Asian J. Research Chem.

4(3): March 2011

ISSN 0974-4169 [Link]

REVIEW ARTICLE

Iron Determination - A Review of Analytical Methods

Bhavna V. Mohite
Department of Biotechnology, Moolaji Jaitha College, Jalgaon - 425001, (MS), India
*Corresponding Author E-mail: mohite_bhavna@[Link], bhavna_v_mohite@[Link]

ABSTRACT:
In nature understanding of biogeochemical processes can be done by determining the concentration of soluble reactive
species. Iron is one of the most reactive elements in environments, and its cycling is coupled to other major
biochemical elements like C, O, S, and P along with trace elements such as heavy metals. Iron determination found a
major boon for the various clinical and agricultural activities so in the last 25 years the range for clinical determination
of iron status has been extended with important methods. This review focuses on conventional as well as recent
analytical methods for iron determination and a comparative account of it.

KEYWORDS: Iron, biogeochemical elements

1. INTRODUCTION: Fe (II) and Fe (III) are relatively small ions with a marked
Iron belongs to third group of cations, which is subdivided propensity to form six-coordinate complexes with ligands
into III A, and III B groups. Iron group or III A group containing O, N, and S. This property, combined with the
includes iron, aluminum, and chromium.1 It is one of the remarkable range of redox potentials covered by iron-
most abundant trace elements; ranking fourth (about 5 % of containing enzymes, accounts for the role of iron in such
the earth) among the elements in the lithosphere, after fundamental reactions as ribonucleotide and dinitrogen
oxygen, Si and Al. Iron is an essential element in cellular reduction as well as in the energy-yielding electron transfer
metabolism and growth because of its importance in major reactions of respiration and photosynthesis. At the same
biological mechanisms such as DNA synthesis, electron time, the chemical properties of iron place limitations on
transport and nitrogen fixation.2 Nature has exploited the cellular accumulation of this element. First, Fe (II) and
bivalent and trivalent status of iron in the context of Fe (III) can act catalytically to generate hydroxyl radicals
electron transport and hence metabolic processes. This that are the most potent oxidizing agents known.6
makes iron a vital element. Iron present in the hydrosphere
in two oxidation states, II and III, which are From blood samples it is now possible to obtain information
thermodynamically stable under anoxic and oxic conditions, on the size of the body iron stores and on possible tissue
respectively.3 iron deficiency by the levels of serum ferritin and serum
transferrin receptor, respectively. These iron measures can
Iron is an essential trace element for the human body and be used together to distinguish between 7
iron deficiency
plays an important role in oxidative energy production. anaemia and anaemia from other causes.
Body iron which transports and uses oxygen in the
Iron determination has major applicability in number of
production of energy is referred to as functional iron and
fields so as a need this review focused on iron
can be found in hemoglobin, myoglobin, iron-dependent
determination methods including the traditional classical
enzymes, and respiratory chain proteins.4 Both inadequate
methods such as spectrophotometry and titrimetry as well
iron supply and excessive iron accumulation in the body
as instrumentation methods such as atomic absorption
lead to increased morbidity.5
spectrometry and recent advanced one like electron probe
microanalysis.

2. History of iron estimation:

Traditionally iron from paper as cellulodic material have
been analyzed by gravimetric, titrimetric ,colorimetry or
flame photometric absorption8.
Received on 16.11.2010 Modified on 30.11.2010 Photometry and atomic absorption spectrometry are offered
Accepted on 01.12.2010 © AJRC All right reserved for determination of iron microquantities in water and food
Asian J. Research Chem. 4(3): March 2011; Page 348-361
products (Table 1)

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Table 1: Characteristics of some iron determination techniques*

Concentration
Technique Reagent Experimental condition
range
Solutions
Fe(III),Ext/Ph9 Eosine + 1,10-phenantroline =9, extraction by CHCl3+(CH3)CO 1-10 g
Fe(II), Ext/Ph10 Bathophenanthroline =4-5, extraction by CHCl3, oxidants are interfering: 1-10 g
Food products
=4-6, interfere Ag(I), Bi(III), Hg(II), Mo(VI), Ni(II),
Ortho-
Fe(II), Ph11 W(VI), CN–, PO43–, not interfere up to 25 g Al(III) As(V), detection limit:10 g
phenanthroline
Mn(II), Pb(II), Cl–, NO3–
Fe(III), S12 - 0,0010-0,02 % wt
Drinking water
Fe(III), Ph13 Sulphosalicylic acid >9 0,10-2,00 mg/L
Fe(II), Ph13 Ortho-phenanthroline =4-5nterfere CN–, NO2–, polyphosphates,10-fold excess of 0,05-2,0
Cr(VI), Zn(II), Co(II) , Cu(II) > 5 mg/L mg/L
Fe(II), Ph13 2,2-dipyridyl =3-9, interfere Ag(I), Bi(III), Co(II), Cu(II), Hg(II), 0,05-2,0 mg/L
Mo(VI), Sn(II), W(VI), Zr(IV)
Fe(III),Ext/Fl14 Ortho-phenylenediamine Extraction of iron by polyathomic alcohols at =8, not 0,05-5,0 mg/L
interfere up to 1 g/L NH3, alkali and alkaline-earth metals,
SO42–, Cl–, up to 200 mg/L Cr(III), PO43–, up to 100 mg/L
Cu(II), up to 50 mg/L F–
Cognacs, grape and fruit wines
Fe(III), Ph15 K4Fe(CN)6 0,5-3,5mg/L
Alcohol products and raw for its making [18]
IVA - = 1-4 0,03-20mg/L
Mineral drinking waters
, Fe(III) Complexon III =2 > 0,5 mg
Nickel, nickel and copper-nickel alloys
Ph, Fe(II) 1,10-phenantroline separation from matrix 0,001-0,1 % wt
or 2,2-dipyridyl
, Fe(II) Titration by K2CrO7 separation from matrix 0,4-6,5 % wt
AAS, Fe(III) - 0,004-6,5 % wt
* Ph – photometry, Fl – fluoroscence, – titrimetry, AAS – atomic absorption spectrometry, Ext/Fl – extraction-fluorometric technique, Ext/Ph -
extraction-photometric technique, IVA - inversion voltammetry16

Iron determination in mineral drinking water is carried out Spectrophotometric method for the determination of iron in
by titrimetric method17. Inversion voltammetry (IVA) is a biological sample is based on release of bound iron and
applied to determination of the element in alcoholic further the released iron is reduced to Fe (II). A color
products and raw materials for its making18. Current reagent solution is added to it and the formed color complex
Russian standard offers photometry, atomic absorption is measured photometrically.20 Spectrophotometric
spectrometry and titrimetry for iron determination in nickel, determination of trace iron (III) after its preconcentration
nickel alloys and copper-nickel alloys19. with a chelating resin is also developed.21

1) Phenanthroline Method:
The limitation of atomic absorption method is rather high
cost of the equipment and consumables. In IVA, oxygenPhenanthroline reacted with iron (II) with red coloured
and organic substances should be removed from solution;
complex formation which was measured at 510nm.
the electrodes in use either contain toxic mercury or require
Total Iron was determined by oxidizing any Iron (II) from
surface regeneration at regular intervals. Titrimetry is
Iron (III) with Persulfate. A Thiocyanate complex of the
usually used only for determination of enough high Iron (III) is formed in a dilute acid solution. A direct
concentration of iron. measurement of the Iron-Thiocyanate is made using a
Advantage of molecular spectroscopy techniques is relative
calibrated spectrophotometer at 474 nm. The Iron (II) is
cheapness of consumables and availability of equipment;
determined by adding a sample to deaerated 1, 10-
fluorimetric ones is much more sensitive, than photometric.
Phenanthroline/Sodium Acetate reagent. Any Iron (II)
present in the sample will form a colored complex with the
3. Iron determination methods: reagent that can be measured on a spectrophotometer at 510
Iron can be analyzed by different methods which were nm. Iron (III) is Calculated by subtracting the g/L Iron (II)
covered in the next part of the review one by one. from the g/L total Iron.22

1. Spectrophotometric determination: A method for the determination of Fe (III) at trace levels

This method is based on number of free metal particles was described. Thus, prior to the spectrophotometric
present in solution. Generally there is decrease in the determination, a preconcentration of the trace amounts of
number of particles during complex formation and so this iron (III) using a chelate forming resin was proposed. A
method can be used to detect formation of complex. strong base anion-exchange resin (Dowex 2X4) loaded with

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Ferron (7-iodo-8-hydroxyquinoline-5-sulphonic acid) was was pushed out into a micro cell for the absorbance
used for Fe (III) preconcentration, at pH 2.2. After measurement at [Link] column was reusable after
desorption with 5 % ascorbic acid in 0.5 M HCl, the analyte washing with ethanol. This method had a detection limit of
(converted from Fe (III) to Fe (II) was determined 0.001mgl-1 can be determined.
spectrophotometrically at 510 nm as Fe (II)-o-
phenanthroline complex. The accuracy of the proposed Preconcentration of iron was done by chelate-forming
method was verified by comparing the obtained results with sorbents. Very efficient systems provided by
those obtained using AAS with the standard addition immobilization of 8-hydroxyquinoline on solid support.
method. The sensitivity of the spectrophotometric method Also, a sulphonic acid derivative of 8-hydroxyquinoline,
(after preconcentration) was 0.01 g Fe (III)/ml. The namely ferron (7-iodo-8-hydroxyquinoline-s-sulphonic acid
recovery for iron (III) at the 7 µg/l level was 97 %.22 loaded on anion exchange resin. Ferron is one of the most
selective chelating agents used in the Spectrophotometric
A procedure has been developed for the determination of determination of Fe (III) at pH [Link] retention on the
iron (III) dimethyldithiocarbamate by converting it into the complexing resin at pH 2, the analyte is desorbed with
iron (II) − bathophenanthroline complex, which is then ascorbic acid, (thereby converting from Fe (III) to Fe (II)
dissolved in acetone-water (1:1), and the absorbance is and determined spectrophotometrically as the Fe (II)-O-
measured at 534 nm against a reagent blank. Beer's law is phenanthroline complex at 510nm.21
obeyed over the concentration range 0.5−20 g mL-1 in the
final solution. The method is sensitive and highly selective A simplified method is described by Barry (1968) in which
and is used for the direct determination of ferbam in a urine is digested and iron content of the residue determined,
commercial sample and in mixtures with various without need for quantitative transfer. A standard of
dithiocarbamates (ziram, zineb, maneb, etc.) and from quantity of acid is used to ash urine to yield a dry residue,
wheat grains. 23 which is reduced with thioglycolic acid, and a pink colour is
developed with bathophenanthroline sulphonate in the
A method for the simultaneous quantification of the various presence of acetic acid-acetate buffer at pH [Link] main
forms ferrous, ferric, soluble and chelated form of iron purpose of ashing to destroy the chelating agents and render
added or endogenous to foods has been developed. The all iron available for colour formation.27
total, elemental, and soluble irons are determined with Ferrozine (PDT disulfonate; 3-[2-Pyridyl]-5, 6-diphenyl-1,
minimal pre-treatment by atomic absorption 2, 4-triazine-4, 4´-disulfonic acid. Na-salt)
spectrophotometry. The iron valences and complexed iron
are measured spectrophotometrically using the Method:
bathophenanthroline reagent. The method was both Stookey (1970) proposed a method for determination of
reproducible and accurate in measuring iron added to plant ferric and total (soluble and insoluble) iron with ferrozine.28
material and to a formulated instant beverage. The The concentration of iron in urine is of interest in
procedure may be applied to determination of possible monitoring desferoxamine therapy for acute iron
changes in the iron-fortified processed foods.24 poisoning29 or iron overload. Atomic absorption
spectrophotometry (AAS), the reference Method for iron
Pizzaro et al (1987) described a quantitative method for measurement30 is usually not available in most routine
determination of iron in stool to monitor consumption of laboratories. Routine automated methods for measuring
iron-fortified milk in infants. The method is simple, fast, serum iron with use of a chromogen do not detect iron in
and inexpensive. Stool samples from infants consuming these samples29, because the complex of desferoxamine and
fortified milk or non fortified milk were ashed, and ashes Fe3+ is not dissociated.
were diluted in hydrochloric acid and reacted with
bathophenanthroline disulphonate.25 Several types of pretreatment of urine samples to dissociate
the deferoxamine-Fe3+complex have been described.29, 31
Kawakubo et al, 2004 developed a visual and micro Given the need for a sensitive, (semi) and routine analysis
spectrophotometric methods based on the extraction of a automated procedure to monitor deferoxamine therapy in
coloured ferroine thiocyanate allowed determinations of urine, we combined a ferrozine-based method for iron
trace iron in fresh water samples26. For the visual method, a measurement (kit 1127683/69 1; Boehrunger Mannheim,
water sample mixed with a reagent solution containing 1, Mannheim, Germany), implemented on an automated
10-phenanthroline, sodium thiocyanate and 0.1M HCl. Iron analyzer (Hitachi 747; Hitachi, Tokyo, Japan), with a
was extracted as pink ferroine thiocyanate with 1ml of 4- pretreatment. Verhasselt et al (1991) optimized and
32
[Link] sample up to 20ml was added compared two types of pretreatment : thioglycolate
29 31
step-by-step, until the colour of the extract was detected trichioroacetate and ascorbate citrate.
visually. Without any special instrument or colour standard,
iron down to 0.001 mg-1 (0.025 g) in a sample can be To measure the recovery of iron, we added FeC13 to normal
determined. For the micro spectrophotometric method, the urine with or without equimolar concentrations of
extract for 20ml of sample was separated by capillary deferoxamine. Urine samples of patients with proteinuria or
suction in a column with acrylic fibers. A part of the extract in desferoxamine therapy were collected. Each sample was

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thoroughly mixed, and aliquots were stored at 4°C and flotation process in which the material of interest is
analyzed within 4 h. In the thioglycolate-trichloroaoetate adsorbed on the surface of gas bubbles on liquid, and
pretreatment, the reagent (0.29 mol/L thioglycolic acid and collected on an upper layer of an immiscible liquid. Solvent
0.61 mol/L trichloroacetate in water) is added to an equal sublation has advantages that the analytes of complexed
volume of fresh urine. In the ascorbatecitrate pretreatment, forms can be directly extracted into a solvent by the
3.5 volumes of reagents (0.48 mol/L ascorbate and 0.57 flotation and determined in the solvent as a concentrated
mol/L citrate in water) were added to 1 volume of fresh state by different analytical methods.
urine. These pretreatments are done at room temperature,
and the pretreated samples are analyzed after 15-60 mm on The method for separation and preconcentration of metals
the automated analyzer. A control urine sample is processed by flotation and solvent sublation, followed by their
the same way for a reagent blank. The linearity was tested spectrophotometric and/or AAS determination, has
of the procedures from 0.41 to 1000 µmol/L iron. In the increased in popularity recent years. A literature survey
absence of deferoxamine, recovery of added iron (95%, at revealed that solvent sublation followed by
1000µmol/L) and linearity (r2 > 0.9999) were excellent sepctrophotometetric and/or AAS determination is rarely
without pretreatment. reported. Ferrozine finds its role for a direct
In urine containing both desferoxamine and iron, iron could spectrophotometric determination of Fe (II),and also gives
not be detected without pretreatment of the samples. high performance in the preconcentration of Fe (II) via a
solvent sublation as ion pairs of [Fe(FZ)3]4- and
Pretreatment with thioglycolate - trichioroacetate yielded tetrabutylammonium ions. Ali Akl et al (2006) reported
poor recovery (19%) and linearity (r2 = 0.81). Pretreatment ferrozine as a complexing agent formed ion pair with
with ascorbate-citrate was superior in both recovery (92%) tetrabutylammonium(TBA+ ) ion as counter ion, oleic acid
and linearity (r2 = 0.9996) and was thus adopted for further (HOL) used as surfactant then (TBA)4[Fe(FZ)3] ion pairs
experiments and routine analysis. The ascorbate citrate were floated by vigorous shaking in the flotation cell and
reagent solution was stable for at least 3 days at 4°C, extracted into methyl isobutyl ketone (MIBK)on the surface
although values for the reagent blank tended to increase. of the aqueous solution. The iron collected in the MIBK
The reagent can be stored for longer periods at – 18°C.32 layer was measured directly by spectrophotometry and/or
flame atomic absorption spectrophotometry.3
A direct method for determination of iron in blood serum Carter (1971) proposed the use of ferrozine a commercially
comprising reacting the blood serum with ferrozine in the available sulfonated ferroin, to the determination of sub
presence of a reducing agent to bring iron ion into divalent microgram levels of iron in human serum.34
form, thiosemicarbazide and hydrochloric acid at a pH
between 1.7 and 2.1, without any addition of buffers and Seligman and Schleicher (1999) compared two methods for
tensioactive agents and determining colorimetrically the determination of serum iron in presence of iron gluconate or
iron content of the specimen by means of the colored iron dextran.35 One method uses an acetate buffer (pH 4.5)
complex formed between iron ion and ferrozine against a with 15 g/L hydroxylamine hydrochloride to release the iron
reagent blank. from transferrin. The released iron reacts with 8.5 g/L
Ferrozine reagent during 15min incubation at 37°C to
In a method for direct colorimetric determination of iron in produce a magenta-colored complex. Another method used
blood serum by reacting the serum iron with ferrozine in the an acetate buffer (0.1 mol/L, pH 4.8) containing ascorbic
presence of acid and a reducing agent to release serum iron acid (56.8 mM/L) and guanidine (6 mol/L) to release
from transferrin and reduce iron ion to divalent form. The transferrin-bound iron. The absorbance is measured at 595
iron-ferrozine reaction carrying out the in the presence of nm, before and after 5-min incubation at 37°C with 36
sufficient thiosemicarbazide to eliminate significant copper mmol/L Ferrene S. In this trial methodology that measure
interference, without any addition of buffer and tensioaction serum iron, particularly those using an ascorbic
agents and at a pH sufficiently low that such elimination of acid/guanidine buffer method, will cause in vitro
copper interference is obtained without significant reaction dissociation of iron bound to gluconate or dextran
of the thiosemicarbazide with iron, said low pH also being complexes.
such that substantially complete formation of a colored
iron-ferrozine complex is obtained.33 2) Pyridyl regent:
The invention concerns a method for the interference-free
There are several possible strategies for enhancing the determination of iron in biological samples; in particular in
overall sensitivity fro metal determination when using a serum in which bound iron is released, the released iron is
metal-chelate technique. In an effort to develop a reduced to Fe (II), a color reagent solution is added and a
preconcentration technique for measuring Fe (II) and total color complex that forms is measured photometrically and
iron in natural water samples, investigations were it also concerns a combination of reagents which is suitable
conducted concerning the use of ferroine as for the interference-free determination of iron especially in
preconcentrating agents for iron. These techniques involve the presence of high amounts of EDTA.
solvent sublation, solvent extraction, and ion exchange.
According to IUPAC definition solvent sublation is a

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The reagents used are composition of reagent 1 and reagent ([Fe total]–[Fe+2]). For the determination of Fe total the
2 containing following components used to determine iron three previous methods were compared with those of
in serum: 20 Atomic Absorption Spectrometry (AAS). 40
Reagent 1 200 mmol/l citric acid
100 mmol/l thiourea The procedure used in the SSA method was as follows:
7% (w/v) Alkylpolyethylene glycol ether Into a 100 mL volumetric .ask, add an adequate volume of
10 mmol/l indium (III) chloride
the sample. Then add 3 mL of 10% (w/v) sulphosalicylic
Reagent 2 150 mmol/l sodium ascorbate
6 mmol/l 3'(2'-pyridyl)-5,6-bis(4- acid solution and complete volume with Ultrapure Millipore
phenylsulfonic acid)-1,2,3- triazine Water. Stir during 2 min and measure de absorbance at 500
monosodium salt nm. Then add 3 mL of ammonia solution 25% (w/v) and
stir during 2 min and measure the absorbance at 425 nm.41,
42
Methods in common use for the determination of
complexed iron in compounds such as haemoglobin,
ferritin, or ferrioxamine depend on a preliminary treatment Squaric acid (1,2-dihydroxy-3,4-diketo-cyclobutene):
to release free iron which is then estimated by a Squaric acid (1, 2-dihydroxy-3, 4-diketo-cyclobutene) is
colorimetric reaction. The release of iron may be used in a specific reaction with Fe (III) for the
accomplished either by dry ashing in a furnace or wet spectrophotometric determination of Fe (III) and total iron
ashing with hot concentrated acid.36 content. The analytical procedure includes mixing
ammonium squarate (40mM), prepared in a phthalate buffer
Both these procedures were time consuming and potentially solution of pH 2.7, with the sample and measuring the
hazardous. A simple technique is described for the release absorbance at 515nm. The method has been successfully
of iron from haemoglobin, ferritin, or ferrioxamine in applied to the determination of iron (III) and the total iron
solution by digestion with an acid-permanganate solution at content after quantitative oxidation of iron (II).43
room temperature. Excess permanganate is reduced with
ascorbic acid, and iron in the resulting clear solution is Suwansaksri et al (2003) compared 3 different methods of
estimated by the method of Young and Hicks (1965) using serum iron determination, nonprecipitating ferrozine
tripyridyl triazine.37, 38 colorimetric method44, precipitating ferrozine colorimetric
method, and iron liquicolor cleaning factor colorimetric
Salicylic acid: agar-based (CAB) method (Human, Taunusstein,
Iron can also be determined by spectrophotometric titration Germany). For a setting with limited resources such as
in which salicylic acid and iron (III) form a deep coloured Thailand, use of the first analytical technique,
complex with a maximum absorption at 525nm; this nonprecipitating ferrozine colorimetric method, is
complex is used as the basis for the photometric titration of recommended because of its low cost and ease of
iron (III) with standard EDTA solution. At pH ~2.4 the performance.
EDTA-iron complex is much more stable (higher stability
constant) than the iron-salicylic acid complex. In the 3) Bis(2-hydroxymethyl-5-hydroxy-4-pyrone-6)ketone:
titration of iron-salicylic acid solution with EDTA the iron- A method has been developed for the direct
salicylic acid colour will therefore gradually disappear as spectrophotometric determination of iron by using the
the end point is approached.39 reddish-orange coloured complex formed in the interaction
The leaching solutions from hydrometallurgical processing of iron with bis (2-hydroxymethyl-5-hydroxy-4-pyrone-6)
45
of copper sulphides have a high concentration of several ketone.
metallic sulphates.
4) Determination of Iron (III) with Amines and
The Fe+2 determination was carried out by 1, 10- Thiocyanate:
phenanthroline method.40 The Fe total determination is Hayashi et al (1986) studied that the coloration of iron (III)-
realized using potassium dichromate volumetric method thiocyanate complexes was enhanced by the addition of
and Atomic Absorption Spectrometry (AAS) technique. amine to acidic solution containing the nonionic
surfactant.46 A coexistence of 0.25% Capriquat
In this sense, in all of these cases the determination of Fe+3 (trioctylmethylammonium chloride) brought about a 1.5-
is not direct and for instance, a quantitative determination fold increase in sensitivity (molar absorptivity:
4 -1
of both oxidation states could be imprecise. Recently, a 2.95×10 mol dm3 cm-1 at 485nm) of the
simple and fast method has been described for the spectrophotometric iron determination when compared to a
quantitative simultaneous determination of Fe+3 and Fe total method containing no amines.
with SSA in acid mine drainages (AMD) and other
solutions.41 5) Potassium thiocyanate:
Some metals, such as iron, will form highly colored
For the direct determination of Fe+3 the method of the SSA complexes when reacted with the thiocyanate ion which
was used, and indirectly Fe+3 also was determined by means resulted in red coloured complex and measured absorbance
o-phenanthroline method and potassium dichromate method at 470nm.

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Fe3+(aq) + 6 SCN-(aq) [Fe(SCN)6]3-(aq) sensitivity, due to its long optical path in addition to
Another simple and rapid calorimetric method is described atomization without the use of chemical flame. For low
for the determination of iron in blood by Wong (1928).47 boiling point metal elements, the analytes can be easily
The method described was as follows, vaporized and effectively introduced into HQC and their
Transfer accurately with an Ostwald pipette 0.5 cc. of blood atomic absorption can be measured. The method required
into a 50 cc. volumetric flask and introduce 2 cc. of iron- only several microliters of sample solution for the
free concentrated sulfuric acid. Whirl the flask to agitate the determination at nanogram level. Sugesaka et al (2001)
mixture for 1 or 2 minutes. Add 2 cc. of saturated potassium proposed a heated quartz cell atomic absorption
persulfate solution and shake. Dilute to about 25cc. with spectrometry coupled with in situ butylation/vaporization
distilled water and add 2cc. of 10 per cent sodium tungstate for the determination of iron and tin.50
solution. Mix, cool to room temperature under the tap and Fe (III) could be determined by AAS with a calibration
then dilute to volume with distilled water. Stopper the flask curve or by the standard addition method used from a
and invert two or three times to effect thorough mixing. solution with Fe (III) concentration under 0.1µg/ml.21
Filter through a dry filter paper into a clean, dry receiving
vessel. Pipette exactly 20cc of the clear filtrate into a large An ultramicro method was described by Olsen et al (1973)
test-tube graduated at 20 and 25cc. Measure into another for determining iron in 50 µl of serum.51 Hot trichloroacetic
similar test-tube exactly 1cc of the standard iron solution acid was used to remove protein and hemoglobin iron, and
containing 0.1 mg of Fe per cc. Add with a graduated 1cc. atomic absorption analysis of 10µl of the supernatant fluid
pipette 0.8 cc of iron-free concentrated sulfuric acid and was carried out with use of a graphite furnace and a
dilute to the 20 cc mark with distilled water. Cool to room specially grooved graphite tube. Results correlate well with
temperature under the tap. To both the unknown and those obtained by a conventional flame atomic absorption
standard 1cc of saturated potassium persulfate and 4 cc of 3 method for iron that requires a 1-ml sample of serum. When
N potassium sulfocyanate solutions were added. Insert a serum was analyzed with no pretreatment, results were
clean rubber stopper, mix, and compare in a Duboscq higher, probably because of hemoglobin iron present in
calorimeter. Calculation - As the 20cc. of filtrate taken serum, even serum with no visible haemolysis.51
represent 0.2cc of the original blood, and the quantity of
standard solution used contains 0.1 mg of Fe, if the reading Brecque et al (1978) proposed an interference-free atomic
is made with the standard set at 20 mm., then 20 divided by absorption method for the determination of total iron in
the reading (R) of the unknown and multiplied by 50 will geological samples.52 The method is based on the
give the number of mg of Fe in 100cc of the blood conventional standard curve technique employing the
examined. 386nm atomic absorption line in the fluoboric-boric acid
20/R X 50 = mg of Fe per 100cc. of blood.47 matrix. Interference studies of aluminum and/or silicon on
the iron determination conclude that these interferences do
Propylene carbonate (4-methyl-1,3-dioxolane-2-one) not exist under the conditions of this method.
simultaneously extracts the 2,9-dimethyl-1,10-
penanthroline chelate of copper(I) and the tri(2-pyridyl)- Olsen et al (1973) developed an ultramicro method for the
1,3,5-triazine(TPTZ) chelate of iron(II) from acetate- determination of iron from in 50 l of serum sample
buffered aqueous solutions. Molecular absorption compared with conventional flame atomic absorption
Spectrophotometric quantification is accomplished by method for iron that requires a 1ml sample of serum.51 In
measuring the absorbance of the iron (II)-TPTZ chelate at this method of analysis hot trichloroacetic acid was used to
458nm and that of the copper (I)-NC chelate at 458nm. The remove protein and hemoglobin iron, and atomic absorption
iron (II)-TPTZ chelate exhibits an absorbance at 458nm that analysis of 10 l of the supernatant fluid was carried out
is 0.123 times its absorbance at 596nm.49 with use of a graphite furnace and a specially grooved
graphite tube.
Berman (1918) devised a method of iron determination
from blood sample in which iron held is split off by the 7) Flame atomic absorption method:
action of concentrated hydrobromic acid.49 The iron was A Flame atomic absorption spectrophotometer (FAAS) has
oxidized and the organic matter was destroyed by been widely used because of its high specificity, while not
potassium permanganate. The resulting solution was mixed being an expensive instrument. However, the direct
with ammonium sulphocyanate in water and acetone and determination of trace concentrations of metals by FAAS is
the colour was compared with the standard iron solution generally difficult because of matrix interference problems
similarly treated. and concentrations below the detection limit of FAAS.
These problems can be overcome by using preconcentration
6) AAS: and separation procedures. For this purpose, various
Tang (1978) used CRA (Carbon rod atomizer) flameless preconcentration/separation methods, such as solvent
with atomic absorption equipment for determining iron extraction, coprecipitation, ion exchange, resin chelation
from solid sample.8 and solid phase extraction have been widely used.
A heated quartz cell (HQC) is an attractive atomizer for A solid phase extraction (SPE) technique has found
atomic absorption spectrometry (AAS) in regard to its increasing applications for the preconcentration of trace

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metal ion and the elimination of matrix interference prior to determination in pharmaceutical product were compared in
AAS analysis. Different solid phase extractors, such as terms of uncertainty budgets by Jurgens et al (2007).58 Non
Amberlite XAD resins, activated carbon, polymeric fibers, linearity, instrument drift, volumetric measurements
and silica gel have been used to preconcentrate trace metal contributes for the uncertainty of the two methods.
ions from various media. Kenduzler and Turker (2002) used
Ambersorb 572 as a solid phase extractor where iron, Saleh et al (2008) proposes a method to determine iron in
manganese and zinc were preconcentrated from a water samples of fish feed and feces using ultrasound in the
matrix as their chelates using Ambersorb 572 as a solid extraction of the analyte and in subsequent quantification
phase extractor.53 Ambersorb 572 was chosen because it has by flame atomic absorption spectrometry.59 The results
a high surface area, durability and purity. obtained with the proposed extraction method allowed
calculating the coefficients of apparent digestibility of iron
The low concentration of iron present in natural waters (as in the food sources, which was not possible when using
gl-1) necessities the selection of a suitable preconcentration results obtained from samples mineralized by acid
procedure. In recent years, microorganisms such as, yeast, digestion.
bacteria, fungi have often been proposed for the
preconcentration and speciation of trace metals. Bag et al 8) Solid phase spectrophotometry:
(2001) proposed preconcentration of Fe (II), separation of Teixeira et al (2002) developed a multivariate calibration
Fe (II) from Fe (III) and flame atomic absorption method, in the reduced range of 560-650nm60, in
spectrophotometric determination of each iron species by association with solid phase spectrophotometry with the
using an adsorbent, Aspergillus niger, immobilized on advantage of rapid determination of the analyte in a mixture
sepiolite.54 Repeated use of the column is possible and without previous masking, preconcentration or separation
relatively stable upto 20 runs. steps.

For the determination of iron in light and heavy petroleum Fe (II), Ni (II) and Zn (II) are retained on 1-(2-thiazolylazo)
fractions by the use of graphite furnace atomic absorption 2-napthol (TAN) immobilized in C18 bonded silica,
spectrometry after sample dilution with methyl isobutyl forming complexes allowing in situ analyte separation. The
ketone, and the use of the method of standard additions are method based on the different reaction/retention ratios of
advised for iron determination by Torre et al (1990).55 the studied ions on the solid support.

A new Schiff base, N,N -bis-(2-hydroxy-5-bromobenzyl)-2- Garcia et al (1995) utilized a cataionic exchanger paper to
hydroxy-1,3-diiminopropane, has been synthesized by retain analytes in solution and, after drying, to analyze
Pehlivan and Kara (2006) for the very sensitive directly by measuring the UV-Vis absorbance of the
determination of Fe (III) and Fe (II) in natural water paper.61 The method was applied to determination of iron
samples.56 The method has also been applied to total iron using its known 1, 10-phenanthroline complex. By this way
determination in sediment samples. In the preconcentration determination of iron at ng/ml level by solid phase
system, the Schiff base reagent was mixed with the samples spectrophotometry after preconcentration on cation
and chelates containing Fe (III). exchange filters was done.

The complexes were then adsorbed on silica gel within a 9) Photoacoustic Spectrometry:
column system. Elution of the adsorbed chelate from the Shida and Masuda (1998) developed a sensitive method
silica gel was performed with a small volume of acetone based on preconcentration on a membrane filter with a
containing 2.5% nitric acid. The iron was measured off-line finely pulverized anion-exchange resin for the
by flame atomic absorption spectrometry. The method can determination of iron (II) in water samples by
be applied to the preconcentration, separation and Photoacoustic spectrometry. Iron reacts with 2-nitroso-5-
speciation of iron. The results were compared statistically (N-propyl-N-sulfopropylamino) phenol (Nitroso-PSAP) to
with those from the standard 1, 10 phenanthroline method form a water-soluble chelate anion, which is adsorbed on
used for iron speciation in water systems. A Student t-test the resin at pH 8.0; the resulting resin is filtered through a
indicated no significant difference between the two membrane filter.62 The resin with the membrane filter is
methods. inserted into cell, and the intensity of the photoacoustic
signal of iron (II) is measured at 826nm. The colour
Yurchenko and Kharenko (2007) showed that sodium intensity of the green iron (II)-Nitroso-PSAP complex on a
dodecyl benzenesulfonate used as the modifier in flame membrane filter was measured visually following the
atomic absorption determination of iron in real samples standard series method. Iron (II) was determined as total
make it possible to make the measurements in a low- iron after reduction with 0.1M ascorbic acid solution,
temperature flame (propane-butane-air type) with increased because the brown iron (III)-Nitroso-PSAP complex was
sensitivity and selectivity and also a lower detection limit also adsorbed on the anion exchange resin solution (ARS).62
for iron.57 The detection limit for iron is Cmin = 0.008
µg/cm3. The fortification of foods, such as cereals, with iron has
UV–Vis spectrophotometric (UV–Vis) and flame atomic been recognized as a worldwide necessity, since a
absorption spectrometric (FAAS) analysis for iron
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 Asian J. Research Chem. 4(3): March 2011

deficiency of this element produces different metabolic The analytical task of iron content measurement in the ink
disorders. Therefore the developments of methods to writing on the surface of an 18th century manuscript by
measure low concentrations in foods has become of electron probe microanalysis (EPMA) using a scanning
increasing importance. In order to achieve higher sensitivity electron microscope equipped with an energy-dispersive X-
and selectivity the use of photothermal (PT) technique is ray spectrometer (SEM/EDS) was done by Virro et al
advantageous. The technique based on a photoinduced (2008) and It was concluded that EPMA using a SEM/EDS
periodical changes in the thermal state of a sample. Light is suitable for at least semi-quantitative determination of
energy, which is absorbed and not lost by reemission, leads iron in the writing of ink-written manuscripts.68
to sample heating, which induces changes in the
temperature-dependant sample parameters. A catalytic kinetic determination of iron was performed by
Khan (2006) by exploiting the reactions of Neutral Red
Delgado-Vasallo et al (2003) reported microphone based (NR).69 The iron catalyzed oxidation of NR with potassium
open Photoacoustic (PA) cell especially designed for bromate was studied kinetically by using fixed time
spectroscopic measurements in liquid systems.63 It renders method. The reaction was followed by measuring the
same results as optical spectroscopy, but several advantages decrease in absorbance at 535nm. The method was applied
such as independence of sample thickness, which allows successfully to the determination of iron in the
measurements over a wider absorbance range. pharmaceutical and synthetic samples. The results showed
good agreement with those obtained by atomic absorption
10) Nondispersive Atomic fluorescence spectrophotometer (AAS) with a detection limit of 0.019
spectrometer(NAFS) µgmL-1.
The analytical potential of the first commercial
nondispersive atomic fluorescence spectrometer (NAFS) 2. Chromatographic method:
with a tantalum coil atomizer for monitoring iron in HCl Chromatography based on partition coefficient can be used
gas has been investigated by Khvostikov. The limit of for separation and determination of iron along with ligand.
detection (LOD) of Fe in a mixture of 1% HCl in Ar was Various ways of chromatographic methods have been used.
found to be 9 ppb (v/v). 64 For the determination of the chelated Fe content in
synthetic Fe3+ fertilizer Garcia Marco et al(2006) reported
11) ELAN DRC (Dynamic reaction cell) ICP-MS: an ion pair HPLC method that allows the determination of
Nixon et al developed an ELAN DRC (Dynamic reaction o,p-EDDHA/Fe3+ in EDDHA/Fe3+ commercial fertilizers.70
cell) ICP-MS that made analysis of iron and copper from
liver tissues much faster and simpler.65 The DRC eliminate Total iron measured by AAS analysis to a detection limit of
the interfering species through a process called chemical 0.1mg/L. The sample as well as standard should be
resolution which used a reaction gas to selectively remove acidified with HPLC to a final concentration of HCL 1M
the interferences (40Ar16O+,40Ar16OH+,23Na40Ar+). DRC (10ml solution + 1ml HCL 11M) and chelated iron
provided lower detection limit therefore use of higher measured by HPLC.71
dilution factors was possible. This reduced the amount of
sample matrix introduced into the instrument, overall A selective determination method for Iron (Fe) ion in tap
routine maintenance, cleaning the interface cones and water has been developed by solvent extraction, followed
changing the pump oil. by reversed phase HPLC with photometric detection in
which The Fe (III) ion was quantitatively extracted into
12) X-ray fluorescence spectroscopy (XRF) and X-ray chloroform over the pH range of 3.2 to 4.3 as 3, 4-dihydro-
absorption near edge structure spectroscopy (XANES): 3-hydroxy-4-oxo-1, 2, 3-benzotriazine (DHOB) chelate.
X-ray fluorescence spectroscopy (XRF) and X-ray The extracted Fe-DHOB chelate was then separated on a
absorption near edge structure spectroscopy (XANES), both phenyl column with an eluent of methanol/water/0.05 M
using synchroton radiation (SR) microbeams can be apply DHOB (40:20:40, v/v) and detected at 500 nm. The
for determination of the chemical state of the elements and detection limit of the Fe ion in 5 mL water was estimated at
also provide the spatial distribution of each element in each 7 ppb.72
oxidative state selectively.66
1) Micellar Electrokinetic Chromatography (MEKC):
Bower (2007) proposed a method of ferrous iron Simultaneous analysis of two analytes is important not only
determination in which Iron (II) oxide in silicate rocks for biochemistry and environmental chemistry, but also in
measured after digestion with HF and HCl by reaction with many scientific and industrial fields. So far atomic
KI and KIO3.67 The iodine produced during the digestion absorption spectrometry (AAS), inductively coupled plasma
was trapped in CCl4 and measured at 512 nm (ICP) and x-ray fluorescent spectrometry (XRF) have been
spectrophotometrically. generally known as analytical methods for metal ions;
however, these methods can not distinguish any difference
in the metal valences.
13) Electron probe microanalysis (EPMA):

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Capillary electrophoresis (CE) is a high performance phenanthroline the conditional redox potential of the Co
separation technique but there are few reports regarding (III)/Co (II) system falls below those of the Cr (VI)/Cr (III)
analysis of oxidation states of metal ions. The use of two and Fe (III)/Fe (II) systems at pH around 1. Therefore,
selective ligands for Fe (II) and Fe (III) was reported by chromium (VI) and iron (III) can be titrated with cobalt (II)
Pozdniakova et al (1997) in which EDTA and 1, 10- in the presence of phenanthroline, alternatively. Firstly, to a
phenanthroline was used with CE. Takagai and Igarashi sample solution containing iron (II) and iron (III), a known
(2003) considered that, if we could use two selective amount of chromium (VI) is added in excess at pH 1, so
ligands for Fe (II) and Fe (III, a distinguishable analysis that Fe (II) can be oxidized to iron (III) in the absence of
could be made.73 The method was attempted using phenanthroline. After the pretreatment, the sample solution
bathophenanthroline derivative and desferioxamine B containing the excess of chromium (VI) and iron (III) (total
derivative as a selective ligands. Thus simultaneous iron) is titrated with a standard cobalt (II) solution in the
determination of Fe (II) and Fe (III) was achieved. presence of phenanthroline. The concentrations of iron (II)
Moreover, when Micellar electrokinetic chromatography and iron (III) can be calculated from the first and second
(MEKC) was performed under the condition of negligible potential breaks at the equivalence points for the titration.75
electroosmotic flow, the Fe-complexes were
preconcentrated by micelle sweeping. With this method, the A known volume of a sample solution, containing less than
resulting iron analysis was capable not only complete 10 mg of Fe (II), was taken into a 25mL flask. Then, 2.5mL
separation, but also a sensitive determination.73 of pH 6.5 buffer solution, 1mL of the masking reagent, and
5.0mL of the CTAB–EBBR (Cetyl trimethyl ammonium
Voltammetry: bromide - Erichrome blue black R and) aggregate solution
Voltammetry is Volt-Am (pero)-Metry where voltage ramp were added. The solution was then diluted to 25mL and
applied to electrodeand current is measured. A voltage ramp mixed well. After 20 min, the absorbances were measured
is applied and the corresponding current measured. Current at 480 and 610m with a 2-cm cell against a reagent blank
flows when a substance is reduced or oxidized on the electrode. without Fe; then, Ac of the ternary complex was calculated.
In case of no electrochemical reaction, no current is generated. For analysis of a solid sample, first its aqueous solution
must be prepared as follows: pulverize 1 g of a sample, add
Stripping voltammetric procedures belong to the most it to a mixture of 5mL of nitric acid and 5mL of 5%
sensitive method of determination of iron traces. The hydroxylamine hydrochloride for 2 h, volatile the excessive
catalytic adsorptive stripping voltammetry (CAdSV) nitric acid and hydroxylamine hydrochloride by boiling
methods, which are based on the adsorptive accumulation and, finally, neutralized to pH 6–7 with 5% NaOH and
of iron-nioxime complex on the hanging mercury drop filter. Then, the filtrate was diluted to 100mL with
electrode (HMDE), followed by a stripping voltammetric deionized water and colored and measured according to the
measurement of the catalytic reduction current of the same procedure as given above.76
adsorbed complex in the presence of sodium nitrite. The
detection limit obtained for a 20s accumulation time was Starke (1963) proposed a method for determination of basic
0.90nM iron.74 iron (III) compounds and free acid in iron (III) solutions by
iodometric titration with the same sample.77 After the iron
3. Titration has been reduced with potassium iodide and liberated
Iron determination by titration can be possible by various iodine was titrated with thiosulphate and determined,
ways including direct titration, back titration, further it was precipitated as iron (II)
potentiometric titration, and complexometric titration. hexacyanocobaltamate(III) which was almost colorless
Salicylic acid and iron (III) ions form a deep colored which did not interfere with the location of the end point of
complex with 525nm as max. This complex can be used as the acid titration.
the basis for the photometric titration of iron with standard
EDTA solution.39 5. Flow injection analysis
Kas and Ivaska proposed a procedure for determination of
ISO 2597-1:2006 specifies a titrimetric method for the concentrations of iron (III) and total iron by sequential
determination of the total iron content of iron ores, using injection analysis.78 The method is based on the strong blue-
potassium dichromate after reduction of the trivalent iron coloured complexes formed between iron (III) and trion.
by tin (II) chloride. The method is applicable to total iron The absorbance of the complexes is measured
contents between 30 % (mass fraction) and 72 % (mass spectrophotometrically at [Link] of iron (II) and
fraction) in natural iron ores, iron ore concentrates and masking of interfering fluoride is simultaneously done by
agglomerates, including sinter products(ISO 2597-1:2006 injecting one zone of hydrogen peroxide and one of thorium
American National Standards Institute (ANSI). (IV) between the sample and reagent zones. The sample
throughput is approximately 17 samples per hour, including
Kuwabara et al (1999) proposed a potentiometric titration three repetitive determinations of each sample.
of Fe (II) and Fe (III).75 The method is based on the effect Sakai et al, 2000 developed a new design of a multi-channel
of 1, 10-phenanthroline on the redox reactions of chromium micro cell for simultaneous analysis of copper, iron and
(VI) and iron (II) with cobalt (II). In the presence of zinc in the flow injection method.79

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Lunvongsa et al (2006) developed a Flow injection analysis to a purple colour. Calculate the percentage of iron in the
method for the sequential determination of iron and copper solid M.83
in the mixture by using oxidation reaction of the N, N-
dimethyl-p-phenylenediamine (DPD) with hydrogen Iron can also be determined by direct titration using
peroxide.80 Sample solutions in the absence and in the variamine blue as a redox indicator and titrating with EDTA
presence of TFTA were used for sequential determination with end point of the titration from blue to grey just before
of iron and copper .One injection is used for the sum of iron end point and final to yellow.39 Iron can be determined by
and copper contents, and the other for iron contents. For redox titration with an oxidant such as KMnO4 or K2Cr2O7
iron determination, TFTA was used as a masking agent for that converts Fe (II) to Fe (III). Potassium dichromate can
copper. The difference in peak heights was used to calculate be used as a primary standard by drying in oven at 150-
the copper concentration. 2000C for two hours to remove any bound water. As the
titration precedes the sample solution will turn green due to
Several spectrophotometric procedures have been used in the presence of Cr3+. The endpoint is reached when the very
flow injection analysis for the determination of Fe (II) and fine yellow colour of the Cr6+ titrant appears. In oxidation-
Fe (III) in many kinds of samples. Most of the regents are reduction titration Fe (II) solutions can be titrated with
solid and produce iron complexes with a limited solubility potassium dichromate solution. After all the Fe (II) has
in water. Consequently, mixed solvent systems or acidic been oxidized, the endpoint of the titration can be
buffers have been used as reagent carrier solutions in the recognized by the colour change (from green to yellow)
flow injection analysis systems. For example, ammonium when excess dichromate ion [Link] can be
diisopropyldithiophosphate in an aqueous-alcoholic determined by redox titration in which conversion of Fe
medium and norfloxacin in ammonium sulfate-sulfuric acid (III) to Fe (II) with a suitable reducing agent and a redox
medium have been used for iron determination. Asan et al indicator (diphenylamine), a colourless compound was used
(2003) described a method for the determination of Fe (III) which was oxidized by dichromate to di-phenyl benzidine
using dimethylformamide (DMF) as a chelating agent in a to form violet coloured compound.84
flow injection system.81 DMF is a sensitive and selective
reagent for Fe (III) for giving a 2:3 (Fe: DMF) complex 7. Electrochemical method:
with a sharp absorption peak at [Link] iron in the The rising importance of fuel ethanol in economic Brazilian
range of 5-90ng ml-1 can be determined with this method. set as an alternative automotive fuel or raw material for
alcohol chemical industry requires the identification and
A flow injection method was described by Yaqoob et al control of its contaminants, due to its potentiality for engine
(2006) for the determination of iron in fresh water based on corrosion effects and passivation of industrial catalyzers.
potassium permanganate chemiluminescence detection via Several organic and inorganic contaminants have been
oxidation of formaldehyde in aqueous hydrochloric acid.82 studied in fuel ethanol by different instrumental methods.
Total iron concentrations are determined after reducing Fe These contaminants can be originated from micro-nutrients
(III) to Fe (II) using hydroxylamine hydrochloride. The used on sugar cane production, during the fermentation
detection limit (three standard deviations of blank) is 1.0 process, such as transport and storage of final product.
nM, with a sample throughput of 120 h−1.
Under the analytical point of view, the use of chemically
Asan et al (2008) proposed a highly sensitive and very modified electrodes (CME) coupled to electrochemical
simple spectrophotometric flow-injection analysis (FIA) methods has presented a considerable increase in the last
method for the determination of iron (III) at low decade, specially for dosage of metallic species.
concentration levels81. The method is based on the
measurement of absorbance intensity of the red complex at Mattos et al (2008) presented a methodology for iron
410 nm formed by iron (III) and diphenylamine-4-sulfonic determination in fuel ethanol using a modified carbon paste
acid sodium salt (DPA-4-SA). Repeatability of the electrode with 1, 10 Phenantroline/nafion.85 An
measurements was satisfactory at the relative standard accumulation time of 5 minutes, such as a 100 mV of pulse
deviation of 3.5 % for 5 determinations of 10 g L−1 iron magnitude (Esw) and frequency (f) of 25 Hz were used as
(III). optimized experimental conditions.

6. Redox determination: The development of the proposed carbon paste-CME for

Iron can be determined by titration of dichromate with iron iron detection employing square wave voltammetry has
(II) using indicators like diphenylamine, diphenylbenzidine, presented good results for determination of total iron in
diphenyl amine sulphonate. The colour change for all three hydroalcoholic medium, allowing this methodology for iron
indicators is green to violet. The method applied as follows: determination in commercial fuel ethanol samples. When
Prepare a standard solution by dissolving 0.4g in water and the official Flame atomic absorption spectroscopy (FAAS)
make up the volume [Link] a flask 0.7g of the iron (II) method was used for comparison; a good correlation was
solid M provided .Add 30ml of diluted sulphuric acid, obtained for iron dosage at fuel sample.85
100ml of water, 7ml of 85% phosphoric acid and 5 drops of
diphenylamine sulfonate indictor. Titrate with dichromate

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Table 2: Summery of various iron determination techniques

Sr. Analytical Principal of method Salient features of method References
No. method
1. Spectrophotom It depends on measuring the amount Trace amounts (~ 10-6M) of Fe (II) can Saleh et al, 200859
etric Analysis of radiant energy of a particular be estimated. Methods include, Ide-Ektessabi and Rabionet,
wavelength absorbed by the sample, - Flame atomic absorption 200566
or measuring the amount of radiant spectrophotometry,
energy of a particular wavelength - X-ray fluorescence (XRF) and X-ray Olsen et al,200751
emitted by the sample absorption near edge structure Delgado-Vasallo et al, 200363
spectroscopy (XANES)
- AAS Teixeira et al, 200260
- Photoacoustic spectrometry Khvostikov, 199764
- Solid phase spectrometry Nixon et al,65
- NAFS Virro et al,200868
- DRC ICP-MS
- electron probe microanalysis Kawakubo et al,200426
Colorimetric methods using Ali Akl et al, 20063,
- Phenanthroline, Wong, 192847
- Ferrozine, Treiber andWolfgang,200120
- Potassium thiocyanate/Persulphate Karamanev et al, 200241
- - Pyridyl reagent Stalikas et al, 200343
- Salicylic acid Wilson and Daniels,196345
- Squaric acid
-Bis (2-hydroxymethyl-5-hydroxy-4-
pyrone-6) ketone.
2. Chrom Separative process for mixtures of Reversed phase ion-pair liquid Garcia-Marco et al, 200670
atography substance based on partition between chromatography detects the free and Susumu et al, 200972
mobile and stationary phase of the chelated Fe at conc. as low as 50 m. Takagai and Igarashi, 20037
sample to be determined, also can be - Gradient ion pair HPLC used for
used for reliable quantitative quantification of chelated Fe.
determination - Reversed phase HPLC
- MEKC
3. Stripping Two step process ,first step consists - The method is based on adsorptive Bobrowski and Krolicka,
Voltammetry of concentrating the ions from the accumulation of a metal complex on a 200274
bulk solution by electrode position hanging mercury drop electrode,
onto an electrode surface and second followed by a stripping voltammetry
step species of interest are
electrolytically stripped from this
electrode back into solution
4. Titration Quantitative chemical analysis by It includes oftenly indirect methods, Kuwabara et al,199975
knowing volume of solution of the - Potentiometric titration Fischer and Peters, 196884
substance to be determined - Redox titration Mendham et al, 200039
- Spectrophotometric titration
5. Flow injection Continuous segmented flow Flow injection method based on the Kas and Ivaska, 200278
analysis sequential analysis complex formation between iron and a Sakai et al, 200079
specific reagent which was measured by Lunvongsa et al, 200680
flow injection analysis Asan et al, 200381
6. Redox The reaction involves change of Iron can be determined by redox titration Robert L,200683
determination oxidation number or transfer of with using oxidant/using redox indicator. Mendham et al, 200039
electrons among the reacting Fischer and Peters, 196884
substances
7. Electro Different compounds can be detected chemically modified electrodes (CME) Mattos et al, 200885
chemical and measure nanoampere currents, coupled to electrochemical methods used
very sensitive to flow rate and for determination of total iron in
temperature hydroalcoholic medium and in
commercial fuel ethanol samples
8. Fluorometric Fluorescence based on strokes shift - Fluorescein isothiocyanate and Thomas et al, 199986,
effect Calcein used as ligand whose Zhu et al, 200287
fluorescence is quenched upon binding to Teixeira et al,200788
iron.
- Energy dispersive X-ray fluorescence
(EDXRF)
9. Chemiluminesc Emission intensity is directly reversed Micellar –mediated
Kyaw et al, 199889
ence proportional to excitation intensity chemiluminescence (RMM-CL)
10. Polarography Components either oxidized or The determinations of both Fe2+ and Fe3+ Peramaki et al, 20090
reduced depending upon the polarity in the same run possible.
of the impressed potential at
characteristic potential which used to
identify the species

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8. Fluorometric measurement: 10. Polarographic Method:

Fluorescence based method on the “Strokes shift” can be When reduction potential of ion is too low, then no
applied for iron determination. Calcein (CA), a fluorescein electrochemical reaction takes place. More metal ions are
isothiocyanate can be used as iron ligand which gives reduced at electrode surface resulting into required
emission on binding with iron. Calcein is an interesting iron reduction potential. This reaction produces current, which is
ligand whose fluorescence is quenched upon binding to measured to estimate metals.
iron. Thomas et al (1999) concluded that calcein (CA) is a
good chemosensor for iron (III) in cells and biological The determination of both Fe2+ and Fe3+ in the same run is
fluids also discussed the use of CA as fluorescent probe for also possible using polarography. Direct current
studying the pools of ferrous and ferric iron.86 They also polarography (nowadays current–sampled (tast)
presented use of CA for measurement of the kinetics of iron polarography) may be used in the determination of two
(II) nutrition of plant cells and physiological study of CA as oxidation states of iron. Pulse Polarographic method is
a nutritional siderophore. more sensitive than ordinary direct current methods.90
Table 2 gives the summary of above discussed methods for
Zhu et al (2002) reported a fluorescence enhancing method iron
for iron (II) based on fluorescein isothiocyanate (FITC)
with iodine to produce a non-fluorescence species result in DISCUSSION:
fluorescence quenching. Iron (II) could compete with Trace metal iron have important role in life, involving a
fluorescein isothiocyanate to react with iodine, resulting in wide spectrum of activities. Thus the determination of trace
fluorescence enhancement of the system.87 iron is becoming increasingly important in environmental
samples as well as in body. In order to access its
Fluorimetric method is much more sensitive, than accumulation or deficiency in biological and water samples,
photometric. But frequently fluorometric and photometric sensitive reproducible and accurate analytical methods are
determination of iron. In various objects is carried out with required.53 This review shows that a wide variety of
application of toxic organic solvents. Therefore search of protocols and techniques are currently used for the
new water-soluble reagents for fluorometric iron determination of iron. In earlier days iron determination
determinations is actual. Dedyukhina et al (2008) used N- was done by titrometric, spectrophotometric method but
aryl-3-aminopropionic acids and their derivatives (AAPA) due to low accuracy now other sophisticated methods were
as the water-soluble organic reagents for determination of applied like atomic absorption spectroscopy, flow injection
iron by fluorimetric way.16 method, voltammetry, electrochemical method,
chemiluminescence etc.
Teixeira et al (2007) developed an analytical method using
energy dispersive X-ray fluorescence (EDXRF) to Most of the methods based on determination of reduced
determine Fe and Cu in gasoline samples.88 In the proposed form (with reagents like dipyridyl, ferrozine,
procedure, samples were distilled and the distillation Phenanthroline) and total concentration. Concentration of
residues were spotted on cellulose paper disk to form a oxidized form was calculated by subtraction of reduced
uniform thin film and to produce a homogeneous and from totalConcentration.91 Some possess the sensitivity to
reproducible interface to the XRF instrument. The limit of measure iron at the picomolar level. However, the lack of
detection (LOD) was found to be 10 and 15 g L-1 for Fe standard cleaning, sampling and filtration protocols and an
and Cu, respectively. appropriate certified reference material has resulted in
possible erroneous reported environmental data.92 Recent
9. Chemiluminescence Method: works focus on siderophores as biological ligands produced
Kyaw et al (1998) examined the development of a hybrid by selective microorganisms which organically complex
method based on the combination of the Reversed with iron but about the rate at which it complex with iron
Micellar–mediated chemiluminescence (RMM-CL) very little is known so there is need to isolate and
detection with online solvent extraction for the iron (III) characterize such biological ligand
determination89. A reversed micelle is a droplet in an
organic solvent of low polarity, where their polar groups are REFERENCES
concentrated into inner aqueous core of the aggregate, while 1. Svehla G. Vogel’s Qualitative Inorganic analysis, Reactions of the
their hydrophobic moieties extend into to the bulk organic cations. G. Svehla (ed.), 7th edition, Longman publishers, New York
solvent. The water pool, formed by water molecules 2006; pp. 109-118
2. Laulhere P J and Briat J F. Iron release and uptake by plant ferritin:
included in the core of reverse micelles, provides a unique effects of pH, reduction and chelation. Biochem.J. 1993; 290: 693-
and versatile reaction field, referred as microreactors. 699
Modification of this method would provide for iron(III) 3. Ali Akl M, Mori Y and Sawada K. Solvent sublation and
analysis only, Fe(II) + Fe(III), or iron(II) and iron(III) in the spectrophotometric determination of iron (II) and total iron using 3-
(2_pyridyl_-5-6-bis (4-phenylsulfonic acid)-1, 2, 4-triazine and
same sample using the hydrogen peroxide cycling by which Tetrabutylammonium bromide. Analytical Sciences. 2006; 22: 1169-
a speciation analysis of iron(II,III) is expected to be made 1174
possible. 4. Bothwell TH, Baynes RD, MacFarlane BJ and MacPhail AP.
Nutritional iron requirements and food iron absorption. Journal of
Internal Medicine. 1989; 226: 357-365

359
 Asian J. Research Chem. 4(3): March 2011

5. Fidler MC. Optimizing the absorption of fortification iron. Ph.D. a comparative investigation of chromogens improved tripyridyltrianne
Thesis, 2003, Swiss Federal Institute of Technology, Zurich micromethod. J Clin Chem Clin Biochem. 1988; 26: 697-703
6. Guerinot ML and Yi Y. Iron: Nutritious, noxious, and not readily 32. Verhasselt B, Delanghe J, Verstraete A and Leroux-Roels G.
available. Plant physiol. 1994; 104: 815-820 Colorimetric quantification of urinary iron during desferioxamine
7. Borch Iohnsen B. Determination of iron status: brief review of therapy. Clinical chemistry. 1991; 40 (3): 497-498
physiological effects on iron measures. Anlyst. 1995; 120(3): 891-893 33. Ceriotti Ferruccio AR. Method and composition for direct
8. Tang LC. Determination of iron and copper in 18th and 19th century determination of iron in blood serum. United States Patent 4308027.
books by flameless atomic absorption spectroscopy. JAIC. 1978; 17: 1981
19-32 34. Carter P. Spectrophotometric determination of serum iron at the
9. Golovina AN and Levshin LV. Chemical Luminiscent Analysis of submicrogram level with new reagent (ferrozine). Analytical
Neorganical Substances. Moscow: Khimiya. 1978 biochemistry. 1971; 40(2): 450-458
10. Korostelev PP. Photometric and Complexometric Analysis in 35. Seligman PA and Schleicher RB. Comparison of Methods Used to
Metallurgy. Moscow: Metallurgy. 1984 Measure Serum Iron in the Presence of Iron Gluconate or Iron
11. GOST (All union State Standard), Foodstuffs. Method for Dextran. Clinical Chemistry. 1999; 45: 898-901
determination of iron. Moscow: Izd. Standartov, 1986. 26928-86 36. Bothwell TH, Baynes RD, MacFarlane BJ and MacPhail AP.
12. Practical Guidelines. Atomic Absorptive Methods for Determination Nutritional iron requirements and food iron absorption. Journal of
of Toxic Elements in Foodstuffs and Food Raw Materials: Internal Medicine. 1989; 226: 357-365
Moscow.1992 37. Young DS and Hicks JM. Method for automatic determination of
13. GOST (All union State Standard), Drinking water. Methods for serum iron. J. Clin. Path. 1965; 18: 98-102
Determination of Total Iron, Moscow: Izd. Standartov, 1972. 4011-72 38. Tennant GB and Greenman DA. Determination of iron in solutions
14. Practical Guidelines. Determination by Flurometric Method of Total containing iron complexes. J. Clin. Path. 1969; 22: 301-303
Iron in Drinking Water and Surface and Ground Springs of Water 39. Mendham J, Denney RC, Barnes JD and Thomas MJK. Titrimetric
Uses: 4.1:064-96), 1996. Moscow: Minzdrav RF analysis In Vogel’s Textbook of quantitative chemical analysis.
15. GOST (All union State Standard), Wines, wine base, cognacs and Pearson education , Singapore. 2000; 411: 422-423
cognac spirits. Method for determination of iron, Moscow: Izd. 40. American public Health Association (APHA). Standard methods for
Standartov, 1973. 13195-73 the examination of water and wastewaters. 1998
16. Dedyukhina EV, Pechishcheva NV, Shunyaev KYu, Osintseva EV 41. Karamanev DG, Nikolov LN and Mamatarkova V. Rapid
and Neudachina LK. Fluorometric determination of iron with some simultaneous quantitative determination of ferric and ferrous ions in
N-aryl-3-aminopropionic acids. Seventh Russian-Israeli Bi-National drainage waters and similar solutions. Minerals Engineering. 2002;
Workshop 2008 15: 341–346
17. GOST (All union State Standard), Drinking Medicinal, Medicinal- 42. Paipa C, Mateo I, Poblete E and Oral T. Comparative study of
Table and Natural Table Mineral Waters. Method for Determination alternative methods for the simultaneous determination of Fe (III) and
of Iron Ions, Moscow: Izd. Standartov, 1978. 23268.11-78 Fe (II) in leaching solutions and in acid marine drainage. Minerals
18. GOST (All union State Standard), Alcohol Production and Raw for its Engineering. 2005; 18 (1): 1116-1119
making. Inversion-Voltammetry Method for determination of 43. Stalikas CD, Pappas Ach, Karayannis MI and Veltsistas PG. Simple
Cadmium, Lead, Zinc, Copper, Arsenic, Mercury, Iron and total and Selective Spectrophotometric Method for the Determination of
sulfur dioxide, Moscow: Izd. Standartov, 2001. 51823-2001 Iron (III) and Total Iron Content, Based on the Reaction of Fe(III)
19. GOST (All union State Standard), Nickel, Nickel and Copper-Nickel with 1,2-Dihydroxy-3,4-Diketocyclo-Butene (Squaric Acid).
Alloys. Method for Determination of Iron Ions, Moscow: Izd. Microchimica Acta. 2003; 142: 1-2
Standartov. 1992. 6689.5-92 44. Suwansaksri J, Sookarun S, Wiwanitkit V, Boonchalermvichian C
20. Treiber W and Weilheim DE. Method and reagent for the and Nuchprayoon I. Comparative Study on Serum Iron Determination
interference-free determination of iron. United States Patent 6274382. by Different Methods. Laboratory Hematology. 2003; 9(4): 234-36
2001 45. Wilson R F and Daniels RC. Spectrophotometric determination of
21. Zenovia M and Eleonora A. Spectrophotometric determination of iron with bis (2-hydroxymethyl-5-hydroxy-4-pyrone-6) ketone.
trace iron (III) after its preconcentration with a chelating resin. J. Fresenius journal of analytical chemistry. 1963; 194(3): 190-194
Serb. Chem. Soc. 2002; 67(10): 669-676 46. Hayashi K, Sasaki Y, Tagashira S, Soma Y, Ichinose T and Akiyama
22. Bleach ECN-2-ML-3260. Spectrophotometric Determination of Iron H. A Sensitive Spectrophotometric Determination of Iron (III) with
(Total) and Iron (II) in “ML” Processing KODAK Motion Picture Amines and Thiocyanate. Analytical Sciences. 1986; 2(5): 457-460
Films, Module 3, Analytical Procedure.H24.03 47. Wong SN. Colorimetric determination of iron and hemoglobin in
23. Malik AK. Direct Spectrophotometric Determination of Ferbam (Iron blood. II. J. of Biological Chemistry. 1928; 409-412
(III) Dimethyldithiocarbamate) in Commercial Sample and Wheat 48. Stephens BG, Felkel HL and Spinell WM. Spectrophotometric
Grains Using 4, 7-Diphenyl-1, 10-phenanthroline. J. Agric. Food determination of copper and iron subsequent to the simultaneous
Chem. 2000; 48 (12): 5808–5811 extraction of bis(2,9-dimethyl-1,10-phenanhroline)copper(I) and
24. Lee K and Ydesdale FMCL. Quantitative determination of elemental, bis[2,4,6-tri(2-pyridyl)-1,3,5-triazine] iron(II) into propylene
ferrous, ferric, soluble and complexed iron in foods. Journal of Food carbonate. Analytical chemistry. 1974; 46 (6): 692
Science. 1979; 44(2): 549-554 49. Berman L. A rapid method for the determination of iron in small
25. Pizzaro F, Amar M and Stekel A. Determination of iron in stools as a quantities of blood. J. of biological chemistry. 1918; 231-236
method to monitor consumption of iron-fortified products in infants. 50. Sugesaka Y, Okamoto Y, Fujiwara T and Kumamaru T. Heated
Am J. Clin Nutr. 1987; 45: 484 - 487 quartz cell Atomic absorption spectrometry coupled with in situ
26. Kawakubo S, Naito A, Fujihara A and Iwatsuki M. Field butylation/vaporization for the determination of iron and tin.
determination of trace iron in fresh water samples by visual and Analytical Sciences. 2001; 17: 667-669
spectrophotometric methods. Analytical Sciences. 2004; 20: 1159- 51. Olsen ED, Jatlow PL, Fernandez FJ and Kahn HL. Ultramicro
1163 Method for Determination of Iron in Serum with the Graphite
27. Barry M. Determination of chelated iron in the urine. J. Clin. Path. Furnace. Clinical Chemistry. 1973; 19: 326-329
1968; 21: 166-168 52. Brecque LJJ, Mendelovici E, Villalba RE and Bellorin CC. The
28. Stookey LL. Ferrozine- A new spectrometric reagent for iron. Determination of Total Iron in Venezuelan Laterites: The
Analytical Chemistry. 1970; 42(7): 779-781 Investigation of Interferences of Aluminum and Silicon on the
29. Yatsooff RW, Wayne BA and Tenenbein M. An objective criterion Determination of Iron in the Fluoboric-Boric Acid Matrix by Atomic
for the cessation of deferoxamine therapy in the acutely iron poisoned Absorption. Applied Spectroscopy. 1978; 32(1): 57-60(4)
patient. Clin Toxicol. 1991; 29: l -10 53. Kenduzler E and Turker AR. Determination of iron, manganese and
30. Liang L, DýHaese PC, Lambert LV and De Broe ME. Direct zinc in water samples by flame atomic absorption spectrophotometry
determination of iron in urine and serum using graphite furnace after preconcentration with solid-phase extraction onto Ambersorb
atomic absorption spectrometry. Analyst. 1989; 114: 143-147 572. Analytical sciences. 2002; 18: 917-921
31. Solem E, Seiffert B, Jalner U, Kaul M, Behnken LI and FrohlichK 54. Bag H, Turker AR, Tuncell A and Lale M. Determination of Fe (II)
Zirker M. Automated determination of serum (plasma) and urine iron: and Fe (III) in water by flame atomic absorption spectrophotometry

360
 Asian J. Research Chem. 4(3): March 2011

after their separation with Aspergillus niger immobilized on sepiolite. Journal of Liquid Chromatography and Related Technologies. 2009;
Analytical sciences. 2001; 17:901-904 32(2)P: 281-292
55. Torre M, Gonzalez MC, Jimenez O and Rodriguez AR. Study of 73. Takagai Y and igarashi S. Simultaneous determination of iron (II)
analytical methods for iron determination in complex organic liquids and iron (III) by micellar elcerokinetic chromatography using an off-
by atomic absorption spectrometry. Analytical letters. 1990; 23(8): line selective complexing reaction. Analytical Sciences. 2003; 19:
1519-1536 1207-1209
56. Pehlivan E and Kara D. Iron speciation by solid phase extraction and 74. Bobrowski A and krolicka A. A novel catalytic adsorptive stripping
flame atomic absorption spectrometry using N, N -bis-(2-hydroxy-5- voltammetric method of iron determination as its 1, 2-
bromobenzyl)-2-hydroxy-1, 3-diiminopropane. Microchimica Acta. Cyclohexanedione dioxime complex. Electroanalysis. 2002; 15,5-6
2007; 158(1-2): 137-144 75. Kuwabara M, Katsumata H, Teshima N, Kurichara M and
57. Yurchenko OI and Kharenko IP. Increasing the sensitivity of atomic Kawashima T. Successive potentiometric titration of Iron (II) and Iron
absorption determination of iron. J. of Applied Spectroscopy. 2007; (III) with cobalt (II) in the presence of 1, 10-phenanthroline.
74(2): 295-299 Analytical Sciences. 1999; 15: 657-660
58. Jurgens J, Paama L and Leito I. The uncertainty of UV–Vis 76. Jian-Fu Z, Si-Qing X, Hong-Wen G and Ya-Lei Z.
spectrophotometric and FAAS analysis for the determination of iron Spectrophotometric titration of iron using Erichrome Blue Black R
in pharmaceutical products. Accreditation and Quality Assurance: and cetyltrimethylammonium bromide. Instrumentation science and
Journal for Quality, Comparability and Reliability in Chemical technology. 2004; 32(1): 77-91
Measurement. 2007; 12(11): 593-601 77. Starke K. Iodometric titration of free acid in iron (III) solutions and
59. Saleh M, Neves R, Moraes P, Lima Dos P, Silva F and Padilha P. determination of basic iron (III) compounds. Anal. Chem. 1963;
Iron determination by FAAS in fish feed and feces after ultrasound- 35(9): 1310-1311
assisted extraction. Sensing and Instrumentation for Food Quality and 78. Kas M and Ivaska A. Spectrophotometric determination of iron(III)
Safety. 2008; DOI 10.1007/s 11694-008-9066-9 and total iron by sequential injection analysis [Link].
60. Teixeira LSG, Costa ACS, Garrigues S and Guardia M. Flow- 2002; 58(6): 1131-1137
injection solid phase partial least-squares spectrophotometric 79. Sakai T, Teshima N, Sakashita M and Ura N. New design of a multi-
simultaneous determination of iron, nickel and zinc. J. Braz. Chem. channel micro cell for simultaneous analysis in the flow-injection
Soc. 2002; 13(1): 54-59 method. Analytical sciences. 2000; 16: 251-252
61. Garcia FC, Checa R, Avidad R and Capitán-Vallvey LF. 80. Lunvongsa S, Tsboi T and Motomizu S. Sequential determination of
Determination of iron at ng/ml level by solid phase spectrophotometry trace amounts of iron and copper in water samples by flow injection
after preconcentration on cation exchange filters. Talanta. 1995; analysis with catalytic spectrophotometric detection. Analytical
42(5): 711-715 sciences. 2006; 22: 169-172
62. Shida J and Masuda I. Photoacoustic spectrometric determination of 81. Asan A, Andac M, Isildak I and Tinkilic N. Flow injection
trace iron (II) after preconcentration on a membrane filter with a spectrophotometric determination of iron (III) using diphenylamine-4-
finely pulverized anion-exchange resin. Analytical sciences. 1998; 14: sulfonic acid sodium salt. Chemical Papers. 2008; 62 (4): 345–349
333-336 82. Yaqoob M, Waseem A and Nabi A. Determination of total iron in
63. Delgado-Vasallo O, Pena JL, Martinez SN, Calderon A, Pena fresh waters using flow injection with potassium permanganate
Rodriguez G, Jaime Fonseca MR and Marin E. Photoacoustic chemiluminescence detection. J. of analytical chemistry. 2006; 61 (9):
determination of iron in corn meal. Analytical sciences. 2003; 19: 917-921
599-602 83. Robert John L. Determination of iron using potassium dichromate:
64. Khvostikov V, Grazhulene S, Golloch A, Telgheder UG and Joosten redox indicator. 2006.
H. Investigation of iron determination in HCl gas using the [Link] [Link]
nondispersive atomic fluorescence spectrometer with a tantalum coil 84. Fischer RB and Peters DG. Redox determination of iron. Quantitative
atomizer. Spectrochimica acta part B: Atomic spectroscopy. 1997; chemical analysis. Philadelphia, 1968; 12-18, 51-65.536-546
52(9-10): 1551-1557 85. Mattos CDS, Carmo DR, Oliveira MF and Stradiotto NR.
65. Nixon DV, Butz J, Eckdahl SJ, Burritt MF, Naubauer KR and Wolf Voltammetric determination of total iron in fuel ethanol using a 1, 10
RE. Determination of copper and iron in liver tissue using the ELAN Fenantroline/Nafion carbon paste-modified electrode. Int. J.
DRC ICP-MS. Application note. Perkin Elmer Sciex instruments. Electrochem. Sci. 2008; 3: 338 – 345
http: //[Link] 86. Thomas F, Sterratrice G, Beguin C, Aman ES, Pierre JL, Fontecare M
66. Ide-Ektessabi A, Rabionet M. The role of trace metallic elements in and Laulhere JP. Calcein as a fluorescent probe for ferric ion. Journal
Neurogenerative Disorders; Quantitative Analysis using XRF and of Biol. Chemistry. 1999; 274(19): 13375-13383
XANES spectroscopy. Analytical Sciences. 2005; 21: 885-892 87. Zhu G, Zhu Z and Qiu L. A fluorometric method for the
67. Bower NW. Simple Spectrophotometric Determination of Ferrous determination of iron (II) with fluorescein isothiocyanate and iodine.
Iron in Twelve French Geochemical Reference Standards. Analytical sciences. 2002; 18:1059-1061
Geostandards and geoanlaytical research. 2007; 8(1): 61-62 88. Teixeira Leonardo SG, Rocha Rodrigo BS,; Sobrinho eledir V,;
68. Virro K, Mellikov E, Volobujeva O, Sammelselg V, Asari J, Paama Guimaries Paulo RB, Pontes luiz AM and Teixeira josanaide SR.
L, Jurgens J and Leito I. Estimation of uncertainty in electron probe Simultaneous determination of copper and iron in automotive
microanalysis: iron determination in manuscripts, a case study. gasoline by X-ray fluorescence after pre-concentration on cellulose
Microchimica Acta. 2008; 162(3-4): 313-323 paper. Talanta. 2007; 72(3): 1073-1076
69. Khan MN. kinetic determination of iron and spectrophotometric 89. Kyaw T, Fujiwara T, Inoue H, Okamoto Y and Kumamaru T.
determination of mercury by exploiting the reaction of neutral red. Reversed micellar mediated luminol chemiluminescence detection of
Ph.D. thesis. Department of Chemistry/ University of Karachi. iron (II, III) combined with on-line solvent extraction using 8-
Pakistan research repository. 2006. quinolinol. Analytical Sciences. 1998; 14: 203-207
70. Garcia-Marco S, Torreblanca A and Lueena JA. Chromatographic 90. Peramaki P, Kumpumaki M, Valimaki I and Heikkainen R.
determination of Fe chelated by Ethylenediamine-N (o- Preliminary studies of iron speciation (Fe2+ and Fe3+) in peat samples
hydroxyphenylacetic)-N’-(p-hydroxyphenylacetic) Acid in using polarography. Analytical Sciences. 2000; 16: 751-756
commercial EDDHA/Fe3+ fertilizers. J. [Link] Chem. 2006; 541: 91. Faulkner SP, Gambrell RP and Ashby SL. Analytical Methods for
380-1386 Iron and manganese determinations in reservoir tailgaters: Laboratory
71. Garcia-Mina J, Cantera RG and Zamarreno A. Interaction of different investigations. 1996; Water quality technical note PD-01:1-10
iron chelates with an alkaline and calcareous soil: A complementary 92. Achterberg Eric P, Holland Toby W, Bowie Andrew R, Mantoura R,
methodology to evaluate the performance of iron compounds in the Fauzi C and Worsfold Paul J. Determination of iron in seawater.
correction of iron chlorosis. Journal of Plant Nutrition, 2003; Analytica Chimica Acta. 2001; 442: 1–14
26(10):1943-1954
72. Susumu I, Shogo F and Youichi F. Selective Determination of Iron
Ion in Tap Water by Solvent Extraction with 3, 4-Dihydro-3-hydroxy-
4-oxo-1, 2, 3-benzotriazine, Followed by Reversed Phase HPLC.

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