FST 606

EXPERIMENT 1: DETERMINATION OF SOLID FAT CONTENT USING THE

PULSED – NUCLEAR RESONANCE (NMR) SPECTROMETER

Prepared by : ERNIE NAJWA NAJIHAH BINTI FAIDI

Student ID : 2017283562

Group members :

1. NURUL FATINI BINTI MOHAMAD HALIMIN

(2017405588)
2. SITI NADIA BINTI JUMPAAN (2017405658)
3. NUR NABILAH BINTI JOHARI (2017405596)

Group : AS2465Q

Prepared for : DR MARYAM BINTI HUSIN

Date of experiment : 13TH MARCH 2019

 Experiment 1: Determination of Solid Fat Content Using the Pulsed – Nuclear Resonance
(NMR) Spectrometer

INTRODUCTION

According to Fiedler, Mallin, & Vesci (2011), Nuclear Magnetic Resonance (NMR)
was observed by Edward Purcell and Felix Bloch, independently, in 1946 when investigating
the behavior of nuclei once placed in an external, uniform magnetic field and subject to a radio
frequency (RF) magnetic field oriented perpendicular to the external field.

OBJECTIVES

1. To understand the principle of pulsed nuclear magnetic resonance

2. To determine the solid fat content of difference food samples (margarine, butter,
shortening) at different set of temperature.

MATERIALS

Pulsed – NMR (minispec MQ20, Bruker, Germany)

Sample tubes (180 mm x 10 mm)

Water bath (different temperature setting)

SFC standards (0%, 30.6% and 72.7% SFC)

Droppers

Stopwatch

Food sample: shortening, margarine, butter

METHOD

1. Standard Operating Procedure For Pulsed NMR

The power for the magnet unit and PC was switched on before pressing ‘1’nat
the back of the electronic controller. The PC was switched on and the ‘Bruker The
 Minispec’ was clicked and the PC was waited to connecting the magnet unit. Next, the
LED on both magnet and electronic unit were observed.
Electric Control Unit

Right LED, Connection status indicator:

ON (green): PC communication to Electronic Control Unit OK

OFF : No communication of the Electronic Control Unit to PC.

Left LED, Status indicator:

ON (green): Electronic Control Unit is working

OFF : Electronic Control Unit not working

Magnet Unit

Right LED, indicates magnet temperature status:

RED: preheating (fast mode)

YELLOW: Transition to normal heating (air heat mode)

GREEN: Temperature OK

Left LED, connection status indicator:

ON (green): Electronic Control Unit to Magnet Unit OK

OFF: No communication between Electronic Control Unit and Magnetic Unit.

The list of recent application was appeared, the top application was the latest, an application
can be chosen from the list or click ‘setup’ and ‘browser’. The ‘Application Pool V5.2 Ratio’
was opened. Then customize was clicked, making sure the SFC analyser and Calibration
permission were checked. OK was clicked again to proceed. After a while, the PC tried to
communicate with the Minispec. The result appeared on the PC when the right LED on the
control PC turned green. The temperature of the magnet unit has been making sure to stabilize
by observing the right LED or clicking the instrument status. The daily check was performed.
2. Sample Preparation
The sample used in this experiment was prepared a day earlier than the actual
experiment. About 10 g fat samples were melted on a hot plate until the fats were clear
with no suspended solids. It was then transferred into a glass NMR tubes while it was
still hot, filling the tubes to a height of 4 cm. The tubes was then capped. No fats was
ensure adhering to the outside of the tubes. Next, the samples were transferred into
water bath at 60°C for 10 min and then at 0°C for 90 minutes interval. The instrument
was then calibrated by measuring three standard sample of 0%, 30.6% and 72.7% solid
respectively. After the holding time ended, the sample was next transferred into 10°C
water bath for 30 minutes. After that, the samples were quickly removed from the water
bath before letting it to dry and transferred to the measurement port on the pulsed –
NMR. The samples was then transferred into 20°C water bath and was let for 30
minutes. The measurement was then repeated for 30°C, 40°C, 50°C and 60°. All the
SFC result were obtained and printed for analyse.
Figure 1.1 shows the flowchart of the method in application of pulsed – NMR.
Figure 1.2 shows the sample preparation steps to determine the Solid Fat Content.
RESULTS

Table 1.1 shows the data obtained by the pulsed – NMR spectrometer.
 Table 1.2 shows the SFC obtained from the data analysis.

Sample Temperature Solid Fat Content (% SFC) Average ± S.D

(°C)
Tube 1 Tube 2 Tube 3

Margarine 10 46.526 47.982 45.429 46.646 ± 1.28

20 18.446 18.695 19.043 18.728 ± 0.30

30 8.52 8.441 8.615 8.525 ± 0.09

40 1.861 2.397 2.291 2.183 ± 0.28

50 0.427 0.134 -0.435 0.042 ± 0.44

60 0.145 -0.053 0.243 0.112 ± 0.15

Butter 10 46.573 46.201 45.39 46.055 ± 0.60

20 24.176 24.221 24.337 24.245 ± 0.08

30 10.853 10.742 11.312 10.969 ± 0.30

40 3.567 4.104 4.127 3.933 ± 0.32

50 0.055 -0.005 -0.041 0.003 ± 0.05

60 0.017 0.146 0.085 0.083 ± 0.06

Shortening 10 72.669 72.962 73.496 73.042 ± 0.42

20 61.138 61.123 61.448 61.236 ± 0.18

30 32.819 32.439 33.082 32.78 ± 0.32

 40 17.159 17.028 17.094 17.094 ± 0.07

50 7.673 8.391 8.667 8.244 ± 0.51

60 -0.038 -0.069 0.09 -0.006 ± 0.08

Figure 1.3 shows the graph of Solid Fat Content versus Temperature

Solid Fat Content VS Temperature

80

70

60
solid Fat Content (%)

50

40

30

20

10

0
10 20 30 40 50 60
-10
Temperature (°C)

Margarine Butter Shortening

CALCULATION

𝑠𝑢𝑚 𝑜𝑓 𝑡𝑟𝑖𝑝𝑙𝑖𝑐𝑎𝑡𝑒 𝑝𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒

Average percentage solid fat content (SFC) =
3

For margarine:

46.526 + 47.982+ 45.429

10°C =
3

= 46.646
 ∑(𝑥1− 𝑥)2
Standard Deviation (S.D) =√
2

For margarine:

∑(𝑥1− 𝑥)2
10°C = √
2

∑(46.526− 46.646)2+(47.982−46.646)2+(45.429−46.646)2
=√
2

= 1.28

DISCUSSION

Nuclear Magnetic Resonance spectroscopy is perhaps one of the most important

scientific development since its first discover in the mid 1940’s. NMR has found a wide range
of applications from solid state physics and materials science to chemical analysis and
biophysics and also in food industries. NMR imaging has begun to attract the attention of food
scientists since the distribution of water within an object can be measured non – invasively and
changes with processing or storage can be followed (Colquhoun, 1993). NMR is divided into
two main types which are Proton NMR and also Carbon NMR. Proton NMR is specialized in
detecting only the hydrogen atom within the molecule meanwhile the carbon NMR is used in
detecting the number and types of carbon atoms within the molecules.

According to Colquhoun (1993), the solid fat content (SFC) and its temperature
dependence are important factors governing the organoleptic properties of fat- based products.
Although fat is an essential component in the human diet the excessive consumption of food
with high fat content has been the major cause of obesity, one of the most serious human health
problems in the modern developed world. Therefore, the fast precise determination of the fat
content in raw and processed foods is an important analytical task (Manhas et al., 2015).

Based on the graph obtained, shortening has the highest Solid Fat Content at the
beginning of the experiment at 10°C followed by butter and margarine. The greater the heat
applied to the fat, the lower the percentage of SFC of the substance. This is due to the
breakdown of the molecule bonding occurred during the heat treatment of the sample.
However, all samples reach almost 0% of SFC at 60°C.

There are some limitation due to the usage of the NMR spectrometer. First and
foremost, the samples desired has to be sure to be a pure compound unless it will altered the
result obtained and lead to errors in the experiment. Thus, the heating of the samples before
running the test is crucial to get only the fat composition of the sample.

CONCLUSIONS

This experiment was carried out to understand the principle applied to Pulsed Nuclear
Magnetic Resonance (NMR) and to determine the Solid Fat Content (SFC) of three different
food sample which are Margarine, Butter and also Shortening. Shortening has the most SFC
content among the sample and SFC is decreasing when heat treatment is applied to the sample.
The objectives of the experiment were successfully achieved.

REFERENCES

Colquhoun, I. J. (1993). NMR Spectroscopy in Food Science.

Fiedler, C., Mallin, D., & Vesci, A. (2011). Pulsed Nuclear Magnetic Resonance to Obtain
Characteristic Times for Mineral Oil and Gycerin, (March), 1–3.

Manhas, F., Pereira, V., Paula, A., Azevedo, J., Pallone, L., & Alberto, L. (2015). Through-
package fat determination in commercial samples of mayonnaise and salad dressing
using time-domain nuclear magnetic resonance spectroscopy and chemometrics. Food
Control, 48, 62–66. https://doi.org/10.1016/j.foodcont.2014.02.028