A universal calibration method for flame atomic absorption analysis

Dr. Joachim Dahmen, Dr. Gerald Fuchs-Pohl, Annelore Bischoff Central Analytical Laboratory, E.Merck, Darmstadt, Federal Republic of Germany Dr. Barbara Pohl, Varian GmbH, Darmstadt, Federal Republic of Germany

Number AA-88 June 1989

Introduction
Flame atomic absorption analysis is a relatively simple and quick method for the determination of a series of elements in the ng/mL range. G.M. Hieftje, USA, states when he was interviewed by the late J.M. Ottaway, United Kingdom, in 1985 (1) : .. I would say that the use of flames in atomic absorption will continue to exist prominently side by side with furnace techniques. My own feeling is that it is wise to use flame whenever possible and furnaces should be used only whenever a flame wont suffice. In flame atomic absorption spectrometry chemical and physical interferences are fewer than in furnaceAAS, nevertheless they exist. Physical interferences are caused by differences in the physical properties (such as density, surface tension, viscosity, temperature) of the sample and the calibration standard solutions. These parameters control the aspiration rate of the nebulizer, the size distribution of the produced aerosol and thus the volatilization behaviour of the dissolved sample solution. Ionization interference, which is also a physical interference, is based on differences in the formation of ions, which are species not detected by normal atomic absorption. This type of interference occurs especially in the determination of the easily ionized alkali and alkaline-earth elements. Chemical interferences are due to the formation of chemical species during nebulization and atomization which alter the atomization efficiency compared to aqueous standard solutions.

Due to the above mentioned interferences, analysis of a complex sample matrix against a calibration graph, which is obtained by measurements of aqueous standard solutions, often leads to systematic errors and incorrect results. Several methods are available to eliminate these interferences and to guarantee more accurate results: (1) The use of standard solutions with similar matrix content. (2) The well known standard additions method. (3) Chemical pretreatment such as matrix isolation. (4) The use of a nitrous oxide/acetylene flame instead of an air-acetylene flame. The performance of most of these methods is very time consuming and thus expensive. A fast universal calibration method was described by Schuhknecht and Schinkel, Analytical Laboratories of Saarbergwerke, FRG (2). They recommended, as a spectroscopic and physical buffer system, an addition of cesium chloride and aluminium nitrate to the samples and standard solutions in the flame emission spectroscopic determination of sodium, potassium and lithium. Cesium was chosen because of its low ionization energy. Its addition leads to a large quantity of free electrons in the flame. The ionization of sodium, potassium and lithium is suppressed and their mutual interferences can then be neglected. 1 The alkaline-earth elements are the main interfering elements in the determination of sodium, potassium

and lithium. This interference is suppressed by an addition of aluminium nitrate, because compounds with a low volatility are found with alkaline-earth elements. Another effect of the addition of cesium chloride and aluminium nitrate is the formation of solutions with unique or almost unique physical properties, i.e. with respect to density, surface tension and viscosity. Therefore, in the case of various matrices a one-point calibration is sufficient to obtain accurate results. A partially-rotated burner with a reduced flame width in the light path, and therefore reduced sensitivity, is often applied to analyze higher analyte concentrations. This emission technique of Schuhknecht and Schinkel (2) is also applied successfully in flame atomic absorption analysis. In 1984 Schinkel (3) extended the method to the flame atomic absorption spectrometric determination of fourteen elements (Ca, Cd, Co, Cr, Cu, Fe, K, Li, Mg, Mn, Na, Ni, Sr, Zn) in a variety of sample types by the use of an addition of cesium chloride and lanthanum chloride. Both recommended buffers (CsCl/Al(NO3)3 and CsCl/ LaCl3) are commercially available as concentrated solutions which can simply be added to the samples and standard solutions. In this paper both methods are applied to the determination of various elements in diet drinks, diet menus and to the determination of magnesium in human serum. Results from different calibration procedures are compared to establish the accuracy of the buffer methods. In the case of the determination of magnesium in human serum an enzymatic method is compared. Equipment All measurements were carried out with a Varian SpectrAA-40 flame atomic absorption spectrometer and Varian hollow cathode lamps. An air-acetylene flame was used for all determinations except for calcium where a nitrous oxide-acetylene flame was also used. Deuterium background correction was applied whenever needed. Reagents All reagents were supplied by E.Merck, Darmstadt, Federal Republic of Germany. a) Titrisol<R> for the preparation of 1 L sodium, potassium and lithium standard solution after Schuhknecht and Schinkel (Merck-No. 9985), concentrations 0.1 g/L respectively, called solution A.

(b) Cesium chloride/aluminium nitrate buffer solution after Schuhknecht and Schinkel (Merck-No. 2037), called solution B. (c) Cesium chloride/lanthanum chloride buffer solution after Schinkel (Merck-No. 16755). (d) Titrisol<R> for the preparation of 1 L magnesium standard solution (Merck-No. 9949), concentration 1 g/L. (e) Titrisol<R> for the preparation of 1 L calcium standard solution (Merck-No. 9943), concentration 1 g/L. (f) Titrisol<R> for the preparation of 1 L zinc standard solution (Merck-No. 9953), concentration 1 g/L. (g) Titrisol<R> for the preparation of 1 L iron standard solution (Merck-No. 9972), concentration 1 g/L. (h) Titrisol<R> for the preparation of 1 L sodium standard solution (Merck-No. 9927), concentration 1 g/L. (g) Titrisol<R> for the preparation of 1 L potassium standard solution (Merck-No. 9924), concentration 1 g/L. Sample Pretreatment For the analysis of powdered diet menus and diet drinks a sample pretreatment is necessary. Sixty millilitres of a mixture of nitric and hydrochloric acid (90 mL HCl 25% +10 mL HNO3 65%) is added to approximately 10 g of the accurately weighed homogenized sample in a 200 mL volumetric flask. With continuous shaking the flask is heated in a boiling water bath for 30 minutes. Then approximately 90 mL deionized water is added, carefully mixed and the digestion continued for another 30 minutes. After cooling the solution is made up to volume with deionized water and centrifuged. The supernatant liquids are diluted to a suitable concentration range. Serum samples are analyzed after one dilution step without any further pretreatment. The concentrated buffer solution is added in the last dilution step (1:10 of the resulting volume). Calibration procedures An example for the preparation of five buffered multielement standard solutions of sodium, potassium and lithium after the method of Schuhknecht and Schinkel (2) in the range from 2 to 10 mg/L is given in Table 1. Solution A is the mixed standard solution, 0.1 g/L of sodium, potassium and lithium respectively. Solution B is the concentrated buffer solution (2). Table 2 presents the resulting composition of the calibration standard solutions.

Table 1 Preparation of buffered calibration standard solutions (2) for Na, K and Li, 100 mL each made up to volume with deionized water.
Solution A Blank Standard Standard Standard Standard Standard 0 2 4 6 8 10 Solution B 10 10 10 10 10 10 Deionized water 90 88 86 84 82 80 (mL) (mL) (mL)

1 2 3 4 5

Table 2 Compositions of the buffered calibration standard solutions Na

Blank Standard 1 Standard 2 Standard 3 Standard 4 Standard 5 (mg/L) 0 2 4 6 8 10

K
(mg/L) 0 2 4 6 8 10

Li
(mg/L) 0 2 4 6 8 10

CsCl
(g/L) 5 5 5 5 5 5

Al (NO3)3.9 H2O
(g/L) 25 25 25 25 25 25

Buffered standard solutions of the other elements are obtained by similar dilution schemes. A mixture of cesium chloride and lanthanum chloride serves as the buffer system [3]. For one-point calibrations, only one standard with a maximum absorbance of 0.5 is used. If more than 0.5 absorbance units are detected the burner is partially rotated to an angle where this maximum absorbance is approximately achieved. In the standard addition mode two known amounts of the element to be determined are added to the diluted, unbuffered samples Results and Discussion 1. DIET MENU AND DIET DRINKS The sample labels of the analysed diet menus and diet drinks are listed in Table 3. Tables 4 - 7 present the results of Fe, Zn, Mg and Ca measurements with three different calibration procedures:

A.

1-point calibration (3), concentrations:

1 mg Fe / 200 mL, 0.2 mg Zn / 200 mL, 1 mg Mg / 200 mL, 1 mg Ca / 200 mL B. C. 5-point calibration buffered as per (3) Standard addition method

For the Ca measurements by standard addition a nitrous oxide-acetylene flame and the less sensitive wavelength of 239.9 nm were used.

Table 3 Sample Labels

Sample No. 1 2 3 4 5 6 7 8 9 Sample Label Blackcurrant drink I Blackcurrant drink II Beef broth I Beef broth II Beef broth III Pineapple drink I Pineapple drink II Orange drink I Orange drink II

Table 4
Fe results using different calibration procedures (mg/200 mL) Wavelength: 248.3 nm, air-acetylene flame Calibration Procedure Sample No. Dilution
1 2 6 7 8 9 1:2 1:2 20:25 20:25 20:25 20:25

A Result
1.0 1.3 0.55 0.53 0.53 0.56

B Dilution
1:2 1:2 20:25 20:25 20:25 20:25

C Dilution
1:5 1:5 1:4 1:4 1:4 1:4

Result
0.95 1.3 0.52 0.51 0.52 0.54

Result
0.98 1.3 0.59 0.54 0.55 0.52

Table 5 Zn results using different calibration procedures (mg/200 mL) Wavelength: 213.9 nm, air-acetylene flame Calibration Procedure Sample No. Dilution
6 7 8 9 20:25 20:25 20:25 20:25

A Result
0.15 0.16 0.16 0.15

B Dilution
20:25 20:25 20:25 20:25

C Dilution
1:2 1:2 1:2 1:2

Result
0.14 0.15 0.16 0.15

Result
0.14 0.15 0.16 0.14

Table 6 Mg results using different calibration procedures (mg/200 mL) Wavelength: 285.2 nm, air-acetylene flame, partial burner rotation.
Calibration

Procedure
C Dilution
1:100 1:100 1:100 1:100 1:100 1:25 1:25 1:25 1:25

Sample No. Dilution

1 2 3 4 5 6 7 8 9 1:10 1:10 1:10 1:10 1:10 1:5 1:5 1:5 1:5

A Result
8.0 9.9 7.9 6.5 6.9 4.6 4.5 4.5 4.5

B Dilution
1:10 1:10 1:10 1:10 1:10 1:5 1:5 1:5 1:5

Result
7.6 10.2 7.5 6.1 6.5 4.7 4.7 4.6 4.7

Result
7.3 9.8 8.2 6.7 7.2 4.4 4.4 4.2 4.5

Table 7 Ca results using different calibration procedures (mg/200 mL) Wavelength: 422.7 nm, air-acetylene flame (method A and B) Wavelength: 239.9 nm, nitrous oxide-acetylene flame (Method C) Sample No. A Dilution
1 2 3 4 5 6 7 8 9 1:50 1:50 1:50 1:50 1:50 1:50 1:50 1:50 1:50

Calibration Procedure B Dilution

1:50 1:50 1:50 1:50 1:50 1:50 1:50 1:50 1:50

C Dilution
1:5 1:5 1:5 1:5 1:5 1:4 1:4 1:4 1:5

Result
45 55 49 47 51 31 31 30 31

Result
45 55 48 47 51 32 31 30 30

Result
49 61 52 49 57 31 34 33 33

The relative standard deviations of five replicate measurements are between 0.5 and 2%. The 1-point calibration procedure (3) is sufficient and its performance is less time consuming than others. The differences between the results obtained with the 1-point calibrations (3) and the results obtained by standard additions never exceed 10%.

In the determination of sodium and potassium the following calibration procedures were compared: A. 1-point calibration (3), concentrations: 1 mg Na / 200 mL, 1 mg K / 200 mL B. 5-point calibration, buffered as in (3) C. 1-point calibration (2) The results are presented in Tables 8 and 9.

Table 8 Na results using different calibration procedures (mg/200 mL) Wavelength: 589 nm, air-acetylene flame, partial burner rotation Calibration Procedure Sample No. Dilution
1. 2 3 4 5 6 7 8 9 1:10 1:20 1:200 1:200 1:200 1:20 1:20 1:20 1:20

A Result
8.6 13.5 164 135 143 14.8 14.6 15.1 15.7

B Dilution
1:10 1:20 1:200 1:200 1:200 1:20 1:20 1:20 1:20

C Dilution
1:20 1:20 1:250 1:250 1:250 1:20 1:20 1:20 1:20

Result
8.5 13.4 158 132 141 15.1 14.8 15.6 16.5

Result
8.8 10.4 160 125 140 14.4 14.8 14.0 14.8

Table 9 K results using different calibration procedures (mg/200 mL) Wavelength: 766.5 nm, air-acetylene flame, partial burner rotation Calibration Procedure Sample No. Dilution
1 2 3 4 5 6 7 8 9 1:50 1:100 1:100 1:50 1:50 1:50 1:50 1:50 1:50

A Result
44 61 53 40 43 44 43 42 42

B Dilution
1:50 1:100 1:100 1:50 1:50 1:50 1:50 1:50 1:50

C Dilution
1:50 1:100 1:100 1:50 1:50 1:50 1:50 1:50 1:50

Result
43 57 49 38 41 46 45 43 44

Result
41 56 50 38 40 48 47 45 46

In the determination of sodium and potassium no significant differences between the calibration procedures were observed, so that the 1-point calibrations (3) or (2) were chosen for future determinations 2. DETERMINATION OF MAGNESIUM IN SERUM For the determination of magnesium in serum a 5point calibration with magnesium concentrations between 1 and 5 mg/L was used. The cesium chloride/lanthanum chloride buffer after Schinkel was added. The burner was rotated to a position where a maximum absorbance of approximately 0.5 was achieved. Ninety-five samples of human serum and control serum were analyzed against the buffered calibration graph. The same samples were analyzed by an enzymatic method and excellent agreement was shown with the atomic absorption results (Figure 1).
Figure 1. Determination of magnesium in serum, results obtained by atomic absorption vs an enzymatic method [mmol/L].

These universal calibration methods of Schuhknecht and Schinkel (2) and of Schinkel (3) are introduced as additional worthwhile tools in every flame atomic absorption laboratory.

Conclusions
The applications described show only a few examples for the applicability of the methods of Schuhknecht and Schinkel (2) and of Schinkel (3). If the analyte is in a medium concentration range, it is possible to obtain accurate results with acceptably short analysis times by the addition of concentrated buffer solutions to all the measured solutions.

References
1. 2. 3. G.M. Hieftje, J. Anal. At. Spectrom. 1, 3, (1986). W. Schuhknecht, H Schinkel, Fresenius Z. Anal. Chem 194, 161, (1963) (in German) H. Schinkel, Fresenius Z. Anal. Chem. 317, 10, (1984) (in German)