OrigiMl Article Ann Clin Biochem 1990; 27: 345-352

Interference by haemolysis, icterus and lipaemia in assays on the

Beckman Synchron CX5 and methods for correction
A G Randall, P Garcia-Webb and J P Beilby
From the Department of Clinical Biochemistry. Queen Elizabeth II Medical Centre. Perth. Western
Australia .

SUMMARY. As part of an evaluation of a Synchron eX5 analyser (Beckman Instru-

ments Inc, Brea, USA) we examined a range of tests for interference from haemolysis,
bilirubin and lipaemia. Tests investigated were urea, creatinine, urate, total protein,
albumin, calcium, total bilirubin, alkaline phosphatase (ALP), aspartate
transaminase (AST), y-glutamyl transferase (GGT) and inorganic phosphate. Two
types of interferences were found. One type is found on other analysers and represents
analytical difficulties with the measurement of that particular analyte. The other type
of interference was a consequence of the bichromatic optical system used on the eX-5.
This latter group includes haemoglobin interference in the measurement of total
protein and inorganic phosphate, and bilirubin interference with the measurement of
total protein, glucose and inorganic phosphate. Lipaemia interfered with total
protein, total bilirubin, inorganic phosphate, urate and glucose. Alternative and
modified methods are proposed to improve the measurement of total protein, glucose,
total bilirubin and inorganic phosphate. The use of the modified methods for glucose,
inorganic phosphate and total bilirubin are limited, at this time, by an error in the
calculation algorithm used by the analyser for two step or triggered chemistries, and
to a lesser extent, by a reduction in sample throughput.
Additional key phrases: automated analysis: bichromatic spectroscopy

Many modern analytical systems and methods
are subject to substantial interference in the pres-
ence of icterus, haemolysis or lipaemia in
samples.' These, and possible interference by
commonly used drugs, should be tested for in
every new method and analytical system prior to
its use in patient care.
The Synchron eX5 system (Beckman Instru-
ments Inc, Brea, USA) is a fully automated
discrete clinical chemistry analyser suitable for
use with samples of serum, plasma, urine and
cerebrospinal fluid. 2 The spectrophotometric
system comprises a multi-wavelength diffraction
grating, a pulsed xenon flash lamp and a
photodiode array detector. Nominal wave-
lengths available for analysis are 340, 380, 410,
470, 520, 560, 600, 650, 670 and 700 nm with a
half band width of 5 nm. To overcome the varia-
tion in the intensity of the light source that occurs
with each flash, the manufacturers have incor-
porated an optical system that allows simul-
taneous measurement of the absorbance of the
chromophore at both a primary wavelength and
a secondary wavelength; the secondary wave-
length acts as a reference and the instrument does
not use a conventional reference system. This
system of bichromatic spectrophotometry defi-
nitely improves the precision of the absorbance
measurements but it also increases the potential
for interference effects. This shortcoming of
bichromatic spectrophotometry with endpoint
absorbance measurements has been recognized
for a long time,' as has the need for the absorban-
ces of potential interferents to be checked at the
primary and the secondary wavelengths to avoid
inaccurate results,"
In this paper we describe the extent of the
interferences we detected in a number of assays
on the eX5 analyser and ways that these inter-
ferences can be minimized or overcome.

Correspondence: Mr A G Randall, Department of Clinical MATERIALS AND METHODS

Biochemistry, Queen Elizabeth" Medical Centre, Nedlands,
Western Australia, 6009, Australia. Plasma concentrations of urea, creatinine, urate,
345
346 Randall, Garcia- Webb and Beilby
total protein, albumin, calcium, total bilirubin,
alkaline phosphatase (ALP), aspartate
aminotransferase (AST), y-glutamyl transferase
(GGT), inorganic phosphate, glucose and choles-
terol were measured on the CX5 using Beckman
Instruments Inc. reagents and according to the
manufacturer's instructions, unless stated other-
wise.
Haemoglobin interference
Packed red cells (5 mL) from heparinized whole
blood was mixed with an equal volume of a
plasma pool. The mixture was lysed by sonica-
tion and centrifuged at 3000 rpm for 30 min. The
supernatant was analysed for haemoglobin on a
Coulter Cell Counter model S-PLUS V (Coulter
Electronics, Hialeah, USA). Sufficient
haemolysate to give a final haemoglobin con-
centration of 5 giL was added to 10mL of the
plasma pool. The spiked plasma was linearly
diluted in the original plasma pool and assayed
as before. No attempt was made to correct results
for the volume of haemolysate added to the pool
because the results are an approximate reflection
of results obtained from haemolysed specimens.
The analytes listed above were measured on each
sample.
Bilirubin interference
Bilirubin (crystalline, from bovine gallstones,
Sigma-Aldrich Chemicals, St Louis, USA)
62·4 mg was dissolved in I mL dimethylsulph-
oxide and added to 2 mL of 0·1 mol/L sodium
carbonate. This solution was then made up to
5 mL with a mixture of one part dimethylsulph-
oxide to two parts 0·1 mol/L sodium carbonate.
Of the resultant stock bilirubin solution, 0·3 mL
was added to 10mL of a plasma pool. The spiked
plasma pool was diluted in a linear series in the
original plasma pool and assayed for the analytes
listed above. The results were corrected for the
dilution effect of the added bilirubin solution.
The possibility of a blank effect due to
dimethylsulphoxide was checked using a blank
solution made by mixing 1mL ofdimethylsulph-
oxide with 2 mL of 0·1 mol/L sodium carbonate
and then adding O· 3 mL of this to 10mL of the
plasma pool. No interference was detected.
Elfect of lipaemia
Eleven lipaemic samples were selected from non-
fasting diabetic subjects and other hospital
patients. These samples were visually graded for
lipaemia from trace to + + such that it was just
possible to see a thick black line through a 1em
diameter tube containing sample with + + grade
of lipaemia.' The analytes listed above, with the
exception of cholesterol and triglyceride, were
measured on each sample before and after
centrifugation at 45 ()()() g for 30 min.
Results for glucose, bilirubin, uric acid, inor-
ganic phosphate and total protein were affected
by the lipaemia and therefore were investigated
further. Lipaemic plasma was pooled and diluted
with normal pooled plasma to yield a lipaemia
grade of + +. This pool was further diluted 1:3,
1: I and 3: I with the normal pool. The three
dilutions, the normal pool and the + + lipaemic
pool were then assayed for glucose, bilirubin,
uric acid, inorganic phosphate and total protein
before and after centrifugation at 45000 g for
30min.
Modified and alternative methods
Total protein
The CX-5 total protein method uses the Biuret
reagent and absorbances are measured at a
primary and secondary wavelength of 560 nm
and 470nm, respectively. At the pH of the
reagent, 12'5, both haemoglobin and bilirubin
absorb at 470 nm but not at 560 nm. The method
for total protein was modified by changing the
secondary wavelength to 670 nm and the analytes
remeasured. The sensitivity and precision of the
modified assay was the same as for the original
assay.
Inorganic phosphate
The CX-5 recommended method for inorganic
phosphate measures the increase in absorbance
caused by the formation of a phosphomolybdate
complex in an acid medium. The primary wave-
length is 340 nm and the second wavelength is
180 nm. The acid reagent and the molybdate
reagent are stored in separate compartments of
the reagent cartridge and mixed in the cuvette
before addition of the sample. Both haemoglobin
and bilirubin, under the reaction conditions,
absorb at 380 nm but not at 340 nm.
The method was changed to a two step or
triggered method with the molybdate reagent
being used to start the reaction with reagents
prepared according to Denegaar." Absorbance
was read before and after the addition of the
molybdate reagent. The first' reading was
therefore effectively a sample blank. This pro-
cedure also works with Synchron CX5 reagents,
using the reagent in compartment A as the
sample diluent and the reagent in compartment B
as the trigger reagent (data not shown).
All results were recalculated from raw absor-
 Interference by haemolysis, icterus and lipaemia 347

bance data with the appropriate second reagent bance data with the appropriate second reagent
volume correction applied to the blank reading. volume correction applied to the blank reading.

Total bilirubin
The recommended CX5 method for bilirubin is a
modification of the Jendrassik and Grof method
in which alkaline tartrate is omitted. Absorbance
of the neutral azobilirubin is read at 560 nm. As
an alternative'. Beckman Instruments Inc. also
supplied a Beckman Dri-STAT Reagent
(enzymatic bilirubin) kit with a preliminary
application to the CXS.
All results were recalculated from raw absor-
bance data with the appropriate second reagent
volume correction applied to the blank reading.
Glucose
The recommended CXS method for glucose is a
hexokinase method with the reagent supplied in
two separate compartments of the reagent
cartridge. The two reagents are mixed in the
cuvette prior to the sample being added. Using
reagent, timing and sample volume information
supplied' a user defined method was
programmed using the reagent in compartment 8
as a trigger reagent,
All results were recalculated from raw absor-
TABLE I. Interference produced by haemoglobin and bilirubin
RESULTS
Haemoglobin interference
Interference due to haemoglobin is shown in
Table I. In our laboratory, specimens subjective-
ly assessed to have a haemoglobin level of 3 gIl
are generally rejected. Interference from haemog-
lobin was deemed significant if the error at 3 gIL
was greater than 2·8 times the within-run stan-
dard deviation we had determined for that
method, using the CX5. Haemoglobin showed
significant interference with the analysis of the
following analytes: total protein, calcium, total
bilirubin, ALP, AST, GGT, glucose, inorganic
phosphate and cholesterol.
Interference by haemoglobin in the measure-
ment of ALP,8.9 AST and GGT IO activity has
been described elsewhere and IS due to mechan-
isms other than the use of bichromatic
spectrophotometry. Interference in the total
bilirubin assay can also be ascribed to reasons
other than the instrumental design.l':" For
example, in the classical Jendrassik and Grof

Analytc Interference

Name Concentration Haemoglobin Bilirubin

at I giL at 100mmoI/L

Albumin 30 giL 0 0
ASl 31 U/L + \·4" 0
ALI' 91 U/L -I·S" 0
Calciun. 2·30 mmol/L -0'()3" 0
Cholesterol 43 mmol/L +0·06" -0'3"
Creatinine 170 ~mol/L 0 -10"
GGT 106 U/I -3" 0
Glucose 7-8 rnrnol/L 0'04" -0·1·
Inorganic phosphate \·4 mmol/l. -0'2" -0,1"
Total bilirubin 17 jlmol/L + 14· na
Total protein 64 gil. -3" - 3"
Triglycerides 1·65 rnrnol/L 0 0
Urate 0·37 mrnol/L +0·01 -0·02·
Urea 12·S mmol/L 0 0
Modified methods:
Inorganic phosphate 1·4 mmol/L +O·OS· 0
Total protein 64 giL +2· -I·
Glucose 7·S mrnol/L nt -0,05"
Total bilirubin 17 jlmol/L -I na

Results are expressed as the effect produced for a final ctllcentration of 1 giL and 100 jlmol/L of haemoglobin and
bilirubin, respectively. Figures are derived from linear regression analysis of results from 10 progressive dilutions
of the spiked pool. The effects observed were approximately linear up to a final haemoglobin concentration of 5 giL
and bilirubin concentration of 600 /-lmol/L. na: not applicable. nt: not tested. ·Interference > 2·S times within run
SD at 3 giL and 300jlmol/L for haemoglobin and bilirubin. respectively'
348 Randall, Garcia-Webb and Beilby
method the use of alkaline tartrate results in the
formation of alkaline azobilirubin which absorbs
at a different wavelength than neutral
azobilirubin. On the CX5 in the absence of alka-
line tartrate, both the neutral azobilirubin
formed and haemoglobin absorb at the primary
wavelength of 560 nm, thus resulting in the
demonstrated positive interference.
Interferences in the measurement of glucose,
cholesterol and calcium, though significant, were
small (Table I). No attempts were made to
modify the methods to reduce these interferences.
Haemoglobin interference in the measurement
of total protein and inorganic phosphate is due to
the bichromatic optics of the CX5. In each case
the interferent absorbs at the secondary wave-
length but not at the primary wavelength, thus
giving a negative interference. The modified
method described here for inorganic phosphate
overcame this interference but, as expected, pro-
duced a small positive interference due to the
high concentration of phosphate in red cells. In
the case of total protein the negative interference
also converted to a small positive interference,
with most of the difference being attributable to
haemoglobin and other extra protein being
added from red cell contents.
Bilirubin interference
Bilirubin interference was tested up to a level of
600 J.LmoIJL. In our laboratory plasma samples
with a bilirubin concentration approaching
600 J.LmolJL are rare, whilst specimens with a
concentration of 300 J.LmolJL are seen several
times a month. Interference from bilirubin was
deemed significant if the error at 300 J.LmolJL was
greater than 2·8 times the within-run standard
deviation for that method. Bilirubin significantly
interfered with creatinine, urate, total protein,
glucose, inorganic phosphate, and cholesterol
(Table I). The interferences noted with
creatinine, urate, and cholesterol are not due to
limitations of the analyser. 1.13.14 Interference with
the total protein, glucose and inorganic
phosphate methods is due to the use of
bichromatic spectrophotometry. As with
haemoglobin interference, in each case bilirubin
absorbs at the secondary wavelength and not at
the primary wavelength.
The described modification of the inorganic
phosphate method removed the interference. The
described total protein method reduced the
bilirubin interference but did not eliminate it
completely. The modified glucose method
reduced interference due to bilirubin.
Interference in Iipaemic samples
Lipaemia at trace levels interferes with the total
protein, total bilirubin, glucose and inorganic
phosphate methods (Table 2). Clarification of
specimens by centrifugation prior to analysis
would appear to be the only way to avoid this
problem with the methods provided by Beckman
Instruments Inc.
Using the described method modifications, in-
terference in total protein measurement is
minimized and interference in inorganic
phosphate, total bilirubin and glucose measure-
ment is removed, at least up to lipaemia levels
described as + +.
The data presented here are the interferences
found for one pooled specimen. This was done
because of practical problems in collecting
sufficient amounts of different lipaemic
specimens. Because the absorption spectrum of
lipaemic specimens will depend on the size and
shape of the lipoprotein particles present we
anticipate that the amount of interference found
for different specimens will vary.
DISCUSSION
The CX5 optical system uses a xenon flash tube
light source. Compensation for variation in flash
intensity is achieved by taking all readings at two
wavelengths. The results are calculated from the
difference in absorbance at the primary and
secondary wavelengths. Bichromatic spectro-
photometry will work well only when the reac-
tion chromogen has a narrow absorption peak
ami the two wavelengths are as close together as
possible.
The approach relies on any interfering sub-
stance absorbing equally at the primary and
secondary wavelength. When this is not the case,
a positive or negative error will result. The absor-
bance spectra of the interferents bilirubin and
haemoglobin depend on the pH of the reaction
and therefore each new method on the CX5
requires careful evaluation to avoid interference.
Methods for removing interference
Wavelength alteration
If an interferent absorbs more at the secondary
wavelength than at the primary, the obvious
solution is to alter the secondary 'wavelength,
providing that this does not effect either the sen-
sitivity or precision of the assay. The Biuret total
protein reaction is monitored at a primary wave-
length of 560 nm and a secondary wavelength of
470 nm. At the reaction pH of 12'5, both
bilirubin and haemoglobin absorb at 470 nm, but
 Interference by haemolysis, icterus and lipaemia 349

TABLE 2. Interference produced with lipaemic samples

Degree of Lipaemia

Analyte ± + ++

Albumin giL 0 0 0
AST U/L 0 0 0
ALP U/L 0 0 0
Calcium mmol/L 0 0 0
Creatinine Jlmol/L 0 0 0
GGT U/L 0 0 0
Glucose mmol/L +0·2· +0-4· +0·7·
Inorganic phosphate mmol/L +0·09· +0'24· +0·36·
Total bilirubin Jlmol/L +6· +10 +20·
Total protein gIL -2· -9· -15·
Urate mmol/L +0·02 +0·04 +0·07·
Urea rnmol/L 0 0 0
Modified methods:
Inorganic phosphate mmol/L 0 0 0
Total protein gIL +1 +3· +5·
Glucose mmol/L 0 0 0
Bilirubin Jlmoi/L 0 0 0

Data are the difference in results on specimens before and after clearing by centrifugation at 45 000 g for 30 min.
The three levels of lipaemia were produced from a pool of lipaemic plasma by dilution in a pool of normal plasma.
·Interference > 2·8 times within run SD.

do not absorb at 560 nm. As the level of the
interferent increases, the absorbance difference
between the primary and secondary wavelength
decreases, thus lowering the apparent total
protein concentration.
Small shifts in the secondary wavelength to
reduce the effects of the interferent are not pos-
sible with the fixed range of wavelengths pro-
vided on the CXS. Were it to be possible, altering
the secondary wavelength to 480 or 490 nm, for
example, would reduce the interference to almost
zero. The Biuret chromogen absorbance peak
covers a range of just under SOO nm to over
600 nm. Altering the secondary wavelength to a
shorter wavelength would worsen interference
due to lipaemia, since lipaemic samples scatter
considerably more light at lower wavelengths
than at higher wavelengths, thus giving an ap-
parent increased absorption. Therefore the
closest available wavelength to the primary that
was not affected by bilirubin, haemoglobin or the
Biuret chromogen was 670 nm. The use of
670 nm as the secondary wavelength does not
satisfy the criterion of having the primary and
secondary wavelengths as close together as pos-
sible, but it does reduce the interference from
haemoglobin, bilirubin and moderate lipaemia.
Using a two step system
Sample blanking can be achieved using a two
step reaction for those reagent systems that can
easily be divided into a sample diluent and a
trigger reagent, and where any interferent is not
altered by the addition of the trigger reagent. In
this type of method the sample is mixed with the
first reagent and an absorbance reading is taken,
which in effect is the sample blank. The second
reagent, which contains the chemicals necessary
to start the reaction, is then added and another
absorbance reading is taken. The difference
between absorbance reading one and absorbance
reading two is then used to determine the result.
However, because the blank absorbance is
taken in a reaction volume less than that for the
test absorbance, it must be corrected ifit is not to
be overestimated. The correction factor is cal-
culated as follows
RlV + SV
VCF = RIV + R2V + SV
where VCF = volume correction factor, RIV =
reagent I volume, R2V = reagent 2 volume, SV
= sample volume.
In a reaction system in which absorbance
increases, not applying the correction factor will
result in a negative error. For reactions in which
the absorbance decreases, a positive error would
occur. There is no indication in the instrument
manuals that this blank correction is made with
the software supplied with the CXS. Com-
munication with the manufacturer in USA has
confirmed that this correction was not available
350 Randall, Garcia- Webb and Beilby
in the software provided, although the manufac-
turer plans to modify the software shortly.
Inorganic phosphate. The phosphomolybdate
complex absorbs at 340 nm and is used as a
measure of phosphate concentration. The secon-
dary wavelength recommended by Beckman
Instruments Inc is 380 nm. Both haemoglobin
and bilirubin absorb at 380 nm and not at
340 nm. However, a change in the secondary
wavelength of approximately 200 nm would be
required to prevent absorption by these substan-
ces at the secondary wavelength. This is contra-
indicated both in theory, and also by the fact that
it would result in more interference from
lipaemia, since lipaemic samples scatter light
more strongly at 380 nm than at about 550 nm.
In the recommended CX5 method, the inor-
ganic phosphate reagent is in two portions, an
acid diluent and an ammonium molybdate
solution. The two are stored separately and
mixed together prior to sample addition. In the
modified method the sample and acid diluent are
mixed, an absorbance reading taken, the molyb-
date reagent is added and another absorbance
reading taken. The first reading is effectively a
sample blank and the difference between the final
reading and the blank reading gives the absor-
bance due to the formation of phosphomolyb-
date.
If the volume correction were not applied, the
blank would be over-estimated. The resultant
error depends solely on the magnitude of the
blank absorbance reading. If the absorbance
reading happens to be zero, the error 1S zero.
With bichromatic spectrophotometry, because
the actual absorbance value used is the difference
between absorbance readings taken at two dif-
ferent wavelengths. it is possible for the blank
reading taken at the beginning of a two step or
triggered reaction to range from positive,
through zero to negative over a range of con-
centration of an interferent.
The volume correction can be done manually
by obtaining raw absorbance data from the
instrument and applying the appropriate calcula-
nons. The error in the modified two step inorgan-
ic phosphate method due to a lack of an adequate
blank calculation, was found to be approximate-
ly + 0·2 mmol/L at a haemoglobin concentration
of 5 gIL. This error is relatively small because the
blank absorbance for inorganic phosphate is
always close to zero.
Total bilirubin. The CX5 total bilirubin method
is based on the method of Jendrassik and Grof
without the addition of alkaline sodium tartrate
and without the application of a blank. Positive
interference from haemoglobin is to be expected
since under the reaction conditions haemoglobin
absorbs at the same wavelength as the
azobilirubin formed in the reaction.
The bilirubin reagents are stored on the instru-
ment as a diazo reagent and as a reagent contain-
ing caffeine, benzoate and acetate. Using these
reagents and defining a method to use the diazo
reagent as a trigger reagent gives a negative inter-
ference for haemoglobin almost equal in mag-
nitude to the positive interference found for the
original method. This effect has been described
previously?" and occurs because oxyhaemo-
globin is converted to methaemoglobin quite
quickly after the addition of the diazo reagent.
The absorbance of methaemoglobin under the
reaction conditions is less than that of
oxyhaemoglobin. As a consequence the absor-
bance due to haemoglobin in the blank reading is
higher than in the test reading. The interference
in the two step modification using CX5 reagents
is thus not a consequence of bichrornatic
spectrophotometry.
An alternative method suggested and provided
by Beckman Instruments Inc was to use
Beckman Dri-STAT Enzymatic Bilirubin. A.
preliminary method application to the CX5 was
provided with the reagents. In this method
bilirubin oxidase is used to convert bilirubin to
biliverdin, The corresponding decrease in absor-
bance at 420 nm (secondary wavelength, 700 nm)
is used as a measure of bilirubin present in the
sample. This method should reduce interference
due to haemoglobin (the quoted interference is
- 5% for each gram of haemoglobin per litre of
sample) and remove interference from lipaemia.
However as it uses a two step system it suffers
from the fact that there is no volume correction
applied to the initial blank reading as described
above. The effect in this case was found to be
much larger than that found for the modified
inorganic phosphate method because of the
blank absorbance was always large.
For example, in the CX-5 method application
the sample voiume is 10 ul.; reagent I volume is
200 JlL; reagent 2 volume is 16 JlL. From these
figures it can be calculated that the volume cor-
rection factor is 0·l}29. In a specimen with a
bilirubin of 24 Jlmoll L and no added haemo-
globin the blank absorbance was 0·04706 and the
reaction absorbance was 0·02406. The blank ab-
sorbance is over-estimated by 0'()()36 which is
15·6% of the difference between the blank and
reaction absorbance. The resultant error is
 Interference by haemolysis, icterus and lipaemia
+ 4 jlmol/L. In the same specimen when the
added level of haemoglobin was 3 gIL the blank
absorbance was 0·36100 and the reaction absor-
bance was 0·31699. The blank absorbance is
over-estimated by 0·0274 which is 62·3% of the
difference between the blank and the reaction
absorbance. The resultant error is + 30 jlmol/L.
Glucose. The recommended CX5 method is a
hexokinase method with the reagents supplied in
two portions, in compartments A and B of the
reagent cartridge. An assumption was made that
the reagent in compartment B could be used as a
trigger reagent. This appears to be true. Using
the reagent in this way produces the same errors
in blank readings as described for inorganic
phosphate and bilirubin. For example in a
specimen with + + lipaemia with a glucose of
10·1mmol/L the error caused by the incorrect
calculation is - 0·5 mmol/L.
Using a full sample blank
This mode of blanking requires a completely
separate tube or cuvette to be used, with the
difference in reagent in the two tubes designed so
that the absorbance in the blank tube accurately
reflects the interferent absorbance in the test
tube. Such an approach appears to be the only
way to apply an appropriate blank to the
Jendrassik and Grof bilirubin method to correct
for haemoglobin interference. The CX5 does not
at present provide for this sort of blanking.
Rate or two point assays
Rate assays, such as for enzymes, and initial rate
assays, such as for creatinine, will only show
interference due to bichromatic spectro-
photometry if there is an absorbance change at
the secondary wavelength during the time the
assay is being monitored. While theoretically
possible, this was not found to be a problem for
the assays described in this publication.
End point assays, in which the initial absor-
bance reading is taken very soon after sample
addition could also be used as a means of reduc-
ing interference. However, the earliest reading
time available on the CX5 is 16s after sample
addition. For most assay systems this is too long
a delay, and therefore this approach was not
considered.
CONCLUSIONS
There are various approaches that can be used to
correct for spectral interference in spectro-
photometric assays: full sample blank in a
351
separate cuvette, a two step or trigger reaction
system, bichromatic spectrophotometry, rate
assays and early reading two point assays. The
CX5 uses bichromatic spectrophotometry for
every analysis but, as has been shown, this has
severe limitations and can actually introduce
problems where they are not usually found. The
CX5 allows the application of a two step trig-
gered reaction system but the calculation algo-
rithm used appears to be flawed. The amount of
error introduced by this will vary with the par-
ticular application. Two step reaction systems
will solve blanking problems for most chemistries
but would also halve the throughput of the
machine for the particular test involved. Full
sample blanking will solve spectral interference
problems but is not an option currently available
on the CX5. The rate assays tested, some
enzymes and creatinine, have no interference
problems relating only to the use of bichromatic
spectrophotometry.
The modified methods described in this paper
will reduce interferences in the measurement of
total protein and inorganic phosphate to a
manageable level. Interference in the glucose and
total bilirubin assays will remain a problem until
the error in the triggered chemistry calculation
algorithm is corrected. That problem should be
resolved by the manufacturer in the relatively
near future. However, decreased sample
throughput remains as a potential problem. A
simple operating compromise would be to use the
standard method for all samples not noted by the
operator to be haemolysed, icteric or lipaemic.
Such samples could then be analysed using the
appropriate modified method if indicated. Used
in this way, the suggested methods would have
very little impact on test throughput and provide
a practical solution to the difficulties posed by
some types of sample.
Acknowledgement
We wish to thank Nola Hebiton for her excellent
technical assistance.
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Accepted/or publication 26 January 1990