Morphine is a natural opium alkaloid first isolated in 1805. It is a powerful pain reliever that acts on opioid receptors in the brain and nervous system. Spectroscopy techniques like IR, NMR, and mass spectrometry are used to characterize the molecular structure of morphine. IR spectroscopy identifies functional groups based on vibrational frequencies. NMR spectroscopy provides information on bonding and stereochemistry. Mass spectrometry identifies molecular mass and fragmentation patterns. Together these techniques allow for full structural elucidation of morphine.

1. GURU JAMBHESHWAR UNIVERSITY OF
SCIENCE AND TECHNOLOGY, HISAR
STRUCTURAL CHARACTERISATON OF MORPHINE
BY USING DIFFERENT TYPES OF SPECTROSCOPIES
SUBMITTED TO- Dr. Sandeep Jain Sir
SUBMITTED BY- Shikha Kamboj
M Pharm 1st Sem
Pharmaceutical Chemistry

2.  CONTENTS: 1. Introduction
2. Classification
3.Physical and chemical properties
4. Mechanism of action
5. Stereochemistry
6. Constitution
7. SAR
8. Spectroscopy
(a) IR
(b) NMR
(c) MS
9. Uses of morphine
10. Adverse effect of morphine

3. INTRODUCTION:-
• 1. INTRODUCTION
• MORPHINE:- It is a natural opium alkaloid, was first
obtained from opium poppy (Papaver somniferum) in 1805
by German pharmacist Friedrich Serturner. This is generally
believed to be the first isolation of an active ingredient from
plant. The term morphine is derived from Greek word
“Morpheus” which means “god of dreams”.
• In opium, it is present in a quantity of 10-23% along with
other substances.
• Morphine is said to be the most powerful pain reliever
medicine.
• It can be administered by mouth, by injection into a muscle,
by injection under the skin, intravenously etc.
• Its maximum effect is reached after about 20 mins when
administered IV and 60 mins when administered by mouth.

4. 2. CLASSIFICATION
1.

5. 2. ACCORDING TO SYNTHESIS
Meperidine

6. 3. PHYSICAL AND CHEMICAL
PROPERTIES OF MORPHINE
• It is solid, white crystalline alkaloid and needle shaped crystals.
• It is odorless and bitter in taste.
• Boiling point is 190°C.
• It is insoluble in water and soluble in alcohol and alkali
solutions.
• Its molecular formula is C17H19NO3 and molecular mass is
• 285.34 g/mol.
• Morphine is laevorotatory.
• SOLUBILITY: When exposed to air morphine sulfate gradually
loses its water of hydration. The drug darkens on prolonged
exposure too.
• DECOMPOSITION: When heated it emits toxic fumes of nitrogen
oxide.
• pH: pH of saturated solution is 8.5

7. 4. MECHANISM OF ACTION
 Morphine is considered the classic opioid analgesic with
which other painkillers are compared. Like other
medications in this class, morphine has an affinity for
delta, kappa and mu opioid receptors.
 Opioids activate 7 transmembrane GPCRs located
presynaptically and postsynaptically along pain
transmission pathways. High densities of opioid
receptors known as mu, delta and kappa are found in
the dorsal horn of the spinal cord and higher CNS
centers.

8. • Opioids cause hyper polarization of nerve cells,
inhibition of nerve firing and presynaptic
inhibition of transmitter release.
• Cellular effects of these drugs involve
enhancement of neuronal potassium
efflux(hyperpolarizes neurons and makes them
less likely to respond to a pain stimulus) and
inhibition of calcium influx (decreases neuro-
transmitter release from neurons located along
the pain transmission pathway).
• Constipation results from activation of opioid
receptors in the CNS and in the GIT.

11. 5. STEREOCHEMISTRY
• Morphine- The molecular formula of morphine is
C17H19NO3 and it is a ‘T’ shaped molecule.
• The three dimensional structure of morphine is
fascinating and in all morphine alkaloids,
phenanthrene nucleus is present. Due to this, these
are also known as phenanthrene alkaloids.
• It consists of five rings, three of which are
approximately in the same plane.
• The other two rings, including the nitrogen one, are
such at right angles to the other trio.
• It also consist of phenolic, ether and alcoholic group
and tertiary amine.

12. • Natural morphine molecule exists as a single
isomer.
• Naturally occurring levorotatory (-) morphine has
5R, 6S, 9R, 13S, 14R configuration.
• It undergoes epimerization at 14position, but not
beneficial to analgesic activity.
• Carbons at 5,6,9,13,14 position are chiral centers.

15. 6. CONSTITUTON
• MOLECULAR FORMULA: C17H1903N
• NATURE OF NITROGEN ATOM: As morphine adds
on one mole of methyl iodide to form a quaternary
salt, this shows that it contains tertiary nitrogen atom.
The tertiary nature of the nitrogen is further confirmed
by Hofmann degradation of codeine derivative which
also reveals the presence of nitrogen in the ring.
• NATURE OF OXYGEN ATOM: 1. Morphine when
acetylated or benzoylated yields the diacetyl or
benzoyl derivative, indicating that the morphine
contains two hydroxyl groups.

16. • 2. Morphine is also soluble in aqueous sodium
hydroxide solution to form monosodium salt
which is reconverted into morphine by passing
CO2 through it. All these facts reveals that one of
the two hydroxyl groups is phenolic in nature.
• PRESENCE OF ETHYLENIC BOND: When codeine
is reduced catalytically in the presence of
palladium, it takes up one molecule of hydrogen,
suggesting that both codeine and morphine
contain one ethylene bond.

17. • PRESENCE OF BENZENE NUCLEUS: Morphine when
brominated yields, a mono-bromo derivative along
with evolution of a molecule of hydrogen bromide,
suggesting that morphine possesses a benzene
nucleus.
• PRESENCE OF PHENANTHERENE: 1) When morphine
is distilled with zinc dust, it yields phenanthrene and
a no of bases suggesting that morphine may contain
phenanthrene nucleus.
• 2) When codeine is treated with methyl iodide, it
yields codeine methiodide. The later compound
when boiled with sodium hydroxide solution, yields
alpha- methylmorphimethine.
• 3) Which on further heating with acetic anhydride
yields an mixture of methyl morphol and
ethanoldimethylamine.

18. • PRESENCE OF THREE OXYGEN ATOMS: 1) B-
methylmorphimethine when heated by water,
yields a mixture of trimethylamine, ethylene and
methylmorphenol. Methylmorphenol when
demethylated by the hydrochloric acid yields
morphenol, a compound which contains one
phenolic hydroxyl group and an inert oxygen
atom.
• 2)

19. • 3) Codeine methiodide : a codeinone methiodide
on heating separately with a mixture of Ac2O-
AcONa gives 3-methoxy-4-acetoxyphenantthrene
an 3-methoxy-4, 6-diacetoxyphenanthrene
respectively. The addition of the acetoxyl group in
position 6 latter indicates the secondary alcoholic
group which further lost as water molecule during
dehydrogenation.

20. 7. SAR (STRUCTURAL ACTIVITY
RELATIONSHIP)
• 1. THE PHENOL MOIETY
• R=H Morphine
• R=Me Codeine
• An aromatic phenyl ring
is essential for activity.
• Any other substitution on phenyl ring diminishes
activity.
• Esterification at C3 decrease the analgesic
activity but increase the anti-tussive activity.

21. 2. THE 6-ALCOHOL
• The alcoholic group at C-6
when methylated, esterified,
Oxidized, removed analgesic
activity as well as toxicity of
the compound increase.
• The saturation of the double bond at C-7 position
gives more potent compound.
• Bridging of C-6 and C-14 through ethylene
linkage gives potent derivatives.

22. 3.MODIFICATION OF 3 NITROGEN
• Nitrogen is essential for the Activity, the removal
of nitrogen Atom results in decreases in
Analgesic activity.
• Double bond at 7 and 8 Position is not important
to binding.

23. • The substitution of certain bulky groups on
nitrogen 17 converts an opioid agonist into an
opioid antagonist. The most important of which
is naloxone.
• Additionally, substitution of certain very bulky
groups on carbon 6 converts naloxone into a
peripherally-selective opioid antagonist i.e.
naloxegel.

24. 4.EPOXIDE BRIDGE
• Removal of epoxide bridge (4,5) in morphine
structure result in the compound that is referred
to as morphinans.
• As the synthetic procedure, only levo isomer
possesses opioid activity while the dextro isomer
has useful antitussive activity, for eg.
Levorphanol and butorphanol.
• Levorphanol is more potent analgesic than
morphine.

25. • Summarized SAR of morphine analogues is given
below:-

26. 8. SPECTROSCOPY
• There are a number of techniques that can help
us to determine the molecular structure,
molecular mass, bond angle, bond length,
functional group etc. of an unknown compound.
• IR spectroscopy
• UV/Vis spectroscopy
• NMR spectroscopy
• Mass spectroscopy
• X-Ray diffraction
• Emission spectroscopy

27. STRUCTURAL CHARACTERIZATION OF
MORPHINE BY USING IR SPECTROSCOPY
• IR spectroscopy is one of the most powerful
analytical techniques which offers the possibility of
chemical identification.
• PRINCIPLE: In any molecule, it is known that atoms or
groups of atoms are connected by bonds. These
bonds are analogs to springs. Because of the
continuous motion of the molecule they maintain
some vibrations with some frequency, characteristic
to every portion of the molecule. This is called the
natural frequency of vibration.
• Applied IR frequency = Natural frequency of
vibration.
• Change in dipole moment

29. The first plane in the case of morphine involves C4,
C9 to C14, O19, O21, O19 do not know enough
about the subject. comprising benzene (R1), oxide
(R2), and carboxylic (R3) rings. The second part
includes C5 to C9, C13 to C7, O20 and N22 and
contains the cyclohexenyl (R4) and ethenamine (R5)
rings. The two planes are nearly perpendicular to
each other and the molecule has a T shape.
The atoms in ring R1 are already coplanar, however
the pentagonal ring R2 is distorted, this is due to
the anti-bonding electron (oxygen) repulsion.
The ring R5 has typical chair form, and ringR4 has a
boat form.

30. This is caused by the 4, 5 ether bridge, which is also
responsible for the rigidity of morphine. The calculated
values match well with the experimental values. For
example in morphine, the optimized bond lengths of C-C of
the rings I made some changes; hope they are ok. R1, R2,
R3, R4, and R5 lie, respectively, between 1.36–145 ˚A,
1.37–1.55 ˚A, 1.33–1.56 ˚A, 1.34–156 ˚A, and 1.52–155 ˚A,
however the respective experimental data lies between
1.36–1.41 ˚A, 1.37–1.46 ˚A, 1.36–1.55 ˚A, 1.36–1.55 ˚A,
and 1.52–155 ˚A. In the case of heroin the optimized bond
lengths of C-C for the rings R1, R2, R3, R4, and R5 lie
between 1.38–1.40 ˚A, 1.38–1.50 ˚A, 1.33–1.55 ˚A, 1.38–
1.56 ˚A, 1.52–1.55 ˚A, and the corresponding experimental
data lies in between 1.36–1.42 ˚A, 1.38–1.51˚A, 1.29–1.55
˚A, 1.36–1.62 ˚A, and 1.52–1.55 ˚A, respectively.

31. As expected the two adjacent bonds of C7=C8 are
slightly shorter than the other bond in ring R3, e.g., the
bond lengths between 14C-8C, 7C-6C, and 6C-20O (23O
in case of heroin) are shorter than the C-C bond of ring
R3.
The bond length between C-C in the ring R5 is slightly
greater than the bond length between C-N in the same
ring of both the molecules. The bond lengths of each C-H
bond in the methyl group are not the same, but the
bond lengths between the hydrogen which is in plane
are .01 ˚A greater than the hydrogen which is out of
plane. Because of ring R2, which is distorted in shape,
some bond is elongated and its adjacent bond is
contracted a bit. The calculated substituent impact on
the C-C-C bond angles of the benzene ring is summarized
in table.

32. The data reflects the well-known trends observed for the
various substituents: morphine becomes heroin when two
O-H groups are attached at the position of C3 and C6, and
in morphine they are replaced by –OCOCH3. This causes
decrease in ortho, and it also results in a small increase in
para in the case of heroin, e.g.,
the calculated value of the ipso bond angle between C2-C3-
4C of morphine is 116◦ which
increases in the case of heroin and becomes 117◦. The bond
angle ortho at position C3 of
morphine is 121◦, and it decreases in the case of heroin and
becomes 119◦. The bond angle
para at position C3 of morphine is 119◦ and, due to a small
increment, in the case of heroin it becomes 119.21◦.

33. Bond lengths between oxygen attached with carbon at
the position 3, 6 are greater than in the case of heroin.
The bond lengths between C3-O19, C6-O20 in the case
of morphine are 1.37 ˚A, 1.42 ˚A, however the bond
lengths between C3-O23, C6-O26
are 1.39 ˚A, 1.44 ˚A in the case of heroin. It was
suggested by Bye (1976) at this
short bond distance resulted from a strong O-H...N
hydrogen bond in the case of morphine.
However, most of the substituents in the present study
have a mixed / character, and
the geometrical parameters of the ring are a result of
the superposition of overall effects.
Based on the above comparison, although there are
some differences between the theoretical
values and the experimental values.

38. STRUCTURAL CHARACTERIZATION OF
MORPHINE BY USING MASS
SPECTROSCOPY
 Mass spectroscopy is an instrumental technique in which
sample is converted to rapidly moving positive ions by
electrons bombardment and charged particles are
separated according to their masses. Mass spectrum is a
plot of relative abundance OR intensity against the ratio
of mass/charge.
 PRINCIPLE: Conversion of neutral molecule into a
charged molecule to a positively charged
molecule.
 Separation of positively charged fragments
formed, based on their masses.

39. • Despite the importance of morphine and its biosynthetic
relatives, the literature dealing with electrospray
ionization mass spectrometry is limited. A number of
studies reported on identification and quantification of
morphine and related metabolites. Accurate mass data
of selected morphinans were obtained by quadrupole,
time-of-flight and FT-ICR mass spectrometry,
respectively.
• The substances under investigation are alkaloids with a
basic nitrogen, which makes them predestined for
electrospray ionization (ESI) in positive ion mode.
• Morphine and codeine show a very similar
fragmentation pattern since codeine contains only one
additional methyl group. Therefore, most fragments
found for morphine were also detected for codeine.

40. • The [M H H2O] ion formed by the loss of water
from position 6 is only found in the ion trap MS2.
The fragments of the type a are formed by
cleavage of the piperidine ring and loss of an
amine (CH2CHNHCH3, m 57). Such a fragment is
found for all morphinans and plays a crucial role
in further fragmentation pathways. Fragments of
the type (a-2H) are not found in case of ion trap
but for triple quadrupole MS/MS. Loss of water
from ion a leads to the key ion b, expulsion of
carbon monoxide to the fragment c. Both
pathways come together in the e-type fragment,
representing a loss of CO and water/MeOH from
the a fragment, respectively.

41. • The ion of type (e-H2O) with the elemental composition of
[C13H9] represents the most prominent fragment ion in
triple quad, and FT-ICR. The (e-CO) fragment is especially
abundant in triple quadrupole MS/MS. Another way leads
independently from [M H] to the key ion of type d.
• Ion trap MS3 of the [M HH2O] ion and ion a results in b,
whereas fragment c is only formed from a. In case of
morphine, MS3 of the base peak at m/z 201 (c) gives rise
to peaks at m/z 183 (e, 100), m/z 173 (c-CO, 18), m/z 165
(e-H2O, 5),m/z 155 (e-CO, 20), m/z 145 (8), and m/z 123
(10), (relative intensities in brackets). While MS4 of m/z
183 (e) results in m/z 165 (e-H2O) and m/z 155 (e-CO),
MS3 of m/z 211 (b) generates m/z 193 (b-H2O), m/z 183
(e), and m/z 165 (e-H2O). MS4 of m/z 183 (e) gives 165 (e-
H2O) and 155 (e-CO), indicating that this ion with
[C13H11O] plays a key role in the fragmentation.

44. fragmented ions m/z ratio Relative abundance (%)
[M H]+ 286 13
a 229 68
b 211 36
c 201 100
d 185 10
Table1: showing relative abundance and m/z ratio of morphine

45. STRUCTURAL CHARACTERIZATION OF
MORPHINE BY USING NMR
SPECTRSCOPY
• Nuclear Magnetic Resonance (NMR) spectroscopy is an
analytical chemistry technique used in quality control and
research for determining the content and purity of a
sample as well as its molecular structure.
• PRINCIPLE: The principle behind NMR is that many nuclei
have spin and all nuclei are electrically charged. If an
external magnetic field is applied, an energy transfer is
possible between the base energy to a higher energy level.
• The energy transfer takes place at a wavelength that
corresponds to radio frequencies and when the spin
returns to its base level, energy is emitted at the same
frequency.

47. • Morphine Systems - The two aromatic protonated
carbons, C-1 (d) and C-2 (d), of morphine (la) and codeine
(lb) were easily differentiated from the olefinic carbons C-
7 (d) and C-8 (d) by the upfield shift of the latter
resonances on reduction of the double bond.
• Carbon 1 and C-2 were distinguished by the larger ortho
effect from the C-3 hydroxyl substituent compared to the
para effect from the C-4 0-alkyl moiety. Likewise, the
even larger ortho effect of the C-3 methoxyl substituent
caused the C-2 resonance in lb (and 2b) to appear upfield
relative to that in la (and 2a). Carbon 7 and C-8 of la (and
lb) were differentiated by the upfield shift of the C-7
resonance on going from la to either 3,6-
diacetylmorphine (IC) or 6-acetylmorphine(ld). The
upfield shift was due to the y effect of the C-6 acetoxyl
group.

48. • The assignment of the C-3 (s) and C-4 (s)
resonances was accomplished by comparing the
compounds possessing a C-3 hydroxyl group to
those containing a C-3 methoxyl group. On
changing the hydroxyl group to the methoxyl
group, substituent constants for methine carbons
in substituted benzenes predict that the C-3
resonance should be shifted downfield. 3-4 ppm
while the C-4 resonance should go upfield. 2 ppm
owing to the larger ortho effect of the C-3
methoxyl substituent.

53. Morphine rule
• A tertiary nitrogen with small alkyl substitution.
• A quaternary carbon
• A phenyl group directly attached to the quaternary
carbon.
• A 2 carbon spacer between the quaternary carbon
an the tertiary nitrogen.

54. 9. USES OF MORPHINE
• Antitussive effect
• Analgesic
• Sedation
• Nausea and Vomiting- Higher dose of morphine
inhibit the vomiting Centre.
• Papillary construction- Miosis.
• Respiration- Resulting in increase in plasma CO2
concentration.
• Heat regulation- It shift the equilibrium point of
heat.

55. 10. ADVERSE EFFECT OF MORPHINE
• CVS: Bradycardia, edema
• CNS: depression, confusion, dizziness,
drowsiness, fatigue, headache
• Endocrine and metabolic: Dehydration
• Gastrointestinal: constipation, nausea, vomiting
• Idiosyncrasy and allergy
• Tolerance and dependence
• Histamine release
• Respiratory depression

56. Recent advancements
• 1. Dual Naked-Eye and Optical Chemo-sensor for
Morphine Detection in Biological Real Samples
Based on Cr(III) Metal–Organic Framework
Nanoparticles:

57. • 2. Vibrational Spectroscopy for Identification of
Metabolites in Biologic Samples.
• The average Raman spectra of five semen
samples (black) (a–e), and the Raman spectra of
blood (f) and saliva (g) spectroscopic signature.