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CLIN.CHEM.34/2, 227-234 (1988)
Determinationof Manganese in BiologicalMaterialsby ElectrothermalAtomic Absorption
Spectrometry:a Review
Fran#{231}ois
Baruthio,1 Oilvier Guliiard,2Joslane Amaud,3 Francis PIerre,1and Richard Zawlsiak4
The great diversityof methodsfor measuringmanganese in
biological materials (serum, plasma, whole blood, urine,
spinal fluid, and hair) reflects the difficulty in measuring
extremelysmallquantitiesof thiselement. Detailedexamination of these methodsdemonstratesthat the one most used
is flameless atomic absorptionspectrometry.In this review
we reportthe differentinstrumentsettingsfor wavelength,slit
width, protection gases, graphite furnaces, type of background correction, amounts measured, and thermal programs. We give detailed recommendationsby various authors for collectingsamples. A thorough descriptionof the
preliminarysteps and the handlingof the specimensamples
is also included:direct determinationwith or withoutdilution,
additionof a matrix modifier or determinationafter ashing,
withor withoutchelation-extractionsteps.The preparationof
the standards,proceduresused, analyticalcriteria(accuracy,
precision,specificity,detectionlimit, linearity),problems (interferences, matrix effects), and reference values and their
physiologicalvariationsare also described.We give a consensus of recommendations concerning the choice of a
method.
Manganese
(atomic number 25, atomic mass 54.938 Da) is
transition metal after iron and tin.
widespread
in nature,
it is never found in the
the most ubiquitous
Although
metallic
state. The most frequently
encountered
valencies
for manganese-containing
compounds are + 2, + 3, and + 7.
Manganese
shares with certain other metals the property
of being essential
to life, but large doses are toxic (1,2). The
estimated daily requirement for adults is 2 to 3 mg. However, exactly how the body assimilates
Mn still remains
unknown. Furthermore, manganese metabolism apparently
varies from individual to individual. Manganese is found in
the body mainly in mitochondria-rich tissues.
Like most transitional metals, Mn is found in various
complex enzymatic compounds (1-3): phosphoenolpyruvate
carboxykinase
(EC 4.1.1.32), pyruvate kinase (EC 2.7.1.40),
pyruvate carboxylase
(EC 6.4.1.1), superoxide dismutase
(EC 1.15.1.1), and arginase (EC 3.5.3.1). It is recognized as a
nonspecific activator of enzymes that require the presence of
a divalent ion. The ion Mn2 is analogous to Mg2, which it
may replace in numerous
Mn is mainly associated
biological
molecules.
with the formation of connective
and bony tissues, with growth and reproductive
functions,
and with carbohydrate and lipid metabolism (4). Manganese
concentrations are controlled by the liver, most of it being
eliminated in bile (4), with a small proportion eliminated
through the urinary tract and superficial body growth such
as hair
(5).
Determination of manganese is of particular
interest with
regard to detecting excessive exposure
to it in the workplace. Chronic intoxication from inhaling Mn at work is
characterized
primarily by psychological and neurological
manifestations [Parkinson-like syndrome (6)1.
Although frequently observed in the animal kingdom, no
manganese deficiency is apparent among human beings.
Nevertheless, a prolidase deficiency, which becomes evident
in certain cases of chronic ulcers, is accompanied
by a
decreased
concentration of manganese in serum and urine
(7).
Pathologically,
the manganese content of serum increases
among patients with liver diseases of whatever etiology (8),
and among hemodialysis patients when the Mn content of
the dialysis liquid is increased (9). For patients with acute
myocardial infarction, the mean serum Mn values do not
significantly differ from the controls (10, ii).
These deficits and overloadings studies have led analysts
to develop methods for Mn determination
in biological
fluids. The analytical difficulties and the very low concentrations to be measured lead to an important dispersion of
values and to a great diversity of methods. The most fully
developed analytical
procedures are in the EAAS field.5
Grenoble Cedex, France.
Methodology
The numerous
analytical techniques include flame atomic
absorption spectrometry (AAS), electrothermal atomic absorption spectrometry (EAAS), neutron activation analysis
(NAA), inductively coupled plasma (ICP), photometry, spectrochemical emission, spectrophotometry,
and fluorometry.
In the AAS method, air-acetylene
is the most frequently
used mixture
(12-23).
Determination
of Mn in serum, whole blood, urine, hair,
tissue, or spinal fluid by NAA is reported
in numerous
publications (24-32).
Laboratoire
d’Analyses, Institut de Chimie Biologique, 11 rue
Human, 67085 Strasbourg Cedex, France.
The authors are members of the Commission for Trace Elements
of the Societe Francaise de Biologie Clinique (chairman: J. Arnaud).
Received August 12, 1987; accepted November 27, 1987.
Nonstandard
abbreviations: AAS, flame atomic absorption spectrometzy; EAAS, electrothermal atomic absorption spectrometry;
NAA, neuron activation analysis; ICP, inductively coupled plasma;
and ns, not specified (in the literature).
1 INRS, Service Epidemiologie, BP 27, 54501 Vandoeuvre
Cedex,
France.
2Centre de dosage d’#{233}l#{233}ments
traces, H#{244}pital
Jean Bernard, BP
577, 86021 Poitiers Cedex, France.
3Laboratoire de Biochimie C, Hopital Nord, BP 217X, 38043
CLINICAL CHEMISTRY, Vol. 34, No. 2, 1988
227
Sometimes ICP is coupled with EAAS. Aziz et al. (33-34)
used this double system for determining
Mn in serum and
body tissues: after the sample is dried, mineralized,
and
atomized in a furnace, the nebulized material is transferred
to the ICP system. However, Black et al. (35) used ICP alone
for serum and tissues, as did Barnes and Fodor (36) for urine
samples alone. Three other methods are not so widespread:
photometry is mentioned by Bonier et al. (9) and Mehnert
(37), fluorometry by Rubio et al. (38), and spectrochemical
emission is only used by Hambidge and Droegenmueller
(39) for plasma and hair and by Yoakum et al. (40) for body
tissues.
From a close examination of the research published to
date, we find the method most commonly used in clinical
chemistry for determining manganese is EAAS.
Operating Conditionsin EAAS
Wavelength and slit width. Hollow-cathode lamps (singleor multi-element)
are the most frequently used, generally
set at the most sensitive wavelength, 279.5 nm. The smallest slit width should be chosen (generally 0.2, or 0.7 nm if
the available energy is not sufficient at 0.2 nm), as determined by the optical characteristics
of each apparatus.
Carrier gas. Although some authors use nitrogen (41-46),
argon is prefered to avoid nitrure generation. Furthermore,
Brodie (47) found that argon offers a greater sensitivity.
Background
correction. Continuum
source background
correction is the most widespread system. Deuterium
arc
lamps are the most frequently used, and in recent times the
Zeeman effect has been used for improved
background
correction (48-53). Graffiage et al. (54) and especially Favier
et al. (55, 56) found that this correction
is absolutely
essential to free the determination from the effect caused by
covolatilized matrix (55). More importantly, background
correction is almost always necessary to avoid errors near
the detection limit.
Furnaces. Pyrolytically coated tubes are the most commonly used (45, 47, 53, 54, 57, 58). According to our
experience, they offer a better sensitivity and higher thermal stability than that of standard tubes. The L’vov pyrolytic platform, generally associated with a matrix modifier, is
used for complex media such as seawater (59,60), but its use
in analysis of biological materials is likely to increase (61,
62). Some manufacturers
suggest the use of graphite “cuptype” cuvettes (48, 51).
Injected volumes. Injected volumes depend on the dimensions of the furnace. They range from 2(52) to 50 ML (54,56,
63, 64); the most common are between 20 and 50 1zL. For
such volume8, a wetting agent is added to the sample, to
spread out the drop in the furnace. Injection of the sample
into the furnace may be manual or automatic. Automatic
injection of course offers the advantage of better within-run
precision. It is recommended to cut the catheter tip regularly to avoid adsorption (56). For small volumes (5 ML),
Manning and Slavin (65) used a special apparatus with a
deposit of the sample on tungsten wire, prepared beforehand.
Graphite furnace settings. Drying depends on the volume
and type of sample. Times range from 5 s for Wei et al. (57)
to 2 miii for Bek et al. (66), with various numbers of stages
(47, 51, 67, 68).
Various charring temperatures
are used (46,54), the most
suitable being 1100 “C (24, 41, 47, 54, 56, 64, 66, 69-72),
with a ramp time of lOs and a hold time of 30 s for a 20-L
sample deposit. No loss of Mn can be observed at this
228 CLINICALCHEMISTRY, Vol. 34, No. 2, 1988
temperature.
For atomization, the most frequently mentioned temperatures are 2400 #{176}C
and 2600 “C (48, 50, 63, 64, 73-75). Fast
heating (0 s ramp time) is recommended
to increase the
atomic concentration in the vapor phase. Narrower and
sharper absorption peaks are observed if the furnace
is
heated very rapidly (76, 77), and the sensitivity and linearity are improved (55).
SpecImen CollectIon
Manganese may be measured
in senun,
plasma, whole
blood, hair, and urine. Its presence in hair is of particular
interest as a good indicator of the current body burden of
manganese (5, 32, 70).
Assay of whole-blood specimens has been preferred to
serum, owing to the strong concentration of Mn in the
erythrocytes (3,26, 78).
Because the concentrations of manganese ordinarily present in the biological materials analyzed are very low (<1
1zmol/L or 1 pmol/g), certain conditions must be met when
samples are collected, to avoid contamination.
Blood
Blood samples must be obtained with sampling devices
free of manganese and this preferably in the morning after
overnight
fasting (9, 39, 79). Some studies mention much
Mn contamination when disposable syringes and stainlesssteel needles are used (69,80). However, other authors have
not noted any contamination from the use of stainless-steel
needles (12). Versieck et al. (81) recommended, however,
that when this procedure is used, the first few milliliters of
blood collected should be discarded. Use of a nonmetallic
catheter is clearly preferred (41,54,69). The use of evacuated containers does not seem to pose any major problems of
contamination (48, 56, 82, 83). For collection and storage,
plastic tubes are most frequently used (polypropylene, polystyrene, or polyethylene),
previously
decontaminated
by
washing in diluted hydrochloric or nitric acid (39, 54, 69,
80). Generally, the samples are stored at -20 “C (39,42,48,
80,84). Glass tubes are not recommended for storage. Hudnik et al. (80) have observed noticeable contamination from
this material, and Grafflage et al. (54) noted adsorption of
manganese by the glass.
If an anticoagulant
is used, it must be rigorously controlled. The studies of Delves et al. (82) have pointed out the
high concentrations of manganese in some heparuns. However, heparin is the anticoagulant most frequently used in
manganese determination (24,39,48,82),
although Muzzarelli and Rocchetti used oxalate (43), Nath (84) recommended
citrate, and Casey et al. used dipotassium EDTA (75).
Urine
Urine collected in normal urination
or, more often, in
over 24 h is acidified and collected in plastic vials
previously washed in acid (14, 69, 85). The specimens are
stored at -20 “C (15).
diuresis
Hair
Hair specimens are taken in sufficient quantities (about
20 mg is needed for good precision) and washed by various
methods (each author has a preference) before determination of manganese. The washing should eliminate adsorbed
elements without, however, desorbing the manganese actually in the hair itself (25, 86). Many different procedures
have been used for washing the hair (87). This difficult
has long been a point of contention,
as emphasized
by Chittleborough
(88). Organic solvents (28, 39, 70, 71),
detergents
(53, 74,89), and ultrasonic treatment with water
(25) have been shown to remove surface contamination
from
hair effectively without extracting the Mn contained in the
shaft, but it is difficult to conclude which wash is the best
(53). We recommend sequential
washings with non-ionic
detergents (such as Triton X-l00), water (several times),
95% ethanol solution, and again with water. After drying at
60#{176}C
the specimens can be stored at 20 “C until determination. The use of a chelating agent should be avoided.
problem
Tissue
Tissue biopsies are carefully soaked several times in a
solution of wetting agent, rinsed with de-ionized water, and
placed in a flask previously washed with a diluted acid (80,
90). The samples may be stored at -20 “C after lyophilization, or not (80, 91).
PreparatIon and Treatment of the Sample
The analysis for manganese
in biological media may be
carried out in two different ways: directly, with or without
dilution and perhaps with the addition of a matrix modifier,
or indirectly, after mineralization
with or without chelation!
extraction.
Direct methods. In EAAS, the introduction of pure biological media limits the risks of contamination
but requires
lengthy temperature
ramps. Interferences,
matrix effects,
and carbon residues may be important in this procedure.
Very small quantities of liquid media may be injected into
the graphite furnace in a pure state without any treatment
or dilution (46,50,54,63,66,
72,92). Tissues may be placed
directly in the graphite furnace after lyophilization (52). For
hair, Alder et al. (71,93) decomposed the sample directly in
the graphite
tube. In direct methods with dilution, deionized water (43, 46, 54, 68, 69, 73, 94, 95), Triton X-l00
(24, 47, 48, 92) or other wetting agents (42, 96), acids (75,
96), animonium compounds (57), or quaternary
amines (51,
97, 98) are commonly used.
Indirect methods. Complex biological substances may be
simplified by acid digestion followed by simple dilution (41,
45, 54, 99), or by acid digestion followed by chelation and
extraction with a solvent (14, 15,46, 54, 100-102). Guided
by our experience we recommend, for urine and blood
serum, direct analysis with dilution, which allows good
practicability and minimizes contaminRtion.
For tissues, three procedures of mineralization
and solubilization have been proposed: solubilization with quaternary
amines (97, 98, 103, 104); acid digestion without extraction
(45, 49, 67, 72, 90, 105); and acid digestion with chelation!
extraction (44, 101).
Hair is dissolved, after washing (as described above), by
acid digestion and Mn is determined after simple dilution
(53,89). Saner et al. (70) dissolved the residue in HC1 after
dry ashing.
(44,49,69), or with the same diluents as for sample dilution
(42, 56). For Favier and Ruffleux (42), Muzzarelli and
Rocchetti (43), and Wei et al. (57), the results obtained with
an aqueous solution and standard additions are comparable.
On the other hand, Smeyers-Verbeke
et al. (64) do not
recommend the use of aqueous solutions because the chemical evolution of manganese depends heavily on the composition of the medium.
Determination
by standard additions. This is the method
preferred by most researchers
(14,43,46,47,58,63,64,67,
75, 90, 92, 96, 107).
Addition calibration. Pleban and Pearson (48), like Boiteau et al. (51), set up a range of standards by additions of
standard to the same medium (serum pool or whole blood
with very low manganese concentrations). For each sample,
the concentration
of manganese is taken directly from a
graph after deducting for each sample the value of a blank
reagent. The two last procedures of standardization
are the
most suitable, taking into account the matrix effects.
PrecIsIon
Within-Run Precision
For whole blood, serum, or plasma: Better within-run
results are obtained when the concentration in manganese
is high: reported CVs range from 1.7% (1019.3 nmol/L) to
12.5% (114.6 mnol/L) (48,51). Favier et al. (56) demonstrate
the importance of prior dilution of the sample: precision was
poor for a non-diluted sample (CV 27%), owing to high
background, and for one diluted 20-fold (CV 33%), probably
owing to low concentration after dilution. The best results
(CV 4%) are obtained with a fivefold dilution of the sample
in 10-fold-diluted ethylene glycol (56).
For urine: The within-run CV is generally <10%. Halls
and Fell (69) found a within-run CV of 11.9% for undiluted
urine and 7.4% for a twofold-dilution of the same urine. The
twofold dilution decreases the background and gives a much
smaller range for analytical
recoveries (69). Lekehal and
Hanocq (14) compared two techniques for treating urine:
extraction with and without prior acid mineralization
with
two different methods of determination (AAS and EAAS);
within-run results were comparable, and CVs ranged between 0.4% and 3%.
For hair: Guillard et al. (53) and Friel and Nguyen (74)
observed, for non-exposed subjects, within-run CVs of 3.9%
and 8%, respectively.
For tissue: The CV ranged between 2% and 5.6% (49, 72).
Between-Run Precision
CVs range from 2.3% (46) to 16% (54) for serum. The
results improve as the concentrations
in manganese
increase: Meissner et al. (73) observed CVs of 5.6, 3.8, and
2.4% for concentrations of 1.8, 18.2, and 182 nmol/L, respectively. For determination in tissue, the between-run CV was
between 6.3 and 18.5% (45,49, 52, 72).
Standards
Accuracy/SpecIficIty
The standard solution of manganese is prepared either
from the pure metal dissolved in sulfuric acid (92) or from a
commercial
standard
solution made from different compounds of manganese, such as MnCl2 (15,52, 106) or Mn02
(66), in water (14), nitric acid (44), or 0.5 mol/L sulfuric acid
recovery ranges between 90 and
56, 58, 69, 74, 78, 101).
Interferences. Matrix effects are significant (69), and they
increase in proportion to the amount of the deposit. The use
of background correction (deuterium lamp or Zeeman effect)
eliminates nonspecific interferences. Smeyers-Verbeke et al.
(64) specify that slow thermal pretreatment results in a
more effective reduction of interference by matrix. In most
(96).
ExternoJ standards. The aqueous standard solutions are
prepared with hydrochloric acid (66, 91, 105) or nitric acid
Recovery.
Analytical
110% (14, 44, 46, 49, 51-53,
CLINICAL CHEMISTRY, Vol. 34, No. 2, 1988
229
cases,
positive
interferences
is calculated
concentration
can be decreased when the
on the basis of the peak areas
Table 1. Reported
(64).
The significance of the interferences seems to be related to
the temperature
chosen for the mineralization.
Bek et al.
(66) observed a high background noise at a charring temperature of 750 “C, and d’Amico and Klawans (63) say this
temperature
must be 800 “C. Alt and Massmann (107) find
1000 “C to be apparently
insufficient to avoid interference
by absorption of molecular sodium chloride (107).
Sodium chloride in concentrations
greater than 10 to 20
mmolJL causes a reduction in the signal (44, 73). As a result,
the following are required: dilution of the sample (73),
extraction (44), addition of arnmonium nitrate, or, as mentioned above, an increase in the charring temperature
from
850 to 1100 “C. Favier et al. (56), like Sekiya et al. (99),
observed
no significant
interferences
by sodium. For
Smeyei-s-Verbeke
et al. (64) only sodium in sulfate compounds increased the signal (by 25%). Balling and Jones (44)
noted a negative interference
from potassium, not observed
by Sekiya et al. (99) or Favier et al. (56). Interferences from
chlorides should be compensated by the background correction (108). There appear to be no interferences from iron,
copper, or zinc (56). Several authors point out negative
interference from calcium and magnesium ions (44,56,109),
which can be eliminated by the addition of EDTA (56, 109)
or by extraction (44). Smeyers-Verbeke
et al. (64) studied
the influence of very high concentrations (18.2 mmol/L) of
calcium and magnesium
in different salts (chlorides, nitrates, phosphates, and sulfates). Calcium and magnesium
salts, according to their different natures, affected the signal
in different ways (from -80% to +50%). With calcium and
magnesium
chlorides,
the signal
reduction
can be explained
by a masking of the manganese
by these salts. SmeyersVerbeke et al. (109) observed an increase in the atomization
speed in the presence of calcium or magnesium nitrate.
Favier et al. (56), however, did not observe any major
interferences in the case of sulfates and phosphates.
Smeyers-Verbeke
et al. (64) demonstrated
the influence of
acids on the signal. Phosphoric acid, 100-fold and 20-fold
diluted (by vol), increases absorption by 15%. On the other
hand, hydrochloric acid in dilutions lower than or equal to
10-fold and sulfuric or nitric acids, 100-fold diluted, have no
effect. The sulfuric or nitric acids in 10-fold or 20-fold
dilutions (by vol) reduce the signal by 20% to 60%.
The volatility of manganese varies according to the concentration and the nature of the salts, so it is necessary to
test an extended series of concentrations
and different
temperature programs in accordance with different matrices. The use of aqueous standards does therefore not seem to
be suited for the determination of manganese in biological
materials.
Detection Limits
The detection
limits observed are reported
in Table 1.
LInearIty
Very few authors mention the upper limit of linearity in
methodologies.
Favier et al. (56) found it to be 910
nmollL for serum, and Pleban and Pearson (48,49) found it
to be 364 nmol!L for serum, 728 nmolJL for whole blood, and
820 mnol/L for solubilized tissues.
their
Reference Values
The data presented in the following tables were obtained
from the available published mean values, which vary
230
CLINICAL
CHEMISTRY,
Vol.34, No.2,
1988
Detection Limit of Various EAAS
Assays of Mn
Detection lImit,
Serum or plasma
nmol/L
Ref. no.
4.55
54
46
18.2
10.92
1.27
5.46
1.45
5.46
1.8
5.46
Blood
5.46
18.2
36.4
3.64
Original
Tissues
0.009 pmol/assay
5.46 nmol/g
0.18
46
92
78
authors’ units
0.626 nmol/g
3.64
nmol/g
0.145 pmol/assay
0.2 pmol/assay
Hair
107
63
48
56
72
41
51
48
nmoi/g
44
90
72
49
52
71
53
from one author to another, especially for serum and
urine (Tables 2 and 3). This discrepancy is due in part to the
sample-collection
conditions, possible sources of contamination, as well as individual
variations
in addition to the
different methodologies
(9, 14, 15, 30,46, 54,57, 107, 110).
For serum, neutron activation leads to low values, while
flame atomic absorption and photometry give high values.
For urine, the methodology seems to play a less-important
role. Values for plasma (Table 2) vary greatly, probably
owing to the type of anticoagulant used. Reported reference
values for manganese
in whole blood and erythrocytes
(Table 2) are more uniform. A 30-fold greater concentration
of manganese in erythrocytes
than in serum may explain
this uniformity of results.
Reference values for manganese in hair are also widely
disparate (Table 3). The effect of hair pigmentation
(see
below and 111) and the method used for washing are the
predominant factors here.
greatly
Physiological VarIatIons
Manganese
in the serum may be subject to a nyctohemeral cycle (42), with a peak at 0800, although Borner et al. (9)
did not observe such a cycle. Manganesemia
does not vary
according to sex (2, 8, 12, 42, 54). Masiak et al. (30),
however, observed slightly higher concentrations in women.
There also does not appear to be any variation in manganesemia in relation to age in adolescents or adults (42, 50,
54). During gestation, manganesemia
increases early, then
stabilizes, reaching normal values for adults at the time of
delivery (42).
The use of birth-control pills leads to an increase in
manganese in urine (112).
In hair, the concentration varies in relation to the pigmentation. A study by Guillard et al. (111) demonstrates
that dark hair is richer in manganese than grey hair. The
concentration of manganese in hair is low at birth (0.19 ±
0.11 g/g dry wt), increases rapidly during the first few
weeks after birth (0.965 ± 0.39 ig/g dry wt), and then
decreases between the sixth week and the third year (0.398
± 0.21 p.glgdrywt)(89).
Table 3. Reference Values for Manganese In Urine,
Cerebrospinal Fluid, Hair, and Liver Tissue
Table 2. Reference Values for Manganese In Blood
Method
Serum
2S
NAA
25c
2
95
NAA
AAS
50
EMS
ns
6
EMS
EMS
EMS
Whole blood
Erythrocytes
ns
EMS
EAAS
EMS
EAAS
EAAS
EMS
Plasma
n
NAA
9
19
ns
Reference values,
nmol/L
10,7 ± 2.2
10 ± 2.5
654
9to127
436±127
9to144
145to318
Method
28
30
12
164±78
43
20
33±12
48
40±11
50
37
4
SO
44±9
33±7.3
20.7 ± 3.7
20.5 ± 3
Photometry
36c
9
27
51
NM
EMS
16
4
36 to 145
10.5 ± 3.3
33 ± 7.3
EMS
13
145to418
MS
NM
NM
NM
MS
10
14
ns
145±18
153 ± 50
200±2
ns
908
206’
34
35
60
68
ns
6.5 to 17.4
73 to 351
148±46
218 ± 164
327
196±80
182±73
164±41
153±47
303 ± 94
78
69
57
10
10
EMS-MS
EMS-MS
EMS
50
ns
50
ns
ns
176
16
Cerebrospinal fluid
NM
EMS
ns
10
NM
ns
NM
3
MS
EMS
20
10
EMS
15
EMS
6
EMS
31
EMS
10
EMS
EMS
30
Hair
26
78
99
113
26
114
115
82
valuss
ns
ns
EMS
83
Ref. no.
nmol/L
38±11
18to182
114
110
102
116
22
19
46
14
15
69
36to76
163±5.3
24±4
163±54
229
181 to 327
ito 48
2to27
nmol/L
19±9.5
12±2
nmol/g (dry wt)
213±64
114
63
114
25
116
89
231±7
58±39
3.4 ± 2
5±0.9c
2.36 ± 091d
53
4.37 ± 0.54
Liver tissue
43
Mean
92
35
± SD, or range. 5Birth. cBrn.
43.6±21.8
70
74
13.47 ± 1.1
nmol/g (dry wt)
21.8 ± 6.5
94±24
unexposed. dwhlte.
101
72
8rown.
48
78
115
1Mean ± SD, or range. bMen. CWomen ‘Childrenupto 14 years. Boys
and girls, 2 monthsto 15 years old. ‘Adults.
Liver tissue showed no variation in concentrations
NM
MS
MS
MS
MS
MS
63
107
26d
n
Urine
54
154to736
18.5 ± 3.4
91 to 364
Rmnos
79
66
46
EMS
EMS
EMS
EMS
EMS
EMS
NM
Ret, no.
related
to age or sex (72).
ConclusIon
The determination of manganese
in biological materials
by EAAS still leaves us with many problems to be solved, as
is illustrated
by the discrepancy
in usual values and in
methods used for treating the sample before it is injected
into the graphite furnace. Certain analytical
parameters
have now been well established and these include wavelength (279.5 nm), slit width (generally 0.2 or 0.7 nm), the
necessity of using a background correction (D2 lamp or
Zeeman effect), and the use of pyrolytic tubes. A wetting
agent (such as Triton X-100) also seems useful. Charring
and atomization temperatures
do vary from author to author, but it seems that 1100#{176}C
and 2400 to 2600 “C are the
most suitable. The importance
of interferences makes us
favor internal standard additions or calibration by standard
additions as opposed to external standards. Precision and
accuracy are generally acceptable, while the limits of detection, values for which vary from author to author, often
approach the normal reference values, especially for biological materials such as serum, plasma, urine, and spinal fluid.
Clinical interest in Mn determination
is not yet well
established in cases of liver disease (cirrhosis), psychological
and neurological disorders (epilepsy, Parkinson-like
syndrome), and in certain cases of chronic ulcers (prolidase
deficiency). On the other hand, Mn determination
is of
special clinical interest in the detection of overloading as a
result of workplace exposure, and among hemodialysis
patients when the content of manganese
in the dialysis
liquid is high. In this case, it may be as important as
aluminum.
References
1. World Health Organization. Manganese. Environmental health
criteria 17. Geneva: WHO, 1981:123.
2. Saric M. Manganese. In: Friberg L, Nordberg GF, Vouk V, eds.
Handbook on the toxicology of metals, 2nd ed. Amsterdam: Elsevier, 1986:354-86.
3. Clegg MS. Lonnerdal B, Hurley LS, Keen CL. Analysis of whole
blood manganese by flamelesa atomic absorption spectrophotometry
and its use as an indicator of manganese status in animals. Anal
Biochem 1986;157:12-8.
4. Leach RM Jr. Metabolism and function of manganese. In: Prasad
AS, Oberleas D, eds. Trace elements in human health and disease.
Volume U. Essential
and toxic elements. New York: Academic
Press,1976:235-47.
5. Schroeder HA, Nason AP. Trace metals in human hair.J Invest
Dermatol 1969;53:71-8.
6. Cotzias GC, Papavasiliou PS, Ginos J, Streck A, Duby S.
Metabolic modification of Parkinson’s disease and of chronic manganese poisoning. Annu Rev Med 197 122:305-26.
7. Larregue M, Charpentier C, Laidet B, et al. Deficit en prolidase
et en manganese.
A propos d’une observation: diagnostic et traitement. Ann Dermatol Venerol 1982;109:667-78.
8. Versieck J, Barbier F, Speecke A, Hoste J. Manganese, copper
and zinc concentrations
in serum and packed blood cells during
acute hepatitis, chronic hepatitis, and posthepatic cirrhosis. Clin
Chem 197420:1141-5.
9. Borner H, Mehnert
E, Bruhn P, Hudec R Serum copper and
manganese concentrations in uremia. Z Urol Nephrol 1983;76:5448.
10. Mehnert E, Huth
Mangankonzentration
Fortbild
(Jena)
JH, Walter F, Hudec R. Das Verhalten der
des Blutserums beim Herzinfarkt. Z Arztl
1982;76:615-7
CLINICAL
CHEMISTRY,
Vol. 34, No. 2, 1988
231
11. Versieck J, Barbier F, Speecke A, Hosts J. Influence of myocardial infarction on serum manganese, copper, and zinc concentrations. Clin Chem 1975;21:578-81.
12. Mahoney JP, Sargent K, Greland M, Small W. Studies on
manganese. I. Determination
in serum by atomic absorption spectrophotometry. Clin Chem 1969;15:312-22.
13. Nath R. Preparation of biological samples and use of standard
reference material in trace analysis. Bull Postgrad Inst Med Educ
Res 1982;16:134-9.
14. Lekehal N, Hanocq M. Determination de traces de manganese
urinaire apres extraction a l’aide de 2,2,6,6-t#{233}tram#{233}thyl
heptane.
3,5-dione et dosage par spectrom#{233}trie
d’absorption atomique. Anal
Chim Acts 1976;83:93-l01.
15. Watanabe
T, Tokunaga R, Iwahana T, Tati M, Ikeda M.
Determination of urinary manganese by direct chelation-extraction
method and flameless atomic absorption spectrophotometry. Br J
hid Med 1978;35:73-7.
16. Berndt H, Messerschmidt J. Schlaufenmethode der AAS/AES
zur Element-analytik
in biologischen Materialien. Eine neue Moeglichkeit zur mikro und Spurenanalyse. Fresenius Z Anal Chem
34. Azi.z A, Broekaert JAC, Lois F. Analysis of microamounts
of
biological samples by evaporation in a graphite furnace and inductively coupled plasma atomic emission spectroscopy.Spectrochini
Acts Part B 1982;37B:369-79.
35. Black MD, Thomas MB, Browner RF. Determination of metal
chelates by inductively coupled plasma atomic emission spectrometry and applications to biological materials. Anal Chem
1981;53:2224-28.
36. Barnes RM, Fodor P. Analysis of urine using inductivelycoupled plasma emission spectroscopy with graphite rod electrothermal vaporization. Spectrochim Acts Part B 1983;38:1191-202.
37. Mehnert E. Diagnostic significance of manganese concentration in blood serum. Current state of knowledge. Z Arztl Fortbild
(Jena) 1982;76:419-21.
38. Rubio 5, Gomez-Hens A, Valcarcel M. Fluorimetric determination of manganese at the nanogram level by catalytic oxidation of
pyridoxal 2-pyridylhydrazone
by hydrogen peroxide. Analyst
1984;109:717-22.
39. Hambidge KM, Droegemnueller W. Changes in plasma and
hair concentrations of zinc, copper, chromium and manganese
1982;311:365-6.
during pregnancy.
Delves HT, Shepherd G, Vinter P. Determination
of eleven
metals in small samples of blood by sequential solvent extraction
and atomic absorption spectrophotometry. Analyst 197 1;96:260-73.
18. Farag RS, El-Aaasar ST, Mostafa MA, Abdel-Rahim EA. Use of
atomic absorption analysis of mineral content of certain animal
hairs in relation to sex (I). J Drug Res 1980;12:217-25.
19. Holler U, Voigt H, Bergert KD. Determination of manganese in
urine by flame atomic absorption spectrophotometry.
Zentralbl
Pharm Pharmakother Laboratoriurosdiagn
1984;123:273-6.
20. Lau HKY, Ashmead H. Trace minerals in human hair by
atomic absorption spectrophotometry;
recovery study. Anal Lett
40. Yoakum AM, Stewart PL, Sterrett JE. Method development
and subsequent survey analysis of biological tissue for platinum,
lead and manganese content. Environ Health Perspect 1975;1O:85-
1975;8:815-24.
21. Lonnerdal B, Keen CL, Ohtake M, Tamura T. Iron, zinc, copper
and manganese in infant formulas. AmJ Dis Child 1983;137:433-7.
22. Pantucek MB. The determination of manganese in urine. Czech
Med 1982;5:225-7.
23. Shearer DA, Cloutier RO, Hidiroglou M. Chelate extraction
and flame atomic absorption spectrometric determination of nanogram amounts of manganese in blood and animal tissue. J Assoc Off
Anal Chem1977;60:155-9.
24. Ward NI, Stephens R, Ryan DE. Comparison of three analytical
43. Muzzarelli RAA, Rocchetti R. Atomic absorption determination
of manganese, cobalt and copper in whole blood and serum, with a
graphite atomizer. Talant.a 1975;22:683-5.
44. Belling GB, Jones GB. The determination of manganese in
small samples of biological tissues by flameless atomic absorption
spectrometry. Anal Chim Acts 1975;80:279-83.
45. Paynter DI. Microdetermination of manganese in animal tissues by flameless atomic absorption spectrometry.
Anal Chem
1979;51:2086-9.
46. Ross RT, Gonzalez JG. The direct determination
of trace
quantities of manganese in blood and serum samples using selective volatilization and graphite tube reservoir atomic absorption
spectrophotometry. Bull Environ Contam Toxicol 1974;12:470-4.
47. Brodie KG. Trace analysis of lead in blood, aluminium
and
manganese in serum and chromium in urine by graphite furnace
atomic absorption spectrometry. Clin Biochem 1984;17:19-26.
48. Pleban PA, Pearson KH. Determination of manganese in whole
blood and serum. Clin Chem 1979;25:1915-8.
49. Pleban PA, Kerkay J, Pearson KH. Polarized Zeeman-effect
flameless atomic absorption pectrometry
of cadmium, copper, lead,
and manganese in human kidney cortex. Clin Chem 198 1;27:68-72.
50. Hoffmann H. Manganese serum levels of epileptics [Letter].
17.
methods for the determination of trace elements in whole blood.
Anal Chim Acts 1979;110:9-19.
25. Fergusson JE, Holzbecher J, Ryan DE. The sorption of copper
(II), manganese (II), zinc (II) and arsenic (ifi) onto human hair and
their desorption. Sci Total Environ 1983;26:121-35.
26. Cotzias GC, Miller ST, EdwardsJ. Neutron activation analysis:
the stability of manganese concentrations in human blood and
serum. J Lab Clin Med 1966;67:836.-49.
27. Versieck J, Cornelis R, Lemey G, De Rudder J. Determination
of manganese in whole blood and serum. Clin Chem 1980;26:531-2.
28. Wiesener W, Grund W. Neutronenaktivierungsanalyse
von
Mangan in menschlichen
Haar und Serum. Isotopenpraxis
1978;14:147-8.
29. Kocsis E, Kovats M, Molnar M. Bestimmung von Mangan in
Blut mittels Neutronenaktivationsanalyse.
Period Polytechn Chem
Eng 1981;25:133-41.
30. Masiak M, Skowron S, Owczarek H, Wodkiewicz L, Stacherzak
J. Serum levels of certain trace elements (Cu, Au, Mn) in healthy
subjects. Acts Physiol Pol 1981;32:537-46.
31. Meurling S, Plantin LO. The concentrations of trace elements
in blood from healthy newborn infants. Acta Chir Scand
1981;147:481-5.
32. Grund W. Manganeinlagerung
in das Kopfhaar des Menschen,
em Maszstab fur die Expositionsbewertung.
Arch Exp Vet Med
(Leipzig) 1980;34:347-51.
33. Aziz A, Broekaert JAC, Lois F. A contribution to the analysis of
microamounts of biological samples using a combination of graphite
furnace and microwave induced plasma atomic emission spectrometry. Spectrochim Acts Part B 1982;37B:381-9.
232
CLINICALCHEMISTRY,Vol.34, No.2, 1988
Obstet Gynecol 1974;44:666-72.
93.
41. Subramanian
KS, Meranger JC. Graphite furnace atomic absorption spectrometry with nitric acid deproteinization for determination of manganese in human plasma. Anal Chem 1985;57:247881.
42. Favier A, Ruffleux D. Physiological variations of serum levels
of copper, zinc, iron and manganese.
Biomed Pharmacother
1983;37:462-6.
Klin Wochenschr
1980;58:157-8.
51. Boiteau HL, M#{233}tayer
C, Ferr#{233}
R, Pineau A. Dosage automatique du plomb, du cadmium, du manganese et du chrome dana le
sang par spectrom#{233}trie
d’absorption atomique Zeeman. Analusis
1983;11:234-42.
52. Rorsman P, Berggren P0. Direct determination of manganese
in microgram amounts of pancreatic tissue by electrothermal
atomic absorption spectrometry. Anal Chim Acts 1982;140:325-9.
53. Guillard 0, Brugier JC, Piriou A, Menard M, Gombert J, Reiss
D. Improved determination of manganese in hair by use of a miniautoclave and flameless atomic absorption spectrometry with Zeeman background correction: an evaluation in unexposed subjects.
Clin Chem 1984;30:1642-5.
B, Buttgereit G, Kubler W, Mertens HM. Die Messung der Spurenelemente Chrom und Mangan im Serum mittels
flammenloser Atomabsorption. Z Klin Chem Kim Biochem
54. Grafllage
1974;12:287-93.
55. Favier A. Misc au point de microdosages des oligo#{233}lements
fer,
cuivre, zinc et manganese par absorption atomique sans flamme.
Role de ces elements dana Ia gestation [Ph.D. thesis]. Univ Pharmac
Sci of Grenoble, France, 1981.
56. Favier
A, Ruffleux D, Alcaraz A, Maljournal B. Flameless
atomic absorption assay of serum manganese. Clin Chim Acts
1982;124:239-44.
57. Wei FS, Qu WQ, Yin F. Direct determination
of ultra-trace
manganese in serum by controlled temperature graphite furnace
method of atomic absorption. Anal Lett 1982;15:721-9.
58. Casey CE, Hambidge KM, Neville MC. Studies in human
lactation: zinc, copper, manganese and chromium in human milk in
the first month of lactation. Am J Clin Nutr 1985;41:1193-200.
59. Carnrick GR, Slavin W, Manning DC. Direct determination of
manganese in seawater with the L’vov platform and Zeeman
background correction in the graphite furnace. Anal Chem
1981;53:1866-72.
60. Slavin W, Carnrick GR, Manning DC. Magnesium nitrate as a
matrix modifier in the stabilized temperature platform furnace.
Anal Chem 1982;54:621-4.
61. Slavin W, Carnrick GR, Manning DC, Pruszkowska E. Recent
experience with the stabilized temperature
platform furnace and
Zeeman background correction. At Spectrosc 1983;4:69-86.
62. Ottaway JM. Recent innovations in electrothermal
atomization. Proc Chem Soc London 1984;21:55-9.
63. D’Amico DJ, Kiawans ilL.Direct microdetermination of manganese in normal serum and cerebrospinal fluid by flameless atomic
absorption spectrophotometry. Anal Chem 1976;48:1469-72.
64. Smeyers-Verbeke J, Michotte Y, Van Der Winkel P, Massart
DL. Matrix effects in the determination of copper and manganese in
biological materials using carbon furnace atomic absorption spectrometry. Anal Chem 1976;48:125-30.
65. Manning DC, Slavin W. The determination of manganese by
electrothermal atomic spectrometry with a graphite furnace at
constant temperature. Anal Chim Acts 1980;118:301-6.
66. Bek F, Janouskava J, Moldan B. Determination of manganese
and strontium in blood serum using the Perkin Elmer HGA 70
graphite furnace. At Absorpt Newsl 1974;13:47-8.
67. Clegg MS, Keen CL, Lonnerdal B, Hurley LS. Analysis of trace
elements in animal tissues. III. Determination of manganese by
graphite furnace atomic absorption spectrometry. Biol Trace Element Res 1982;4:145-56.
68. Graifiage B, Buttgereit G, Kubler W. Use of flameless atomic
absorption for the measurement of trace elements in human serum.
Verh Dtsch Ges Inn Med 1974;80:1710-3.
69. Halls DJ, Fell GS. Determination of manganese in serum and
urine by electrothermal atomic absorption spectrometry. Anal
Chim Acts 1981;129:205-11.
70. Saner C, Dagoglu T, Ozden T. Hair manganese concentrations
in newborns and their mothers. Am J Clin Nutr 1985;41:1042-4.
71. Alder JF, Samuel AJ, West TS. The single element determination of trace metal in hair by carbon furnace atomic absorption
spectrometry. Anal Chim Acts 1976;87:313-21.
72. Hudnik V, Marolt-Gomiscek M, Gomiscek S. The determination of trace metals in human fluids and tissues. Part 1: Estimation
of “normal values” for copper, zinc, cadmium and manganese in
blood serum and liver tissues. Anal Chim Acta 1984;157:143-50.
73. Meissner D, Drescher V, Schmidt LII. Determination
and
clinical significance of manganese. Zentralbl Pharm Pharmakother
Laboratoriumsdiagn
1982;121:432-7.
74. Friel JK, Nguyen CD. Dry and wet-ashing techniques compared in analyses for zinc, copper, manganese, and iron in hair. Clin
Chem 1986;32:739-42.
75. Casey CE, Jacobs Goodall MA, Hambidge KM. Atomic absorption spectrometry
of manganese
in plasma.
Clin Chem
1987;33:1253-4.
76. Lundgren G, Lundmark L, Johansson G. Temperature controlled heating of the graphite tube atomizer in flameless atomic
absorption. Anal Chem 1974;46:1028-31.
77. Thou NG, Frech W, De Galan L On the relationship between
heating rate and peak height in electrothermal atomic absorption
spectrometry. Spectrochim Acts 1984;39B:225-36.
78. Weissman N, Babich-Armstrong M. Determination of manganese in human whole blood [Abstract]. Clin Chem 1980;26:1023.
79. Versieck J, Barbier F, Speecke A, HoeteJ. Normal manganese
concentrations
in human serum. Acts Endocrinol 1974;76:783-8.
80. Hudnik V, Marolt-Gomiscek
M, Gomiscek S. The determination of trace metals in human fluids and tissues. Part 3: Contamination in sampling of blood serum and liver tissue and their stability
on storage. Anal Chim Acts 1984;157:303-11.
81. Versieck J, Speecke A, Hoste J, Barbier F. Determination of
manganese, copper and zinc in serum and packed blood cells by
neutron activation analysis. Studies on the metabolism of trace
elements. Z Klin Chem Klin Biochem 1973;11:193-6.
82. Delves HT, Clayton BE, Bicknell J. Concentration of trace
metals in the blood of children. Br J Prey Soc Med 1973;27:100-7.
83. Favier A, Ruffleux D. Determination of manganese in serum,
normal values and physiological variations. J laker Metab Dis
1983;6:90.
84. Nath R. Preparation of biological samples and use of standard
reference material in trace analysis. Bull Poatgrad Inst Med Educ
Res 1982;16:134-9.
85. Barnes RM, Fodor P, Inagaki K, Fodor M. Determination of
trace elements in urine using inductively coupled plasma spectroscopy with a poly(dithiocarbamate) chelating resin. Spectrochim
Acts Part B 1983;38B:245-57.
86. Salmela S, Vuori E, KilpioJO. The effect of washing procedure
on trace element content of human hair. Anal Chim Acts
1981;125:131-7.
87. Assarian GB, Oberleas D. Effect of washing procedures on
trace-element content of hair. Clin Chem 1977;23:1771-2.
88. Chittleborough G. A chemist’s view of the analysis of human
hair for trace elements [Review]. Sci Total Environ 1980;14:53-75.
89. Collipp PJ, Chen SY, Maitinsky S. Manganese
in infant
formulas and learning disability. Ann Nutr Metab 1983;27:488-94.
90. Bonilla E. Flameless atomic absorption spectrophotometric
determination of manganese in rat brain and other tissues. Clin
Chem 1978;24:471-4.
91. Hudnik V, Marolt-Gomiscek
M, Gomiscek S. The determination of trace metals in human fluids and tissues. Part 2: The
homogeneity of liver tissue for sampling. Anal Chim Acts
1984;157:183-6.
92. Tsalev DL, Langmyhr FJ, Gunderson N. Direct atomic absorption spectrometric determination of manganese in whole blood of
unexposed individuals and exposed workers in a Norwegian mangaaese alloy plant. Bull Environ Contam Toxicol 1977;17:660-6.
93. Alder JF, Samuel AJ, West TS. The simultaneous multielement analysis of hair: a nonparametric
method for evaluating
the ability of the data to distinguish between individuals. Anal
Chim Acts 1977;94:187-91.
94. Halls DJ. Applications of graphite furnace atomic absorption
spectrometry in clinical analysis. Anal Proc 1981;18:344-6.
95. Askari A, Long CL, Blakemore WS. Zinc, copper, iron and
manganese in blood and blood products. Nutr Support Serv
1983;3:28-9.
96. Bourdon R, Galliot M, Prouillet F. Dosage du cuivre, du plomb,
du manganese, du bismuth, du cadmium et de l’or dana les liquides
biologiques par spectrom#{233}trie
d’absorption atomique sans flanime.
Ann Biol Clin 1974;32:413-22.
97.
Dorner WG. Aufschluszmethoden
Fachz Lab 1982;26:750-4.
in der Spurenanalyse. GIT
98. Dorner WG. Application of a tetraalkylammomum
hydroxide
as a solubilizer in trace analysis. G1T Fachz Lab 1983;27:833-4.
99. Sekiya T, Tanimura H, Hkiasa Y. Simplified determination of
copper, zinc, and manganese in plasma and bile by flameless atomic
absorption spectrometry. Arch Jpn Chir 1981;50:729-39.
100. Cholak J, Hubbard DM. Determination of manganese in air
and biological material. Am md Hyg Assoc J 1960;21:356-60.
101. Sumino K, Hayakawa K, Shibata T, Kitamura S. Heavy
metals in normal Japanese
tissues. Arch Environ Health
1975;30:487-94.
CLINICAL
CHEMISTRY,
Vol. 34, No. 2, 1988
233
102. Van Ormer DG, Purdy WC. The determination of manganese
urine by atomic absorption
spectrometry.
Anal Chim Acts
1973;64:93-105.
103. Pfueller U, Fuchs V, Golbs S. Ebert E, Pfeifer D. Solubilization to prepare biological samples for determination of heavy metals
by flameless atomic absorption spectrophotometry. Arch Exp Vet
Med 1980;34:367-72.
104. Goldberg WJ, Allen N. Determination
of Cu, Mn, Fe, and Ca
in six regions of normal human brain by atomic absorption spectroscopy. Clin Cheni 1981;27:562-4.
105. Smeyers-Verbeke J, Segebarth G, Massart DL The determination of copper and manganese in small biological samples with
graphite
furnace absorption spectrometry.
At Abeorpt Newsl
1975; 14:153-4.
106. Ikeda M, Kaneko I, Watanabe T, Ishihara N, Miura T. An
automated system for the determination of lead in blood, manganese in urine and nickel in waste water. Am md Hyg Assoc J
1978;39:226-32.
107. Alt F, Massmann H. Atomabeorptionspektrometrische
Bestimmung von Spuren-elementen (Mn-Co) in Seren. Fresenius’ Z
Anal Chem 1976;279:100-2.
108. Suzuki M, Ohta K, Yamakita T. Elimination of alkali chloride
interference with thiourea in electrothermal atomic absorption
spectrometry of copper and manganese. Anal Chem 1981;53:9-13.
109. Smeyers-Verbeke J, Michotte Y, Massart DL. Influence of
in
234
CLINICAL CHEMISTRY, Vol. 34, No. 2, 1988
some matrix elements on the determination of copper and manganese by furnace atomic absorption spectrometry.
Anal Chem
1978;50:1O-2.
110. Ajemian RB, Whitman NE. Determination of manganese in
urine by atomic absorption spectrometry.
Am Ind Hyg Assoc J
1969;30:52-6.
111. Guillard 0, Gombert J, Barriere M, Reiss D, Piriou A.
Manganese concentration in the hair of greying (“salt and pepper”)
men reconsidered [Letter]. Clin Chem 1985;31:1251.
112. Klinger G, Dawczynski H, Winnefeld K, et al. Zur Bestimmung von Spuren-elemente im Serum (Fe,Cu,Zn) und im Urin (Fe,
Cu, Zn, Mn) sowie von Carnerproteinen bei hormonaler Kontrazeption. Z Arztl Fortbild (Jena) 1980;74:1077.-84.
113. Capel ID, Pinnock MB, Williams DC, Hanham 1W. The serum
levels of some trace and bulk elements in cancer patients. Oncology
1982;39:38-41.
114. Cotzias GC, Horiuchi K, Fuenzalida
5, Mena I. Chronic
manganese poisoning clearance of tissue manganese concentrations with persistence of the neurological picture. Neurology
1968;18:376-82.
115. Versieck J, Barbier F, Hosts J, Cornelis R, Do Rudder J.
Manganese in serum [Letter]. Talanta 197623:745.
116. Bergert XI), Voigt H, Holler U. Detection of exposure in
welders by determining manganese contents of biological materials.
Z Gesamte Inn Med 1982;37:504-7.