Report on

Synthesis and Analysis of Superparamagnetic Iron Oxide

Nanoparticles (SPIONs)
By
Mudabar Ahmad Mubashar #1329975
Muhammad Qamaruddin #1404155
Students of
Master of Nano Science and Nanotechnology
Universität Siegen
Under kind supervision of
Dr. Johannes Weber
Submitted to
Prof. Dr. Jörn Schmedt auf der Günne
Dr. Johannes Weber
Table of Contents
1. Introduction ......................................................................................... 1
1.1 Iron Oxide Fe3O4 ....................................................................... 1
1.2 Sol-Gel Technique..................................................................... 1
1.3 XRD Powder Diffraction .......................................................... 4
1.4 Fundamental principal of XRD ................................................. 4
1.5 Bragg law and Diffraction ..........................................................5
1.6 Strengths and Limitations of X-ray Powder Diffraction (XRD)7
1.7 Crystallography and Scherrer Equation ..................................... 8

2. Experimental Setup and Synthesis ....................................................... 9

2.1 Chemicals ................................................................................... 9
2.2 Synthesis of Fe3O4 nanoparticles ................................................9

3. Results and discussion ....................................................................... 13

3.1 Determination of Nanoparticles size ....................................... 15

3.2 Lattice Constant ........................................................................16

4. Conclusion .......................................................................................... 17

5. Reference… ......................................................................................... 17
List of Figures

Figure 1 Schematic representation of Sol-Gel method ........................ 2

Figure 2 Working principle of X-Ray Powder Diffraction ............... 5

Figure 3 Experimental setup .............................................................. 9

Figure 4 Nitrogen flow setup ......................................................... 10

Figure 5 Solution of Iron oxide nanoparticles ................................. 11

Figure 6 (a) Concentrated Iron oxide NP (b) Dried Iron oxide NP . 12

Figure 7 Sample holder for XRD with reference and sample ......... 12

Figure 8 Phosphor imaged results ................................................... 13

Figure 9 X-Ray Diffraction Pattern graph ....................................... 14

List of Tables

Table 1 Particle sizes measurement table ....................................... 15

Table 2 Table of lattice constant at different plane ......................... 16

Chapter 1. Introduction.
1.1 . Fe3O4
Fe3O4 (magnetite) named iron (II, III) oxide is a chemical compound which
is one of most widely used superparamagnetic iron oxide nanoparticles(SPIONs).
Magnetic nanoparticles are a type of particles which we can manipulate using a
magnetic field. Such particles mostly contain magnetic elements such as Iron,
Nickel and Cobalt and their compounds. Superparamagnetic is a type of
magnetism which appears in small ferromagnetic or ferrimagnetic nanoparticles,
which shows paramagnetic behavior above Curie temperature. When there is no
external magnetic field then the magnetic properties of particles appear to be zero,
but upon external magnetic field, they show magnetization like paramagnet.
However, the magnetic susceptibility of these particles is much higher than
paramagnets. SPIONs have many attractive applications like to be used in
biomedicine, magnetic resonance imaging, data storage etc. [1] Nanoparticles can
be defined as particles having at least one dimension less than 1 micron are
Nanoparticles. Fe3O4 exists in nature as a mineral magnetite, hematite is its other
name. It contains both Fe2+ and Fe3+ ions. It is extensively used as a black pigment
which is manufactured rather than being produced from naturally occurring
mineral as the shape and particle size can vary by the method of its production.
Fe3O4 is an electrical conductor with a conductivity significantly higher (x 106)
than Fe2O3, and this is ascribed to electron exchange between the Fe II and FeIII
centers.[2]

1.2 . Sol-Gel Technique

The sol-gel processing techniques have also been extensively used for
synthesis of metal oxide nanoparticles. This method has good control over
textural and surface properties of the materials. Sol gel method undergoes few
major steps to synthesis desired nanoparticles which are hydrolysis, condensation
and calcination and drying process. The synthesis of metal oxide starts by
hydrolysis of metal precursor to produce metal hydroxide solution and them
1
immediate condensation leads to the formation of three dimentional gel. The gel
is the allowed to dry, depending on the mode of drying the gel is then converted
into Xerogel or Aerogel. A Sol is a colloidal or molecular suspension of solid
particles of ions in a solvent. A gel is semi-rigid mass that forms when the solvent
from the sol starts evaporation and the ions left behind starts to join together in
continuous network. The precursor for synthesizing these colloids consists of ions
of metal alkoxides. The most widely used are tetramethoxysilane (TMOS), and
tetraethoxysilane (TEOS) or inorganic salts such as sodium silicate (Na 2Sio3) in
the presence of mineral acid (e.g HCl) or base (e.g NH3) as catalyst and to control
pH. [3]
a. Hydrolysis
b. Condensation
c. Growth of particles

Figure 1. Schematic representation of the Sol-Gel method [5]

2
a. Hydrolysis
During hydrolysis, the addition of water results in the replacement of [OR]
group with [OH-] group. Hydrolysis occurs by an attack of oxygen on silicon
atoms in silica gel. Hydrolysis can be accelerated by adding a catalyst such as
HCl and NH3. Hydrolysis continues until all alkoxy groups are replaced by
hydroxyl groups. Hydrolysis occurs by an attack of oxygen contained in the water
on the silicon atom. The general reaction equation is shown below.

M-O-R + H2O → M-OH+ R-OH (Hydrolysis)

b. Condensation
Polymerization to form M-O-M bonds occurs by either a water producing or
alcohol producing condensation reaction. The end result of condensation
products is the formation of monomer, dimer, cyclic tetramer, and high order
rings. The rate of hydrolysis is affected by pH, reagent concentration and H2O/Si
molar ratio (in case of silica gels). Also, aging and drying are important. By
control of these factors, it is possible to vary the structure and properties of sol-
gel derived inorganic networks.
M-OH + HO-M → M-O-M + H2O (water condensation)
M-O-R + HO-M → M-O-M +R-OH (alcohol condensation)

c. Growth and Agglomeration

As the number of M-O-M bonds increases, the molecules aggregate in the
solution, where they form a network, a gel is formed upon drying. The water and
alcohol are driven off and the network shrinks. At values of pH of greater than 7,
and H2O/Si value ranging from 7 to 5. Nanoparticles are formed above a pH of 7,
Growth stops when the difference in solubility between the smallest and largest
particles becomes indistinguishable. Larger particles are formed at higher
temperatures. Zirconium and Yttrium gels can be similarly produced. [4]

3
1.3 . What is X-ray Powder Diffraction (XRD)?

X-ray powder diffraction (XRD) is the rapid analytical technique which

primarily used for identification of phase of the crystalline material. It’s provide
the information of unit cell parameter as well as positions of atoms. The analyzed
material is finely ground, homogenized and average bulk composition is
determined.
1.4 . Fundamental Principles of X-ray Powder Diffraction (XRD)
In 1912, Max von Laue discovered that crystalline substances behaved as
3D (three-dimensional) diffraction gratings for X-ray wavelengths as the spacing
of planes in the crystal lattice. Now, for the study of atomic spacing and
crystalline structures, X-ray diffraction is the ordinary technique.

It is based on constructive interference of monochromatic X-rays on a crystalline

sample. With the help of cathode ray tube, these X-rays are produced, filtered to
generate monochromatic radiations, collimated to concentrate and then directed
towards the sample. If the condition i.e. Bragg's Law (nλ=2d sin θ) is satisfied,
then constructive interference (and diffracted ray) is produced when these
incident rays hit the sample. This law shows the relation of the wavelength of
electromagnetic radiation with an angle of diffraction and lattice spacing in a
crystalline sample. Then these diffracted X-rays are detected, processed and
counted. Through the range of 2θ angles, all possible diffraction directions of the
lattice should be attained due to the random orientation of the powdered material.
The obtained diffractogram pattern can be analyzed and compared with standard
reference patterns.

X- ray powder diffraction method is based on the generation of X-rays. These

X-rays are directed at the sample, and the diffracted rays are collected. A key
component of all diffraction is the angle between the incident and diffracted rays.
Powder and single crystal diffraction vary in instrumentation beyond this. [6]
4
Figure 2 shows the component of X-ray powder diffraction.

Figure 2. Working principle of X-Ray Powder Diffraction [7]

1.5 . Bragg’s Law and Diffraction

We can explain Bragg’s Law with the following equation:

nλ = 2dsinθ

Where n is an integer, variable λ is the wavelength of the incident beam, d is

the spacing between net layers of crystal and θ is the angle of incident.

Only when the Bragg’s Law is valid then diffraction occurs. In 1915 a noble
prize was awarded to Bragg for determining crystal structure ZnS, NaCl, and
diamond.

5
How Does Diffraction work?

►Wave interaction with a single particle

● Incident beams uniformly scattered in all directions.

► Wave interacting with solid

● In some directions, scattered beams interfere constructively to produce

diffracted beams.

► Crystalline Material

● Regular diffraction pattern produced by a regular pattern of

crystalline atoms.

● This diffraction pattern gives information about crystal structure. [6]

Why we use Powder X-Ray Diffraction?

We use Powder XRD due to the following:

i. It can measure the average spacing between the rows and layers of
net planes.
ii. It can find the crystal structure of unknown materials.
iii. It can determine the orientation of single grain or crystal.
iv. It can measure the size, internal stress and the shape of a small region
of crystals.
v. It can measure the thickness of multilayers and thin films.
vi. It can also find either the compound is solvated of not.
vii. It is used to access the degradation of natural and synthetic minerals.

6
1.6 . Strengths and Limitations of X-ray Powder Diffraction
(XRD)

➢ Strengths

• It is powerful and rapid (< 20 min) technique for identification of an

unknown mineral

• In most cases, it provides an unambiguous mineral determination

• Minimal sample preparation is required

• XRD units are widely available

• Data interpretation is relatively straight forward

➢ Limitations

• The homogeneous and single-phase material is best for identification of an

unknown

• Must have access to a standard reference file of inorganic compounds (d-

spacings, hkl)
• For mixed materials, the detection limit is ~ 2% of the sample

• For unit cell determinations, indexing of patterns for non-isometric crystal

systems is complicated

• Peak overlay may occur and worsens for high angle 'reflections'. [6]

7
1.7 . Crystallography and Scherrer Equation

Crystallography is the branch of science which describes the bonding

arrangement of atoms in crystalline solids and geometric structure of crystal
lattices, crystallography is mostly based on the analysis of the diffraction of X-
rays.

The arrangement of surface or a crystal plane may be described that how

the plane intersects the main crystallographic axes of solid. Miller indices (h,
k, l) are a crystallographic notation of planes determined by integers h, k and l.
Using lattice parameters, d-spacing and peak position of XRD.

The lattice constant for cubic crystal can be found by this equation.
𝑎0
𝑛𝜆 = 2 𝑠𝑖𝑛 𝜃ℎ𝑘𝑙
√ℎ +𝑘 2 +𝑙 2
2

In X-ray diffraction and crystallography, Scherrer formula is used to

determine the size of sub-micrometer particles or crystallites in a solid peak by
a diffraction pattern. This formula is used for determination of the size of
particles of crystals in the form of powder [8].

𝑙𝑛2 𝜆 1
𝐿 = 2√ · ·
𝜋 𝐹𝑊𝐻𝑀 cos⁡(𝜃)

8
Chapter 2.
Experimental Setup and Synthesis.
2.1 Chemicals

Ferrous Chloride FeCl2, Ferric chloride hexahydrate FeCl3 6H2O, Sulfuric acid
H2SO4, Ammonia solution NH4OH (w%= 25%), Oleic acid C18H34O2, Decane.

2.2 Synthesis of Fe3O4 nanoparticles

In our experiment, we did a synthesis of superparamagnetic iron oxide

nanoparticles by the Sol-Gel method. This is the simplest chemical method for
synthesis of the iron oxide. The experimental setup consists on sealed precision
glass (KPG) stirrer, gas supply and gas inlet, three necks round bottom flask,
thermometer, reflux condenser, oil bath and hot plate and nitrogen flow system,
some clamps and metal rods and some plastic pipes for gas and water flow. The
KPG stirrer was used for uniform mixing of the solutions, it consists of an
electric motor with the control unit to provide continuous controlled
stirring.The experimental setup and adjustment of apparatus are shown in
Figure 3.

Figure 3: Experimental setup

9
 To start the Sol-Gel synthesis process firstly 100 ml of demineralized
water was degassed for 15 minutes in 500 ml three-neck flask with water
pump and was flushed for several minutes with nitrogen gas. The whole
process was done during continuous N2 gas flow to prevent oxidation of Fe3O4
into Fe2O3. Figure 4 demonstrates the visual arrangement of the experimental
setup as well as the N2 gas flow process.

Figure 4. Nitrogen flow setup

Then 2.74 g FeCl2 and 11.66 g FeCl3 6 H2O were dissolved in the flask and
the mixture could heat up to 85 oC under continuous stirring. When the
required temperature was reached then to maintain the pH level, 13.5 ml of
ammonia solution (w%=2.5%) was added quickly (because any delay of
solution addition can also effect on particle size). The chemical reaction of
Fe3O4 formation is shown below.

2 FeCl3 + FeCl2 + 8 NH3 + 4 H2O → Fe3O4 + 8 NH4Cl

When the color changes to black 5 ml oleic acid was added to prevent
agglomeration of particles. Afterward, the heating source was removed and
allow to cool down. Figure 5 shows the overview of the solution while cooling
down.
10
 Figure 5. Solution of Iron oxide nanoparticles

After cooling down the solution, by using permanent magnet

precipitates were separated from the aqueous phase and was filled in a
separate beaker. This gel-like precipitate was washed two times with cold
water and two times with hot water and then allowed to dry overnight in the
oven.

Next day we took our dried sample and we added Decane dropwise in
0.5 g of particles in sample vial with stirring using a permanent magnet at the
bottom of beaker until the particles completely soaked up the solvent. When
the mixture started showing viscous behavior then the addition of decane was
stopped. Figure 6 demonstrates our precipitate before and after drying.

11
 (a) (b)
Figure 6. Precipitate Fe3O4 (a)wet (b) dried

we used 0.5 g sample for characterization in powder X-ray diffraction. For X-

ray diffraction, first, we placed our sample very carefully on sample holder we used
mylar foil and grease to stick our nanoparticle on the surface of the holder. The sample
of SiO2 was used as a reference sample. The overview of the sample holder is shown
in Figure 7.

Figure 7. Sample holder for XRD with reference and sample

12
 After sample preparation, we put our sample holder in X-Ray diffractor
as shown in above picture. Then we put the camera with film inside at its
place and after closing all gates carefully by making sure from the green light.
We started increasing current and voltage step by step with an interval of 10
seconds as mentioned on the chart attached on apparatus. Shutter was open
after the tube voltage and current was increased (to 45 kV and 35 mA
respectively). By seeing the red light we make sure the diffractometer was
turned ON successfully. Similarly, after measurement, we also turned our
apparatus in standby mode by going down step by step, each step with an
interval of 10 seconds. Then we removed the film in a completely dark room
by turning OFF all the light producing devices and put the film in Phosphor
imaging scanner. The scanned sample result are shown in Figure 8 (upper two
blackish bars are for Fe3O4 and the lower one was of silicon dioxide) were
then used to generate diffraction pattern which was compared with a database
to find our material.

Figure 8. Phosphor imaged results

Chapter 3.
Results and discussion.
To find out the size of our nanoparticles we did a characterization of them
by using X-Ray diffraction technique. After experimental measurements,

13
calculations are needed to be done to find our results. The calculations are given
below. The wavelength of (our apparatus) monochromatic X-ray is 1.5418 Å. The
obtained results with at 35mA and 45kV for 15 minutes were used to compare
with the standard database to find out our material by our diffraction pattern peaks
with closely and best-matched database diffraction peaks. Which was matched
with the standard Fe3O4 pattern. Our X-Ray diffraction pattern is shown in Figure
9.

Figure 9. X-Ray Diffraction Pattern graph

Then in our obtained X-ray diffraction graph pattern we find out and named our
diffraction peaks of (220), (311), (400), (511) and (440).

14
 3.1. Determination of Nanoparticles size
To determine the size of the iron oxide nanoparticles we used Scherrer
equation, where 𝝀 is the incident X-ray wavelength (1.5418 Å), and Full Width
Half Maxima (FWHM) is the peak width at its half intensity in radians.

𝑙𝑛2 𝜆 1
𝐿 = 2√ · ·
𝜋 𝐹𝑊𝐻𝑀 cos⁡(𝜃)

𝑙𝑛2 1.54 · 10−10 1

𝐿220 =2 √ · · = 19.03⁡𝑛𝑚
𝜋 7.68 · 10−3 cos⁡(15°)

Similarly, for we calculated other values and plotted a table 1.

Plane 2𝜽 (°) FWHM (°) FWHM NP size (nm)

(hkl) (radians)

(220) 30.1 0.44 7.68x10-3 19.03

(311) 35.3 0.51 8.90x10-3 16.47
(400) 42.9 0.4 6.98x10-3 21.11
(511) 56.9 0.71 0.01257 12.05
(440) 62.5 0.79 0.01177 12.3

Table 1. Particle sizes measurement table

Thus, the average size of our synthesized Magnetite Fe3O4 nanoparticles is

16.19 nm.

15
 3.2. Lattice Constant.

We can find the lattice constant (a0) by the given formula,

𝑎0
𝑛𝜆 = 2 𝑠𝑖𝑛 𝜃ℎ𝑘𝑙
√ℎ2 + 𝑘 2 + 𝑙 2

Where n=1 and 𝜆=1.5418 Å and the values from Table 1 as well as figure 10 we
put in the above formula.

Plane (hkl) Incidence angle 𝛉𝐡𝐤𝐥 (°) Lattice constant (𝐚𝟎)

Å

(220) 15.05 8.36

(311) 17.65 8.42
(400) 21.45 8.42
(511) 28.45 8.40
(440) 31.25 8.38

Table 2. Table of lattice constant at different plane

Hence, the lattice constant of the Fe3O4 nanoparticle is 8.39 Å.

Fe3O4 has a face-centered cubic structure which is based on O2- ions and are
closely packed along 111 directions. Fe3O4 has a cubic inverse spinel structure
which consists on a closed packed array of oxide ions in which half of the
octahedral sites are occupied by Fe2+ ions and Fe3+ are placed on remaining evenly
split octahedral and tetrahedral sites.
16
 Chapter 4. Conclusion.

SPIONs have useful application specially in medical for drug delivery. We

prepared Fe3O4 magnetite by using Sol-Gel method approach, which is an effective
method used for synthesis of nanoparticles. The size, morphology, magnetic
properties are controllable by varying pH, ionic strength, temperature, reaction time
and by type of salts used. After successful synthesis of Fe3O4 nanoparticles we
determined its magnetic properties by using permanent magnet. After drying and
grounding we used powder X-Ray Diffraction technique for characterization. The
comparison with standard database of our diffraction peaks confirmed the material
Fe3O4. We also calculated and identify FWHM and crystal planes and from them by
using Scherrer equation we found the average particle diameter which was around
16 nm. The lattice parameters found using Bragg´s law equation was 0.839 nm with
face-centered cubic crystal structure.

5. Reference.

[1] Y. Lu, Y. Yin, B. T. Mayers, and Y. Xia, “Modifying the Surface Properties
of Superparamagnetic Iron Oxide Nanoparticles through A Sol−Gel Approach,”
Nano Lett., vol. 2, no. 3, pp. 183–186, Mar. 2002.
[2] X.-M. Li, G. Xu, Y. Liu, and T. He, “Magnetic Fe3O4 Nanoparticles: Synthesis
and Application in Water Treatment,” p. 11.
[3] B. G. Rao, D. Mukherjee, and B. M. Reddy, “Chapter 1 - Novel approaches
for preparation of nanoparticles,” in Nanostructures for Novel Therapy, D. Ficai and
A. M. Grumezescu, Eds. Elsevier, 2017, pp. 1–36.
[4] E. Kemnitz and J. Noack, “The non-aqueous fluorolytic sol–gel synthesis of
nanoscaled metal fluorides,” Dalton Trans., vol. 44, no. 45, pp. 19411–19431, 2015.
[5] L. L. Hench and J. K. West, “The sol-gel process,” Chem. Rev., vol. 90, no. 1,
17
pp. 33–72, Jan. 1990.
 [6] A. G. Marangoni, M. F. Peyronel,University of Guelph, “X-Ray Powder
Diffractometry,” AOCS, Mar. 2013.
[7] Lodovico, “XRD su Polveri,” PhysicsOpenLab, 24-Jul-2018.

[8] P. Scherrer, Nachrichten Von Ges. Wiss. Zu Gott. Math.-Phys. Kl. 1918,
1918, 98–100.

18
19