General Instruction for Writing Report for Continuous

Guru Nanak Institute of Pharmaceutical Science and

Technology
157/ F, Nilgunj Road, Panihati, Kolkata 700114

Title of Work: Structure elucidation of phytochemicals

Paper Code: R21_PT512T

Paper Name: Pharmacognosy and Phytochemistry II

Report Submitted for the Evaluation of Continuous Assessment II

Submitted by Name: Sovan Sarkar

Roll No: 186012111012

Program: B.Pharm

Semester: 5
 Introduction

Phytochemicals are chemicals. are chemicals produced by plants through primary or secondary
metabolism of plant origin. They generally have biological activity in the plant host and play
a role in plant growth or defence against competitors, pathogens, or predators.

Phytochemicals are generally regarded as research compounds rather than essential nutrients
because proof of their possible health effects has not been established yet. Phytochemicals
under research can be classified into major categories, such as carotenoids and polyphenols,
which include phenolic acids, flavonoids, stilbenes or lignans. Flavonoids can be further
divided into groups based on their similar chemical structure, such as anthocyanins, flavones,
flavanones, isoflavones, and flavanols. Flavanols are further classified as catechins,
epicatechins, and proanthocyanins. In total, between 50,000 and 130,000 phytochemicals have
been discovered.

Phyto chemists study phytochemicals by first extracting and isolating compounds from the
origin plant, followed by defining their structure or testing in laboratory model systems, such
as in vitro studies using cell lines or in vivo studies using laboratory animals. Challenges in
that field include isolating specific compounds and determining their structures, which are
often complex, and identifying what specific phytochemical is primarily responsible for any
given biological activity.
Chromatographic fingerprinting and marker compound analysis

An herbal medicine's (HM) chromatographic fingerprint is an analysis of the extract's common

chemical components to identify pharmacologically active and/or chemical characteristics.
This chromatographic profile should be featured by the fundamental attributions of “integrity
” and “fuzziness” or “sameness” and “differences” so as to chemically represent the HM inve
stigated. It is suggested that with the help of chromatographic fingerprints obtained, the
authentication and identification of herbal medicines can be accurately conducted (integrity)
even if the amount and/or concentration of the chemically characteristic constituents are not
exactly the same for different samples of this HM or, the chromatographic fingerprints could
demonstrate both the “sameness” and “differences” between various samples successfully.
HPLC fingerprinting includes recording of the chromatograms, retention time of individual
peaks and the absorption spectra (recorded with a photodiode array detector) with different
mobile phases. Similarly, GLC is used for generating the fingerprint profiles of volatile oils
and fixed oils of herbal drugs. Furthermore, the recent approaches of applying hyphenated
chromatography and spectrometry such as High-Performance Liquid Chromatography–Diode
Array Detection (HPLC–DAD), Gas Chromatography–Mass Spectroscopy (GC–MS),
Capillary Electrophoresis- Diode Array Detection (CE-DAD), High-Performance Liquid
Chromatography–Mass Spectroscopy (HPLC–MS) and High-Performance Liquid
Chromatography–Nuclear Magnetic Resonance Spectroscopy (HPLC–NMR) could provide
the additional spectral information, which will be very helpful for the qualitative analysis and
even for the on-line structural elucidation.

Liquid chromatography

a. Liquid chromatography is a preparative high performance liquid chromatography. There are

basically two types of preparative HPLC. One is low pressure (typically under 5 bar) traditional
PLC, based on the use of glass or plastic columns filled with low efficiency packing materials
of large particles and large size distribution. A more recent form PLC, Preparative High
Performance Liquid Chromatography has been gaining popularity in pharmaceutical industry.
In preparative HPLC, larger stainless-steel columns and packing materials. The examples of
normal whereas in analytical work the goal is to get information about the sample. Because its
higher column efficiencies and faster solvent velocities permit more difficult separation to be
conducted more quickly.
b. Liquid Chromatography- Mass Spectroscopy (LC-MS)
In Pharmaceutical industry LC-MS has become method of choice in many stages of drug
development. Recent advances include electro spray, thermos spray, and ion spray ionization
techniques which offer unique advantages of high detection sensitivity and specificity, liquid
secondary ion mass spectroscopy, later laser mass spectroscopy with 600 MHz offers accurate
determination of molecular weight proteins, peptides. Isotopes pattern can be detected by this
technique.
c. Liquid Chromatography- Nuclear Magnetic Resonance (LC-NMR)
The combination of chromatographic separation technique with NMR spectroscopy is one of
the most powerful and time saving method for the separation and structural elucidation of
unknown compound and mixtures, especially for the structure elucidation of light and oxygen
sensitive substances. The online LC-NMR technique allows the continuous registration of time
changes as they appear in the chromatographic run automated data acquisition and processing
in LC-NMR improves speed and sensitivity of detection.

Gas chromatography
a. Gas Chromatography Fourier Transform Infrared spectrometry
Coupling capillary column gas chromatographs with Fourier Transform Infrared Spectrometer
provides a potent means for separating and identifying the components of different mixtures.
B. Gas Chromatography-Mass Spectroscopy
Gas chromatography equipment can be directly interfaced with rapid scan mass spectrometer
of various types. The flow rate from capillary column is generally low enough that the column
output can be fed directly into ionisation chamber of MS. The simplest mass detector in GC is
the Ion Trap Detector (ITD). In this instrument, ions are created from the eluted sample by
electron impact or chemical ionisation and stored in a radio frequency field; the trapped ions
are then ejected from the storage area to an electron multiplier detector. The ejection is
controlled so that scanning on the basis of mass-to-charge ratio is possible. The ions trap
detector is remarkably compact and less expensive than quadrupole instruments. GC-MS
instruments have been used for identification of hundreds of components that are present in
natural and biological system.

Supercritical Fluid Chromatography (SFC)

Supercritical fluid chromatography is a hybrid of gas and liquid chromatography that
combines some of the best features of each.
This technique is an important third kind of
column chromatography that is beginning to find use in many industrial, regulatory and
academic laboratories.
SFC is important because it permits the separation and determination of
group of compounds that are not conveniently handled by either gas or liquid
chromatography.
As it is Inapplicable or contain no functional group that makes possible detection by
the spectroscopic or electrochemical technique employed in LC.
 Conclusion

It can be said that phytochemicals and their structure are determined by various methods like
NMR spectroscopy Gas chromatography and Liquid chromatography which will be very
helpful for the qualitative analysis and even for the on-line structural elucidation to get a better
knowledge and discover new phytochemicals in nature.
 Reference

o Nahar, Lutfun, and Satyajit D. Sarker. "Automated structure elucidation of

phytochemicals." Trends in Phytochemical Research 1.3 (2017): 109-110.

o Matsuda, Fumio, et al. "Mass spectra-based framework for automated structural

elucidation of metabolome data to explore phytochemical diversity." Frontiers in plant
science 2 (2011): 40.

o Doughari, James Hamuel. Phytochemicals: extraction methods, basic structures and

mode of action as potential chemotherapeutic agents. Rijeka, Croatia: INTECH Open
Access Publisher, 2012.

o Dagnaw, Wondu, and Alemayehu Mekonnen. "Preliminary phytochemical screening,

isolation and structural elucidation of chloroform leaf extracts of Maesa
lanceolata." Chem. Int 3.4 (2017): 351.

o Ingle, Krishnananda P., et al. "Phytochemicals: Extraction methods, identification and

detection of bioactive compounds from plant extracts." Journal of Pharmacognosy and
Phytochemistry 6.1 (2017): 32-36.
Plagiarism Report