Full text of DOI:10.1002/etc.5620210613

Environmental Toxicology and Chemistry, Vol. 21, No. 6, pp. 1206–1212, 2002
2002 SETAC
Printed in the USA
0730-7268/02 $9.00 .00
᭧
ϩ
ANTIFUNGAL ACTIVITY OF SOME TRITYL-BASED SYNTHETIC DYES
G
YULA
†Plant Protection Institute, Hungarian Academy of Sciences, Budapest, Hungary
‡Institute of Chemistry, Chemical Research Centre, Hungarian Academy of Sciences, P.O. Box 17, 1525 Budapest, Hungary
OROS,† TIBOR CSERHATI,‡ and ESTHER FORGACS*‡
(
Received 21 March 2001; Accepted 18 October 2001)
Abstract—The fungicidal activity of 10 trityl dyes and six reference compounds was determined on 36 fungal strains, and the data
matrix was evaluated separately by principal component analysis (PCA) and by spectral mapping technique (SPM). The dimen-
sionality of the maps of the principle component loadings and variables and the selectivity maps were reduced to two by varimax
rotation and by nonlinear mapping. Calculations proved that both the strength and selectivity of the fungicidal activity of trityl
dyes considerably depended on the chemical structure of the dye and on the type of fungi. Both PCA and SPM were suitable for
evaluation of the antifungal activity of dyes; however, the strength and selectivity of the fungicidal effect can be separated only
by SPM. Due to its advantageous application parameters, use of SPM in future quantitative structure-activity relationship studies
is highly recommended.
Keywords—Antifungal activity
Trityl-based synthetic dyes
Mathematical-statistical methods
INTRODUCTION
lationships between the columns and rows of any data matrix
without being one of the dependent variables [15]. Because
PCA is versatile and easy to carry out, it has been applied
often in many fields of research [16,17]. The method clusters
the elements of the matrix with simultaneous consideration of
the strength and selectivity of the effect under investigation;
that is, it is not suitable for data evaluation when separate
calculations of the strength and selectivity of the effect are
required.
The spectral mapping technique (SPM), another multivar-
iate mathematical-statistical method, has been developed for
separation of the strength and selectivity of the effect [18].
The SPM divides the information into two matrices using the
logarithm of the original data. The first matrix is a vector
containing the potency values related to the overall effect (in
the present study, the overall antifungal effect of dyes and the
overall sensitivity of fungi toward dyes). The second matrix
(i.e., selectivity map) contains the information related to the
spectra of activity (in the present study, the selectivity of the
antifungal effect of dyes and the selectivity of the response of
fungi toward dyes) [19].
Various synthetic dyes have been frequently employed in
agrochemistry [1,2], in up-to-date food science and technology
[3,4], and in numerous industrial processes [5,6].
The amount of synthetic dyes produced in the world is not
exactly known. It is assumed to be more than 10,000 ton/year.
Accurate data regarding the quantity of dyes released in the
environment are also not available. A loss of 1 to 2% in pro-
duction and of 1 to 10% in use may be a fair estimate. Because
of large-scale production and use of synthetic dyes, these sub-
stances are now serious environmental pollutants [7] and health
risk factors [8]. The impact and fate of dyes discharged in the
environment have been widely discussed [9,10]. Because a
considerable number (several thousands) of synthetic dyes are
used that exert various toxicological effects, it is understand-
able that knowledge regarding their effects on the environment
and health hazards related to their application is still incom-
plete. Synthetic dyes released in industrial effluents and waste-
waters can contaminate soil and decrease the growth of com-
ponents of soil microflora, such as nitrogen-fixing cyanobac-
teria [11] and the gram-negative soil bacterium Myxococcus
xanthus [12], decreases the growth of Bacillus subtilis [13],
etc. It has further been established that exposure to alternating
current makes Escherichia coli susceptible to basic dyes [14].
The assessment of large data matrices containing a high
number of columns and rows (as in the present study, which
involves the antifungal activity of 10 synthetic dyes and six
reference compounds on 36 fungi) is cumbersome or impos-
sible with the application of linear regression analysis, as is
generally used for quantitative structure-activity relationship
studies. A considerable number of multivariate mathematical-
statistical methods have been developed and successfully em-
ployed to facilitate elucidation of the maximal information
present in such matrices. Principal component analysis (PCA),
a multivariate method, makes possible assessment of the re-
The objectives of the present study were measurement of
the antifungal effect of some trityl dyes and reference com-
pounds, assessment of the data by PCA and SPM, and com-
parison of the results of PCA and SPM by stepwise regression
analysis [20].
MATERIALS AND METHODS
Common as well as International Union of Pure and Ap-
plied Chemistry names of trityl dyes are listed in Appendix
1, and their chemical structures are shown in Figure 1. Each
trityl dye was a commercial product and was used as received.
The selection of this set of trityl dyes was motivated by their
markedly different chemical structures (with and without qua-
ternary amino groups, etc.) and by the fact that they are ex-
tensively used in various large-scale industrial processes. Clo-
trimazole (XI), fenarimol (XII), nuarimol (XIII), triarimol
(XIV), blasticidin-S (XV), and trichotecin (XVI) were em-
* To whom correspondence may be addressed
(forgacs@cric.chemres.hu).
1206

Antifungal activity of some trityl-based synthetic dyes
Environ. Toxicol. Chem. 21, 2002
1207
Fig. 2. Similarities and dissimilarities among the antifungal activities
of trityl dyes and reference compounds as shown by a two-dimensional
nonlinear map of principal component loadings (no. of iterations, 116;
Ϫ
maximal error, 6.29
ϫ
10 2). Roman numbers refer to trityl dyes and
reference compounds in Appendix 1 and Materials and Methods,
respectively.
tophane (2 g/L), yeast extract (1 g/L), Na-␤-glycerophos-
phate·5H2O (0.6 g/L), MgSO4·7H2O (0.5 g/L), KH2PO4 (0.25
g/L), Na2HPO4·7H2O (0.25 g/L), KCl (0.15 g/L), and
FeSO4·5H2O (0.01 g/L). The potato broth was prepared from
peeled tubers (200 g) cut in pieces (ϳ1 ϫ 1 ϫ 1 cm), cooked
in distilled water (1 L) for 20 min, and then filtered through
a Perlon meshwork (no. 500; Leverkusen, Germany). The fil-
trate was made up to 1 L with distilled water. The broth was
solidified with 12 g/L of agar-agar (PDA) and sterilized at
Fig. 1. Chemical structures of trityl dyes. Roman numbers refer to
trityl dyes in Appendix 1.
120
Њ
C for 20 min. The inoculated agar slants were incubated
C. For the production of conidia, fungi (species 1–
at 21
Ϯ 2Њ
34) were grown on malt agar slants containing NaNO3 (3 g/
L), KCl (0.5 g/L), KH2PO4 (0.5 g/L), K2HPO4 (0.5 g/L),
MgSO4 (0.5 g/L), pyridoxine-HCl (1 mg/L), thiamin-HCl (10
mg/L), riboflavin (1 mg/L), nicotinamide (20 mg/L), and citric
acid (0.2 g/L). Inocula were prepared by suspending conidia
of vegetative cells (105 cell/ml) in sterile, distilled water con-
ployed as reference compounds with well-established antifun-
gal activity. Unless otherwise noted, 0.2 mM stock solutions
of test compounds were prepared in analytical-grade methanol
and stored at 0 to 5ЊC in the dark. Analytical-grade mineral
salts, glucose, amino acids (Reanal Fine Chemicals, Budapest,
Hungary), vitamins (Chinoin Pharmaceutical, Budapest, Hun-
taining glucose (2 g/L) and Tween௢ 40 (0.2 g/L; ICI Americas,
gary), Na-␤-glycerophosphate (Sigma-Aldrich, Budapest,
Bridgewater, NJ, USA).
Hungary), bacteriological agar no.1, Trypton T (L43) (Oxoid,
Basingstoke, UK), yeast extract (L21; Oxoid, Basingstoke,
UK), and malt extract broth (Sigma-Aldrich) were used for
preparing the media.
Oomycetes (species 35 and 36) were maintained on green
pea agar (GPA). The GPA was prepared by adding PDA (11
g/L), glucose (10 g/L), Trypton T (2 g/L), yeast extract (0.5
g/L), Na-␤-glycerophosphate·5H2O (0.5 g/L), KCl (0.25 g/L),
The fungi tested and their taxonomical positions are listed
according to Hawksworth et al. [21] in Appendix 2. All test
organisms listed in Appendix 2 were taken from the collection
of the Plant Protection Institute of the Hungarian Academy of
Sciences (Budapest, Hungary). The test species represent var-
ious taxonomic groups as well as modes of habitation (sap-
rotrophes, species parasitizing predominantly by necrotrophic,
hemibiotrophic, and biotrophic modes on whole plants or aerial
or underground parts of plants) [22].
and MgSO4 (0.15 g/L) to the broth made from green peas. The
broth was prepared from sugar peas (450 g) cooked in 1 L of
distilled water for 20 min, then treated in the same way as the
potato broth. For both species, 4-d-old colonies, grown on
cellophane film on the surface of GPA plates, were gently
triturated in green pea broth, and these suspensions (50 mg
mycelium/ml) were used as inoculum.
For antifungal activity tests, the PDA was inoculated with
fungus (105 propagules per 20 ml of medium), and a layer
(depth, 5 mm) was dispensed into Petri dishes (diameter, 90
mm). Filter paper discs (diameter, 5 mm) impregnated with
Saprophytic and facultative parasitic fungi (species 1–34)
were maintained on agarized potato broth amended with tryp-

1208
Environ. Toxicol. Chem. 21, 2002
G. Oros et al.
Ϫ
6
Table 1. Antifugnal effect of trityl dyes as the diameter of inhibitory zones (mm) caused by 10 M of dyes
Dyes
Fungi
I
II
III
IV
V
VI
VII
VIII
IX
PS%b
1
2
3
4
5
6
7
8
9
0
10
0
11
7
14
0
37
15
30
7
31
8
34
2
22
14
29
22
23
0
27
13
22
31
37
0
0
8
0
0
0
0
6
5
0
5
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
7
0
0
5
0
7
0
6
15
0
0
0
0
0
0
7
0
0
25
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
3.5
0
0
0
0
0
0
5
0
0
6
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1.0
47.3
33.6
39.4
56.8
57.3
26.6
85.1
33.2
100.0
60.6
53.9
62.2
28.6
45.6
56.8
66.4
39.8
51.0
39.4
57.3
71.0
56.8
52.3
61.4
28.6
33.6
39.0
58.5
66.8
81.7
58.1
51.9
70.5
40.7
36.5
31.5
0
0
0
5
0
0
0
0
5
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
15
30
0
0
0
0
0
0
18
0
11
34
34
10
17
29
23
30
33
30
20
30
34
25
13
35
35
32
5
38
29
41
5
5
5
12
29
47
32
22
38
7
10
6
79.6
33
34
37
40
21
60
49
73
45
54
28
62
42
40
30
37
45
33
34
43
32
42
34
31
27
45
47
57
35
40
46
35
37
25
135.2
9
15
30
20
30
31
35
21
37
33
25
21
37
35
16
5
39
30
43
5
5
6
35
31
43
30
19
38
5
44
18
42
21
35
20
31
18
18
26
23
20
19
14
17
26
22
14
18
15
27
25
32
20
30
27
18
19
13
74.6
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
PA/%c
6
7
11
11
15
5
14
15
10
11
0
9
0
11
47
11
30
9
18
18
14
14
15
9
7
9
81.6
8
38.5
0
6.9
6.0
a Roman and Arabic numerals refer to trityl dyes and fungi species in Appendices 1 and 2, respectively.
b PS%
ϭ
ϭ
potential sensitivity of species toward trityl dyes expressed as a percentage of the most sensitive species.
potential activity of trityl dyes toward species expressed in percent of the most active trityl dyes. Least significance difference (LSD5%
c PA%
)
ϭ
1.90 (Fcalc. ϭ 1346; F5% ϭ 1.42).
Ϫ
6
10 M of test compounds were placed centrally on the agar
plate (one disk per dish), and growth-inhibition zones were
and selectivity of the test species toward dyes were scaled by
SPM, and the potential sensitivity of species toward trityl dyes
and potential activity of trityl dyes toward species were cal-
culated as percentages (with the highest sensitivity and highest
activity being 100%).
measured after incubation at 22
Ϯ 1ЊC for 48 h.
For anti-oomycete tests, the GDA medium was inoculated
with Pythium irregulare or Phytophtora cactorum mycelium
suspension (1 ml/dish) and a layer of the medium (depth, 5
mm) was dispensed into Petri dishes (diameter, 90 mm). Filter
paper disks (diameter, 5 mm) were impregnated with 106 M
of test compounds and placed centrally on the agar plate (one
disk per dish). The growth inhibition zones were measured
The PCA was employed for assessment of the similarities
and differences in 15 test compounds and 36 species simul-
taneously, taking into consideration simultaneously the
strength and selectivity of the antifungal activity. The columns
in this matrix are the activity data of test compounds, and the
rows are the species listed in Appendix 2. The variance ex-
plained was set to 95%. To calculate the strength and selectivity
of the antifungal effect of test compounds and the strength and
selectivity of the sensitivity of test organisms, SPM was carried
out on the data matrix used for PCA and on the transversed
matrix. Because evaluation of the multidimensional maps of
principle component (PC) loadings, PC variables, and selec-
tivity (i.e., spectral) maps calculated by SPM is difficult, the
dimensionality of the maps was reduced to two by the non-
linear map technique (NLMAP). Theoretically, NLMAP can
reduce the dimensionality of any multidimensional data matrix
without submitting the data to PCA. According to our previous
after incubation at 18
Ϯ 1ЊC for 72 h.
All experiments were carried out at least three times.
Fisher’s probe was applied for calculation of the least signif-
icant difference values of response data. Because phenolphtha-
lein (compound X) showed no antifungal activity, it was omit-
ted from subsequent calculations.
The data matrices used for multivariate analyses consisted
of the antifungal activities (inhibitory zones in mm).
Cluster analysis (unweighted pair group average method)
was employed for elucidation of the similarity of fungi in their
responses toward dyes (with the 36 test species and 10 dyes
being the observations and variables, respectively). Sensitivity

Antifungal activity of some trityl-based synthetic dyes
Environ. Toxicol. Chem. 21, 2002
1209
Table 2. Antifungal effect of reference compounds as the diameter of
culated by the first and second co-ordinates of the two-di-
mensional nonlinear map of PC variables, the corresponding
potency values, and the first and second co-ordinates of the
Ϫ
6
inhibitory zones (mm) caused by 10 M of reference compoundsa
Reference compounds
two-dimensional nonlinear selectivity map (5
ϫ 36 matrix).
Fungi
XI
12
34
22
23
23
58
5
40
42
60
16
15
17
44
18
5
XII
XIII
XIV
XV
XVI
The SPM was carried out on the original matrix.
Comparison of the results of PCA and SPM was motivated
by the following considerations: Simultaneous or separate ap-
plication of PCA and SPM for the evaluation of multidimen-
sional data matrices is only justified when the information
content of the results is different. It was assumed that the
presence or absence of significant correlations is a good in-
dicator of the difference in the results. Presumably, it can be
employed as a suitable validation parameter for such types of
comparison.
1
2
3
4
5
6
7
8
9
11
19
0
21
13
38
5
41
70
81
6
13
0
8
5
15
23
25
24
43
23
20
14
42
5
37
71
82
14
25
5
5
41
44
19
13
25
34
14
31
85
11
11
11
33
5
27
8
20
41
30
5
13
15
15
13
14
5
40
43
21
12
5
19
30
47
38
51
8
25
26
40
9
49
50
74
21
19
17
6
10
23
16
44
5
43
82
89
19
25
7
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
PA%b
RESULTS AND DISCUSSION
30
0
0
45
5
5
The antifungal effects of trityl dyes and reference com-
pounds are compiled in Tables 1 and 2, respectively. The co-
efficient of variation of the parallel measurements ranged from
0 to 7.2%, depending considerably on the compound and the
species. It exceeded 4% in only 45 of the 360 cases. The
highest value was achieved in the measurement of the effect
of methyl green against Trichoderma harzianum. No signif-
icant differences were observed among the parallel determi-
8
34
42
90
38
0
25
26
23
27
31
5
65
18
28
18
19
26
15
0
43
49
90
47
7
34
35
39
17
25
5
69
14
42
16
31
43
17
0
5
5
29
83
34
5
20
30
11
15
39
6
40
13
36
8
20
0
0
32
34
35
90
46
0
25
39
37
18
28
5
61
13
43
20
6
28
18
55
85
27
17
35
30
22
10
15
0
33
22
21
11
0
nations (Fcalc.
ability of the measurements. The sensitivity of species toward
dyes showed significant differences ( calc. ϭ 91.5, 5% ϭ 1.54),
ϭ 0.34, F10% ϭ 2.71), which verifies the reli-
F
F
indicating that the various dyes exert different impacts on the
fungi in the environment. None of the species showed complete
tolerance, and Alternari solani, Colletotrichum graminicola,
and Mycophaerella tulasnei displayed the lowest sensitivity.
In general, saprotrophic and dominantly necrotrophic species
were less sensitive than hemibiotrophic and biotrophic ones,
and species parasitizing mainly the underground parts of plants
were less sensitive than those parasitizing on the aerial parts.
The antifungal activity of dyes also showed marked dif-
27
7
0
21
72.0
35
0
15
83.0
7
83.4
7
0
95.2
16
91.8
100.0
ferences (Fcalc.
ϭ 3,828, F5% ϭ 2.46), indicating that various
a Roman and Arabic numerals refer to reference compounds and fungi
dyes may exert different impacts on the fungi in the environ-
ment. Cationic dyes were more active than the sulfone deriv-
atives or those without alkyl substituents. The activity of mal-
achite green even surpassed that of the reference compound
with the most potential activity.
species in Materials and Methods and Appendix 2, respectively.
ϭ potential activity of reference compounds toward species in
percent of reference compound XIII.
b PA%
experience, however, NLMAP for any original data matrix
requires considerable computer time. Moreover, the resulting
maps are distorted, and the data points are not well distributed
on the map, irrespective of the composition of the original
matrix. It must be emphasized that the conclusion drawn is
based only on some practical examples and is not the result
of theoretical considerations; therefore, its generalization may
lead to serious misunderstanding of the results. The iteration
for NLMAP was carried out to the point at which the difference
The results of cluster analysis showed that neither the tax-
onomical position of a species nor its mode of habitation re-
lated to its sensitivity toward trityl dyes (cluster not shown).
The individual clusters contained species belonging to differ-
ent higher taxa and species with different modes of habitation.
The results of PCA indicated that seven hypothetical (i.e.,
background) variables explained the overwhelming majority
of total variance with only 9.22% loss of information, sug-
gesting that the 15 original variables can be reduced to seven
theoretical variables. Both dyes and reference compounds have
high loadings in more than one PC component, indicating that
they show different effects when the strength and selectivity
of their antifungal activity are taken into consideration si-
multaneously.
The two-dimensional nonlinear map of PC loadings is pre-
sented in Figure 2. Neither trityl dyes nor the reference com-
pounds form clear-cut clusters on this map, which suggests
that the antifungal activity of dyes and reference compounds
deviate considerably. The effect of triarimol and trichotechin
was similar, whereas the points of the other reference com-
pounds are located at the opposite end of the map. The two-
Ϫ
8
between the last two iterations was lower than 10 . The PC
loadings were also evaluated by varimax rotation around two
axes.
The similarities and dissimilarities between the results of
PCA and the two SPMs were assessed by stepwise regression
analyses. For matrix A, linear correlations were calculated by
the first and second co-ordinates of the two-dimensional non-
linear map of PC loadings (nlmap1 and nlmap2), the first and
second rotated PC loadings around two axes, the corresponding
potency values (P), and the first and second co-ordinates of
the two-dimensional nonlinear selectivity map (spmap1 and
spmap2, 7
ϫ 36 matrix). The SPM was carried out on the
transversed matrix. For matrix B, linear correlations were cal-

1210
Environ. Toxicol. Chem. 21, 2002
G. Oros et al.
Fig. 4. Similarities and dissimilarities among the selectivities of the
antifungal activities of trityl dyes and reference compounds as shown
by a two-dimensional nonlinear selectivity map (no. of iterations, 195;
Ϫ
maximal error, 3.02
ϫ
10 2). Roman numbers refer to trityl dyes and
reference compounds in Appendix 1 and Materials and Methods,
respectively.
Fig. 3. Similarities and dissimilarities among the sensitivities of fungi
toward trityl dyes and reference compounds as shown by a two-di-
mensional nonlinear map of principal component variables (no. of
Ϫ
iterations, 101; maximal error, 5.32
species in Appendix 2.
ϫ
10 2). Numbers refer to fungi
3 support our previous qualitative conclusions. The strength
of the overall antifungal activity of dyes shows high variation
among the dyes, proving again that their biological activity
strongly depends on their chemical structure. Interestingly, the
potency values of dyes are commensurable with those of ref-
erence compounds, indicating that trityl dyes accumulated in
the environment may cause serious pollution. The response of
fungi toward dyes also exhibits considerable diversity, indi-
cating that some fungi can tolerate this class of environmental
pollutants, with the sensitivity depending on the character of
the fungi.
The dyes form a concise cluster on the two-dimensional
nonlinear selectivity map (Fig. 4), indicating that their selec-
tivity is more similar than their overall biological activity.
The two-dimensional nonlinear selectivity map of species
is given in Figure 5. Fungi do not form clear-cut clusters
according to their taxonomical position on the selectivity map,
which proves again that the taxonomical position does not
dimensional nonlinear map of PC variables is shown in Figure
3. Fungi did not form well-defined clusters according to their
taxonomical position. This finding supports our previous con-
clusion, according to which the strength and selectivity of the
sensitivity of fungi toward trityl dyes and reference compounds
do not depend on their taxonomical position. However, it can-
not be excluded that the modes of action by trityl dyes and
reference compounds are different, and that the distribution of
the fungi on the map reflects not only the different sensitivities
of fungi but also the different modes of action of the com-
pounds.
The potency values of individual synthetic dyes and fungi
calculated by SPM are compiled in Table 3. The data in Table
Table 3. Strength of antifungal activity of trityl dyes and reference
compounds and sensitivity of fungia
Dyes and reference compounds
No. Potency No. Potency No. Potency No. Potency
I
II
III
IV
68.17
V
144.33 IX
10.67 XI
12.17 XII
6.17 XIII
1.83 XIV
146.83 XV
147.50 XVI
176.83
168.33
127.33
162.33
140.83 VI
239.17 VII
131.83 VIII
Fungi
No. Potency No. Potency No. Potency No. Potency
1
2
3
4
5
6
7
8
9
47.51
60.16
62.74
73.59
76.69
67.13
44.93
98.89
112.83
10
11
12
13
14
15
16
17
18
147.17
71.00
89.08
74.10
92.44
54.22
46.99
51.38
75.14
19
20
21
22
23
24
25
26
27
132.71
80.30
45.44
104.31
78.75
65.32
51.90
66.87
44.93
28
29
30
31
32
33
34
35
36
104.05
70.23
84.95
74.36
60.68
85.21
52.41
24.27
36.66
Fig. 5. Similarities and dissimilarities among the selectivities of sen-
sitivities of fungi toward trityl dyes and reference compounds as
a Potency values (arbitrary units) are calculated by the spectral map-
ping technique. Roman and arabic numbers refer to trityl dyes in
Table 1 and to reference compounds in Materials and Methods and
fungi species in Appendix 2, respectively.
shown by a two-dimensional nonlinear selectivity map (no. of itera-
Ϫ
tions, 164; maximal error, 5.12
in Appendix 2.
ϫ
10 2). Numbers refer to fungi species

Antifungal activity of some trityl-based synthetic dyes
Environ. Toxicol. Chem. 21, 2002
1211
Table 4. Coefficients of correlations of linear relationships between the first and second co-ordinates of the two-dimensional nonlinear map of
principal component (PC) loadings and variables (nlmap1, nlmap2), first and second rotated PC loadings (var1, var2), the potency values (P), and
first and second co-ordinates of the two-dimensional nonlinear selectivity map (spmap1, spmap2). Italicized data show significant correlations
Trityl dyes and reference compounds (
n
ϭ
15, r99.9% ϭ 0.7603)
nlmap1
nlmap2
var1
var2
P
spmap1
nlmap2
var1
var2
0.0438
0.6759
0.5162
0.4229
0.7373
0.6559
Ϫ
Ϫ
Ϫ
Ϫ
0.7930
0.6880
0.2951
0.5242
Ϫ
Ϫ
0.1226
0.1544
0.4033
0.8469
P
0.7885
0.6495
0.0876
spmap1
spmap2
0.4802
0.0368
Ϫ
0.7110
Ϫ
ϭ
Ϫ
Ϫ0.4097
Fungi (
n
36, r99.9% ϭ 0.5189)
nlmap1
nlmap2
P
spmap1
0.1074
nlmap2
P
spamp1
spmap2
0.1392
0.8288
0.7495
0.0423
0.0480
0.6244
0.2122
0.5873
0.4035
Ϫ
dyeing processes of synthetic textiles. European Water Pollution
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6. Petek J, Glavic P. 1996. An integral approach to waste minimi-
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17:169–189.
7. Sahu RK, Acharya NN, Mishra RN. 1987. Cytomorphological
effects of some dyes released with paper mill effluents of Allium
cepa root meristems. Indian Bot Rep 6:73–79.
8. Gatto C, Milanick MA. 1993. Inhibition of the red cell calcium
pump by eosin and other fluorescein analogs. Am J Physiol 246:
C1577–C1586.
9. Calin C, Miron M. 1995. Where to the dyers? The textile finishing
of the 2000 year. Ind Text (Bucharest) 46:140–147(in Romanian).
10. Hunger K. 1995. The organic pigments and their effects on tox-
icology and environment. Pitture e Vernici Europe 71:30–32(in
English and Italian).
define the selectivity of the antifungal effect of trityl dyes and
reference compounds.
The correlation coefficients of linear relationships between
the first and second co-ordinates of the two-dimensional non-
linear map of PC loadings and variables, the first and second
rotated PC loadings, the potency values, and the first and sec-
ond co-ordinates of the two-dimensional nonlinear selectivity
map are compiled in Table 4. No significant correlation was
found between the first and second co-ordinates of rotated PC
loadings, with nonlinear and spectral maps proving the real
orthogonality of each calculation method. Significant relation-
ships were, however, found between the co-ordinates, which
verifies the similarity of the methods.
11. Pandey KD, Kashyap AK. 1992. Induction of mutation in the
It can be concluded from the data that trityl dyes show
marked antifungal activity; therefore, they may be considered
as environmental pollutants when accumulated in groundwa-
ters and soil. Various multivariate methods, such as PCA and
SPM, combined with nonlinear mapping and varimax rotation
can be successfully employed for the study of their antifungal
effect. Because of its capacity to separate the strength and
selectivity of the antifungal effect, SPM is highly recom-
mended.
cyanobacterium Anabaena doliolum. Folia Microbiol (Prague)
37:377–380.
12. Arnold JW, Shimkets LJ. 1988. Inhibition of cell-cell interactions
in Myxococcus xanthus by Congo red. J Bacteriol 170:5765–
5770.
13. Ogawa T, Shibata M, Yatome C, Idaka E. 1988. Growth inhibition
of Bacillus subtilis by basic dyes. Bull Environ Contam Toxicol
40:545–552.
14. Shimada K, Shimara K. 1987. Effect of alternating current ex-
posure on the resistivity of resting Escherichia coli B cells to
crystal violet and other basic dyes. J Appl Bacteriol 62:261–269.
15. Mardia KV, Kent JT, Bibby JM. 1979. Multivariate Analysis.
Academic, London, UK.
16. He´berger K, Lopata A. 1998. Assessment of nucleophilicity and
electrophilicity of radicals and of polar and enthalpy effects on
radical addition reactions. J Org Chem 63:8646–8653.
17. He´berger K, Go¨rge´nyi M. 1999. Principal component analysis of
Kova´ts indices for carbonyl compounds in capillary gas chro-
matography. J Chromatogr A 812:21–31.
Acknowledgement—This study was supported by a grant from the
Ministry of Environmental Protection (HP-206).
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G. Oros et al.
APPENDIX 1
Common and International Union of Pure and Applied Chemistry (IUPAC) names of trityl dyes
No. of
dyes
Common name
IUPAC name
4-((4-Aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl)benzenamine monohydrochloride
I
II
Pararosaniline
Brilliant green
N
N
N
-[4-[[4-(diethylamino)phenyl]phenylmethylene]-2,5-cyclohexadiene-1-ylidene]-
nium sulfate
-[4-[[4-(dimethylamino)phenyl)phenylmethylene]-2,5-cyclohexadiene-1-ylidene]-
anaminium chloride
-[4-[bis[4-(dimethylamino)phenyl]methylene]-2,5-cyclohexadien-1-ylidene]-
nium chloride
N
-ethyl-ethanami-
-methyl-meth-
N-methyl-methami-
III
IV
V
Malachite green
Crystal violet
Methyl green
N
4-[[4-(Dimethylamino)phenyl][4-(dimethylimino)-2,5-cyclohexadiene-1-ylidene]methyl]-
N-ethyl-
N,N-dimethylbenbenzaminium bromide chloride
VI
VII
Bromthymol blue
Cotton blue
3Ј,3-Dibromothymolsulfonphthalein
Aminomethyl[[4-((sulfophenyl)imino]-2,5-cyclohexadien-1-ylidene)methyl]-benzenesulfonic acid
disodium salt
VIII
IX
Brilliant Blue R-250
Brilliant Blue G-250
Phenolphthalein
N
-[4-[[4-[(4-ethoxyphenyl)-amino]-phenyl][4-[ethyl[(3-sulfophenyl)methyl]-amino]-phenyl]
methylene]-2,5-cyclohexadien-1-ylidene]-
inner salt (monosodium salt)
N-ethyl-3-sulfo-benzenemethanaminium
N
-[4-[[4-((4-ethoxyphenyl)-amino]phenyl][4-[ethyl[(3-sulfophenyl)methyl]-amino]-2-methylphen-
yl]methylene]-3-methyl-2,5-cyclohexa-di-en-1-ylidene]-
inner salt (monosodium salt)
N
-ethyl-3-sulfobenzenemethanaminium
X
2-[Bis(4-hydroxyphenyl)methyl]benzoic acid
APPENDIX 2
APPENDIX 2
List of species, their origin, and taxonomical position according
to Hawksworth et al. [21]a
Continued
No.
Species
Source
Tomato
Rice
Apple
No.
Species
Source
19 Verticillium albo-atrum Reinke&Berthold
20 Magnaporthe grisea (Hebert) Barr
21 Trichotecium roseum (Pers.:fr.) Link (mi-
tosporic)
Mucorales (Oomycetes)
1
2
Mucor racemosus Fresenius
Rhisopus stolonifer (Ehrenberg:Fries)
Air
Sunflower
Lind
Phyllacorales (Sordariomycetes)
Saccharomycetales (Saccharomycetes)
22 Colletorichum coccodes (Wallr.) Hughes
23 Colletotrichum lindemuthianum (Saccar-
do&Magnus) Briosi&Cavara
24 Colletotrichum dematium (Pers.:fr.) Grove Soya
25 Colletotrichum graminicola (Cesati) Wil-
Tomato
Bean
3
Saccharomycetes cerevisiae (Hansen)
Baker’s yeast
Eurotiales (Eurotiomycetes)
4
5
Aspergillus niger van Tieghem
Penicillium oxalicum Currie&Thom
Onion
Cucumber
Wheat
son
26 Colletotrichum musae (Berk.&Curt.) v.
Banana
Pleosporales (Dothideomycetes)
Alternaria solani (Ellis&Martin) Sorauer
Arx
6
Potato
27 Myrothecium roridum Tode:fr. (mitospor-
Primula sp.
Tobacco
Dothideales (Dithideomycetes)
ic)
28 Thielaviopsis basicola (Berke-
ley&Broome) Ferraris
7
8
Didymella applanata (Niessl.) Sacc.
Botrydosphaeria rhodina (Berk.&Curt.)
von Arx
Raspberry
Coconut
Diapothales (Sordariomycetes)
29 Cryphonectria parasitica (Murrill) M.Barr Chestnut
9
Venturia inaequalis (Cooke) Winter
Aderhold apple
emend.
Heliothiales (Leotiomycetes)
10 Ascochyta pisi Lib.
Pea
11 Septoria lycopersici Speg.
Tomato
Tomato
Air
30 Botrytis aclada Fresenius
31 Botryotinia fuckeliana (de Barry) Whetzel Grape
Onion
12 Fulvia fulva (Cook) Ciferri
13 Mycophaerella tulasnei (Janz.) Land
14 Cladosporium cucumericum Eliis&Arth.
Ustilaginales (Ustilaginomycetes)
32 Schroeteria decaisneana (Boudier) De
Toni
Cucumber
Viola tricolor
Hypocreales (Sordariomycetes)
15 Nectria haematococca Berk.&Br.
16 Fusarium graminearum Schwabe
17 Fusorium oxysproum Schlecht.:fr.f.sp. ly-
cospersici (Sacc.) Snyder&Hansen
Potato
Maize
Maize
33 Ustilago maydis (de Candolle) Corda
34 Sorosporium cenchri P. Hennings
Maize
Cenchrus sp.
Pythiales (Oomycetes)
35 Pythium irregulare Buisman
36 Phytophtora cactorum
(Lebert&Cohn) Schroeter
Pea
Apple
18 Trichoderma harzianum Rifai
Soil
a Species 1 and 2, 3
(phyla) Zygo-, Asco, Basidio- and Oomycota, respectively.
Ϫ31, 32 and 33, and 35 and 36 belong to divisions