FLAME PHOTOMETRY

Flame photometry is an atomic emission technique which may be regarded as the simplest of atomic spectroscopic methods and is very similar to the flame test which is applied for detection of alkali metals. Flame photometry is good only for elements that are easily excited and do not require very high temperatures (Na, K, Li, Ca are the most widely determined atoms by this technique) [1]. The flame excites the atoms to cause light emissions. These emissions are discrete lines that are characteristic of the elements in the flame. The photometer part has prisms or gratings so you can adjust it to read only certain wavelengths - the same as the element you want to detect and measure. Or it has a detector that can measure a variety of emission lines at once. The quantity of light you measure is proportional to the quantity of the element in the flame. Calcium, alkali metals, etc - anything that can be excited by a flame. That may be propane, acetylene/oxygen or even hydrogen/oxygen flame.

APPLICATION OF FLAME PHOTOMETRY IN BIOLOGY AND MEDICINE

1. Flame photometer for use with Serum and urine (biological samples);the BWB XP and BIO flame photometer:
There are two main differences between industrial use and instruments meant for serum and urine (we prefer the term biological over clinical). Yes, the sample size is one of them. Biological samples can be much smaller so the nebuliser needs to be configured to minimise the aspiration rate. The other difference is that the calibration curves are built-in permanently into the instrument. The BWB-XP can be used with biological uses. It just has not been optimised for that specific market. As an industrial instrument the BWB-XP has many options and settings allowing the user to configure it in many ways. The BWB-BIO will not have such an array of settings but will constrain the user to follow certain protocols. The reason is that this market does not want the user to be able to change those protocols[2].

2. The determination of sodium and potassium in biological fluids with the dual channel ultramicro flame photometer :
A modification of a dual-channel ultramicro flame photometer is described. It was built for analysis of very small amounts of biological fluids including that from renal tubules

and determines potassium and sodium simultaneously in amounts as low as 2 moles. The author's experience in the use of the modified instrument is described. It should be generally useful for analysis wherever the sample supply is very limited[3].

3. THE APPLICATION OF FLAME PHOTOMETRY TO SODIUM AND POTASSIUM DETERMINATIONS IN BIOLOGICAL FLUIDS
The need for a rapid, accurate method to determine quantitatively Na and K present together in biological fluids has been met by the procedures to be described. The necessity of separating these elements prior to their determination by the various chemical methods usually employed has led to procedures which are often prohibitively tedious and time-consuming.The recent development of flame photometry has now made possible physi-cal methods of analysis in which chemical separation of Na and K is unnecessary . Although we are dealing with Na and K determinations only, the principle of flame photometry originally used by Lundegardh and modified and called the air-acetylene flame method by Cholak and Hubbard can theoretically be applied to the determination of all the alkaline metals. At present the available commercial instruments employ a rela-tively low temperature flame which activates the ions rather than the metals themselves; the sensitivity of the photocells is such that the lines of the metals themselves would not be of sufficient intensity to measure, and suitable optical filters for other metals are not readily available[4].

4. The determination of sodium and potassium in blood serum and urine by means of the flame photometer
A method is described for the determination of sodium and potassium in blood serum and urine by means of the flame photometer. Sodium is determined by diluting serum or urine 100 times with distilled water, by spraying the solution obtained into a flame fed by a mixture of butane and air and measuring the intensity of the yellow sodium light with a selenium barrier-layer photocell. Potassium is determined by diluting serum 25 times and urine 100 times with distilled water and measuring the intensity of the red K doublet, which is excited when these solutions are sprayed into the flame, with the help of a gasfilled caesium cell with D.C. amplifier. The potassium and sodium content of the serum of 35 normal persons was determined in this way[5].

5. Determination of potassium in red blood cells using unmeasured volumes of whole blood and combined sodium/potassium-selective membrane electrode measurements.
Recent studies suggest that the measurement of intracellular potassium concentrations in red blood cells (RBC-K) can be a marker for assessing the risk, development, and treatment of hypertension. In this work, the combined use of miniature potassium- and sodiumselective membrane electrodes is evaluated as a simple means to determine RBC-K.

The proposed method requires two separate sets of electrode measurements: (i) (ii) potassium and sodium concentrations in the plasma phase of an unmeasured volume of a whole blood sample . determination of potassium and sodium concentrations in the same sample of blood after complete hemolysis by ultrasonic disruption of the RBC membranes. The dilution of sodium concentration after hemolysis can be used to determine hematocrit (Hct) (volume of red cells per unit volume of blood) of the blood[6].

(iii)

The concentration of potassium within the red blood cells (RBCs) is then calculated using the measured change in potassium levels before and after RBCs lysis and the hematocrit level determined from the sodium electrode measurements and/or a conventional centrifugation method. Good correlation for RBC-K between the proposed method and traditional flame photometry is observed for animal blood samples that possess the range of potassium levels found within human RBCs (80-120 mM)[7]. However, when potassium is much lower than that found in human RBCs (known to occur for certain animal species), the Hct measured by the sodium electrode method is falsely low, compared to traditional spun hematocrit values, because of an increased level of sodium within the RBCs, necessitating use of spun Hct levels to assess RBC-K accurately. It is envisioned that this new approach could be further miniaturized into a single-use disposable cartridge type electrode system that would enable rapid point-of-care screening of RBC-K levels in human subjects[8].

6. Direct Determination of Potassium in Human Blood Serum by Flow Injection Flame Photometry with Automatic Dilution
A method was developed for the automatic quantitative determination of potassium in serum, by flow injection analysis. The serum sample is directly introduced in the flow. The dilution with deionized water stream is done automatically and sequentially in the system to reach the adequate concentration to be introduced in the flame photometer. The results achieved with this method, for serum samples obtained in the Hospital of the University, had been compared with those values of the Hospital laboratory that uses manual dilution and flame photometry. The results obtained by the two methods agree within at least 5%. About 60 samples/hour can be easily analyzed by the method proposed in this work[9].

7. The Flame Photometer as Engine of Nephrology

In the 1940s, the flame photometer made possible for the first time relatively simple and quick measurements of sodium and potassium in serum and urine. During World War II, it unexpectedly fell into the hands of John P. Peters of Yale University, who sought to understand water and electrolyte physiology and apply such knowledge to patient problems[10]. Pupils and young associates of Peters would seed the early nephrology divisions and training programs in the United States; the flame photometer was essential to their work and that of their trainees, both Americans and international visitors. Hyponatremia and the syndrome of inappropriate antidiuretic hormone secretion became the attribute disorders of nephrologists. Invention of a microflame photometer fostered the revival of

micropuncture and transport studies. In the 1960s, the flame photometer was linked to Leonard Skeggs' sequential automated analysis system, leading to enormous numbers of routine measurements of electrolytes. The growing number of nephrologists, then based mostly at teaching hospitals, thus found plentiful instances of sodium and potassium abnormalities to address. The autoanalyzer also catalyzed use of the anion gap, another emblem of nephrology in its early decades. Not only ideas and theories, but also the usually invisible machinery, enable the growth of a knowledge base and formation of a scientific discipline or medical specialty. the flame photometer did not itself shape the agenda of nephrology, but it allowed the most influential group of progenitors and their progeny to explore normal function and bring a strongly physiologic imperative to their daily work with patients.

8. USE OF THE FLAME PHOTOMETER FOR ANALYSES OF BLOOD:

Barnes described a flame photometer and various authors have since analyzed the sources of errors in the technique and made various suggestions for eliminating them .These investigators have been primarily concerned with the design and operation of the instrument, the interference of one substance with another, and similar problems. The purpose of the present investigation was to determine the most accurate and most convenient method of preparing blood for flame photometric analyses of Na and K and to demonstrate what corrections should be applied to compensate for errors introduced in preparing the blood[11].

9. Glass electrode measurements of blood sodium and potassium

A practical, simple procedure for the estimation of (Na+) and (K+) in undiluted whole blood or serum is presented. The method uses cation-responsive glass electrodes and is based on a graphic simplification of equations and on calibration steps for deriving the necessary constants. When one electrometer is used, the procedure requires about the same time as standard flame photometry; with two electrometers, it is faster. Values for (Na+) and (K+) in our series of 38 normal young adults compare well both absolutely and in terms of variability with recently published flame photometric measurements in protein-free samples. In a random series of hospital patients, occasional striking divergences suggest that ion binding as a factor in disease warrants further investigation[12].

REFERENCES:
1. Barnes, R. B., Richardson, D., Berry, J. W., and Hood, R. L., Id. and Eng. Chem., AnaL Ed., 17, 605 (1945). by guest, on April 9, 2012 www.jbc.org Downloaded from R. R. OVERMAN AND A. K. DAVIS 649 2. Berry, J. W., Chappel, D. G., and Barnes, R. B.,Ind. and Eng. Chem., Anal. Ed., 18, 19 (1946). 3. Lundegardh, H., Die quantitative Spektralanalyse der Elemente, Jena, pt. 1 (1929) ; pt. 2 (1934). 4. Lundegardh, H., Lantbruks-HGgskol. Ann., 3, 49 (1936). 5. Lundegardh, H., and Phillipson, T., Lantbruks-HBgskol. Ann., 6,249 (1938). 6. Cholak, J., and Hubbard, D. M., Ind. and Eng. Chem., Anal. Ed., 16,728 (1944). 7. Instruction manual for operation o f model 18 flame photometer, Perkin-Elmer Corporation, Glenbrook (1945). 8. Peters, J. P., and Van Slyke, D. D., Quantitative clinical chemistry, Methods, Baltimore, 741 (1932). 9. Andes, J., and Eaton, A. G., Synopsis o f pathological chemistry, St. Louis (1941). 10. Levinson, S. A., and MacFate, R. P., Clinical laboratory diagnosis, Philadelphia, 2nd edition (1943). 11. Kracke, R. R., and Parker, F. P., Textbook o f clinical pathology, Baltimore, 2nd edition (1940). 12. Todd, J. C., and Sanford, A. H., Clinical diagnosis by laboratory methods, Philadelphia, 10th edition (1943).