SEMINAR ON

X-RAY CRYSTALLOGRAPHY
Presented By
Mounik Rout
M.Pharm (Ph.technology)
Guided By
Dr. Sasmita Kumari Acharjya

ROLAND INSTITUTE OF PHARMACEUTICAL SCIENCES

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 CONTENT
INTRODUCTION
PRODUCTION OF X-RAY
BRAGG’S LAW
INSTRUMENTATION
DIFFERENT X-RAY DIFFRACTION METHODS
APPLICATION OF X-RAY DIFFRACTION

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INTRODUCTION
X-ray crystallography is a non-destructive technique for determining the molecular structure
of a crystal .

X-ray crystallography uses the principle of X-ray diffraction to analyze the sample .

We can get 3d structure or any sample because it rotates in the sample cell and faces the X-ray
beam in different directions .

This technique helps in analyzing 3d crystal structure of biological materials .

Based on the radiations, X-ray spectroscopy is categorized in 3 type :

1. X-ray diffraction
2. X-ray Absorption
3. X-ray Fluorescence 3
.

 Diagram of x-ray passing through sample

DIFFRACTED RADIATION
FLUORESCENCE RADIATION
INCIDENT RADIATION
TRANSMITTED(Absorbed) RADIATION
Sample

Diffracted radiation –It is the most accurate method of analysis with high degree of
specificity than any other X-ray spectroscopy method .
X-ray diffraction(XRD) is the diffraction of incident radiation produced by crystalline sample
according to the atom present in it . 4
X-ray Absorption :-
When incident beam is passed through the sample then some fraction of X-ray photons are
absorbed .
 So the no of photons absorbed by the sample is used to measure the concentration of the
sample .
This is similar to any other absorption method like UV-Visible / IR spectroscopy by giving
the information about the absorbing material in the sample .

X-ray Fluorescence :-
When incident beam is passed through the sample then the electrons of the atoms of sample
gets excited by absorbing some energy , when those electrons come to ground state from
exited state they emitt some radiation which have longer wavelength than incident beam .
By measuring the wavelength and intensity of the generated radiation analyst can perform
qualitative as well as quantitative analysis .

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Schematic diagram of diffraction pattern

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Production of X-ray
When high velocity of electrons will strike on metal target then X-ray will produce .
It can explain by Bohr’s atomic model .
e¯
e¯

K SHELL

L SHELL e¯

M SHELL

N SHELL

Bohr’s atomic model

Energy of the outer shell is higher than inner shell .
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Principle :-
If one high velocity electron will strike the atom then it knock one electron completely from
that atom and get out from that by producing a void space .

An electron from higher shell falls on the void space .

It will release energy in the form of X-ray because of coming from higher orbit to lower
orbit .

The energy of released X-ray will be equal to the difference in the energy between 2 shells .

Ex-ray = EL – EK (if e¯ falls from L shell)

=EM – EK(if e¯ falls from M shell)
=EN – EK(if e¯ falls from N shell)
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 Theory of production of X-ray:-
The high velocity eˉ will strike the anode material in a discharge tube , which leads to
production of X-ray .
eˉ
X-rays

Atomic nuclei

The striking eˉ interact with the strong electric field of the atomic nuclei constituting the
anode material , which results deacceleration of the striking eˉ and release of energy due to
loosing of kinetic energy and radiation of photons which are responsible for production of X-
rays .

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10
The process by which photons are emitted by an eˉ is known as ‘Bremsstrahlung’ which
means deacceleration of radiation .

Initial energy of striking eˉ = Ei

Energy for X-rays(Photons) =Ex-rays
Final energy of eˉ after deacceleration = Ef
Ef = Ei - Ex-rays
Ei = Ef + Ex-rays

If all initial kinetic energy (Ei) will convert into X-rays ,then velocity of the eˉ become 0 .

So Ef = 0 ,and Ei = Ex-ray

This is the condition which will give the X-rays of highest energy and low wavelength .
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Bragg’s law
X-ray diffraction based on Bragg’s law:-
A C

B D
θ O θ
α
θ
d X θ Parallel planes of crystal surface
Z
θ θ
β d= Inter planner distance
Y
Θ= Glancing angle
C , D = Diffracted X-rays
A ,B = Incident X-rays

When the X-ray is incident onto a crystal surface, its angle of incidence θ, will reflect with
the same angle of scattering, θ. And, when the path difference, d is equal to a whole number,
n, of wavelength, constructive interference will occur.
Bragg’s equation = n λ = 2dsin θ , where n is order of diffraction 12
Derivation of the equation :-
As per the fig. when X-ray falls on the crystal at angle θ then some rays will reflect from upper
plane at same angle θ .
After extrapolating the reflection line then found that there are 2 angle :-
<XOY=θ , <ZOY =θ
And AO=BX , CO=DZ so the path difference
will be XY+YZ ----- 1
Path diff. is defined as an integral multiple of
wavelength =n λ----- 2
So that n λ=XY+YZ---- 3
Taking Sin θ = L L= Length of perpendicular
H H= Hypotenuse
So in ∆XOY , Sin θ= XY
XY=OY×SIN θ OY
=>XY= d × SIN θ----- 4
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In ∆ZOY , SIN θ= ZY
OY
ZY=OY×SIN θ
SIN θ values = 0 30 45 60 90
ZY= d×SIN θ---- 5 0 ½ 1/√2 √3/2 1
Putting equ.5 in equ.3
n λ= d×SIN θ + d×SIN θ
n λ= 2d×SIN θ ----- 6------ Bragg’s equation
where n = Order of diffraction
λ = wavelength , θ =Glancing angle
d = interplanar distance
Bragg’s equation gives the relationship between:-
1.wavelength of X-rays
2. Interplanar distance in crystal planes
3.Angle of reflection

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X-ray spectrometer instrument
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INSTRUMENTATION:-
.

X-ray tube

Collimator Monochromator Sa
m ple
Anode

e¯ Be window
Cathode
Focusing cup Recorder

Filament heating circuit Detector

Amplifier
Tungsten filament

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Working & Instruments :-
High voltage electric current supplied to heat the tungsten filament to emitt electron from
cathode .

Electrons from cathode striking on anode forms X-ray which will go through Beryllium
window towards the sample through a Collimator reducing undesirable radiation .

Then it passes to Monochromator which gives required X-ray beam to the sample .

Sample have the crystal surface which diffract the X-ray beam to detector made up of
photographic type or counter types .

Detector detects the beam intensity and proceed the signal to amplifier .

Amplifier amplifies the given signal and then the signal proceeds to the recorder screen .
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X-ray tube :- It is a large vaccum tube containing a heated cathode of Tungsten filament & a Cu
or Mo(Molybdenum) operated at higher voltage up to 60kv .
Collimator :- It passes narrow beam of X-ray by arrangement of 2 closely packed metal plates
separated by a small gap .
Monochromator:- It has 2 type :
1.Filter type – X-ray beam will partially monochromatize passing required radiation by
absorbing undesirable radiation .
2.Crystal type – It is made up of suitable crystalline materials like-NaCl , Quartz , etc.
Detector :- 1. Photographic method – A film is exposed and developed in X-rays passed through
the sample .
2. Counter Method – 1.Geiger Muller counter
2.Propertional Counter
3.Scintillation Detector
4.Solid-state semi-conductor Detector
5.Semi-conductor Detector
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Detailed image of Bragg’s X-ray spectrometer
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Bragg’s X-ray spectrometer method
Bragg designed a spectrometer to measure the intensity of x-ray beam following Bragg's
equation i.e. n λ = 2dsin θ

Crystal(sample)
θ
θ

Pb slit
…… … CH3Br
…… …
…… …
G
…… …
…… …
Ionization chamber

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Working:-
High voltage current is applied on the tungsten filament generating heat which helps the
cathode to emitt electron .
Later the electron strikes on the anode surface producing X-rays which passes through
Beryllium window to the sample.
Collimator placed right after Be window absorbs undesirable radiation and passes required
X-ray beam towards monochromator .
Monochromator converts polychromatic beam to monochromatic beam and then passes it to
sample surface.
The crystalline sample reflect the beam at glancing angle to ionization chamber through lead
slits .
The reflected X-rays ionizes Ch3Br gas to allow current flow in the chamber by electrodes .
Galvanometer present outside to measure the ionization current, which is require to form a
peak in respective to the intensity of X-ray reflected by the crystal .
The ionization current is measured for different values of glancing angle . A graph is drawn
between the glancing angle and ionization current.
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.

Intensity
θ θ

θ θ

θ θ
θ1 θ2 θ3
Reflection by Crystal Graph

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.

Photographic film Cylindrical Camera

Crystal

Rotating shaft

Motor

Rotating crystal method 23

Working
X-ray is generated in the X-ray tube and X-ray beam is made Monochromatic .
Monochromatic radiation will fall on the crystal Mounted on a shaft which can be rotated at
uniform angular rate .
Shaft will rotate the crystal at slow rate so that planes of crystal surface coming successfully
onto their reflecting positions .
The incident X-ray beam will diffract in many angle and fall on photographic film and some
rays will be transmitted .

Parallel to plane In a rotated angle

In case of any rotated angle of the crystal plane the diffraction of X-ray occurs and directed it
into photographic film and no change in case when plane is parallel to the incident X-ray .
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Each diffracted radiation produces a spot inside the photographic film present in camera .

Photographic film will be fixed perpendicular to the incident ray beam inside the cylindrical
camera .

There is 2 type of photography :- 1.Complete rotation

2.Oscillation method

Complete rotation :- series of complete revolution will takes place .

-Each plane in the crystal diffracts 4 times during rotation of 360º .

Oscillation Method :- Crystal is oscillated through an angle of 15º to 20º .

- The photographic plate is also moved accordingly .
- This method can be use to determine the size of unit cell in crystal25 .
Powder crystal analysis instrument

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POWDER CRYSTAL OR DEBYE METHOD
This technique is a rapid analytical technique used for identification of a crystalline material by
converting it into fine powdered particles .

Cylindrical camera

Spot formation on film

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Unwrapped film
Working :-
Powdered sample is taken in a Capillary tube inside the axis of the camera .
The transmitted beam will pass through a hole to minimize the fogging due to direct beam.
On the photographic film diffraction and reflection pattern will be recorded .
Powder particles having different orientation of plane and they will show different pattern of
cone shape whose interaction with photographic film takes place .
From the shape and distance from 2 similar pattern the identification can be done .
And the equation to get this is l= 2πr × Θ or θ = 360l
360º 4πr

Here θ = Incident angle , Reflection/diffraction =2 θ(∵ of taking 2 angle in that cone)

r = radius of the film , circumference= 2πr .
Corresponds to the scattering angle = 360º
l = Length of arc of a circle or length of any 2 similar pattern .

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APPLICATION
Used for the identification of unknown crystalline materials .
For characterization of crystalline materials .
Polymer characterization can be done .
Used to identify impurity .
Determine the unit cell dimensions .
Measurement of sample purity .
Used to carried out drug excipient incompatibility study .
Uses to analyze 3d crystal structure of biological materials .
The powder method is used to determine the value of the lattice parameters accurately .
Now a days often used to probe specific ways in how the structure of a material , drug will
interact in certain environments .

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1. REFERENCES
2. Critchley, Liam. (2020, October 16). The Applications & Principles of X-
Ray Crystallography.
3. AZoM. Retrieved on March 10, 2022 from
https://www.azom.com/article.aspx?ArticleID=18684.
4. https://youtu.be/HjKI4Kh1RgU
5. Critchley, Liam. 2020. The Applications & Principles of X-Ray
Crystallography. AZoM, viewed 10
6. March 2022, https://www.azom.com/article.aspx?ArticleID=18684.
7. Instrumental methods of analysis –Willards, 7 th ed . ISBN: 9788123909431
8. Modern analytical techniques in failure analysis of aerospace, chemical,
and oil and gas industries, ISBN 9780081001172

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THANK YOU

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