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6
Adulteration and Evaluation of Crude Drugs of Natural Origin
Popat Mohite, Abhijeet Puri
AETs St. John Institute of Pharmacy and Research, Palghar, Maharashtra, India
6.1 Introduction
In numerous countries, herbal remedies are extensively used
for treating illnesses since various drugs are known to have
adverse effects [1]. Substances derived from plants, known as
herbal medicines, are utilized for medicinal purposes due to
their naturally occurring phytochemical compounds. There
have been instances of chemical contamination and undisclosed synthetic medications detected in some herbal treatments, despite the market for natural medicines growing.
The Indonesian Food and Drug Administration released a
press statement in 2020 about a particular case of herbal
medication that contained synthetic pharmaceuticals that
had not been notified [2]. Herbal medicines shouldn’t include
artificial ingredients or the outcomes of medical isolation,
per their regulations. Some instances of tampered herbal
remedies include the presence of unreported substances,
including sildenafil in the herbal extract [3].
The use of medicinal plants and herbal remedies has been
around since the beginning of human civilization. For thousands of years, traditional medical practices like Ayurveda,
Traditional Chinese Medicine, and Native American healing
have relied on the healing properties of various plants.
Nowadays, there has been a renewed interest in these natural remedies due to an increased awareness of their potential
health benefits and a desire for more comprehensive healthcare options that take the whole person into account [4]. The
recent resurgence of interest in herbal medicine has raised
an important issue – the genuineness and excellence of natural crude drugs. Raw plant materials or their extracts,
known as crude drugs, serve as the basis for herbal medicines and contain active components [4]. Being derived
straight from nature, crude drugs are naturally prone to
variations in composition, quality, and safety. The use of
crude drugs presents a significant challenge because of the
problem of adulteration. Adulteration refers to the intentional or unintentional mixing of authentic plant materials
with foreign or lower-quality substances. This practice can
compromise the effectiveness of herbal remedies and also
pose serious health risks to consumers [5]. Embarking on a
journey to explore the complex realm of impurity and the
assessment of natural crude drugs is the central theme of
this chapter. We will delve into the historical significance of
herbal medicine, emphasizing its use throughout the ages
and the origins of modern herbalism in ancient traditions.
Moving forward, we’ll shift our attention to the present-day
landscape, highlighting the renewed interest in herbal remedies as alternative or complementary therapies to traditional medicine.
The chapter further addresses adulteration, which represents a pervasive challenge in the herbal medicine industry.
It discusses the motivations behind adulteration, ranging
from economic gain to the unavailability of genuine plant
materials. Various types of pollutants are explored, including foreign plant materials, contaminants, and substandard
or low-quality herbs. This discussion underscores the importance of quality control (QC) and standardization in the
herbal medicine supply chain. Recognizing that addressing
the adulteration problem necessitates implementing robust
evaluation methods, we delve into the analytical techniques
employed to detect contaminants and authenticate crude
drugs. This includes high-performance liquid chromatography (HPLC), gas chromatography-mass spectrometry
(GC-MS), thin-layer chromatography (TLC), spectroscopy,
microscopy, and more. These techniques are crucial for
ensuring herbal products’ accuracy, safety, and quality [6].
102 6 Adulteration and Evaluation of Crude Drugs of Natural Origin
Furthermore, we shed light on specific case studies
where these analytical methods have been effectively
employed to identify adulteration and evaluate crude
drugs. These cases are practical examples of how scientific
tools and methodologies can be harnessed to tackle adulteration in herbal medicine.
The chapter also explores the role of regulatory bodies
and pharmacopeia’s in setting standards and guidelines for
herbal medicines. It delves into the challenge of harmonizing international regulations, given the global nature of the
herbal product supply chain. An essential aspect of this
chapter is the examination of the prospects and emerging
trends in herbal medicine, with a particular emphasis on
the utilization of advanced technologies and interdisciplinary approaches to address adulteration and enhance the
safety and efficacy of herbal products. This covers the
application of big data, the incorporation of artificial intelligence, and the investigation of new biomarkers and indicators. The findings demonstrate the development of the
analytical procedure; we discovered that methods based on
spectroscopy and chromatography are still often employed
to identify chemical adulterants.
Provided in this chapter is a comprehensive summary of
the challenges and opportunities that exist in the field of
impurity and the assessment of crude drugs derived from
natural sources. It presents a comprehensive outlook on
the subject by blending historical background, modern
perspectives, analytical methods, case studies, and prospects. The aim is to equip readers, including researchers,
healthcare professionals, regulatory authorities, and consumers, with a greater comprehension of the intricacies
associated with herbal medicines and the measures
required to guarantee their genuineness, quality, and
safety. Ultimately, this understanding contributes to the
ongoing development and responsible use of herbal remedies in the ever-changing healthcare landscape.
unscrupulous suppliers and manufacturers seek to cut
costs by substituting expensive or rare botanicals with
cheaper alternatives. This substitution can range from the
use of lower-quality plant materials to the addition of contaminants or adulterants that mimic the appearance of
genuine herbs.
Another driving force behind adulteration is the lack of
regulatory oversight and standardized QC measures. As the
herbal medicine market continues to expand, especially in
the absence of rigorous oversight, opportunities for adulteration multiply. In some instances, the unavailability of
authentic herbs due to environmental, seasonal, or geopolitical factors can inadvertently lead to adulteration [7].
6.2.1.1 Types of Poisonous or Deleterious Adulterants
Adulterants in herbal drugs can take on various forms,
including:
1. Foreign plant materials: these can be plants from
entirely different species or those closely related to
the authentic herb, making visual differentiation
challenging.
2. Contaminants: contaminants like pesticides, heavy
metals, and microorganisms may find their way into
herbal products, posing health risks to consumers.
3. Substandard or low-quality herbs: herbs that do not
meet quality standards or have been improperly stored
can compromise the overall quality of the herbal
product.
4. Pharmaceutical drugs: in some cases, adulteration
involves the inclusion of conventional pharmaceutical
drugs, which can have unpredictable and potentially
harmful interactions with other medications.
5. Adulteration with banned or restricted substances:
this type of adulteration can introduce dangerous or
controlled substances into herbal products, posing
health and legal risks [8].
6.2 Adulteration of Herbal Drugs
6.2.1 Poisonous or Deleterious Substances
The use of herbal drugs and traditional remedies has a rich
and enduring history, spanning centuries and crossing cultural boundaries. The appeal of herbal medicines lies in
their perceived natural origins and holistic approach to
health. However, in the contemporary context, the herbal
medicine industry faces a formidable challenge – the adulteration of herbal drugs with poisonous or harmful substances. This insidious practice threatens the integrity of
herbal products and, more critically, the health and safety
of consumers. Adulteration of herbal drugs is not a recent
phenomenon. It has roots in economic motivations, as
6.2.2 Filth and Foreign Matter of Adulteration
The adulteration of herbal drugs with filth and foreign matter represents a distressing and persistent challenge within
the herbal medicine industry. While the use of herbal remedies is rooted in the desire for natural, holistic, and safe
healthcare alternatives, the infiltration of these products
with impurities, contaminants, and undesirable foreign substances threatens the safety and efficacy of herbal drugs [8].
6.2.2.1 Types and Examples
Filth and foreign matter adulteration can manifest in various forms, and examples abound in the herbal medicine
market. One prevalent form of adulteration is the inclusion
6.3 Types of Adulteration 103
of foreign plant materials, which can range from entirely
different species to closely related botanicals. For instance,
in the case of St. John’s Wort, the genuine herb, Hypericum
perforatum, has been adulterated with closely related
Hypericum species, such as Hypericum maculatum, to cut
costs. This substitution can be challenging to detect by visual inspection alone, leading to the inadvertent use of adulterated products [9].
Another form of adulteration involves the introduction
of contaminants. Contaminants can include heavy metals, pesticides, mold, bacteria, and other microorganisms
[10]. For example, herbal products originating from
regions with lax environmental regulations may contain
high levels of heavy metals like lead, arsenic, and mercury. Such contaminants pose significant health risks to
consumers and undermine the perceived safety of herbal
medicine.
Substandard or low-quality herbs represent yet another
facet of filth and foreign matter adulteration. Herbs that do
not meet established quality standards, often due to
improper cultivation, harvesting, or storage practices, may
find their way into herbal products. As an example, poorquality Echinacea species are sometimes substituted for
premium Echinacea species in products intended to boost
the immune system. This substitution can result in diminished therapeutic effects [11].
mycotoxins, harmful compounds that can cause various
health issues. For instance, the herbal supplement
Echinacea has been found to be contaminated with
Aspergillus, presenting a health hazard for consumers who
expect immune-boosting benefits from the product. Fungal
contamination, when overlooked, can lead to respiratory
distress and other health complications [14].
Mold contamination is equally problematic, as molds
can proliferate on improperly dried or stored herbal materials. Aloe vera gels, which are utilized for their purported
skin-healing properties, have been found to harbor mold
contamination. This can lead to skin irritations and allergic
reactions when applied, directly contradicting the intended
therapeutic effects [15].
These examples underscore the urgency of addressing
microbiological contamination in herbal drugs. Consumer
safety is at stake, and the intrinsic appeal of herbal remedies as natural, holistic healthcare solutions should not be
undermined by the invisible threat of microbial adulteration. Regulatory bodies, QC measures, and advanced testing methodologies are essential in safeguarding the
integrity of herbal medicine and the well-being of those
who turn to these remedies in pursuit of health and
wellness.
6.3 Types of Adulteration
6.2.3 Microbiological Contamination
Microbiological contamination of herbal drugs represents
a concealed yet potent threat to the safety and efficacy of
these natural remedies. While the allure of herbal medicine lies in its perceived natural origins, the presence of
harmful microorganisms, such as bacteria, fungi, and
molds, poses serious health risks to consumers [12]. This
form of adulteration often eludes detection, making it
imperative to shed light on the issue.
6.2.3.1 Examples of Microbiological Contamination
Microbiological contamination can manifest in various
ways within herbal drugs. A common scenario involves the
presence of bacteria like Salmonella and Escherichia coli
(E. coli), both of which can lead to severe gastrointestinal
infections when ingested. For example, a study detected
Salmonella contamination in powdered Kratom, a herbal
product known for its stimulating and pain-relieving
effects. This discovery highlights the potential health hazards associated with microbiological contamination in
herbal drugs [13].
Fungal contamination is another critical concern.
Aspergillus species, which are ubiquitous in the environment, can proliferate on herbal materials and produce
The debasement of an article is a simple explanation for
adulteration. The main motivation behind intentional adulteration is usually commercial, with the aim of increasing
profits. Various reasons may contribute to this, including
the high price and scarcity of the drug in the market [16].
Adulteration can be done intentionally or accidentally, as
depicted in Figure 6.1.
The drug can undergo various types of changes, such as
decay, blending, refinement, replacement, degradation,
and corruption. Decay refers to a decrease in the drug’s
quality, while blending happens when one substance is
mistakenly or carelessly added to another. Refinement is a
deliberate and purposeful type of contamination. On the
other hand, replacement happens when an alternative
material replaces the original medication. Subpar medications are referred to as inferiority, and the growth of
microbes causes spoiling [17].
6.3.1 Intentional/Deliberate Adulteration
Intentional adulteration is driven by the desire for commercial gains, especially when there is a limited supply of
drugs but a high demand. This involves deliberately substituting a herbal drug with lower-quality products, either
104 6 Adulteration and Evaluation of Crude Drugs of Natural Origin
Adulterants Added
Microorganisms that can
cause diseases, Residue
from harmful pesticides,
Rodent droppings, and
Larvae present in the
food.
Unintentional
Adulteration:
Ascribed to recklessness,
apathy, unawareness, or
inadequate infrastructure
and sanitation in all
stages of food processing
from farm to table.
Figure 6.1 Types of adulteration.
Adulterants Added
Food additives and
preservatives. Ripening
agents. Pesticides may be
added as well. Some
other contaminants
include sand, marble, and
chalk.
Adulteration
Types
Intentionally or
accidentally adding
metallic substances
such as Iron, Copper
that Leads to metallic
adulteration.
Adding or removing
substances from food
or intentionally
altering its natural
properties is known
as intentional
adulteration.
Adulterants have been
introduced,
including arsenic,
lead from water
sources, mercury from
effluents, and tin from
cans.
wholly or partially. Adulterants are used, which may or
may not have any chemical or therapeutic potential,
because they resemble the authentic herb. One method of
adulteration is substituting the plant material with
exhausted drugs, which means using the same plant material without the active constituents [18]. This practice is
commonly seen in volatile oil-containing materials. When
these materials are dried, they lose their essential oils but
still look like the original drug. Foreign materials, such as
fragments of the same plant, artificial artifacts, stones,
sand, and artificial inferior principles are employed to
replace the lost oils. Intentional adulteration occurs when
unethical producers and traders add lower-grade materials
to food products [19]. They do this to make more money
and create a false image of superiority. Intentional or economically motivated adulteration is a serious concern in
the food industry. Manufacturers often employ chemical
additives like urea and melamine to enhance a product’s
essential nutrients, but this practice can lead to the depletion of these nutrients. Additionally, they introduce various substances like flour, starch, roasted barley powder,
chalk powder, sugar, water, oils, sand, chicory, ergot, milk,
stone, brick powder, ground papaya seeds, molasses, and
other ingredients to augment the volume of a food product.
This type of adulteration poses a significant threat as it
reduces the nutritional value of the food and introduces
foreign substances that can be harmful to consumers. Some
specific food items, such as honey, milk, apple juice, olive
oil, orange juice, saffron, and coffee, are more likely to be
targeted due to their higher economic value [20]. Regulatory
bodies and consumers must be vigilant in detecting and
preventing such adulteration practices to ensure the safety
and integrity of our food supply. Using a different substance in place of the original one is called substitution,
which is another type of adulteration. Due to deforestation
and the loss of certain species, adulteration and substitution are frequent in the trade of raw medicinal plant
resources [21]. This has led to concerns about the authenticity of numerous herbal medicinal products. Mixing
unintended herbs with adulterated herbs can lead to
adverse reactions. Scientific studies have shown that identifying these adulteration techniques without the necessary
microscopic analysis is difficult.
6.3.2 Unknown or Incidental Adulteration
The presence of adulteration is often linked to the improper
hygienic conditions observed in the production and handling of crude drugs. While producers, traders, and retailers
may not have the intention of purposely adding adulterants,
it’s crucial to remember that the techniques used in processing, storing, transporting, and marketing the products can
inadvertently result in contamination [21]. The inclusion of
any substance that is not naturally found in the product is
considered extraneous and qualifies as an adulterant. Some
examples of the contaminants that can be included in this
6.3 Types of Adulteration 105
are residual pesticides from preservatives, mercury from
effluents, rodent droppings, lead from water, cans, and
other similar substances. The execution of unintentional
adulteration occurs due to several factors, including discrepancies in vernacular names, limited understanding of
reliable sources, similarities in color and form, negligent or
incorrect collection practices, inadequate storage, and subpar preparation [22].
6.3.3 Metallic Contamination
The global concern regarding the presence of elevated
heavy metal concentrations in herbal remedies is a serious
issue that can potentially jeopardize human health, particularly when concentrations surpass established thresholds. The problem of susceptibility has been particularly
prominent in traditional medical therapy, and it has been
associated with various health disorders. It has been determined that herbal formulations possess considerable
amounts of heavy metals, specifically cadmium (As), mercury (Cd), arsenic (Hg), copper (Cu), and lead (Pb) [23].
Heavy metals, due to their low renal excretion rates and
inability to be easily metabolized by the body, can accumulate in soft tissues and cause adverse effects, even at very
low concentrations. The toxic effects on the body are a
direct result of their interference with normal biochemical
and metabolic processes. Several health problems have
been associated with the excessive consumption of dietary
heavy metals and cardiac dysfunction, such as decreased
immunity, impaired psychology, fetal malformation, and
neurological behavior [24]. Traditional remedies from various parts of the world commonly incorporate heavy metals
as a deliberate practice [25]. For this reason, it would be
misleading to describe the existence of heavy metals in traditional remedies as “contamination.” Traditional Chinese
medicine has a history of using heavy metals for various
health conditions, such as utilizing mercury sulfide known
as “cinnabaris,” mercury chloride known as “calomel,” or
mercury oxide known as “hydrargyri oxydum rubrum.”
Despite the skepticism of allopathic medical practitioners
regarding the therapeutic use of mercury, traditional medicine practitioners hold a different viewpoint [26].
These metals become biocompatible when combined,
in accordance with long-standing customs, with organic
substances made from plant extracts. The detoxification
of metals during the processing of herbo-metallic formulations has been the subject of recent scientific studies,
which have demonstrated specific processes involved in
this phenomenon [27]. Despite the fact that heavy metal
salts are commonly included in African traditional medicines, there is currently no established body of knowledge explaining how these substances effectively mitigate
toxicity. Nevertheless, it should be noted that the nonexistence of evidence for such a theory does not imply
that it does not exist. Although the presence of heavy
metals in African traditional remedies has been identified, there is a scarcity of research examining the metals
commonly utilized in African traditional medicine or
exploring the motivations behind their utilization [28].
One example of a commonly used metallic adulterant is
the addition of lead chromate to turmeric in order to
enhance its natural color. However, this practice is highly
harmful as it can lead to health issues, such as anemia,
abortion, paralysis, and brain damage [29].
6.3.4 Adulteration in Synthetic and Artificial Substances
The original substances can undergo adulteration if artificial materials are utilized to bring about changes. Many
times, these materials are crafted in a way that resembles
the look of various drugs. For example, a particular example would be if a shipment of ergot from Portugal was combined with tiny amounts of flour dough, which were
subsequently shaped and colored to precisely mimic the
original product. Basswood is chopped to resemble nutmegs in size and form, then added to them as an adulterant
to trick customers [30]. As a replacement for coffee berries,
compressed chicory is utilized. The substitution of paraffin
wax for beeswax and the use of artificial invert sugar in
place of honey are common practices. Due to their morphological resemblance to the authentic drug, cheaper and
natural substances that have no relationship to the genuine
article are frequently utilized as substitutes [31]. While the
substitutes may bear a resemblance, it is crucial to understand that they lack the identical chemical makeup or therapeutic potency found in the original medication. Ailanthus
leaves are a viable alternative to Belladonna leaves, and saffron can be blended with dried flowers of Carthamus tinc-
torius, to give you an idea. An alternative to almonds can be
found in the form of peach kernels and apricot kernels [8].
There are instances where synthetic pharmaceutical principles are utilized with the aim of improving the market
and therapeutic value of a drug. Lemon oil is enhanced
with the addition of citral, whereas balsam of Peru is
enriched with benzyl benzoate. One noteworthy finding
about fluid or disorganized medications is that legitimate
drugs are often mixed with market trash [32]. As an illustration, one can mix pieces of mixed limestone with asafoetida, mix lead shot with opium, amber-colored glass
with colophony, and mix white oil with coconut oil. The act
of incorporating rodent fecal matter into cardamom seeds
is an adulterant that poses a danger. Apart from the commonly mentioned practices, it is worth mentioning that the
106 6 Adulteration and Evaluation of Crude Drugs of Natural Origin
utilization of synthetic chemicals can be an additional
method to augment the properties of crude drugs [33]. An
effective way to mask cumin seeds is by utilizing grass
seeds that have been tinted with charcoal dust, whereas for
coriander powder, mixing it with dung powder is recommended. The inclusion of washing soda in powdered sugar
and other food items is known to have the potential to
induce intestinal disorders [7].
6.4 Adulteration in Medicinal Plants
The cure of diseases is greatly aided by the significant
role that medicinal plants play in many countries.
Nowadays, it is essential to prioritize quality assurance
for medicinal plant products being sold in the local market. The prevalence of adulteration and misconduct in
local medicinal markets poses a significant risk to customers’ health [6].
6.4.1 Reasons for Adulteration
Whether due to a lack of knowledge or a deliberate intent
to deceive, adulteration can occur by substituting the original plant material with a different one that closely resembles it in smell or appearance. The material that is
substituted has the potential to be a species that is similar
and could potentially cause harm to human health.
Consequently, from the first stage of plant material collection to the last stage of product production, it is critical to
maintain a high degree of attentiveness throughout the
entire process [34].
less fragrant bark of Canella winterana and the original substance [36]. Just like that, the seeds of Mucuna pruriens (L.)
can also be included in the discussion. In the District of
Columbia, there have been cases of adulteration found in certain members of Papilionaceae due to their superficial resemblance. The best example of an unknown authentic plant
that showcases an exceptional resemblance in morphology is
M. pruriens. Similar papilionaceae seeds have been mixed
into the product, causing adulteration. Adulterants that are
frequently used include Mucuna utilis, which is sold as the
white variety, and Mucuna deeringiana, which is sold as the
bigger variety [37]. Besides Mucuna cochinchinensis, Indian
markets also offer Canavalia variso and Canavalia ensiformis
for sale. With a length of up to 1cm, the authentic seeds display a stunning mosaic pattern of black and brown colors on
their surface. In terms of size, M. deeringiana and M. utilis are
larger, with measurements ranging from 1.5 to 2 cm. When it
comes to their coloration, M. deeringgiana stands out with a
lackluster black color, while showcases a more vibrant white
or buff color. The seeds of M. utilis can be identified by their
buff or white color, while the seeds of M. deeringiana are dull
black. Hence, it is crucial to authenticate morphologically
similar drug materials to prevent any potentially serious or
harmful effects [38]. To provide an example, the act of adulterating species can lead to a wide range of detrimental effects
on those who consume them. The validity of the aforementioned statement still applies even when Cinnamomum
verum J.Presl bark is combined with Cinnamomum malabatrum (Burm.f.). J.Presl. The plant Cinnamomum cassia con-
tains a naturally occurring flavoring substance called
coumarin, which is known to cause hepatotoxicity and is present at a concentration of 1% [39].
6.4.2 Adulteration Caused Because of the Similar Morphology
In this scenario, the act of adulteration involves incorporating plants as adulterants. These plants may or may not have
any therapeutic or chemical properties, but they do bear a
physical resemblance to the genuine medicinal plant [35].
Tejpat, which are the aromatic leaves of Cinnamomum
tamal, are exchanged for spices. They also have carminative
properties and are used to treat diarrhea, effectively relieving
colic pain. Cinnamomum obtusifolium (Roxb.), nees, also
known as the Indian bay leaf, closely resembles Cinnamomum
tamala and is commonly used as an adulterant of cinnamon.
One another instance of adultery is Cinnamomum verum,
also known as true cinnamon, whose dried bark is commonly
used to add flavor to a variety of foods, including sweets,
cakes, biscuits, and even pickles. The reason for the adulteration is the close resemblance between the dense, tough, and
6.4.3 Adulteration Caused Because of Confusion in Vernacular Names
The presence of confusion arises when there are instances
of vernacular names being shared by multiple species, as
well as situations where various vernacular names refer to
a single species. This confusion leads to misunderstandings
and increases the likelihood of adulteration. Fumaria parv-
iflora, known as parpata in Ayurveda, is a plant that holds
significance in this traditional medicinal system. The term
“In Siddha” is used to refer to a specific plant called Mollugo
pentaphylla. Because of the similarity in nomenclature
between these two herbs, they are often mixed together,
adulterated, or substituted in traditional medical systems
[40]. The confusion in vernacular names is a significant
factor that leads to this specific type of adulteration. Both
F. parviflora Lam. and M. pentaphylla L., known as parpata
or parpadagam, are herbs that are commonly used and
sold. The local name “parpata” or “parpadagam” is used to
6.5 Methods of Detection of Adulterants and Evaluation of Medicinal Herbs 107
refer to M. pentaphylla L. in some regions of Southern
India, whereas, in Northern India, suppliers of herbal
products use this name to refer to Fumaria parvi-flora Lam.
By examining their leaves and stems, it is quite easy to
identify these two herbs [41]. The leaves of M. pentaphylla
are not only simple but also small, while the stem is characterized by its pale yellow to light brown color, thinness,
and wiry texture. The shape of the leaves in F. parviflora
Lam. is different as compared to Cassia angustifolia Vahl,
which is also significant when it comes to this kind of adulteration [42]. C. angustifolia, commonly known as senna, is
an important medicine. The stem is distinguished by its
dark brown-to-black color. Senna, which is scientifically
referred to as C. angustifolia, is a commonly recognized
medicinal plant that holds great importance. It is extensively utilized for the treatment of digestive disorders, constipation, asthma, depression, and various skin ailments.
C. angustifolia Vahl. is preferred to treat various diseases.
Herbal markets in these countries may contain broken aerial parts of Cassia obtusifolia L., an adulterant plant, which
is also sold under the name of senna [43].
6.4.4 Insufficient Basic Understanding of the Real Plant Source
Ayurveda recognizes Nagakesar as a vital drug with great significance. Regarding authenticity, Mesua ferrea is the source
we can rely on. Nevertheless, it is important to note that the
presence of Calophyllum inophyllum flowers is the cause of
adulteration in market samples. The lack of awareness
regarding this issue extends to suppliers. The presence of a
two-celled ovary is a distinguishing characteristic of authentic flowers, making them easily identifiable, unlike spurious
flowers, which typically have a single-celled ovary [43].
European markets engage in the cultivation and sale of
H. perforatum. This species is not easily found in India due to
limited availability. On the other hand, there is a species
called Hypericum patulum that is found in abundance in
Indo-Nepal and is sometimes mislabeled as H. perforatum.
Identifying the taxonomic classification of the market sample is made easy by the presence of the whole plant, including its flowers. When examining the anatomy of the stem of
H. perforatum, it becomes evident that the transverse section
showcases a phloem that is compressed and thin, a pith that
is hollow, and a notable absence of calcium oxalate crystals.
Conversely, H. patulum exhibits certain traits, including a
wider phloem, a pith that is somewhat hollow, and the existence of calcium oxalate crystals. Additionally, a major factor
contributing to adulteration is the lack of caution exhibited
by herbal collectors or suppliers during the collection of
herbal drugs [44]. Parmelia perlata (Huds.) is another example that can be considered. Ach, which is commonly known
as Shaileya, finds its usage in the Unani, Siddha, and
Ayurveda systems of medicine. The samples that were marketed demonstrated a mixture with other species, including
Parmelia cirrhata and Parmelia perforate. On the other hand,
the nature of the thallus makes it simple to identify the original plant [45].
6.5 Methods of Detection of Adulterants and Evaluation of Medicinal Herbs
6.5.1 Taxonomic Deciding Adulteration of Medicinal Plants
Humans formulated one of the crucial steps in ensuring
quality maintenance, which is to evaluate the drug using a
taxonomic approach. Medicinal plant taxonomy classification system as a means of expediting the recognition of disparities and analogies. Medicinal plants are commonly
acquired by professionals lacking botanical or taxonomical
expertise. Likewise, unrefined medications obtained from
the market are frequently presumed to be the labeled plant
material, without undergoing rigorous botanical identification techniques [46]. The significance of accurately identifying plants for scientific purposes cannot be emphasized
enough, as it is the sole method of linking ethnobotanical
knowledge with literature-based biological and chemical
information. However, prevalent synonyms for flora in literature present a substantial obstacle, as certain titles cannot align with their scientific equivalents in conventional
writings. Plants’ scientific names are difficult to determine
due to vernacular names and the potential for a single local
name to refer to multiple species. The presence of multiple
vernacular names and the attribution of a single local
name to several species adds to the uncertainty around
identifying plants [47]. A pure drug called Flower of
Afsantin, which is derived from Artemisia absinthium L. of
the Asteraceae family, may be contaminated with
Helichrysum graveolens Sweet, which also belongs to the
Asteraceae family, but can be distinguished based on its
taxonomical classification. Alongside, six other species
from the Asteraceae family, including babooneh
(Matricaria recutita), Anthemis wiedemanniana, Anthemis
nobilis, Tanacetum persicum, Tanacetum parthenium,
Tripleurospermum disciforme, and Microcephala lamellata). The Badranjbuyeh (Melissa officinalis) containing six
other species (i.e. Hymenocrater calculus, Dracocephalum
moldavica, Hymenocrater bituminous, Hymenocrater elegans, Hymenocrater platystegius, and Asperugo procumbens) were found to be the most adulterated or substituted
[48]. Another example can be Tulasi: the source of Ocimum
108 6 Adulteration and Evaluation of Crude Drugs of Natural Origin
sanctum is contaminated with Nirgundi, which is obtained
from the botanical source Vitex negundo. Similarly, Saraca
asoca leaves are contaminated with false drug Ashoka
leaves, which come from Polyanthia longifolia [49].
6.5.2 Morphological Analysis
In the morphological investigation of herbal plants,
researchers carefully examine and analyze the physical
attributes of specific plant parts, such as leaves, seeds,
fruits, rhizomes, flowers, stems, and the plant’s overall
structure. The plant or extract typically has a distinct
appearance that allows for easy identification. The examination of the structural makeup of an unrefined drug is
known as morphology, while the depiction of said structure is referred to as morphography [50]. A crucial initial
step in taxonomy, it assists in identifying medicinal plants
based on their unique features. Additionally, this examination is exceptionally advantageous in differentiating the
actual plant from its imposter. Various morphological features have been proposed by scientists as a means to identify different medicinal plants. They argue that a collector
with a basic understanding of the medicinal plant’s morphology can gather it more precisely [51]. Crucial characteristics include the fragmented surfaces of quassia wood,
cascara bark, quillaia, and cinchona. The study identified
the scientific name for the tea plant as Camellia sinensis
(L.). Kuntze, due to its morphological characteristics, is
considered a valuable medicinal species. A similar research
was conducted on Trachystemon orientalis (L) G. Don to
examine its morphology [51].
6.5.3 Microscopic Analysis
To effectively analyze and describe medicinal plants, one
must carefully observe their intricate anatomy and study
their pollen. Genuine plants can be distinguished from
adulterants by examining their anatomical features,
including the shape and type of stomata, trichome category, epidermal cell shape, and structure. Through the
study of pollen, known as palynology, botanists and taxonomists can differentiate between authentic and fake
plant species. By employing scanning electron microscopy
(SEM) and light microscopy (LM), researchers can
precisely verify the identity of medicinal plant species
through their distinct pollen grain characteristics [52].
Distinguishing between authentic and counterfeit plants
relies on analyzing various pollen attributes, including
shape, type, colpi length, sculpturing nature, and surface
ornamentation. By using this method, a more thorough
analysis of a drug can be conducted, enabling the identification of organized drugs based on their distinctive
histological features. Its main purpose is to qualitatively
evaluate the efficacy of prepared medicinal plants,
whether they are in their complete or pulverized forms.
Each plant has its own distinct tissue feature, and a microscope is essential for confirming the structural specifics of
drugs obtained from plants [53]. To achieve accurate
results, one can employ a range of reagents or stains to differentiate cellular structure. Lignin can be easily identified by applying a drop of phloroglucinol and concentrated
applied, the N/50 iodine solution can cause starch and
hemicellulose to take on a blue hue. From the wavy medullary rays of cascara bark to the glandular trichomes of
mint, each plant has unique characteristics that set them
apart. Sclereids and calcium oxalate crystals are absent in
powdered cloves, unlike powdered clove stalks. The process includes the use of techniques, such as microscopic
linear measurements, determination of leaf constants,
and quantitative microscopic evaluation [54]. Linear
measurements encompass various aspects, such as the
size of starch grains, the length and width of fibers, and
the presence of trichomes. Leaf constants are determined
by examining factors, such as stomatal number, stomatal
index, vein islet distribution, veinlet termination number,
and palisade ratios. The stomatal number indicates the
density of stomata per square millimeter of the leaf’s epidermis. Bengal gram flour, known as besan, is a widely
utilized ingredient in Indian cooking. Regrettably, due to
high demand, unscrupulous traders dilute besan flour by
adding other legume flours like pea or lathyrus. When
examined under a microscope, the seed testa macrosclereids of these three legumes are easily distinguishable due
to their unique shapes and sizes [55]. The macrosclereids
from Cicer arietinum, which are the main constituent of
besan flour, have a mean length of 155.6 microns and are
longer, with a bent end. Conversely, macrosclereids from
Pisa sativum and Lathyrus sativus are shorter, with flat
ends and a mean length of 61.8 and 72 microns, respectively, and have a different morphology. Furthermore, the
seed testa macrosclereids of other edible legumes also display dissimilarities. Consequently, the besan flour was
found to contain macrosclereids after being examined
under a microscope [56]. ExtCell walls, starch grains, cell
contents, trichomes, calcium oxalate crystals, fibers, and
vessels are among the features that have been extensively
researched [57]. Surinam quassia lacks calcium oxalate,
while cascara bark contains features, such as uniseriate
medullary rays, crystal fibers, and wavy medullary rays,
unlike frangula bark, where stone cells are absent.
Varieties of aloes can be distinguished by the presence or
absence of pith in rhizomes and roots, as well as the warty
trichomes of senna and the presence or absence of aloin
6.5 Methods of Detection of Adulterants and Evaluation of Medicinal Herbs 109
crystals. Clove stalk powder is composed of sclereids and
calcium oxalate crystals, which are not found in cloves
themselves [58]. Rauwolfia serpentina is often adulterated
with Rauwolfia densiflora, Rauwolfia micrantha, and
Rauwolfia perakensis species of roots; these can be seen
sprawling out from the ground, intertwining with each
other. Bamboo, belonging to the Poaceae family, has guard
cells that resemble dumbbells in shape. Under bright-field
microscopy, when observed, guard cells resembling kidneys with two subsidiary cells arranged in parallel rows
when examining leaf fragments of this product. The presence of this diacytic stomata hinted that the product might
actually be a carnation, not a Poaceae plant, as originally
claimed. The confirmation of this was achieved through
the utilization of plastidic markers matK and rbcL. To
ensure accuracy, I acquired diverse accessions from multiple botanical gardens and commercial sources of this
genus, rigorously verifying their taxonomy. Through
microscopic examination of the fully developed leaves
from these plants, the samples were authenticated to
belong to the species Dianthus chinensis L. [59].
6.5.4 Organoleptic Analysis
Organoleptic analysis allows us to evaluate drugs by
observing their appearance, smell, and texture. This
encompasses techniques, such as assessing hue, aroma, flavor, dimensions, form, and unique attributes like texture.
The appearance alone of a plant or extract is often sufficient for identification. If not, there might be a noticeable
scent or flavor associated with the plant or extract.
Recognizing a plant or extract can often be done simply by
looking at it, as it has unique visual characteristics. In case
this is not sufficient, the plant or extract may also have a
distinct taste or scent [50]. Organoleptic analysis is a basic
yet natural form of analysis, relying on human senses.
Morphology is the field of study that examines the physical
structure of a crude drug, analyzing its characteristics and
properties. Essential characteristics include the rough, jagged surfaces found in quassia wood, cinchona, quillaia,
and cascara barks. The aroma of umbelliferous fruits and
the sweetness of liquorice are notable. The Rauwolfia’s
wavy shape, the pungent taste of capsicum and ginger, the
brown hue of cinnamon, and the scent and flavor of spicedrugs like asafoetida, black pepper, nutmeg, caraway, and
cumin are crucial organoleptic features. Talka gum serves
as an alternative to acacia gum and can be distinguished by
its color and shape. The gum is frequently fragmented,
with some pieces having a brownish hue, while others are
colorless [55]. Acacia gum, on the other hand, is predominantly white or yellow in appearance. Bael fruits can be
substituted with mangosteen fruits, which have a dark
exterior and wedge-shaped radiating stigmas. The differences in the morphological features of Cuprea bark
(Remijia pedunculata) and Cinchona can be easily identified. Bloodroot, with its dark reddish-brown color, is often
added to Hydrastis to deceive others due to its color, while
Hydrastis has a distinctly yellowish hue. Rheum rhaponti-
cum and Chinese rhubarb can be easily differentiated due
to their distinct size differences [54].
6.5.5 Qualitative and Quantitative of Phytochemical for Detection of Contaminants
Chemical assays, instrumental analysis, and qualitative and
quantitative chemical testing are among the various chemical techniques used in substance evaluation. Chemical evaluation methods also encompass the processes of isolating,
purifying, and identifying active constituents. The identification of various phytoconstituents, including alkaloids,
glycosides, and tannins, is conducted through qualitative
chemical tests. Within their respective chapters, the text
presents procedures for conducting identification tests of
various phytoconstituents [60]. Copper acetate can be utilized as a means of identifying colophony as an adulterant
within balsams, resins, and waxes. Similarly, Holphen’s and
Baudouin’s tests serve as methods for detecting cottonseed
and sesame oil adulterants in olive oil. Drug evaluation can
also benefit from the use of chemical treatment methods,
including ester value, acid value, acetyl value, and saponification value. Assays for various constituents, such as alkaloids, resins, volatile oils, glycosides, vitamins, and others,
are conducted through chemical assays. Different substances are tested for their total alkaloid content, such as
ipecacuanha–belladonna herb, for total and non-phenolic
alkaloids, nux vomica for the alkaloid strychnine, jalap for
resin, and cod liver oil for vitamins. The obtained results
have the ability to determine the presence of an inferior or
exhausted drug [61]. Additionally, proving the absence of a
tested component can suggest that a useless item has been
completely replaced. The process of instrumental analysis
involves using chromatographic and spectroscopic methods
to determine the chemical groups present in phytoconstituents. Chromatographic techniques include paper chromatography, TLC, HPTLC, gas chromatography, and HPLC.
Spectroscopic analysis employs techniques, such as ultraviolet (UV), nuclear magnetic resonance (NMR), and mass
spectroscopy (MS).
6.5.6 Establishment of Fingerprint Profiles
DNA fingerprints are produced via the amplification of
chromosomal DNA and can be likened to barcodes due to
their patterned appearance. They serve the purpose of
110 6 Adulteration and Evaluation of Crude Drugs of Natural Origin
distinguishing one person from another. Precise identification is essential when dealing with closely related plant
species. Nevertheless, these methods are constrained by
the diverse chemical composition and quantities found in
different species, which growth conditions, harvesting,
and storage durations can influence [62]. Due to the multitude of compounds in each herb, it is unfeasible to qualitatively or quantitatively analyze all the compounds of
interest, thereby complicating the task of detecting their
presence or absence. These challenges are widely
acknowledged, yet DNA fingerprinting remains an imperative tool for confirming the authenticity of botanicals.
Chemical and molecular markers are utilized to authenticate herbal drugs. This involves scrutinizing the plants’
distinctive biomolecules and genetic composition [63].
Analyzing herbal plants’ unique genetic makeup is the
authentication process through molecular markers. This
method is superior to other taxonomic markers due to its
accuracy, efficiency, and independence from environmental factors, specimen age, and physiological conditions. DNA-based techniques provide a reliable and
cost-effective means of testing millions of samples of
medicinal plants, making it an ideal tool for safety monitoring and QC [64]. Genetic markers, which are short
sequences of nucleotides (genes) found on chromosomes,
can distinguish between cells, individuals, and even species. DNA fingerprinting can detect mini- or microsatellites, which are small repeating segments of DNA that
show more diversity between individuals, and create a
unique pattern that can be used for identification purposes [65]. The plant’s unique DNA sequences allow for
identification through molecular markers by employing
techniques depicted in Table 6.1. This technique can differentiate between the original plant and its adulterant
with ease. In the past decade, a technique known as DNA
barcode has been proposed as a universal molecular tool
for species identification. Symbolically, the DNA barcoding method illustrated the process by which an infrared
scanner accurately identified a product by analyzing the
distinct interspecies boundaries [65]. The authentication
of herbal plants has been facilitated through the establishment of different barcodes, including matK, rbcL,
psbA-trnH, and ITS2. The utilization of this molecular
technique proved to be highly advantageous, as it not only
confirmed obscure species but also yielded significant
results in powdered herbal samples [65].
The verification of Angelica species, referred to as Jeonho
in Korean and Qianhu in Chinese, necessitates the utilization of a SCAR marker. The identification is achieved by
employing particular primers, such as a 273bp amplicon
primer for Anthriscus sylvestris, a 363bp amplicon primer
for both A. decursiva and Peucedanum praeruptorum, and
Table 6.1 DNA markers commonly used in plant identification.
Sr. No. Commonly used molecular markers
1. Random amplified polymorphic DNA (RAPD)
2. Simple sequence repeats (SSR)
3. Single nucleotide polymorphism (SNP)
4. Inter simple sequence repeats (ISSR)
5. Loop-mediated isothermal amplification (LAMP)
6. Restriction fragment length polymorphism (RFLP)
7. Sequence characterized amplified regions (SCAR)
8. Amplified fragment length polymorphism (AFLP)
145bp and 305bp amplicon primers exclusive to P. praerup-
torum. AFLP markers are used to authenticate and identify
both genuine and adulterated samples of Zanthoxylum can-
thopodium and Zanthoxylum oxyphyllum [65]. DNA-based
markers such as ISSR and RAPD are utilized to authenticate the leaves of Ocimum sanctum, Ocimum basilicum, and
Ocimum gratissimum species [66]. Leaves of Mentha piperita, Mentha citrata, Mentha requienii, Mentha spicata,
Mentha arvensis were discriminated using the Random
Amplified Polymorphic DNA (RAPD) technique [67]. The
Vidari plant in Ayurveda, scientifically known as the
Ipomea mauritiana, is identified using two types of DNA
markers – RAPD and SCAR. The SCAR marker is specific to
I. mauritiana and generates a 323bp amplicon. The RAPD
marker, which produces a 600bp amplicon, is utilized to
determine the genetic diversity present in various plant species that are at risk of endangerment [68]. RAPD markers
have been employed in examining the genetic diversity of
Solanum genus, comprising Solanum melongena and
Solanum violaceum. The application of RAPD markers has
allowed the identification of genetic variations among eight
Zingiber officinale varieties, which have high yield potential. Citrus volkameriana, Citrus sinensis, and Citrus reticu-
lata can be distinguished using PCR markers [65].
6.5.7 Multiple Marker-based Fingerprint Profiles for Detection of Adulterants
In spite of significant progress, the ongoing difficulty of
performing DNA barcoding on herbal preparations containing multiple components persists. An assessment of
the precision of outcomes should always be carried out
through method validation. A meticulous analysis of the
complete chemical composition of a plant specimen is
involved in fingerprinting techniques. This intricate profile can be used in combination with multivariate data
analysis or chemometrics to extract a shared pattern that