Norhasmillah Abu Hassan

CONTENT
Learning Objectives
Introduction
Basic Considerations
Methods
Comparison of Methods
Conclusion
Questions
Reference
Learning Objectives
At the end of this topic, student is expected to be able to:
• Differentiate essential and toxic mineral
• Describe the importance of mineral analysis for food
• Describe the methods for mineral analysis and how to select
the method
• List the advantages and disadvantages of the methods
Introduction
• Mineral is considered as micronutrient
• Mineral element exist in food due to:
i. Soil -> where plant has been grown
ii. Composition of the diet of animal from which food is produced
• Major element >100 ppm
• Trace element <100 ppm
• Some elements are essential & some are toxic
Introduction (cont.)
Ultra Trace Toxic element to
Macro Minerals Trace Minerals
Minerals body
Adult require Require in mili or
Require little
more than 100mg microgram per To be avoided
amount
per day day
calcium,
iron, iodine, zinc,
phosphorus,
copper, chromium,
sodium, vanadium, tin, mercury, lead,
manganese,
potassium, nickel, arsenic and cadmium and
molybdenum,
magnesium, boron aluminum
fluoride, selenium
chlorine, and
and silica
sulphur
 Note: Excessive amount of fluoride & selenium in dietary levels are harmful
Introduction (cont.)
Why Analysis of Minerals in Food?
• Implementation of NLEA – Nutrition Labelling & Education Act of 1990 lead
to analysis of minerals
• Minerals have their own function to our body:
i. Sodium: control hypertension
ii. Iron : preventing anaemia
• The content must be analysed/controlled

• Fortification of some foods are allowed

• Minerals are added for functionality : flavour & preservative
Introduction (cont.)
Methods for Mineral Analysis
• Traditional method
i. Titrimetric
ii. Colorimetric
iii. Ion selective electrodes

iv. Gravimetric titration Currently less use in industry

v. Redox reaction

• Modern method
i. Atomic Absorption Spectroscopy (AAS)
ii. Inductively Coupled Plasma - Atomic Emission Spectroscopy (ICP-AES)
Basic Considerations
• Nature of Analysis
• Sample Preparation
• Interferences
Basic Considerations (cont.)
Nature of Analysis
• Separation of minerals from food matrix:
• Specific procedures
i. Complexometric titrations
ii. Precipitation titrations
• Non specific procedures

• Non specific procedures require specific measurement such as

• Colorimetry
• Ion selective electrodes
• AAS
• ICP-AES
Basic Considerations (cont.)
Sample Preparation
• Why?
• To ensure well mixed & representative sample
• To make sample ready

• Problem?
• Contamination during sample preparation (grinding, contaminated
glassware, etc)
• Way to avoid contamination
• Use non metallic element
• Glassware cleaned with pure water
• Usage of reagent blank
Basic Considerations (cont.)
Interferences
• Interference factor :
• pH
• sample matrix
• temperature
• other analytical condition
• reagents

• Way to remove
• isolate sample mineral / remove interfering mineral
• use selective precipitations / ion exchange resins

• Standard curve
• develop when interferences are suspected
• represents the analysis to the sample mineral when analysing the food
Methods
1. EDTA Complexometric Titration
2. Precipitation Titration
3. Colorimetric Methods
4. Ion-Selective Electrodes
Methods (cont.)
EDTA Complexometric Titration
Principles
• EDTA – ethylenediaminetetraacetate forms stable complexes with
numerous mineral ions
• Stability increases with valence of ion
• Calmagite & Eriochrome Black T (EBT) are the indicators - changes colour
from pink to blue
• pH effect the complexometric EDTA titration
• 10 < pH titration < 12 for best result
• Ammonia buffer pH 10 is use for titration of magnesium and calcium
Methods (cont.)
• Procedure: Hardness of Water Using EDTA Titration
• Calmagite is the best indicator with addition of buffer solution + neutral
magnesium salt
• Calcium in the sample replaces the magnesium bound to EDTA
• Free magnesium binds to Calmagite
• Pink magnesium-Calmagite persists until all calcium has been titrated with
EDTA
• Excess EDTA removes magnesium from Calmagite  produces blue
endpoint
Methods (cont.)
• Applications of EDTA
• Major application – testing calcium plus magnesium as indicator
for water hardness
• Suitable for determining calcium in ash of fruits and vegetables
• Test strips impregnated using Calmagite and EDTA is available for
easy application  dipped into water to test for hardness
Methods (cont.)
Precipitation Titration
Principles
• Titration reaction is insoluble precipitate
• Mohr method  direct / forward titration
• formation of orange-coloured solid
• Volhard method  indirect / back titration method
• excess of standard solution of silver nitrate is added to chloride
containing sample solution
• excess silver then back-titrated using standardize solution of
potassium or ammonium thiocyanate with ferric ion as indicator
• Form dark red colour solution at the end point
• Calculation – subtract the excess silver from the original silver content
Methods (cont.)
Precipitation Titration (cont.)
Procedures
• Mohr Titration of Salt in Butter
• Titrating chloride ion with silver
• Orange endpoint occurs when all chloride ion is complexed
• Boiled water is use in reagent preparation to avoid interferences from
carbonates in water
• Volhard Titration of Chloride in Plant Material
• Water must be boiled to avoid interferences from carbonates
• Solubility product of silver carbonate is much less than silver chloride
• Result  Chloride weight must multiplied by 1.648 to obtain salt weight
Methods (cont.)
Calculation for moisture content, ash content and salt
content
• Moisture content in percentage;
𝑚𝑎𝑠𝑠 𝑜𝑓 𝑤𝑒𝑡 𝑠𝑎𝑚𝑝𝑙𝑒 − 𝑚𝑎𝑠𝑠 𝑜𝑓 𝑑𝑟𝑦 𝑠𝑎𝑚𝑝𝑙𝑒
× 100
𝑚𝑎𝑠𝑠 𝑜𝑓 𝑤𝑒𝑡 𝑠𝑎𝑚𝑝𝑙𝑒

• Ash content in percentage;

𝑚𝑎𝑠𝑠 𝑜𝑓 𝑎𝑠ℎ𝑒𝑑 𝑠𝑎𝑚𝑝𝑙𝑒
× 100
𝑚𝑎𝑠𝑠 𝑜𝑓 𝑑𝑟𝑦 𝑠𝑎𝑚𝑝𝑙𝑒

• Salt (Cl) content of the original sample;

𝑚𝑜𝑙 𝑜𝑓 𝐴𝑔 𝑎𝑑𝑑𝑒𝑑 = 𝑚𝑜𝑙 𝑜𝑓 𝐶𝑙 𝑖𝑛 𝑠𝑎𝑚𝑝𝑙𝑒 + 𝑚𝑜𝑙 𝑆𝐶𝑁 𝑎𝑑𝑑𝑒𝑑
Methods (cont.)
• Applications
• Suitable for foods high in chlorides
• To be considered for chloride detection : for added salt in processed food
i.e cheeses and meats
• Automatic titration system commonly use to measure salt content of
potato chips (Mohr Titration)
• Test strip adaptation allows rapid quantification of salt in food product &
accurate to ±10% over a range of 0.3 – 10% NaCl in food products
Methods (cont.)
Colorimetric Methods
• Principles
• Chromogens  form coloured product when reacted with compound of
interest
• The product will be quantified by absorption of light at specific wavelength
 Beer’s Law ; linear relationship between concentration and absorption
• Concentration of mineral is determined from standard curve during
analysis
• Samples must be treated to avoid minerals from organic complexes inhibit
their reactivity with chromogen
Methods (cont.)
• Procedures: Colorimetric Determination of Iron in Meat
• Iron content of foods can be quantified spectrophotometrically
• Absorption of light at 562nm is converted to iron concentration in the
sample via regression equation generated from a standard curve
developed during analysis
• In iron; the method coupled with method heme iron to determine ratio of
heme iron to total iron
• Ferrozine reagent only react with ferrous ion not ferric
• Ascorbic acid is added to convert ionic iron to detectable ferrous form
Methods (cont.)
• Applications
• For detection and quantification of variety
minerals in food
• A viable method to AAS and other detection
methods
• Very specific & can be performed in the
presence of other minerals
• Robust & often immune to matrix effects
• Minimal effort & expenses but precise &
accurate similar to result obtained by
experienced personnel using AAS
Methods (cont.)
Ion-Selective Electrodes (ISE)
• Principles
• pH electrode is one of the example of ISE
• Ion-selective sensor is placed as bridging electrode between two reference
electrodes to produce constant & reproducible potential
• Inside surface of ion-selective sensor is in contact with negative reference
electrode (anode) via a filling solution having a constant concentration of
sample ion
• Outside surface of ion-selective sensor is contact with positive ref
electrode (cathode) via sample solution having concentration of sample
ion
Methods (cont.)
• General Methodology
• sample ion attached to pH meter set on mV scale
• consider the ISE performance when selecting electrode, design sampling and
analysis protocols
• for ISE analysis, electrode performance is effected by interfering ions
• activity coefficient be kept constant in samples and a calibration standard
Methods (cont.)
• ionic strength adjustment (ISA) buffers used to adjust samples and standard to
the same ionic strength
• electrode and solution must be kept at constant temperature and allow
sufficient time for all samples and standard to equilibrate
• allow sufficient time for electrode to stabilize before taking a reading
Methods (cont.)
• Electrode Calibration and Determination of Concentration
• Calibration curve
• allows large number of samples to be measured rapidly
• a series of solutions of known concentration plotted on semilog paper
against standard concentration

Example of ISE calibration curves

Methods (cont.)
Standard addition method
• limited samples analysis
• does not permit development of calibration curve
• eliminates complex and unknown background effects that cannot be
replicated when developing a calibration curve using standards
• concentration of active species in the original sample is calculated as
CO = C∆
(10 ∆E/S – 1)

CO : original concentration of sample ion (mol/L)

C∆ : change in sample ion concentration when spike was added (mol/L)
E : difference in potential between the two readings (V)
S : 0.059 / number of electrons in the ½ reaction of the sample ion
Methods (cont.)
• Endpoints titrations
• using species that form a precipitate or strong complex with the sample
ion
• ISE detects titrant species  T –type titration curves results from the
large increase in titrant activity detected at equivalence point
• ISE detects sample ion  S-type titration curves results from removal of
sample ion activity at equivalence point
• Sample concentration is calculated from titrant volume to reach
equivalence point and stoichiometric relationship between titrant species
and sample ion
Example of T-type titration curve
Methods (cont.)
• Application
• Example application of ISE:
• salt & nitrate in processed meat
• salt content of butter and cheese
• calcium in milk
• sodium in low-sodium ice cream
• carbon dioxide in soft drinks
• potassium and sodium levels in wine
• nitrate in canned vegetables

• Versatile measuring tool

• Major advantage of ISE

• able to measure many anions and cations directly
• analyses are independent of sample volume when making direct
measurement
Methods (cont.)
• Major disadvantage of ISE
• inability to measure below 2-3ppm
 electrode response time is slow at low level of
measurement
 some electrodes have had high rate of premature failure /
possible excessive noise characteristics
Comparison of Methods
• Factor effecting the choice of methods for mineral analysis
• Performance regarding accuracy
• Sensitivity
• Detection limit
• Specificity
• Interferences

• Other factor
• Cost per analysis
• Equipment availability
• Analytical time compared to analytical volume
Conclusion
What have we learnt from this topic?
• Compare macro mineral, trace mineral, ultra trace mineral, toxic mineral
• Importance of analysis; nutritional value, toxicological potential, labelling
• Basic consideration; nature analysis, sample preparation and interference
• Methods ; EDTA Complexometric Titration, Precipitation Titration,
Colorimetric Methods, Ion-Selective Electrodes
• Factors of selecting a method
Reference
• Nielsen S.S (2010). Food Analysis. 4th edition. Springer
Science

Useful video
• https://www.youtube.com/watch?v=ORmO23Ui5E4