typhi). When coupled with C and Tm and O. sanctum leaf extract exhibited
synergistic activity against S. typhi isolates. Eugenol (1-hydroxy-2-methoxy-4-
allylbenzene), the active ingredient in O. sanctum, has been demonstrated to
contribute significantly to Tulsi’s antibacterial therapeutic properties.
Tulsi solvents and water extracts exhibited antibacterial activity against
multidrug-resistant bacteria S. aureus, with MIC values ranging from 1.56 to
6.25 mg ml
1
, while multidrug resistant Klebsiella pneumoniae and
Escherichia coli exhibited higher values (6.25–25 mg ml
1
). The antimicrobial
activity of Tulsi extract against Streptococcus mutans was discovered.
Eugenol, methyl eugenol, linalool, 1, 8-cineole, and Tulsi oils have all been
shown to be extremely cytotoxic toward Candida species.
106
6.9.2 Anticancer
An ethanolic extract of O. sanctum has been shown to inhibit the formation
of Sarcoma-180 solid tumors in mice and to increase their life expectancy.
The anticancer activity of O. sanctum in the Lewis lung carcinoma animal
model validated this conclusion. Ursolic acid is naturally anti-cancerous.
O. sanctum shields DNA from radiation damage. O. sanctum is quite useful
for treating a variety of tumorigenesis stages. The treatment of mice with
Solid Sarcoma-180 tumors with O. sanctum aqueous and alcoholic extracts
leads to a co nsiderable reduction in tumor size.
107
Additionally, Tulsi extract
has anti-inflammatory and radical scavenging characteristics, which are
crucial for limiting the growth of some types of malignant cells. Ursolic acid
and curcumin found in Tulsi are anti-inflammatory, cyclooxygenase in-
hibitory, and antioxidant. O. sanctum increases glutathione synthesis and
storage in mice and glutathione-S-transferase activity by approximately 78%.
The oil of O. sanctum is an effective chemopreventive agent against the de-
velopment of fibro sarcoma in Swiss albino mice following subcutaneous
injections of 20-methylcholanthrene. When given to mice, this resulted in an
increase in survival and a delay in the onset of tumors. When mice were
given oil, the end product lipid peroxidation, liver enzymatic activity, and
malondialdehyde levels were drastically altered in contrast to untreated mice
that received 20-methylcholanthrene injections.
108
On LNCaP prostate cancer cells, Tulsi extract induces apoptosis. After 24
and 48 hours of treatment with various doses of 70% ethanolic extract of
Tulsi, LNCaP prostate cancer cells exhibit cytotoxicity. When combined with
docetaxel (DTL), the flavonoid vicenin-2 (VCN-2) inhibits prostate cancer
(CaP). VCN-2 efficiently inhibits proliferation, angiogenesis, and apoptosis
in CaP cells (PC-3, DU-145 and LNCaP). In vitro, VCN-2 inhibits the EGFR/
Akt/mTOR/p70S6K pathway, as well as c-Myc, cyclin D1, cyclin B1, CDK4,
PCNA, and hTERT. Tulsi extracts also have anti-ulcerogenic activity in rats
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treated with pyloric acid and aspirin. Upon acute and chronic adminis-
tration, the OSL extract decreased the ligated and pyloric ligated ulcer in-
dexes, as well as free and total acidity. Pre-treatment with the medication for
seven days enhanced mucous secretion and decreased acid discharge. OG
inhibits the growth and progression of breast cancer and functions as a
natural inhibit or of metalloproteases found in matrices.
109
6.9.3 Hepatoprotective Activity
By acting as a component of the detoxifying system, it stimulates the removal
of dangerous compounds and works as a hepatoprotective agent. The study
showed that when an alcoholic extract of Tulsi was administered orally, it
protected albino rats’ livers from paracetamol, carbon tetrachloride, and
antituberculosis drug-induced liver impairment. The level of enzymes de-
creased when O. sanctum extract was used to reduce the number of enzymes
in male albino rats weighing 100–150 g of the Wistar strain (5–6 weeks). The
Biometry Research Unit of the Indian Statistical Institute discovered tha t
when albino rats were given Tulsi extract orally for six days, it had a hepa-
totonic effect on paracetamol and carbon tetrachloride. When coupled with
silymarin, Tulsi extract exhibits a substantial hepatoprotective effe ct.
110
Tulsi extracts were found to be efficient in preventing oxidative stress in
the liver by boosting glutathione peroxidase and catalase levels. They were
also found to be effective in preventing hepatic steatosis. Eugenol, carvacrol,
ursolic acid (UA), -caryophyllene, and rosmarinic acid were found to have
anti-inflammatory, gastrointestinal, and hepatoprotective effects. The oil has
anti-inflammatory properties as a result of its combined suppression of
arachidonate metabolism and antihistaminic activity. Antipyretic efficacy is
a result of prostaglandin inhibition, and the oil also acts as a peripheral
analgesic. The oil was found to be helpful against formaldehyde or adjuvant-
induced arthritis in rats, as well as turpentine oil-induced joint edema.
111
6.9.4 Immunomodulatory Activity
When rats were treated with distilled extracts of fresh leaves, changes in the
humoral immune response were observed. These changes were ascribed to
processes such as antibody production, tissue responses, and the release of
hypersensitive mediators of specific organs. It has been demonstrated that
seed oil can affect both cellular and humoral immune responses. GABA-
mediated immunomodulatory effects have been reported. Tulsi has been
demonstrated to enhance cellular as well as humoral immunity. O. sanctum
aqueous extract revealed immunotherapeutic potential in bovine sub-
cutaneous aqueous extract infusions. Additionally, the aqueous extract of
O. sanctum reduces the total number of bacteria, increases the number of
neutrophils and lymphocytes, and has a high phagocytic capacity. Mediratta
et al. examined the immunomodulatory effects of O. sanctum seed oil on a
variety of immunological markers in healthy and stressed mice.
112
As a
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