1

Proposed
Chemical
Markers for
Zingiberis
officinalis
Mejia, JN
Viacrucis, JD
Dagalea, FM
Ph Ch 230
 2

REPORT OUTLINE
PART 1 INTRODUCTION

PART 2 NEGATIVE MARKER

PART 3 THERAPEUTIC & CORRELATIVE MARKER

PART 4 CHEMICAL FINGERPRINT

CHEMICAL
3

MARKERS
Selection of chemical
markers is crucial for
the quality control of
herbal medicines and Authenticity of Harvesting the best Evaluation of post-
genuine species quality raw materials harvesting handling
ideally, chemical
markers should be the
therapeutic
components of herbal
medicines. Assessment of intermediates Detection of harmful or
and finished products toxic ingredients

INTRODUCTION
CHEMICAL
4

MARKERS

Retrieved from: https://www.researchgate.net/publication/337153441 (online access).

Ramawat, Kishan. (2019). An Introduction to Biodiversity and Chemotaxonomy.
The European Medicines Agency (EMEA) defines
chemical markers as chemically defined
constituents or groups of constituents of a
herbal medicinal product which are of interest
for quality control purposes regardless whether
they possess any therapeutic activity.

Ideally, chemical markers should be unique

components that contribute to the therapeutic
effects of a herbal medicine. As only a small
number of chemical compounds were shown to
have clear pharmacological actions, other chemical
components are also used as markers. The
quantity of a chemical marker can be an indicator

Reference:
of the quality of a herbal medicine

INTRODUCTION
CHEMICAL
Chemical markers shared by various herbal medicines in 5
the Chinese Pharmacopoeia (2005 English version)

MARKERS

Li, Shonglin et. al.,. (2008). Chemical markers for the quality control of herbal
At present, some herbs do not have

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2488332/ (online access).

markers for quality control. According
to the Chinese Pharmacopoeia (2005
edition), only 281 out of 551 herbs
have one or two chemical markers for
quality control. A total of 282 chemical

medicines: an overview. Retrieved from:

markers have been listed for
qualitative or quantitative analysis of
herbs. Moreover, many herbal
medicines share the same
chemical markers for quality

Reference:
control.

INTRODUCTION
CHEMICAL
6

MARKERS

Li, Shonglin et. al.,. (2008). Chemical markers for the quality control of herbal
According to Li, Shonglin et. al.,. (2008) in

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2488332/ (online access).

their research “Chemical markers for the
quality control of herbal medicines: an
overview” they suggested a new
classification of eight categories for
chemical markers, namely (1) therapeutic
components, (2) bioactive components,

medicines: an overview. Retrieved from:

(3) synergistic components, (4)
characteristic components, (5) main
components, (6) correlative components,
(7) toxic components, and (8) general
components used with fingerprint

Reference:
spectrum.

INTRODUCTION
 7

NEGATIVE
MARKER
Negative marker 8

• Complete and detailed botanical descriptions of plant parts are important for accurate identification of voucher
specimens. Asarum canadense (wild ginger) root has similar taste and smell to ginger rhizome but contains
aristolochic acid.

• Aristolochic acids are a family of carcinogenic, mutagenic, and nephrotoxic phytochemicals commonly found in the
flowering plant family Aristolochiaceae (birthworts). The family Aristolochiaceae includes the genera Aristolochia and
Asarum (wild ginger).

• In April 2001, the Food and Drug Administration issued a consumer health alert warning against consuming botanical
products, containing aristolochic acid. The agency warned that consumption of aristolochic acid-containing products
was associated with "permanent kidney damage, sometimes resulting in kidney failure that has required kidney
dialysis or kidney transplantation.

• In August 2013, two studies identified an aristolochic acid mutational signature in upper urinary tract cancer patients
from Taiwan. The carcinogenic effect is the most potent found thus far, exceeding the amount of mutations in
smoking-induced lung cancer and UV-exposed melanoma. Exposure to aristolochic acid may also cause certain types
of liver cancer.

NEGATIVE MARKER
Botanical description of Z. 9

officinale

Source: Zingiber officinale Rosc. in Flora of China @ efloras.org" eFlora. Missouri Botanical Garden, St. Louis, MO & Harvard
University Herbaria, Cambridge, MA., 2002. Web. Accessed March 2021.

NEGATIVE MARKER
Fresh Ginger vs. Wild Ginger 10

NEGATIVE MARKER
 11

THERAPEUTIC
AND
CORRELATIVE
MARKER
 12

Therapeutic components possess direct

therapeutic effects of a herbal medicine.
They may be used as chemical markers for
both qualitative and quantitative
assessments.

Originated from the bulbs of Fritillaria species

(family Liliaceae), Bulbus Fritillariae (Beimu)
is commonly prescribed as an antitussive and
expectorant in Chinese medicine practice.
Five different Bulbus Fritillariae derived from
nine Fritillaria species are documented in the
Chinese Pharmacopoeia. Isosteroidal
alkaloids of Bulbus Fritillariae, including
verticine, verticinone and imperialine, were
identified as the major therapeutic
components that account for the antitussive
effect.

THERAPEUTIC COMPONENTS
 13

Correlative components in herbal

medicines have close relationship with
one another. For example, these
components may be the precursors,
products or metabolites of a chemical or
enzymatic reaction. Correlative
components can be used as chemical
markers to evaluate the quality of herbal
medicines originated from different
geographical regions and stored for
different periods of time.

According to the Chinese Pharmacopoeia

(2005 edition), psoralen and isopsoralen
are used as chemical markers for
assessing the quality of Fructus
psoraleae (Buguzhi).

CORRELATIVE COMPONENTS
GINGER
14

Ginger (Zingiber officinale) is a common and

widely used spice. It is rich in various chemical
constituents, including phenolic compounds,
terpenes, polysaccharides, lipids, organic
acids, and raw fibers. The health benefits of
ginger are mainly attributed to its phenolic
compounds, such as gingerols and shogaols.

Accumulated investigations have demonstrated

that ginger possesses multiple biological
activities, including antioxidant, anti-
inflammatory, antimicrobial, anticancer,
neuroprotective, cardiovascular protective,
respiratory protective, antiobesity, antidiabetic,
antinausea, and antiemetic activities.

THERAPEUTIC & CORRELATIVE COMPONENTS

 GINGEROL
15

• Gingerol, properly as [6]-gingerol, is a phenol

phytochemical compound found in fresh ginger that
activates spice receptors on the tongue.

• Molecularly, gingerol is a relative of capsaicin and

piperine, the compounds which are alkaloids, though
the bioactive pathways are unconnected. It is normally
found as a pungent yellow oil in the ginger rhizome,
but can also form a low-melting crystalline solid.

• This chemical compound is found in all members of

the Zingiberaceae family and is high in concentrations
in the grains of paradise as well as an African Ginger
species.

• In a pre-clinical meta-analysis of gingerol compounds

anticancer, anti-inflammatory, anti-fungal, antioxidant,
neuroprotective and gastroprotective properties were
reported, which include studies in-vitro and in-vivo.

THERAPEUTIC & CORRELATIVE COMPONENTS

 SHOGAOL
16

• Shogaols are pungent constituents of ginger similar in chemical

structure to gingerol. The most common of the group is [6]-shogaol.
Like zingerone, it is produced when ginger is dried or cooked.
Moreover, shogaol (and gingerol) are converted to other
constituents when heat is applied over time, which is why ginger
loses its spiciness as it is cooked.

• In fresh ginger, gingerols are the major polyphenols, such as 6-

gingerol, 8-gingerol, and 10-gingerol. With heat treatment or long-
time storage, gingerols can be transformed into corresponding
shogaols. After hydrogenation, shogaols can be transformed into
paradols.

• More research in recent years has suggested that [6]-shogaol has a

strong anti-coughing effect, and could help reduce blood pressure.
Additionally, it may have some anti-allergic effects, as it has been
shown to inhibit the release of histamine from mast cells, a chemical
whose effects manifest as the allergic response to an allergen. An
anti-fever effect has also been noted.

THERAPEUTIC & CORRELATIVE COMPONENTS

GINGEROL VS. SHOGAOL
17

THERAPEUTIC & CORRELATIVE COMPONENTS

FRESH VS. DRIED GINGER
18

THERAPEUTIC USE FOR

DRIED GINGER IS
NOT SPECIFIED

THERAPEUTIC & CORRELATIVE COMPONENTS

APPLICATION OF GINGEROL AND
19

SHOGAOL AS CHEMICAL MARKERS

Lead • In a pre-clinical meta-analysis of gingerol

compounds for
compounds anticancer, anti-inflammatory,
anti-fungal, antioxidant, neuroprotective and

specific
gastroprotective properties were reported,
which include studies in-vitro and in-vivo.

therapeutic • More research in recent years has

suggested that [6]-shogaol has a strong

indication anti-coughing effect, and could help reduce

blood pressure. Additionally, it may have

specific for some anti-allergic effects, as it has been

shown to inhibit the release of histamine

fresh and dried

from mast cells, a chemical whose effects
manifest as the allergic response to an

ginger
allergen. An anti-fever effect has also been
noted.

THERAPEUTIC & CORRELATIVE COMPONENTS

APPLICATION OF GINGEROL AND
20

SHOGAOL AS CHEMICAL MARKERS

https://www.researchgate.net/publication/230131376_Gingerol_Decreases_after_Processing_and_
Zhang X. et al., (2006). Gingerol Decreases after Processing and Storage of Ginger. Retrieved at:
Quality • Gingerol is one of the most abundant constituent in fresh
ginger but it found to decrease during thermal processing

evaluation of and post harvest storage and shogaols even more

pungent than gingerols, is virtually absent in fresh

herbal products
ginger and is derived from gingerol during long term post
harvest storage.

• Fresh cooked ginger and processed ginger products were

Assessment and extracted with methanol and analyzed by HPLC to study
changes in the levels of [6]-, [8]-, and [10]-gingerols after

optimization of

Storage_of_Ginger(online access)
the products had been cooked and processed, and during
storage of the products. Sensory tests evaluated the

processing
intensity of pungency in different processed products.
Cooking and processing of decreased (p 0.05)

methods
gingerol levels, but blanching and freeze-drying had

Reference:
no effect.

THERAPEUTIC & CORRELATIVE COMPONENTS

APPLICATION OF GINGEROL AND
21

https://www.researchgate.net/publication/230131376_Gingerol_Decreases_after_Processing
SHOGAOL AS CHEMICAL MARKERS

Zhang X. et al., (2006). Gingerol Decreases after Processing and Storage of Ginger.
Stability test of
• Shogaols are artifacts formed during storage or through
excess heat, probably created by a dehydration reaction of

proprietary
the gingerols. The ratio of shogaols to gingerols sometimes
is taken as an indication of product quality.

products • Gingerols in all products degraded gradually with storage.

Ginger powder had a lower degradation rate than the paste
stored at 4°C.

_and_Storage_of_Ginger(online access)
FRESH DRY
Gingerol Shogaol Paradol

Retrieved at:
Reference:
THERAPEUTIC & CORRELATIVE COMPONENTS
APPLICATION OF GINGEROL AND
22

SHOGAOL AS CHEMICAL MARKERS

• A Shim-pack XR-ODS (2.2 μm particle size) high-
speed, high-separation column can be use for
analysis. Table 1 shows the analytical conditions.
A column washing stage was added to elute out
the impurity components, which are strongly
retained in the column during analysis using an
actual sample under these conditions. Fig. 2
shows the chromatograms for a standard solution
of 6-gingerol and 6-shogaol (100 mg/L each
component, methanol solution).

• After the pretreatment shown in Fig. 3, the ginger

was analyzed under the analytical conditions in
Table 1. Fig. 4 shows the resulting chromatogram.
This method achieved fast analysis of 6-gingerol
and 6-shogaol in ginger.

THERAPEUTIC & CORRELATIVE COMPONENTS

CHEMICAL MARKERS
23

Quality control of herbal medicines aims to

ensure its quality, safety and efficacy.
Chemical markers are pivotal in the current
practice of quality control. Chemical markers
should be used at various stages of the
development and manufacturing of a herbal
medicine, such as authentication and
differentiation of species, collecting and
harvesting, quality evaluation, stability
assessment, diagnosis of intoxication and
discovery of lead compounds. Lack of chemical
markers remains a major problem for the
quality control of herbal medicines. In many
cases, we do not have sufficient chemical and
pharmacological data of chemical markers.

THERAPEUTIC & CORRELATIVE COMPONENTS

 Gualtiero Simonetti (1990). Stanley Schuler (ed.). Simon & Schuster's Guide to Herbs and Spices. Simon & Schuster, Inc. 24

 Li, Shonglin et. al., (2008). Chemical markers for the quality control of herbal medicines: an overview. Retrieved from:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2488332. (online access).

 Mao QQ, Xu XY, Cao SY, Gan RY, Corke H, Beta T, Li HB, (2019). Zingiber officinale Roscoe. Foods. 8 (6): 185. doi:10.3390/foods8060185.
PMC 6616534. PMID 31151279.

 Mao QQ et al., (2019). Bioactive Compounds and Bioactivities of Ginger (Zingiber officinale Roscoe). Retrieved at:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6616534. (online access).

 Ramawat, Kishan. (2019). An Introduction to Biodiversity and Chemotaxonomy. Retrieved from:

https://www.researchgate.net/publication/337153441 (online access).

 Semwal RB, Semwal DK, Combrinck S, Viljoen AM. (2015). Gingerols and shogaols: Important nutraceutical principles from ginger.
Phytochemistry. 117: 554–568. doi:10.1016/j.phytochem.2015.07.012. PMID 26228533

 Shimazu Co. (2019). Fast Analysis of Gingerol and Shogaol in Ginger. Retrieved at: https://www.shimadzu.com/an/service-support/technical-
support/technical-information/lc-ap/n9j25k00000bd27z.html. (online access).

 US Department of Health and Human Services. (2001). FDA warns consumers to discontinue use of botanical products that contain
aristolochic acid. http://www. fda. gov/Food/DietarySupplements/Alerts/ucm096388. htm.

 Zhang X. et al., (2006). Gingerol Decreases after Processing and Storage of Ginger. Retrieved at:
https://www.researchgate.net/publication/230131376_Gingerol_Decreases_after_Processing_and_Storage_of_Ginger. (online access).

 Google images. Retrieved at: https://www. google.com

REFERENCES
 25

Chemical component may have more than one

attribute, a component may belong to multiple
categories. To know these multiple categories, a
CHEMICAL chemical fingerprint analysis is proposed.
FINGERPRINT

ADVANTAGE
It indicates the overall quality

DISADVANTAGE
Mass data analysis
 26

HOW CHEMICAL FINGERPRINT WORKS?

• Separation of the constituents in the extract in an elution pattern
which is governed by how they interact with the stationary and
mobile phase

By utilizing chemical fingerprinting the presence or absence of marker

compounds characteristic of the target species may be rapidly ascertained,
quantified and referenced against the established range if standardized or
quantified plant extracts are being considered.

CHEMICAL FINGERPRINT
 27

HOW CHEMICAL FINGERPRINT ARE DETECTED?

The technique was originally applied to colored compounds but nowadays

the development of powerful analytical tools has facilitated the detection of
almost any plant natural product:

• absorbance in UV light
• refractive index
• light scattering properties or by electrochemical detection
• Thin Layer Chromatography (TLC)
• Gas Chromatography (GC)
• Liquid Chromatography (LC)
• High Pressure Liquid Chromatography (HPLC)

CHEMICAL FINGERPRINT

HIGH PRESSURE LIQUID CHROMATOGRAPHY (HPLC)
HPLC – DAD system was performed in a ginger sample for its fingerprint analysis
Column: Waters Symmetry C18 column
Elution system: water and acetonitrile
Equilibration time: 10mins
Injection Volume: 3uL per sample
Flow rate: 1mL/min
Column temperature: maintained at 27°C
DAD detector: 230nm
CHEMICAL FINGERPRINT
28
With the use of HPLC-DAD, it is often
difficult to identify the chemical
compounds without the authentic
standards. In order to resolve this
problem, LC–MS/MS was performed
to obtain information about the
molecular mass in order to identify the
chemical composition of a ginger
extract with and without the reference
standards.
 29

HIGH PRESSURE LIQUID CHROMATOGRAPHY (HPLC)

In this figure, each representative

chromatogram of ginger extract from
each origin is depicted. The different
profiles of ginger from each origin was
compared with their retention time and
peak intensities.

CHEMICAL FINGERPRINT
 30

LIQUID CHROMATOGRAPHY (GC)

All chromatographic peaks were identified and confirmed using LC-MS/MS experiment
using an HPLC system coupled to a TQ-S triple quadruple MS

m/z: 100-1000 (ESI)

Capillary voltage: 3 kV
Source Temperature: 150°C
Desolvation Temperature: 500°C
Equilibration time: 10mins
Cone gas: Nitrogen, 150 L/h
Desolvation gas: Nitrogen, 800 L/min
Collision gas: Argon, 0.15 mL/min

CHEMICAL FINGERPRINT
 31

In the study of Yudthavorasit et al (2014), using HPLC-based

fingerprinting method allows the effective judgement of ginger
from different samples sites. High recognition (97%) and
satisfactory predictive ability (91%) proved that HPLC, LC-MS is a
powerful, practical tool that is suitable for tracing unclassified
ginger sample, that could address the quality control and food
authenticity concerns regarding ginger origin.

**Chemometric Analysis. In herbal fingerprint analysis, principal

component analysis (PCA) was commonly the unsupervised applied tool
for the exploratory analysis.

CHEMICAL FINGERPRINT
 32

CHEMOMETRIC ANALYSIS. Principal Component Analysis (PCA)

PCA is a well-known conventional experimental data analysis technique

which is used as a primary step in many fingerprint studies. PCA is applied
to determine underlying information from multivariate raw data by
transforming and reducing the dimensions of the original data matrix for
samples and variables into a product of two matrices, scores (T) and
loadings (P), while containing the same information as of the original data.

CHEMICAL FINGERPRINT
 33

THIN LAYER CHROMATOGRAPHY (TLC)

Compounds exhibiting conjugation show
absorbance in shortwave UV light (254 nm) and
thus appear as dark quenching bands against a
fluorescent green background on TLC plates
equipped with fluorescent indicator. It can be
observed that the different raw materials varied in
their citral content whilst the raw material used to
manufacture the defective batch of lemongrass
strong infusion was deficient in citral. Treatment of
the TLC plate with vanillin-sulfuric acid reagent
also confirmed these findings with citral appearing
as a dark purple band of varying intensity
depending on the raw material batch

CHEMICAL FINGERPRINT
 34

 Chemical Fingerprinting of Botanical Materials – A Case Study. 2014. Ransom Natural. Retrieve: May 2021. Chemical
Fingerprinting of Botanical Materials – A Case Study | Ransom Naturals Ltd

 Li S, Han Q, Qiao C, Song J, Cheng CL, Xu H. 2008. Chemical markers for the quality control of herbal medicines: an
overview. Chinese Medicine, 3(7).

 Ma Q, Chen X, Zhang K, Yao D, Yang L, Wang H, Bulemasi S, Huang J, Wang J. 2020. Chemical Fingerprint Analysis for
Discovering Markers and Identifying Saussurea involucrate by HPLC Coupled with OPLS-DA. Journal of Analytical
Methods in Chemistry. https://doi.org/10.1155/2020/7560710

 Prasad S. Tyagi AK. 2015. Ginger and Its Constituents: Role in Prevention and Treatment of Gastrointestinal Cancer.
Gastroenterology Research and Practice. http://dx.doi.org/10.1155/2015/142979

 Semwal RB, Semwl DK, Combrinck, Vilijoen AM. 2015. Gingerols and shogaols: Important nutraceutical principles from
ginger. Phytochemistry, 117:554-568

 Yeap YSY, Kassim NK, Ng RC, Fe GCL, Yazan LS, Musa KH. 2017. Antioxidant properties of ginger (Kaempferia
angustifolia Rosc.) and its chemical markers. International Journal of Food Properties, 20(51):51158-51172.

 Yudthavorasit S, Wongravee K, Leepipatpiboo N. 2014. Characteristic fingerprint based on gingerol derivative analysis for
discrimination of ginger (Zingiber officinale) according to geographical origin using HPLC-DAD combined with
chemometrics. Food Chemistry, 158:101-111.

REFERENCES
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