Information about the solutions for flame photometry

Status: August 2003

Specifications subject to change!
1. General information and measuring principle

Light emissions of alkali and alkaline earth metal elements are measured with flame photometry.
To this purpose, the measuring substance diluted in water is distributed as a fine aerosol into a non-luminous flame using
an atomizer. During this process, the flame lights up due to the excitation of the individual atoms.
The more atoms available, the greater the amount of radiated light. Therefore, the concentration of the measuring
solution can be calculated by measuring the brightness of the flame.
Every element has a characteristic light color, i.e., it radiates specifically defined wavelengths, determined by the
structure of the atom.
As several elements are available in the sample, the flame radiates a light mixture. With the aid of optical filters, the light
from the desired element is isolated and transferred to a photodiode which converts the light into an electrical signal.
This signal is amplified and digitalized. With the aid of the connected computer, the sample concentration is calculated,
displayed and printed out.
Flame photometry is a relative analytical method, i.e., the measuring signal of the unknown sample is compared with the
signal of a known standard solution and the sample concentration is calculated based on this.
As the concentration of Na, K and Ca is too high for direct determination and the matrix would disturb the measurement
considerably, all samples are diluted automatically 1 + 50.
Eppendorf flame photometers operate according to the reference line method. All samples are diluted with a diluting
solution which contains lithium as a reference element. Every change in atomization, air pressure and gas pressure has
approximately the same effect on the measuring element (Na, K, Ca) and reference element (Li).
During standard adjustment, the lithium concentration is measured and set to "1". A correction factor is calculated from
every lithium modification for correction of the measurement values for each appropriate sample.
The curvature of the calibration graph is stored in the computer for every element and every measured value is corrected
in accordance with the calibration fitting.
Spectral interference (e.g. from Na with K and Ca when using the device with acetylene) is immediately mathematically
corrected through the simultaneous measurement of the elements.
The zero point is measured once. The standard adjustment is carried out three times.
The current average of the measurements is displayed. Abnormal results are not taken into account. More exact
calculation foundations for determining the average can be supplied upon request.

Users can be referred to the following publications, among others, for information on the usage of solutions and
standards in connection with the newer EFOX, EFIX and EFUX photometers:
H. Schlebusch, B. Dürkop, I. Paffenholz
Evaluation des Flammenphotometers EFOX 5053
GIT Labor-Medizin 14 (1991) Seiten 179 – 184
GIT-Verlag Darmstadt
W. G. Guder, G. E. Hoffmann
Analytische und medizinische Aspekte der Elektrolytbestimmung am Flammenphotometer
Ergebnisse einer Evaluierung des EFIX 5055
GIT Labor-Medizin 15 (1992) Seiten 14 – 18
GIT-Verlag Darmstadt
T. Hoff, E. Gurr
Elektrolytbestimmungen mit Hilfe des Flammenatomemissionspektrometers EFUX 5057 in Kombination mit dem
Vielkanalanalysator Hitachi 717
GIT Labormedizin 18 (1995) Seiten 320 – 324
GIT-Verlag Darmstadt
Additional information concerning sample preparation, function, etc., can be found in the respective flame photometer operating instructions. Solutions, standards and devices
whose manufacture or sale were stopped more than 10 years ago are not described here.

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 Information about the solutions for flame photometry

2. Overview of standards and solutions

The following standards and solutions are relevant in connection with the older, no longer produced Eppendorf flame
photometers such as
FCM 6341- 6343, MFM 6350 – 6351, AFM 5051 – 5052,
and the newer flame photometer automated systems
EFOX 5053 – 5054, EFIX 5055 – 5056 and EFUX 5057:

2.1
Serum standard 100 ml Order number: 0030 357.000

The serum standard was offered prior to January 2001 exclusively in a three-pack (3 x 100 ml) with the
order number 0030 311.000.

In this sub-chapter we also present superordinate information for all other flame photometry standards and solutions in
addition to the special information applying only to this standard.

The serum standard is used with the EFOX, EFIX and EFUX devices for the standard adjustment for the determination of
– sodium
– potassium
– calcium (only with acetylene; does not apply to propane)
– lithium used in the serum.

In the case of the older FCM, MFM and AFM devices, the serum standard serves exclusively for the standard adjustment
for the determination of lithium in the serum. For the FCM, the serum standard 1 + 20 is used diluted for the Li determi-
nation. In the case of MFM and AFM, the 1 + 20 dilution takes place in the device. The dilution is carried out with a
ready-to-use blank value solution system (see Ch. 2.5).
The EFOX, EFIX and EFUX devices require the serum standard for the standard adjustment in the serum program and for
the lithium determination. The error ( approx. 0.8 %) resulting from the modified concentration of the Li signal in the
guideline is automatically corrected for the standard by the program. The standard is automatically diluted 1 + 50 in the
EFOX, EFIX and EFUX devices. For the standard adjustment, the standard is used undiluted in special positions in the
device. For the determination of Na, K and Ca, the automatic dilution in the device is carried out using a Li dilution
solution (ready-to-use lithium stock solution; see Ch. 2.4). For the Li determination with EFOX, EFIX and EFUX, the
automatic dilution (1 + 50) in the device is carried out with the standby solution (ready-to-use system blank value
solution; see Ch. 2.5).
If the serum standard is measured as a sample in the EFOX, EFIX or EFUX , the result will be values for Na, K and Ca that
are approx. 0.8 % too low. This is caused by the modified guideline signal when measuring the standard as a sample
(see Ch. 2.4).

The following manufacturer specifications apply for the serum standard:

Element name Element symbol Value (mmol / l) Max. deviation

Sodium Na 143.5 0.3 %

Potassium K 3.84 0.5 %

Calcium Ca 2.50 0.5 %

Lithium Li 2.00 0.5 %

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 Information about the solutions for flame photometry

A reference standard using Suprapur® chemicals (Merck) is referred to. The volumes for reference standards are
gravimetrically tested. The scales used are checked using standard weights. The elements are present in the standards
as chloride. Calcium carbonate serves as the basic material in the case of calcium. Lithium carbonate for lithium. A minor
surplus of heated hydrochloric acid is used for the transformation to the chloride form.
The standard has a useful life of 18 months. This durability is attained with the help of a preservative. The expiration date
is printed on the label as year and month. The standard can be stored at room temperature or in the refrigerator.
Accidentally frozen standard must be completely thawed and then thoroughly mixed prior to usage.
Removal by "pouring from the bottle" can minimize the risk of contamination of the standard.
A batch-related certificate of quality for the serum standard can be requested through the hotline. A safety data sheet
can be downloaded from this homepage.
You can find further information on the serum standard in the operating instructions of the relevant flame photometer.

The reference method value is taken as a control material reference value for the flame photometric measurements. Due
to the high degree of agreement between the flame photometric measured values and the reference method values, the
values published by INSTAND for the normal range also apply for the measurements carried out with our flame photo-
meters. The software of the new flame photometers provides the possibility to identify measured values outside of the
normal range for serum/plasma. In the case of EFOX, EFIX and EFUX, the following factory settings are valid for the
normal range:

Element Warning limits Repeat limits Unit

Sodium 135 – 150 130 – 155 mmol / l

Potassium 3.5 – 5.0 3.0 – 5.5 mmol / l

Calcium* 2.1 – 2.6 2.0 – 3.0 mmol / l

Lithium** 0.5 – 1.0 0.3 – 1.5 mmol / l

* The calcium total is determined for the flame photometry determination.

** In the case of lithium we are referring to the monitoring of the therapeutic range.
The manufacture and sale of the serum standard takes place in observance of the regulations of guideline 98/79/EG
concerning In vitro diagnostics.

2.2
Serum standard stock solution 500 ml Order number: 0030 310.004

Serum standard stock solution is used exclusively in connection with the old FCM, MFM und AFM flame photometers for
the standard adjustment in the serum program.
Serum standard stock solution contains Na, K and Ca in the same concentrations, basic materials and manufacturing
limits as the serum standard described in Ch. 2.1. The serum standard stock solution contains no lithium. It is not
possible for the old flame photometers to automatically carry out a Li correction for the standard. Thus, in the case of the
old flame photometers, it is necessary that only this standard be used for the electrolyte determination in the serum for
the standard adjustment. The evaluation of the measured values by curve adjustment (characteristic), averaging and
correction of spectral interference on the old devices is comparable to the evaluation with the newer devices.
A batch-related certificate of quality and a safety data sheet can be requested for the serum standard stock solution.
The manufacture and sale of the serum standard stock solution satisfies the specifications of guideline 98/79/EG for
In vitro diagnostics.

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 Information about the solutions for flame photometry

2.3
Urine standard 100 ml Order number: 0030 358.007

The urine standard was offered only as a three-pack (3 x 100 ml) with the order number 0030 313.003 prior to
January 2001.
The urine standard is used with the FCM, MFM and AFM devices for the standard adjustment for the determination of
– sodium and
– potassium in urine.

The urine standard is used with the EFOX, EFIX and EFUX devices for the standard adjustment for the determination of
– sodium
– potassium
– calcium in urine.

The following manufacturer specifications apply for the urine standard:

Element name Element symbol Value (mmol / l) Max. deviation

Sodium Na 143.5 0.5 %

Potassium K 100 0.5 %

Calcium* Ca 5.00 0.8 %

One reference standard is taken as a reference point when using Suprapur® chemicals (Merck). For reference standards
the volumes are gravimetrically tested. The scales used are controlled with standard weights. The elements are present
in the standards as chloride. Calcium serves as the basic material for calcium carbonate. A minor surplus of heated
hydrochloric acid is used for the transformation to the chloride form.
The standard has a useful life of 18 months. This durability is attained with the help of a preservative. The expiration date
is printed on the label as year and month. The standard can be stored at room temperature or in the refrigerator.
Accidentally frozen standard must be completely thawed and then thoroughly mixed prior to usage.
Removal by "pouring from the bottle" can minimize the risk of contamination of the standard.
A batch-related certificate of quality for the urine standard can be requested through the hotline. A safety data sheet can
be downloaded from this homepage.

You can find further information on the urine standard in the operating instructions of the relevant flame photometer.
The reference method value is taken as a control material reference value for the flame photometric measurements.
Statements form the Rick "Lehrbuch der klin. Chemie" concerning the normal values of urine:

Potassium 35 – 80 mmol / 24 hours

Sodium 100 – 220 mmol / 24 hours

Calcium* 3.2 – 8.2 mmol / 24 hours

* In the case of flame photometrical determination, the total dissolved calcium is determined. If abnormal Ca salt is also to be recorded, this is added to the
solution by leavening with HCl.

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 Information about the solutions for flame photometry

Because the flame photometer supplies a measured value in the mmol / liter unit, the result should be multiplied by the
total urine volume (Unit: liter / 24 hours).

Example:

Measured value EFOX: 35 mmol K / Liter

Total urine volume: 1.2 Liter / 24 hours
Calculation: 35 mmol K / l x 1.2 l / hours
Result: 42 mmol K / 24 hours

The manufacture and sale of the urine standard satisfies the specifications of guideline 98/79/EG concerning
In vitro diagnostics.

2.4
Lithium stock solution 500 ml Order number: 0030 306.007

The lithium stock solution has a lithium chloride content of 500 mmol Li / l. The maximum fluctuation range of the
Li content is 4.4 mmol Li / l. The solution has a pH value of approx. 1.1. The solution contains 0.2 % of the non-ionogenic
tenside Triton X-100. The useful life of the lithium stock solution is 18 months. The expiration date is printed on the label
as year and month. The solution can be stored at room temperature or in the refrigerator.
The lithium stock solution is diluted 1 + 100 by the user with demineralised or distilled water (recommended conductivity
of the water < 2 µS / cm at 20°C). The diluted lithium solution attained in this way is then used in the devices for the
dilution of the standard and samples in the serum and urine program. The diluted lithium solution has a Li content of
approx. 5 mmol / l. It is important that the same diluted Li solution is always used throughout one analysis run. Or in other
words, if the diluted Li solution is changed, the zero and standard adjustments must be repeated for the new diluted
Li solution.
The signal of the diluted lithium solution is used as the reference element for quotient generation (see Ch. 1).
Samples containing lithium influence the signal of the guideline and thus also the accuracy of the determination of NA, K
and Ca via the quotient generation as follows:
0.5 mmol Li / l erniedrigt die Na, K und Ca-Messwerte um ca. 0.2 %
1.0 mmol Li / l erniedrigt die Na, K und Ca-Messwerte um ca. 0.4 %
2.0 mmol Li / l erniedrigt die Na, K und Ca-Messwerte um ca. 0.8 %
The deviation of 1.0 mmol Li / l is within the range of normal coincidental measured value fluctuations (imprecision) of the
flame photometer. The dispensation of Li salt as an anti-depressant generally occurs at between 0.5 to 1.5 mmol Li / l.
Greater concentrations of Li are toxic.
Serum plasma with a basis of Li heparinate contains considerably less than 0.5 mmol Li / l . In case it is necessary to use
Li-heparinate plasma, the Li content of the Li heparinate tube batch should be requested from the relevant manufacturer
and tested with the Li program of the flame photometer.
The manufacture and sale of the lithium-stock solution satisfies the specifications of guideline 98/79/EG concerning In
vitro diagnostics.
A batch-related certificate of quality for the lithium stock solution can be requested over the hotline. A safety data sheet
can be downloaded from this homepage.

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 Information about the solutions for flame photometry

2.5
System blank value solution 500 ml Order number: 0030 340.000

The system blank value solution superseded the wetting agent solution 0030 307.003 as of November 1996.
The old and new solutions contain the same wetting agent in an unchanged concentration. The earlier sorbic acid
preservative has been replaced by an isothiazoline-based preservative.
The system blank value solution is diluted 1 + 250 by the user with demineralised or distilled water (recommended
conductivity of the water < 2 µS / cm at 20° C) for usage with the flame photometer. The dilution process should be
carried out approx. 24 hours prior to usage in order to enable the degasification of the water. The 1 + 250 dilution is
referred to as a "standby solution".
The system blank value solution contains 1 % of the non-ionogenic tenside Triton X-100.
The undiluted solution has a useful life of 18 months. The expiration date is printed on the label as year and month.
The solution can be stored at room temperature or in the refrigerator.
The standby solution is used for the serum standard and for sample dilution when undertaking lithium determination in
the serum, as well as for the zero adjustment for Li determination. Sufficient pre-rinsing with the standby solution prior to
Li determination is important. This is regulated automatically in the EFOX, EFIX and EFUX devices.
During measurement pauses in the urine or serum program, the standby solution is taken in by the atomizer of the flame
photometer. The permanent taking in of liquid is of decisive importance for maintaining a constant atomizer temperature
in order to prevent drift effects. In addition, due to the wetting agent content, the solution has a purifying effect within the
tube system of the flame photometer.
The manufacture and sale of the system blank value solution satisfies the specifications of guideline 98/79/EG for In vitro
diagnostics.
A batch-related certificate of quality can be requested over the hotline. A safety data sheet can be downloaded from this
homepage.

2.6 Non- IVD reagents

The following cleaning and disinfection solution is not categorized as an IVD reagent:
Detergent D 100 ml Order number: 0030 359.003

Prior to January 2001, detergent D was exclusively available as a three-pack (3 x 100 ml) with the order number
0030 334.000.
Detergent D will be automatically dispensed in diluted form (dilution with standby solution) as a cleaning solution at the
end of an analysis run with EFOX – EFUX and taken in by the atomizer.
Detergent D is used during maintenance work to boil out the atomizer or gas mixing chamber. Detergent D is especially
good for removing protein deposits.
Disinfection concentrate 100 ml Order number: 0030 341.007

The disinfection concentrate is used prior to maintenance and service work for the disinfection of surfaces and cannula.
It therefore also serves a protective function for the user.
A safety data sheet for detergent D and the disinfection concentrate can be downloaded from this homepage.
0030 306.937-00/0903

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Phone: 800-645-3050 · Fax: 516-334-7506 · E-mail: info@brinkmann.com · Internet: www.brinkmann.com