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1,2,4-Triazinones
Overview
The triazinones form a small group of herbicides developed in the 1970s
for the pre- and post-emergence control of grasses and broad-leaved
weeds. Both metribuzin and metamitron inhibit photosystem I1 and are
similar to the s-triazines in this respect. They are not acutely toxic but
metribuzin causes liver toxicity at high doses in mice. There are many
published papers of the fate of metribuzin and this is reflected in the
length of the entry in comparison with that for metamitron. Both herbicides are deaminated photochemically, chemically and biochemically,
Direct conjugation of metribuzin (and probably of metamitron) occurs in
plants and this plays a role in selectivity.
The S-methyl group of metribuzin (compared with the methyl group of
metamitron) is the most important distinguishing feature between the
two herbicides. Much of the metabolism of metribuzin, including conjugation, bound residue formation and hepatotoxicity, is preceded by an
oxidative bioactivation (S-oxidation) to a relatively reactive sulfoxide.
Conjugation with homoglutathione (cf. glutathione in animals) and the
catabolism of the homoglutathione conjugates can be expected to afford
several plant-unique metabolites.
An excellent review of all aspects of metribuzin has been published by
Hatzios and Penner (1988).
References
Hatzios, K.K. and Penner, D. (1988) in Herbicides: Chemistry, Degradation and Mode
of Action, 2nd edition, Vol. 3, eds. Kearney, P.C. and Kaufman, D.D., Marcel
Dekker, New York, pp. 191-243.

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Metamitron
Uses Metamitron is used pre- and post-emergence for the control of
grasses and broad-leaved weeds in sugar and red beets, fodder beet,
mangold and certain strawberry varieties.
Mode of action The herbicide is an inhibitor of photosynthesis. It is
absorbed mainly by the roots but also by the leaves.

Common name

Metamitron

Chemical name (IUPAC) 4-Amino-4,5-dihydro-3-methyl6-phenyl-1,2,4-triazin-5-one

CASRN

41394-05-2

Molecular formula

CIOHlON4O

Molecular weight

202.2

Chemical structure
NdNH2
1

Water solubility

1700 mg 1-'(20 "C)

KO,

Vapour pressure

8.6 x

Pa (20 "C)

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Metabolic pathways
Little has been published on the fate of metamitron. It is readily degraded
by aqueous photolysis. Deamination occurs in soils and plants and it is
readily metabolised and eliminated by mammals. Detailed metabolite
analyses have not been reported.
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Chemica1 degradation
Metamitron is very stable to acids but is decomposed by strong base
(pH >lo); its half-lives at 22 "C are: 410 days (pH 4),740 hours (pH 7) and
230 hours (pH 9). Photodecomposition on soil surfaces and in water is
very rapid, resulting in deamination to deamino-metamitron (3-methyl6-phenyl-l,2,4-triazin-5-onef
2, see Scheme 1)(Cox et al., 1996).

Scheme 1 Degradation of metamitron by aqueous photolysis, in watersediment, soils and plants (sugar beet).

Degradation in soils
Degradation of metamitron in water-sediment systems is rapid, partly
due to the photochemical deamination referred to above. Sediment
particles retard the degradation by light screening but also contribute to
degradation via organic matter content (presumably by microbial processes) (Cox et d.,1996).
Most of applied metamitron is degraded in soil over 4-6 weeks. Soil
column experiments to study leaching after several ageing periods
(Brumhard et al., 1987)showed that 18%mineralisation to COzoccurred in
a sandy soil and 26% in a loess soil over 105 days ageing (21 "C and 40%
moisture holding capacity). Leaching of metamitron was not extensive
and it was classed as having "medium" mobility.

Metabolism in plants
The major metabolite in sugar beet is 2, i.e. deaminated metamitron
(PM)*

Metabolism in animals
Orally administered metamitron is rapidly absorbed and eliminated by

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mammals. Almost quantitative elimination occurs equally in urine and

faeces in 48 hours (PM).

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References
Brumhard, B., Fuhr, F. and Mittelstaedt, W. (1987) Proc. Br. Cvop Prof. Con$ - Weeds,
585-592.
Cox, L., Hermosin, M.C., Cornejo, J. and Mansour, M. (1996) Chemosphere, 33,
2057-2064.

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Metribuzin
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Uses Metribuzin is used pre- and post-emergence for the control of

grasses and broad-leaved weeds in soyabeans, potatoes, tomatoes, sugar
cane, alfalfa, asparagus, maize and cereals.
Mode of action The herbicide is an inhibitor of photosynthesis (photosystem 11). It is absorbed mainly by the roots but also by the leaves and is
translocated in the xylem.

Common name

Metribuzin

Chemical name (IUPAC) 4-Amino-6-fert-butyl-4,5-dihydro3-methy1thio-1,2,4-triazin-5-one

CASRN

21087-649

Molecular formula

C*HI,N40S

Molecular weight

214.3

Chemical structure

0
N
.

, + , ~U
B

" N L !

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Water solubility

1050 mg 1-1(20 "C)

KOC

0.6-3 1.7

Vapour pressure

5.8 x

Log KO,

1.575

PK.3

1(base)

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Metabolic pathways
An excellent review of all aspects of metribuzin has been published by
Hatzios and Penner (1988).Many studies of the fate of metribuzin, mostly
in plants, have been reported. Biomimetic experiments addressing
hydrolysis and oxidation have proved helpful in elucidating pathways
and metabolite structures. Deamination is a major pathway, particularly
under environmental conditions. Conjugation, either directly at the amino
group or after S-oxidation, is important quantitatively and it is also
important in determining the selectivity of the herbicide.

Chemical degradation
Metribuzin is stable to dilute acids and bases at 20 "C. Half-lives at pH
values 1.2,4,7 and 9 are, respectively: 6.7 hours (37 "C) and (at 70 "C) 38,
47 and 8 days. The hydrolysable bond is that between the triazinone ring
and the methylthio group, affording a hydroxy derivative which is seen as
the tautomer (2) (Frear et al., 1985).Prolonged hydrolysis leads to deamination to compound 3. The methylthio group is liable to oxidation (for
example with m-chloroperbenzoic acid) to form the sulfoxide (4); this is
also labile to hydrolysis (or oxidation) to form 2 (Bleeke and Casida, 1984).
Oxidative deamination also occurs with m-chloroperbenzoic acid to yield
deaminated metribuzin (5). Compound 5 is also labile to S-oxidation to
form 6, which is readily hydrolysed to 3.
Aqueous photolysis of metribuzin is very rapid with a half-life of <1
day and this clearly contributes to the half-life of <7 days in natural pond
water (PM).Soil surface photolysis (natural light) proceeds with a half-life
of 14-25 days. The major product of photodegradation is deaminated
metribuzin (5). The reaction requires the combined effect of light and
solvent and the proton in product 5 is derived from the water under
aqueous photolysis conditions (Parlar and Korte, 1979). Minor metabolites were reported to be dimers and azo compounds formed by radical
combination reactions. The chemical and photocheJnica1 reactions of
metribuzin are illustrated in Scheme 1.

Degradation in soils
The DT,, of metribuzin in soil is 1.5-4 months. The compound is
strongly bound to soils of high organic matter content but it is more
mobile in sandy soils. In a study of eight soils, the absorption coefficient
varied between 0.56 for a very sandy loam and 31.7 for a soil containing
60% organic matter (Sharom and Stephenson, 1976).The effect of pH on
absorption to soil has been the subject of much debate (Hatzios and
Penner, 1988) but there is evidence that maximum absorption occurs at
pH 4-5 (Ladlie et al., 1976). The balance between non-biological and

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0
1

Ph

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H+

H+

[OI
3

[OI

0
Bu~#~.N

H2

N &O
H

Scheme 1 Chemical and photochemical transformations of metribuzin

(H+,acid; [O],oxidant; ph, light).

biological degradation of metribuzin and their kinetics under various

conditions are reviewed by Hatzios and Penner (1988). Under cold, dry
conditions where microbial degradation is minimised, degradation may
still occur. The products of such reactions have been identified as 2, 3
and 5 (Scheme l),all of which would be predicted from the hydrolysis
and oxidation chemistry of metribuzin.
[14C]Metribuzinwas mineralised to 15-20/0 14C02in 91 days in the surface layer of a silty loam (Moormanand Harper, 1989).Only 5% I4CO2was
detected in the sub-surface (>10cm). The difference correlated with
reduced microbial population at the lower level. Substantial amounts
of bound residue were formed in both the surface and sub-surface
layers.
A study of the evolution of 14C0, from 14C-ring-labelled metribuzin
under various conditions (Ladlie et al., 1976) demonstrated that mineralisation of metribuzin occurred and that sterilisation with NaN, inhibited
the mineralisation, i.e. it was substantially a microbial process. Metabolites
2, 3 and 5 were detected under laboratory conditions (Sharom and
Stephenson, 1976)and under field conditions (Webster and Reimer, 1976).
Studies by Schumacher (1974) showed that the heterocyclic ring could be
cleaved by soil microorganisms. The release of 14C0,from C3 label was
faster than that from C5 label. The triazinone ring thus appears to be more
labile than the relatively resistant s-triazine ring (see atrazine, etc.).

Metabolism in plants
The metabolism of metribuzin has been studied in several crop plants,
including soyabean, sugar cane, tomatoes, potatoes, wheat and lentil and in
weeds, including barnyard grass, American nightshade, pitted morning
glory, entireleaf morning glory and hemp sesbania (Hatzios and Penner,

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1988).The major primary metabolic reactions are (i) deamination, (ii) sulfoxidation and (iii) demethylthiolation. These latter two processes are
closely related; metribuzin is also directly conjugated with glucose via its
free amino group. The primary metabolites are also subject to conjugation
with glucose, malonic acid and glutathione (homoglutathione). The
metabolic pathways of metribuzin in several crop plants are shown in
Scheme 2 which is derived from the review of Hatzios and Penner (1988).
0
Bu~+~.N

But,$N.NH-

H2
"NASM,

'NAShomoG

gluc- ma1

"NASM,

8(tom,soy)

0
BU~+~

N
. H

"NASMe
1

Bound
Residues

H
3(tom,soy,sug,wh)

Scheme 2 The metabolism of metribuzin in important crop plants showing

species differences (soy, soyabean; sug, sugar beet; tom, tomato; wh, wheat).

Deamination (to form 5 ) is important to the phytotoxicity of metribuzin.

A study of two tolerant and two susceptible soyabean cultivars (Fedtke
and Schmidt, 1983; Fedtke, 1991) showed that deamination was more
rapid, both in vivo and in vitro in the tolerant varieties. High rates were
observed in the stem and this led to less metribuzin arriving at the leaves
of tolerant plants. The reaction is catalysed by peroxisomal preparations
from soyabean leaves and the amino group is liberated as ammonia. The

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in vitvo preparations required the addition of glutathione or ascorbate

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(reducingconditions) for maximum rates (Fedtke, 1991).This complicates

in vitro / in vivo comparisons.
S-Oxidation to metabolite 4 is a major reaction in soyabean (Frear et al.,
1985).The sulfoxides 4 and 6 are not important as terminal residues but
they are prerequisites for the conjugation of metribuzin metabolites with
homoglutathione. A minor metabolite in soyabean and other plants is 2.
This is almost certainly formed via the sulfoxide 4, i.e. S-oxidation is
important in the apparent hydrolysis of metribuzin. Deamination of this
metabolite affords 3 which is one of the terminal residues of metribuzin
(in soyabean and other plants).
Conjugation provides the bulk of the terminal metabolites in plants.
Metribuzin-N-glucoside (7)has been unequivocally identified in tolerant
soyabean and tomato cultivars. This is rapidly acylated to form the
malonyl-glucoside conjugate (8) (Frear et al., 1983, 1985). Glucosylation
is a detoxification reaction and it is considered to play some part in
the tolerance of certain tomato cultivars (Smith et al., 1989). Unlike
0-glucosides, N-glucosides tend to be resistant to hydrolysis by glucosidases and therefore they can appear as terminal metabolites. Oligosaccharide conjugates up to tetrasaccharides and larger have also been found
in soyabean (Falb and Smith, 1987). N-Glucosidation has been shown to
be enzyme-catalysed and it requires UDPG as the donor co-factor (Frear
et al., 1983).
Conjugation with homoglutathione is the major pathway for metribuzin metabolism in soyabean seedlings (Frear et aI., 1985).Also found in
this study as minor metabolites were the conjugates 7, 8 and 9 (the Nmalonyl conjugate of 2). All of these conjugates were identified unequivocally by MS and chemical methods. The homoglutathione pathway to 10
appears to be an important alternative pathway. Soyabean was found to
have very low levels of metribuzin N-glucosyltransferasewhen compared
with tomato (Frear et al., 1985). The resulting metabolite (10) would be
expected as a terminal metabolite but it will also be a precursor of the
numerous metabolites derived from the catabolism of glutathione (and
homoglutathione) conjugates in plants. For a short review of these, see
atrazine and the s-triazine overview. It is known that the polar metabolites of metribuzin form a very complex mixture of sugar and amino acid
conjugates (Falb and Smith, 1987).
The fact that tridiphane, an inhibitor of glutathione transferases, synergises the phytotoxicity of metribuzin to soyabean under field and growth
room conditions (Gaul et al., 1985) suggests that this conjugation (and its
precursor S-oxidation) are important in limiting the action of the
herbicide.
The electrophilic reactivity of sulfoxides 4 and 6 almost certainly leads
to the formation of plant-bound residues by their spontaneous reaction

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with protein thiol groups and other nucleophilic centres in plant polymers. Bound residues can account for up to 40% of the terminal residue in
soyabean (Dupont and Khan, 1992).It is considered (Hatzios and Penner,
1988) that differential metabolism or differential rate of metabolism is
the predominant factor in the selectivity and tolerance to metribuzin.
Differential uptake and translocation are thought to play minor roles.
Metabolism in animals

[14C-carbonyl]Metribuzin was dosed orally to rats and the excreted

products were examined over five days (Bleeke and Casida, 1984). No
ring cleavage occurred (as indicated by the expiration of *C02).The major
metabolite (urine) was deamino-metribuzin mercapturic acid (12, see
Scheme 3) which accounted for about 20% of the dose. This Nacetylcysteine conjugate is derived from the glutathione conjugate 11.

0
H2

15 N%otein

11

16

Scheme 3 The metabolism of metribuzin in animals (rat and mouse).

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Other polar urinary metabolites were eliminated but none accounted for
>5% of the dose and they were not identified. Only 1% or less of the
radioactivity was eliminated (in faeces) as the parent compound. Other
faecal metabolites were not identified but they were shown not to be the
aminotriazinones 2 or 4 or the triazinones 3,5 or 6.
Intraperitoneally injected metribuzin caused a fulminant centrilobular
hepatic necrosis in mice. The effect is exacerbated by the induction of
hepatic monooxygenase (with phenobarbitone) and alleviated by the
oxygenase inhibitor piperonyl butoxide. Mice were treated with [14Ccarbonyl]metribuzin to address this problem. The total yield of mercapturic acid was similar to that in rats but was distributed between
metabolites 12 and 14, the latter being formed via the glutathione conjugate 13.Apparently deamination was not as efficient as in the rat (or the
pathway 1 to 4 to 13 was more efficient in the mouse). This difference was
also seen at the level of isolated liver microsomes. The routes to mercapturic acid in vivo and to glutathione conjugates in vitvo were shown by
microsomal oxidation and biomimetic oxidation (3-chloroperbenzoic
acid) in the presence of N-acetylcysteine or 3,4-dichlorobenzenethiol.The
presumed S-oxide intermediates 4 and 6 reacted spontaneously with the
thiols.
Sulfoxide 4 was shown to react spontaneously also with bovine serum
albumin. The product is depicted as a general protein conjugate (15)in
Scheme 3. Similarly 6 may afford deaminometribuzin-modified protein
(16). Thus the observed reaction with tissue proteins in mice is initiated
by sulfoxidation but limited by competitive glutathione conjugation. At
higher doses of metribuzin, glutathione depletion occurs and uncontrolled reaction of the sulfoxides with tissue components precipitates
metribuzin hepatotoxicity (Bleeke et al., 1985).
Clearly many minor metabolites remain to be identified and the quantitative aspects of these studies are not clear from the original publication,
but the important features of metribuzin metabolism (Scheme 3) have
been identified.
References
Bleeke, M.S. and Casida, J.E. (1984) J. Agric. Food Chem., 32,749-755.
Bleeke, M.S., Smith, M.T. and Casida, J.E. (1985) Pestic. Biochem. Physiol., 23,
123-130.
Dupont, S. and Khan, S.U.(1992) J. Agric. Food Chem., 40,890-893.
Falb, L.N. and Smith, A.E. (1987) Pestic. Biochem. Physiol., 27,165-172.
Fedtke, C. (1991) Pestic. Biochem., 31,175-183.
Fedtke, C. and Schmidt, R.R. (1983) in Pesticide Chemistry: Human Werfare and the
Environment, Vol. 3 (eds. Matsunaka, S., Hutson, D.H. and Murphy, S.D.),
pp. 177-183, Pergamon Press, Oxford, UK.

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Frear, D.S., Mansager, E.R., Swanson, H.R. and Tanaka, F.S. (1983) Pestic.
Biochem. Physiol., 19,270-281.
Frear, D.S., Swanson, H.R. and Mansager, E.R. (1985) Pestic. Biochem. Physiol., 23,
56-65
Gaul, S., Ezra, G., Stephenson, G.R. and Solomon, K.R. (1985)Abstr. Weed Sci. Soc.
Am., 25, 75.
Hatzios, K.K. and Penner, D. (1988) in Herbicides: Chemistry, Degradation and Mode
of Action, 2nd edition, Vol. 3, eds. Kearney, P.C. and Kaufrnan, D.D., Marcel1
Dekker, New York, pp. 191-243.
Ladlie, J.S., Meggitt, W.F. and Penner, D. (1976) Weed Sci., 24,477481.
Moorman, T.B and Harper, S.S. (1989) 1. Enuiron. Qual., 18,302-306.
Parlar, H.and Korte, F. (1979) Chemosphere, 8,797-807.
Schumacher, R.W. (1974) Diss. Abstr. Int., B35,1485.
Sharom, M.S. and Stephenson, G.R. (1976) Weed Sci., 24,153-160.
Smith, A.E., Phatak, S.C. and Emmatty, D.A.(1989) Pestic. Biochem. Physiol., 35,
284-290.
Webster, G.R.B. and Reimer, G.J. (1976) Weed Res., 16,191-196.

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