Reference TOTL-WI-18

Office Egypt

ASTM E394- 09 Issue n° / Date 00 / 01 Novamber 20

Iron in Trace Quantities Using the 1.10 Author / Approver M.Saleh / M. Abd Elrazek

Phenanthroline Method Page n° Page 1 of 8

Reviewed & Approved By Author

Mohamed Abd Elrazek Mohamed Saleh Name

TOTL Manager Technical team leader Position

Signature
05/11/2020 01/11/2020 Date

Control of Amendments

Issue Amendments Echoed

Date Reason for amendments
No. Summary Pages
0.0 01.11.2020 First issue - -
1.0
2.0
3.0
4.0
5.0

Review Frequency: Annually

1
 Reference TOTL-WI-18

Office Egypt

ASTM E394- 09 Issue n° / Date 00 / 01 Novamber 20

Iron in Trace Quantities Using the 1.10 Author / Approver M.Saleh / M. Abd Elrazek

Phenanthroline Method Page n° Page 2 of 8

:Contents
1.0 Scope……………………………………………………………………………3
2.0 Referenced Documents ………………………………………………..3
3.0 Summary of test method………………………………………………3
4.0 Significance and Use …………………………………………………….3
5.0 Interferences ……………………………………………………………….4
6.0 Reagents and materials …………………………………………….…5
7.0 Sampling ……………………………………………………………………..5
8.0 Apparatus…………………………………………………………………….5
9.0 Calibration……………………………………………………………………6
10.0 Procedure ………………………………………………………………….6
11.0 Calculation …………………………………………………………….....7
12.0 Report ……………………………………………………………………….7
13.0 Precision and Bias ……………………………………………………..8

2
 Reference TOTL-WI-18

Office Egypt

ASTM E394- 09 Issue n° / Date 00 / 01 Novamber 20

Iron in Trace Quantities Using the 1.10 Author / Approver M.Saleh / M. Abd Elrazek

Phenanthroline Method Page n° Page 3 of 8

1.0 Scope
1.1 This test method covers the determination of iron in the range from 1 to 100 µg.
This test method is intended to be general for the final steps in the
determination of iron and doesn`t include procedures for sample preparation.
1.2 This test method is applicable to samples whose solutions have a PH less than 2.
It is assumed that the PH is adjusted to within this range in the sample
preparation.

2.0 Referenced Documents

2.1 ASTM standards.
-D1193 Specification for Reagent water
-E60 Practice for Analysis of Metals, Ores and Related materials by
Spectrophtometer.
-E180 Practice for Determining the Precision of ASTM methods.
-E200 Practice for Preparation, Standardization and storage of chemicals.

3.0 Summary of Test Method:

This test method is based upon a photometric determination of the 1,10-phenanthroline
complex with the iron (II) ion. The sample is dissolved in a suitable solvent and the iron is
reduced to the divalent state by the addition of hydroxylamine hydrochloride. The color
is then developed, by the addition of 1,10-phenanthroline. After a short reaction period,
the absorbance of the solution is measured at approximately 510 nm using a suitable
photometer. The absorbance of the solution, once the color is developed, is stable for at
least several months.

4.0 Significance and Use:

This test method is suitable for determining trace concentrations of iron in a wide
variety of products, provided that appropriate sample preparation has rendered the iron
and sample matrix soluble in water or other suitable solvent. This method assumes that
the amount of color developed is proportional to the amount of iron in the test solution.
The calibration curve is linear over the spectrum range.

3
 Reference TOTL-WI-18

Office Egypt

ASTM E394- 09 Issue n° / Date 00 / 01 Novamber 20

Iron in Trace Quantities Using the 1.10 Author / Approver M.Saleh / M. Abd Elrazek

Phenanthroline Method Page n° Page 4 of 8

5.0 Interferences
Table 1 and Table 2 show the effects of various cations and anions on the determination
of 2.0µg/g(ppm) iron. If the maximum level of 500 µg/g (ppm) doesn`t interfere, it is
very likely that the ion will not interfere in any quantity. The data were obtained under
slightly different conditions than those specified in the present test method, but the
interferences should be similar.

4
 Reference TOTL-WI-18

Office Egypt

ASTM E394- 09 Issue n° / Date 00 / 01 Novamber 20

Iron in Trace Quantities Using the 1.10 Author / Approver M.Saleh / M. Abd Elrazek

Phenanthroline Method Page n° Page 5 of 8

6.0 Reagents and Materials

6.1 purity of reagents, it is intended that all reagents shall conform to the
specifications of the committee on Analytical Reagents of the American
Chemical society.
6.2 purity of water, unless otherwise indicated, references to water shall be
understood to mean type II reagent water as defined in specification D1193.
6.3 Hydroxylamine Hydrochloride solution (100g/L): Dissolve 10 g of hydroxylamine
hydrochloride (HONH2 HCL) in approximately 60 ml of water, filter and dilute to
100 ml.
6.4 1.10-phenanthroline solution (3g/L): Dissolve 0.9 g of 1.10-phenanthroline
monohydrate in 30 ml of methanol and dilute to 300 ml with water.
6.5 Ammonium Acetate –Acetic Acid solution: Dissolve 100 g of ammonium acetate
(CH3 COONH4) in about 600 ml of water , filter, add 200 ml of glacial acetic acid
to the filtrate, and dilute to 1 L with water.
6.6 Iron Standard Solution ( 1 ml =0.01 mg Fe) . Dissolve 0.1000 g of iron wire in 10
ml of hydrochloric acid and 1 ml of bromine water. Boil until the excess bromine
is removed. Add 200 ml of HCL, cool, and dilute to 1 L in a volumetric flask.
Dilute 100 ml of this solution to 1 L. As an alternative, the standard iron solution
may be prepared by weighing exactly 0 .7022 g of iron(II) ammonium sulfate
hexahydrate (FeS04 (NH4)2SO4.6 H20) in 500 ml of water containing 20 ml of
sulfuric acid and diluting to 1 L with water. Dilute 100 ml to 1 L.

7.0 Sampling
Because this is a general test method for final steps in determining of iron.

8.0 Apparatus:
Photometer capable of measuring light absorption at 510 nm and holding a 5-cm or 1-
cm cell.

5
 Reference TOTL-WI-18

Office Egypt

ASTM E394- 09 Issue n° / Date 00 / 01 Novamber 20

Iron in Trace Quantities Using the 1.10 Author / Approver M.Saleh / M. Abd Elrazek

Phenanthroline Method Page n° Page 6 of 8

9.0 Calibration
9.1 By means of suitable pipets or a buret, transfer 0 (reagent blank), 2, 4, 6, 8, and
10 ml, respectively, of the standard iron solution to each of six 100-ml, glass –
stoppered graduated cylinders. These cylinders contain 0, 20, 40, 60, 80 and
100µg of iron, respectively. Dilute the contents of each cylinder to 80 ml with
water. Develop the color and measure the absorbance of each calibration
standard.
9.2 Plot, on linear graph paper, the micrograms of iron as a function of absorbance.

10.0 Procedure
10.1 Wash all the tools that will be used in the method by hot water around two
times well, then wash the same tools by distilled water many times.
10.2 Check if there is any contamination with iron by using the indicators
involved in the test method such as (2 ml hydroxylamine, 5 ml Ammonium
Acetate Buffer, 5 ml 1,10-phenanthroline). If you find red color, there is an iron
contamination. So, you should wash the tools again very well.
10.3 Add 2 ml hydroxylamine hydrochloride, 5ml ammonium acetate buffer, 5 ml
1,10-phenanthroline to a 100 ml measuring flask.
10.4 Weight the sample a weigh which include iron from 1 to 100 µg, according
to the type of sample, in a measuring flask 100 ml. Then complete using distilled
water.
10.5 Repeat 9.3 and 9.4 to prepare a blank solution with distilled water and any
chemical used in preparation of the sample.
10.6 Allow the sample solution and reagent blank to sit at room temperature for
a minimum of 15 min.
10.7 Measure the absorbance of each sample solution at approximately 510 nm
in a 5-cm cell using a suitable photometer. Use a matched 5-cm cell filled with
the reagent blank to set the instrument at zero absorbance or 100%
transmittance.

6
 Reference TOTL-WI-18

Office Egypt

ASTM E394- 09 Issue n° / Date 00 / 01 Novamber 20

Iron in Trace Quantities Using the 1.10 Author / Approver M.Saleh / M. Abd Elrazek

Phenanthroline Method Page n° Page 7 of 8

11.0 Calculations
11.1 calculate the iron content of the sample as follows:
B
Iron µg/g (ppm) =
W
Where:
B = micrograms of iron found
W= grams of sample

11.2 If the slope of the calibration line is calculated, the micrograms of iron B,
may be calculated as follows:
S
B= A×( )
L
Where:
A= absorbance
B= micrograms of iron
S= slope of the calibration line
L= cell path length (cm)

12.0 Report
Report the iron content to the nearest 0.01 µg/g(ppm).

13.0 Precision and Bias

13.1 Repeatability: the coefficient of variation for a single determination has
been estimated to be the amount shown in the table 3 at the indicated degrees
of freedom. The 95% limit for the difference between two such runs is the
amount shown in table 3.
13.2 Reproducibility: the coefficient of variation of results obtained by analysts
on different laboratories, has been estimated to be the amount shown in table
3. The 95% limit for the difference between two such averages is the amount
shown in table 3.
13.3 Bias: the bias of this test method hasn`t been determined due to the
unavailability of suitable reference material.

7
 Reference TOTL-WI-18

Office Egypt

ASTM E394- 09 Issue n° / Date 00 / 01 Novamber 20

Iron in Trace Quantities Using the 1.10 Author / Approver M.Saleh / M. Abd Elrazek

Phenanthroline Method Page n° Page 8 of 8

13.4 Laboratory precision:

The coefficient of variation of results obtained by the same analyst on different

days has been estimated to be the amount shown in table 3 at the indicated
degrees of freedom. The 95% limit for the difference between two such
averages is the amount shown in table 3.