INTRODUCTION:

Herbal medicine refers to the use of herbs, herbal materials, and medicines containing plant parts or mixtures of plant parts as active substances. Plant materials such as leaves, bark, flowers, roots, fruits, and seeds are used to make these herbs.¹

Herbal Technology is a broad term that incorporates a wide range of approaches to exploiting plants' diverse abilities for human benefit. This field now has five primary branches, including medicinal plants, natural dyes, bio insecticides, bio fertilizers, and biofuel, while more may be added in the future.²

Herbal medicines are more widely used in health promotion and treatment of chronic diseases than in cases of life-threatening illnesses. However, the use of traditional remedies is increasing when conventional medicine is not effective in treating diseases, such as advanced cancer and when dealing with new infectious diseases.³

Tinospora cordifolia, also known as "Guduchi" in Sanskrit, is a versatile, large, evergreen, evergreen tree that grows in the highlands and belongs to the Menispermaceae family. The male flowers are collected in raceme or racemose panicles, while the female flowers are isolated.^1,4 Summer and winter extend the flowering period. Alkaloids, steroids, diterpenoid lactones, aliphatics, and glycosides, among other active components found in the plant, have been divided into various parts of the plant body, including the root, stem, and leaves, stem, and the whole plant. Recently, this plant is of great interest to researchers worldwide because of its reported medicinal properties such as anti-diabetic, anti-periodic, anti-spasmodic, anti-inflammatory, anti-arthritic, anti-oxidant, antiallergic, anti-stress, anti-leprotic, anti-malarial, hepatoprotective, immunomodulatory and anti-neoplastic activities. In this review, we focus on:

1. The reported genetic diversity in the Plant

2. Biological roles reported in humans and animals and active components from the plant.

3. Biological roles reported in humans and animals.

Tinospora includes a wide range of phytochemicals, including alkaloids, phytosterols, glycosides, and a variety of other substances. Tinospora cordifolia contains Cne, tinosporaside, jatrorrhizine, palmatine, berberine, tembeterine, tinocordifolioside, phenylpropene disaccharides, choline, tinospora acid, tinospora, tinosporin, and tinosporide.⁵

Ayurveda is an ancient medical system practiced in Sri Lanka and its origin dates back to around 3000 years ago, in Vedic times. The main ingredients of Ayurvedic medicine are plant leaves, flowers and roots, and other parts of plants such as bark. In ancient, the Ayurveda doctors themselves collect medicinal plants to prepare medicines.^6,7,8 Today only a few Ayurveda doctors follow the ancient methods. Because the manufacturing of Ayurveda medicines has become a marketing factor. Today the plants are collected from forest areas or gardens. Those who collect them are not professionally trained in identifying accurate medicinal plants.⁹

Fig. Macroscopy of T. cordifolia

Identification Method:

1. Shape

Any object had a special shape, in other words had a contour, a good shape descriptor must be invariant to scaling, rotation, translation, and reflection. Shape representation divided into two types: boundary- based” and “region-based. In plants recognition was based on leaf boundaries. Elliptic Fourier and chain code were computed, the system used to identify plant species^1,9,10

2. Texture:

There is no exceptional definition of texture because it is extremely complex. We can recognize texture by our eyes, but we cannot be able to describe i.e., simple definition of image texture is a surface attributes that can use to recognize objects Texture was described by the number of “primitives’ ‘and the spatial layout of these “primitives”. Texture is the most important technique used to quantify the patterns in images. It can describe the arrangement of the surface. Texture explains the spatial arrangement of “intensities” or “colour” in an image.^1,9,10

3. Colour:

Colour is used every day to distinguish among objects. Colour plays an essential role for flower analysis than for leaf analysis. Methods that used for colour representation were: “Colour moments” which were simple, “colour histograms” which were robust to translation and rotation, “colour coherence vector”, and “colour correlogram”, two kinds of histogram can be used, “global” and “local”. In colour histogram, edge histogram and area were used for medicinal plant species identification^1,9,10

4. Macroscopic:

Watery, soft, with long, upright, airy stems from the branches. Bark warty, creamish white or gray brown, wood, soft, perforated. The dry sample is 5 to 10cm long, conical pieces, light weight, light bark and paper, dull, dark brown, wood with long upper ridges, and lightly divided into wedge-shaped pieces in opposite sections. Pieces that are difficult to break when completely dry and can only be torn twisted, odorless, and tasteless.^1,9,10

5. Microscopic:

TS show cork, cortex and vasculature. The cork comprises of an outer zone of thick walled brownish compressed cells and an inner zone of thin - walled colorless, tangentially arranged cells. Due to some lenticles cork tissue is broken. Cortex is wide. The outer zone of cortex consists of 3 - 5 rows of irregularly arranged tangentially elongated chlorenchymatous cells and the cells situated towards inner side are polygonal in shape and filled with abundant starch grains. Starch grains are simple, ovoid or ovoid elliptical, occasionally compound of 2 to 4 components. Several secretary cells found scattered in the cortex. Pericycle fibres containing a single prism in each chamber. Vascular zone composed of discrete vascular strands with 10 - 12 or more wedge - shaped strips of xylem, externally surrounded by semi - circular strips of phloem, alternating with wide medullary rays. Phloem consists of sieve tube, companion cells and phloem parenchyma of polygonal and tangentially elongated cells. Some of the cells of phloem parenchyma contain calcium oxalate crystals, cambium is of 1 to 2 layers, and xylem consists of vessel elements, tracheids, parenchyma and fibres. Vessel elements cylindrical in shape bearing bordered pits. Medullary rays 15 to 20 cells wide. Pith mostly made up of large thin - walled cells containing starch grains. The presence of discrete vascular strands in the mature stem of Tinospora cordifolia is one of the different anomalous secondary structures found in Menispermaceae.^1,9,10,12,13

Authentication Parameters:

1. Authentic samples:

Tinospora cordifolia (Genuine sample) was procured from wild region and authenticated from Herbarium.¹⁴

2. Morphological Properties:

Microscopical evaluation deals with the identification of various characters of tissues, cells and cell contents by microscope. Methods of preparing specimens of crude materials for microscopical studies vary depending on the part used like leaf, stem, root, bark, flower, fruit and also on the nature of the material i.e. Entire, cut or powdered.^14,15

3. Preparation of sections:

Drugs that are difficult to cut, boiled for 20 to 30 minutes in water. Take the opposite or opposite part with the razor. Thin materials such as leaves, soft stems or flat seeds are inserted into the potato space and the sections are taken with a regular blade. The portion was placed on a slide, filtered with chloral hydrates, covered with a glass cover and visible under a microscope.^16,17

4. Powder microscopy:

To test the properties of the powder, a sufficient amount of powdered substance (fruit or leaves) in chloral hydrate on a slide was heated on a low flame or hot plate for a short time, covered with a glass cover and viewed under a microscope (4, 10, 20 or 45 X). Solid and solid tissue dispersion: The material was cut into small pieces and transferred to a test tube containing 4 ml of nitric acid and heated to boil. Later, powder potassium chlorate was gently added, warmed and allowed to react leading to tissue breakdown. When it was completely clear, pressure was applied with a glass rod to completely disperse the tissues. The material was allowed to settle, contaminating liquids and solvents repeatedly spilled with water until the acid was removed. The goods are transferred to a slide, with a drop of glycerol added, covered with a glass cover and visible under a microscope.^18,19,20

5. Microscopic test Starch:

For examining the presence of starch, the specimen was taken in I2 wherein starch turns blue. Aleuronic grains – For examining the presence of aleuronic grains a specimen was prepared in I2 and aleuronic grains stained yellow. Fixed oil -For examining the presence of fixed oil, specimen was stained with Sudan red resulting in the droplet of fixed oil to become pink colored.

EXTRACTION METHODS:

1. Microwave assisted extraction:

T. cordifolia dry stems were crushed and screened using a 24 mesh sieve for MAE. A 500mL conical flask was filled with 20 gramme of powdered medication. A total of 200 millilitres of ethanol-water at an 80 percent (v/v) concentration was added. The combination was well mixed and stored for a period of time to allow the medication to absorb the solvent. When the flask was maintained in the microwave oven and treated for microwave procedure, bumping of the solvent was minimised, and extraction was improved. After using the center composite design, the best-suited combination was discovered. The extraction time was set at 3 minutes, and the irradiation power was set to 480 W. The conical flask was removed from the oven once the extraction was done. A suitable amount of solvent was added to create a solution, which was then filtered. After that, the extract was concentrated in a water bath, and the % yield of the extract was determined.^18,19,20

2. Supercritical Fluid Extraction:

Due to a rising need for natural methods that do not introduce any leftover organic compounds, supercritical fluid extraction has emerged as an appealing separation approach for the food and pharmaceutical sectors. It uses fluids in well-defined conditions of pressure and temperature and when those conditions are reached, the fluids manifest very interesting properties like high diffusivity, low viscosity, and a density close to that of liquids. In static extraction, a fixed amount of supercritical fluid interacts with the plant material. The extraction vessel containing the plant material is pressurized with the chosen fluid at a certain temperature. After the extraction is completed, a valve is open allowing the extract to be removed by decompression into a trap. Usually a static extraction is followed by sometime of dynamic extraction in order to improve the extraction efficiency.^19,21

3. Ultrasound Assisted Extraction:

This method uses ultrasounds in order to allow the intensification of the extraction process from the plant raw material when used in combination with other techniques like hydro distillation and solvent extraction. Ultrasounds are essentially sound waves with frequencies higher than the upper audible limit of the human hearing. The plant raw material is immersed in water or solvent and at the same time it is subjected to the action of the ultrasound. This waves will induce mechanical vibrations on the walls and membranes of the plants allowing a faster extraction. The size of the particles, or the milling degree of the plant material, is a critical element in these types of operations. When compared to more traditional procedures, it has several benefits, including increased extraction efficiency and rate, lower extraction temperature, and a wider spectrum of solvents that may be utilized. Finally, the features of ultrasound, which can include intensification of mass transfer, cell rupture, improved solvent penetration, and capillary effect, are further advantages of this technology.

Isolation and Purification Technique¹⁷

· General isolation techniques:

Simple extraction processes using organic solvents of various polarities, water, and their combinations are still used in the majority of isolation operations. Maceration, percolation, Soxhlet extraction, ultrasound- assisted extraction, and turbo-extraction are some of the techniques used.

· Maceration:

This is a straightforward extraction procedure with the drawbacks of a long extraction time and low extraction effectiveness. It has the potential to be utilized to extract thermo labile components

· Percolation:

Because percolation is a continuous process in which the saturated solvent is continually replenished by new solvent, it is more efficient than maceration.

· Decoction:

There are a lot of water-soluble contaminants in the decoction extract. The extraction of thermolabile or volatile components cannot be done using decoction.

· Reflux extraction:

Reflux extraction is more effective than percolation or maceration, because it takes less time and consumes less solvent. It can't be utilized to extract thermo labile natural compounds since it's too hot.

· Soxhlet extraction:

The benefits of reflux extraction and percolation are combined in the Soxhlet extraction process, which uses the principles of reflux and syphoning to constantly extract the herb with new solvent.

· Hydro distillation and steam distillation:

The extraction of volatile oil is generally done using hydro distillation (HD) and steam distillation (SD). In both HD and SD, certain natural substances decompose.^23,34

· Chromatographic Techniques:

1. Thin Layer Chromatography (TLC):

In natural product research, TLC is the most often used planar chromatographic technology. This is the simplest and most cost-effective procedure for analyzing, isolating, and establishing the parameters for column chromatography. The stationary phase is usually silica or alumina (more polar) while the mobile phase typically organic solvents (less polar).

This situation is categorized as normal phase chromatography. In contrast to this, reverse phase TLC is available, in which the stationary phase is alkyl bonded silica or alumina (less polar) and the mobile phase is polar solvent like water, alcohol etc.²⁶

2. High Performance Thin Layer Chromatography (HPTLC):

The preparation of HPTLC has plate adsorbent sizes of between 5 and 7 microns and coating layer of between 150 and 200 microns, which is thicker than TLC. Mobile phase is pumped over the plates using steady pressure. HPTLC is also used in combination with spectroscopic techniques to maximize analytical potential of these techniques .These high performance layers are pre-coated plates coated with a sorbent of particle size 5-7 microns and a layer thickness of 150-200 microns.

The reduction in thickness of layer and particle size results in increasing the plate efficiency as well as the nature of separation. HPTLC plates are substantially more expensive (4- to 6-times more) than normal plates but are an efficient alternative when high sensitivity, accuracy and precision are required in situations demanding high performance²⁷

3. High Performance Liquid Chromatography (HPLC)

This techniques is used for separation, quantification and identification of inorganic and organic solutes in industrial, environmental, biological or pharmaceutical samples It is based on solute interactions in the mobile phase (typically polar mixtures of water and other solvents) and the stationary phase's densely packed solid particles (usually non-polar particles like C18). For analyte elution through the column to the detector, a high pressure of between 250 and 400 bars is necessary.

Diode Array Detector (DAD) is one of the most common detectors that measure analyte spectra. This approach is appropriate for substances that are thermally labile and cannot be vaporized, and it may be used in conjunction with gas chromatography to analyses samples.^3,28,29

4. Column Chromatography:

Chromatography is a laboratory technique for separating a mixture in chemical analysis. The combination is dissolved in a mobile phase fluid (gas or solvent) that transports it through a system (column, capillary tube, plate, or sheet) on which a stationary phase material is fixed. The stationary phase has varied affinities for each of the mixture's elements. Depending on their interactions with the stationary phase’s surface sites, various molecules stay on the stationary phase for longer or shorter periods of time. As a result, they split because they travel at different apparent velocities in the mobile fluid. The split is predicated on the mobile and stationary phases being partitioned differently. Differences in the partition coefficient of a compound cause differential retention on the stationary phase, which affects separation.^30,31

Purification Techniques for Isolated Phytoconstituents:

1. Ion-exchange chromatography:

The dialyzed sample was loaded on Hi Trap DEAE FF (1ml, 7 × 25mm) column (GE Healthcare), pre- equilibrated with 50mM Tris–HCl buffer, pH 8.0. Controlling the flow rate and fraction size of elution with an Akta purifier (GE Healthcare) linked system. A 5ml loop was used to introduce the sample into the column. For both binding and elution, a flow rate of 1ml/min is maintained. The bound proteins were eluted with a NaCl linear gradient (0–1 M NaClw/v) in the same buffer after the column was rinsed with equilibration buffer. The first peak, obtained at a concentration of 0.10 M NaCl, was pooled and concentrated using an Amicon Ultra 3 K instrument (Merck Darmstadt, Germany).^31,32,33

2. Gel filtration chromatography:

The Superdex 200column linked to the Akta purifier was loaded with a concentrated protein sample (1ml) (GE Healthcare, USA). At a rate of 0.5ml/min, the column was equilibrated with 50mM Tris–HCl buffer, pH 8.0. Unicorn management (version 5.0) was used to examine the elution profile for absorbance at 280nm vs elution volume (ml).^31,32,33

3. Gel electrophoresis:

The molecular weight of proteins is determined by SDS-PAGE as defined by Laemmli (Laemmli 1970). SDS- PAGE is made from a combination of slab gel using 12% (w / v) acrylamide and 0.02% (w/v) bisacrylamide in a separation gel and 5% (w/v) acrylamide and 0.16% (w/ v) Bisacrylamide in the packing gel. The gel bath was 0.375M Tris – HCl, pH 8.8. The electrode bath contained 0.192 M glycine and 25mM Tris – HCl, pH 8.3. The Coomassie blue brilliant blue G-250 was used for dyeing gels. Molecular weight was determined using molecular mass measurements (10-180 kDa).^31,32,33

4. Mass spectrometry:^2,8

The RGA2 band was released from SDS-PAGE and identified using matrix-assisted laser desorption/ ionization time of flight (MALDI-TOF) system (Kratos analytical, a shimadzu group company in Japan) using 337nm pulsed UV laser, a 1.7m aircraft tube, and a curved field reflector. A detailed description of the MALDI-TOF method can be found elsewhere. Significant spectra, high concentrations compared to the mass/electric (m/z) mono-isotopic ion charge calculated by MASCOT distiller software version1.1.2.0 (Matrix Science, London, UK).^31,32,33

Introduction Different Methods Used for Bioactive Compound Extraction, Isolation, and Purification:

Because of their antioxidant characteristics, bioactive substances can be employed as food additives. Antioxidants are compounds in meals that help to minimize oxidative stress. Because of their stability and general availability, synthetic antioxidants are widely employed; nonetheless, they have been linked to mutagenesis and carcinogenic consequences, prompting researchers to look for antioxidants isolated from plant matrices.

The extraction of bioactive substances is influenced by a number of parameters, including the extraction process, raw materials, and extraction solvent (Tiwari, 2015). There are two types of techniques: conventional and non- conventional. Organic solvents, heating, and agitation are all required in traditional procedures. Soxhlet, maceration, and hydro distillation are examples of this sort of procedure. Modern techniques, also known as non- conventional techniques, are green or clean procedures since they require less energy and employ organic solvents, both of which are good for the environment.^7,10,33,34

Soxhlet, maceration, and hydro distillation are the most common traditional extraction procedures for bioactive chemicals. A tiny quantity of dried sample is placed on the apparatus where the solvent goes through in the Soxhlet procedure.

1. Techniques of Isolation and Purification of Bioactive Molecules from Plants:

Purification and separation of bioactive chemicals from plants is a technology that has recently experienced significant advancement. On the one hand, this new approach allows researchers to keep up with the development and availability of many complex bioassays, while on the other hand, it provides accurate isolation, separation, and purification procedures. When looking for bioactive chemicals, the objective is to design a method that can screen the source material for bioactivity, such as antioxidant, antibacterial, or cytotoxicity, while also being simple, specific, and fast.^1,35

2. Purification of the Bioactive Molecule:

Using paper thin-layer and column chromatography technologies, several bioactive compounds have been identified and purified. Because of their ease, economy, and availability in diverse stationary phases, column chromatography and thin-layer chromatography (TLC) are still widely utilized. The most useful materials for separating phytochemicals are silica, alumina, cellulose, and polyamide. Plant materials include a high concentration of complex phytochemicals, making separation challenging. As a result, utilizing numerous mobile phases to increase polarity is beneficial for highly valued separations. By column chromatography, thin-layer chromatography has long been employed to analyses the fractions of chemicals. Some analytical methods have been employed to separate bioactive compounds, such as silica gel column chromatography and thin-layer chromatography (TLC).^23,24,34

3. Structural Clarification of the Bioactive Molecules:

Data from a variety of spectroscopic methods, including UV-visible, Infrared (IR), Nuclear Magnetic Resonance (NMR), and mass spectroscopy, is used to determine the structure of particular substances. The fundamental premise of spectroscopy is that electromagnetic energy is passed through an organic molecule, which absorbs some but not all of it. A spectrum can be created by measuring the amount of electromagnetic energy that is absorbed. The spectra are unique to each of a molecule's bonds. The structure of the organic molecule may be determined using these spectra. For structural clarity, scientists often utilize spectra from three or four regions—ultraviolet (UV), visible, infrared (IR), radio frequency, and electron beam.^23,24,34

4. UV-visible spectroscopy:

In both pure and biological mixtures, UV-visible spectroscopy may be used for qualitative investigation and the identification of certain classes of substances. Because aromatic compounds are potent chromospheres in the UV region, UV-visible spectroscopy is used for quantitative study. UV-visible spectroscopy can be used to identify natural substances. Anthocyanins, tannins, polymer dyes, and phenols are examples of phenolic chemicals that produce iron complexes that may be identified by ultraviolet/visible (UV-Vis) spectroscopy. Furthermore, it was discovered that spectroscopic UV-Vis methods are less selective and just provide information on the composition of the overall polyphenol content. Total phenolic extract (280nm), flavones (320nm), phenolic acids (360nm), and total anthokyanids were all determined using UV-Vis spectroscopy (520nm). When compared to other approaches, this technique is not time-consuming and has a lower cost.^23,24,34

5. Spectrometry for Chemical Compounds Identification:

In mass spectrometry, organic molecules are blasted with electrons or lasers and transformed to charged ions, which are very energetic. The relative abundance of a fragmented ion is shown against the mass/charge ratio of these ions in a mass spectrum. The relative molecular mass (molecular weight) of a molecule may be estimated with great accuracy using mass spectrometry, and an accurate molecular formula can be established using information of where the molecule has been broken. Bioactive compounds from the pith were previously extracted and purified using bioactivity-guided solvent extraction, column chromatography, and high-performance liquid chromatography (HPLC). The formation of a bioactive molecule was observed using UV, IR, NMR, and mass spectroscopy methods. Molecules can also be hydrolyzed and their output can be identified. When using tandem mass spectrometry (MS), mass spectrometry provides a wealth of information on the structure of objects. As a result, combining HPLC with MS produces much better results. As a result, when pure quality is not available, the combination of HPLC and MS allows rapid and reliable identification of chemical components in therapeutic plants. LC/MS has recently become very popular in phenolic chemical research. Due to its high ionization efficiency in phenolic compounds, electrospray ionization (ESI) is a recommended source.^23,24,34

Methods for standardization of herbal drug:

All aspects that contribute to the quality of herbal drugs should be considered in standardization methods, including sample identity, organoleptic evaluation, pharmacognostic evaluation, volatile matter, quantitative evaluation (ash values, extractive values), phytochemical evaluation, xenobiotics test, microbial load testing, toxicity testing, and biological activity. The phytochemical profile is particularly important since it has a direct impact on the action of herbal medications.

The body of knowledge and control required to produce material of tolerable consistency is referred to as standardization. This is accomplished by using quality assurance procedures in agricultural and industrial operations to reduce the inherent variance in natural product composition.

Phyto-therapeutic drugs are usually sold in the form of standardized liquid, solid (powdered extract), or viscous formulations. Maceration, percolation, or distillation are used to make them (volatile oils). Fluid extracts are made with ethanol, water, or a combination of ethanol and water. Evaporation of the solvents employed in the raw material extraction process produces solid or powered extracts. To boost therapeutic efficacy, several phytotherapeutic substances are highly concentrated.

A pharmacopoeia is used to determine the identity, purity, and quality of herbal medications. Analytical, physical, and structural standards for herbal medications are prescribed by the Pharmacopoeia. Because of the greater diversity and changes in their chemical makeup or characteristics, a thorough identification and testing of crude pharmaceuticals is critical in herbal formulation procedures.

Authentication of crude drug materials includes: stage of collection, parts of the plant collected, identity such as phytomorphology, microscopical and histological analysis (characteristics of cell walls, cell contents, starch grains, calcium oxalate crystals, trichomes, fibres, vessels, and so on), leaf constants such as palisade ratio, vein islet number, vein termination, stomatal number, and stomatal index, and leaf constants such as

In general, herbal formulations may be standardized graphically so that the medicament can be made with raw materials acquired from various locations, and the chemical effectiveness of different batches of formulation can be compared. The formulations with the highest clinical effectiveness will be chosen. To choose the final completed product and confirm the whole production process, all of the usual physical, chemical, and pharmacological criteria are tested for all batches.

Standardization is an important criterion for assuring herbal medication quality control. All procedures implemented during the production process and quality control that contribute to a reproducible quality are referred to as "standardization."

Standardization ensures that one or more active ingredients and marker chemicals are present. Plant environment and genetic variables may have a major impact on the biochemical components of plant extracts, despite the fact that plants are still the most plentiful and cost-effective source for medication development.

The word "standardization" refers to the process of confirming a drug's identification as well as determining its quality and purity. In light of the commercialization of formulations based on medicinal plants, quality control criteria for diverse medicinal plants utilized in indigenous systems of medicine are becoming increasingly important. Standardization promotes innovation by providing standardized processes and trustworthy data, which reduce time in the invention process, and by making it simpler to share ground-breaking ideas and information about cutting-edge approaches.^3,4,7,10,34

Importance of standardization:

The process of establishing and producing standards based on the agreement of many stakeholders such as corporations, users, interest groups, standards bodies, and governments is known as standardization or standardization.

Standardization is a scientific technique for improving the validity and dependability of research.

This allows you to compare values between variables of various kinds. When you compare data to something else, it takes on new significance.

Needs of Quality control and standardization of herbal products can be summarized as

1. Technology and the notion of standardization were quite different when traditional medicines were invented.

2. The identity of plant material may have altered during the last thousand years due to a dynamic process of evolution.

3. As a result of commercialization, obtaining authentic raw materials has become difficult.

4. Due to time and environmental circumstances, botanical properties may have changed.

Adulteration/substitution:

Herbal treatments have been shown to be contaminated with other plant matter and conventional drugs. The necessity for effective quality control of herbal medicines has been emphasized by reports of herbal products devoid of recognized active components.

Estimation of Parameters Limits Used for Standardization:

Physical Parameters Include:

· Colour,

· Odor,

· Appearance,

· Clarity

· Viscosity

· Moisture Content

· Ph

· Disintegration Time

· Friability

· Hardness

· Flowability

· Flocculation

· Sedimentation

· Settling Rate

· Ash Values.

Chemical Parameters Include:

· Limit tests

· Chemical tests

· Chemical assays etc.

Histological parameter studies are:

1. Leaf constant:

· Palisade ratio

· Vein islet number

· Vein termination

· Stomatal number

· Stomatal Index.

2. Trichomes.

3. Microscopy.

4. Taxonomical identity.

5. Foreign matter.

6. Organoleptic evaluation.

7. Ash values and extractive values.

8. Moisture content determination.

9. Chromatographic and spectroscopic evaluation.

10. Heavy metal determination.

Who Guidelines for Qualitative Standerdization:

The subject of herbal drug standardization is massively wide and deep. The guidelines set by WHO Can be summarized as follows:-

1. A reference to the drug's identity. Botanical evaluations include sensory characteristics, foreign organic materials, microscopy, histology, histochemistry, quantitative studies, and more.

2. Refers to the drug's physicochemical properties. Chromatographic fingerprints, ash values, extractive values, moisture content, volatile oil and alkaloid tests, quantitative estimation techniques, and so forth.

3. A list of pharmacological parameters, biological activity profiles, bitterness values, hemolytic index, astringency, swelling factor, foaming index, and other relevant information.

4. Toxicological information, including pesticide residues, heavy metals, microbiological contamination (total viable count), and pathogens such as E. coli, Salmonella, Pseudomonas aeruginosa, Staph aureus, and Enterobacteria, among others.

5. Microbial contamination is number five.

6. Contamination with radioactive material.

CONCLUSION:

In short, plant extracts have shown strong antioxidant potential in both in vitro and in vivo, as well as in extracts can be considered a good source of natural antioxidants and antimicrobials. Polyphenol extraction of plants using fast and efficient techniques is the least expensive method due to reducing the amount of solvent used, in addition to avoiding the need for long-term extraction times compared to the normal output method. In addition, natural bioactive compounds have has been found to interfere with and prevent all forms of cancer. Flavonoids have been shown to act as anti-tumor agents (benign, melanoma) that activate free radicals (i.e., OH, ROO). In fact, many studies have shown that flavonoids play a number of important roles including mutagenic, cell damage, as well as carcinogenic, due to their acceleration of various aging factors. More than antioxidant activity, inhibiting cancer growth by phenolic compounds depends on the number of basic cellular methods. More extensive research related to these compounds will improve drug testing in the field of carcinogenic disease prevention.

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Received on 21.02.2022 Modified on 16.04.2022

Res. J. Pharmacognosy and Phytochem. 2022; 14(3):195-203.

DOI: 10.52711/0975-4385.2022.00035