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Continuous Flow Analysis

Continuous-Flow Analysis ( CFA ) is an analysis technique that automatically carries out wet chemical analysis
processes. The process steps of photometric- analytical methods are processed in a continuous flow of sample
and reagents in the intended sequence.

Manifold total phosphorus on the

CFA multiparameter analyzer

Such an analysis method and its time sequence are usually built on a hardware method manifold , matched to the
concentration range to be measured and the type of sample. The composition of the samples to be measured
(sample matrix) and existing interfering components can make a user-specific structure necessary for the same
determinations.

The CFA analysis system draws in liquid samples from an automatic sampler, forwards each sample to the
existing method manifolds and divides them into aliquots for the simultaneous execution of all established
determination methods in each sample.

Today, CFA analysis methods are state of the art in various application areas as ISO standards, such as water
quality, soil quality (extracts), tobacco (extracts) or food analysis. In the DIN EN ISO standards for water
quality [1] there is the umbrella term flow analysis , under which the CFA technology is jointly managed with the
related FIA technology.

Note: Other standards use the term “continuous flow analysis” in the German translation, which can be
misunderstood as the analysis of a flow. The use of unclear names makes it much more difficult to find specific
information through search engines.

The CFA technique works with regular subdivision (segmentation) of the continuous reaction stream by means
of gas bubbles (air, N2). To differentiate it from the FIA ​technique (flow-injection analysis, injection-flow
analysis), CFA is also referred to in the literature as segmented flow analysis [2] SFA . The segmentation in the
CFA technique enables even more complex analytical process steps to be strung together without the carryover
(dispersion, carry-over) between successive samples becoming inadmissibly large. Modern microflow [3]Even
with complicated methods, CFA systems achieve a sample throughput of 30 to 40 samples per hour, even if the
passage from the sampler through all process steps to the detector takes 10 to 25 minutes. This means that the
CFA analyzer simultaneously feeds 6 to 12 samples through the analytical system without one sample
influencing the next sample in an unacceptable manner.

Table of Contents
1 Details on the technique of a single CFA method
2 examples
3 Other applications
4 History of CFA analysis technology
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5 sources
6 Lileratur
7 Weblinks

More about the technique of a single CFA method

Flow diagram for nicotine in

tobacco extracts with an
integrated dialysis unit

The structure of a wet chemical analytical method on the CFA analyzer is shown in a flow diagram and
documented for the user. In the picture shown you have to imagine a constant flow from left to right. By using a
multi-channel peristaltic pump, the ratio of sample and continuously conveyed reagents to each other is always
constant, regardless of the speed of the pump. The absolute amount of the individual solution is defined by the
pump hose used; the color coding has been established worldwide for flowrated pump tubes for over 40
years . In the case shown of a 12-position peristaltic pump for a CFA Microflow Analyzer, the respective dosing
tubes give the stated amount in µl / min (number).

The method manifold ( Analytical Manifold ), a term that has been established over the decades, includes all
components for performing the analytical method steps that are located between the peristaltic pump (=
numbers) and the measuring cell (= colorimeter). This can be a module with injectors, mixing and reaction
spirals, but also an incubator (heating bath), a dialysis unit, a flow microdistillation module, a UV digestion unit
or a high-temperature hydrolysis unit.

The sequence of the analytical treatment stages of the method manifold can be used as a reaction pathare
designated. The continuous flow is regularly segmented at the beginning of this reaction section by means of gas
bubbles (air, N2), depending on the technology, every 1–2 seconds. The reagents are metered into the ready-
segmented stream, that is, into the individual segments flowing past the respective metering point after a time
determined by the structure. The regular segmentation of a sample sucked in for 60 seconds every 1 second now
means that in this case (40 samples / h, 2: 1), in principle, a 60-fold repeated measurement is carried out for each
sample. The CFA reaction line can be imagined as a conveyor belt, with a reaction tube every second, the
contents of which are then precisely added to further solutions that run through a matrix separation (dialysis)
through a digestion unit, through a reaction stage with incubation, and so on. At the end of the analytical
process, these reaction solution segments flow continuously through a measuring cell, whereby the air
segmentation is usually removed beforehand (debubbling).

The continuous signal of the photometric detection now shows as a graphic the course of the measured values ​of
the successively arriving segments, all of which have run through the entire analytical sequence independently
of one another and in the same way. This makes the reproducibility visible on the photometer graphicof the
overall analytical system, at the moment and also for each sample individually at the time of its individual
run. This unique property of the CFA technique, to show the stability of an analytical system and to make the
plausibility of a single measurement fundamentally comprehensible, is the reason for the decade-long unbroken
success of the CFA technique in the routine chemical laboratory. The second flow analysis technique, FIA, as an
unsegmented technique, does not offer this quality information.

CFA analyzers are used in today's analytical laboratories for ion analysis and conventional parameters mostly as
a system for the simultaneous determination of 2 to 5 parameters in each sample (perception of the author). With
50 samples per hour and 4 parameters simultaneously, i.e. 200 determinations per hour, the CFA technique
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outperforms even large discrete analyzers in sample throughput , with higher analytical sophistication and
significantly lower costs.

Examples

Continuous flow analyzer for

phosphate and nitrate
determination

In the case of total cyanide, acid is first added to the sample and the complex cyanide is split up using UV,
then the cyanide is distilled in flow-through, then the distillate forms a color with reagents, the strength of
which is measured. [4] The run through the analytical system takes about 15–20 minutes, depending on the
conditions.

In the case of total phosphorus, acid is first added to the sample and organophosphorus is converted using
UV light, then the sample is hydrolyzed at 95 ° C to convert inorganic phosphorus compounds, the
orthophosphate then present forms a color with reagents, the strength of which is measured. [5] The run
through the analytical system takes about 20-25 minutes.

In the case of soil extracts, the samples for the determination of ammonium [6] or nitrate nitrogen are first
passed through a dialyzer, which separates the interfering sample matrix (color, humic substances ...), then
diffused ions react with the reagent to give a color which is measured. The run to the detector takes 6–12
minutes, depending on the method. With a sample throughput of 60 / h, there are 6–12 samples in transit
through the analytical system

In the case of fertilizer digestion solutions, the samples are first diluted 1:10 to 1:20 in a dilution loop on
the manifold, then dialyzed and then measured by means of a color reaction. The run to the detector takes
about 8-14 minutes.

Other uses
In addition to the described application of CFA technology for performing photometric analysis processes, there
are also applications with other detectors such as fluorimeters, flame photometers or sensor technology. CFA is
used as a transport technology, which enables the rapid transport of samples one after the other through sample
preparation, without inadmissible concentration carry-over from one sample to the next.

History of CFA analysis technology

The biochemist Dr. Leonard T. Skeggs (1918-2002) first built a prototype of an automatic analyzer for his
laboratory samples in Cleveland, Ohio in 1951. The revolutionary idea was - not to try to move samples
individually from one work step to the next, as was previously the case - but to connect the individual stations
with a thin hose system and to pump the samples through sequentially. In order to prevent the samples from
mixing with one another, air bubbles were introduced at regular intervals. Hence the term "segmented flow
analysis", also CFA (Continuous Flow Analysis).
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Originally nobody was interested in commercialization until Edwin C. "Jack" Whitehead, who had founded
Technicon Instruments Corporation with his father in 1939, bought the development in 1954. The AutoAnalyzer
was fully developed in 1957 and was the first commercially available analysis system to be offered. This laid the
foundation for "automatic chemical analysis. This machine, later called AutoAnalyzer I, was originally capable
of one, later two parameters in parallel, eg glucose and BUN (urea nitrogen) at a rate of initially 20 samples per
hour determine.

The samples were taken from the autosampler with 40 positions and mixed one after the other with the
corresponding reaction solutions. The quantity measurement was effected by choosing different pump tubing
diameters and thus flow rates in the peristaltic pump. Mixing took place in glass spirals, protein fractions were
then removed in a dialyzer and the color reaction was completed in a heating bath. The concentration of the
analyte was determined in a photometer and could be read on the scale of the recorder (initially with a
logarithmic scale, later linearized).

Technicon AutoAnalyzer I with

functional diagram

The next step was the combination of several methods (typically 12) in one device and the optimization of the
components, which increased the sample frequency of the individual determinations to 60 per hour; the
Technicon SMA 12/60 (Sequential Multiple Analyzer) was created, which has become a standard system for
clinical laboratory analysis. At the same time, the AutoAnalyzer II was created, which was presented in 1972
and brought this form of automation to industrial laboratories. In 1979 Technicon and with it the AutoAnalyzer
lines were sold to Revlon.

Since its development, the CFA technique has flourished for decades as “the” standard technique in clinical
analysis. Countless publications on photometric, chemical CFA processes exist from the 1960s to the 1990s. The
device technology developed from the first generation autoanalyzer to the widespread macroflow technology and
to compact and fast microflow CFA systems.

At the end of the 1970s, the FIA ​technique was developed from the CFA principle, omitting segmentation, which
has become well established in the university sector for individual determinations and simpler procedures. For
larger routine laboratories with several simultaneous determinations, considerable sample throughput and
security against the influences of a fluctuating sample matrix, the CFA technology still offers the right solution
today. Low costs for consumables, fast processing of sample series and simple quality management guarantee
that this technology will continue to be used in the future.

sources
1. ↑ Determination of nitrite nitrogen and nitrate nitrogen and the sum of both by flow analysis (CFA) and
spectrometric detection, ISO 13395:1996.
2. ↑ Segmented Flow Analysis, Encyclopedi of Analytical Science, Vol 8, Academic Press.
3. ↑ The use of Microcontinuous Flow Analysis and FIA in Water Analysis, Straka M., International
Laboratory, 09-1990, S. 33.
4. ↑ Water quality – Chemisches Analyseverfahren Determination of total cyanide and free cyanide by
continuous flow analysis, ISO 14403:2002(E), First edition 1. März 2002.
5. ↑ Determination of ortho phosphate and total phosphorus contents by flow analysis, Part 2: Method by
continuous flow analysis (CFA), ISO 15681-2.

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6. ↑ Determination of ammonium by flow analysis and spectrometric determination, EN ISO 11732.

literature
L. T. Skeggs Jr.; Analytical Chemistry, 38 (1966), 32A
Edwin C. Whitehead; Technicon Corporation Worldwide - Review and Outlook; AutoAnalyzer
Innovations; Technicon Symposium 1978; Vol. 1, page 22ff
Morris H. Shamos; The Development of Laboratory Automation; Technicon International Symposium on
Advances in Automated Analysis; April 1974, Sydney
L Lewis, ‘Leonard Tucker Skeggs – a multifaceted diamond’, Clinical Chemistry, 27/10 (1981), pp 1465-
1468
Coakly, William A.: Handbook of Automated Analysis: Continuous Flow Technique, Marcel Dekker Inc.,
1981, ISBN 0824713923
R. Stanley; A multidimensional approach to analytical science; Journal of Automatic Chemistry, Vol. 6,
No. 4; Oct-Dec. 1984, Seite 175ff.

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