Acute Lethality

The first toxicity test perforned on a new chemical is acute toxi- city. The LD50 and other acute
toxic effects aredetermined after one or more routes of administration (one route being oral or
tJ intendecl route of exposure) in one or more species. The species most often used are the
rnouse and rat, but sometimes the rabbit and dog are employed. Studies are performed in both
adult male and female animals. Food is often withheld the night before dos- ing. The number of
animals that die iri a14-day period after a sin- gle dosage is tabulated. In addition to mortality
and weight, daily examination of test animals should be conducted for signs of in- toxication,
lethargy, behavioral modifications, morbidity, food con- sumption, and so on.Acutetoxicity tests
(1) give a quantitative es- timate of acute toxicity (LD50) for comparison with other substances,
(2) identify targer organs and other clinical manifesta= tions of acute toxicity, (3) establish the
reversibility of the toxic sponse, and (4) provide dose-ranging guidance for other studies.
Determination of the LD50 has become a public issue because of increasing concern for the
welfare and protection of laboratory animals. The LD50 is not a biological constant.Many
factors in- fluence toxicity and thus may alter the estimation of the LD50 in any particular study.
Factors such as animal strain, age and weight, pe of feed, caging, pretrial fasting time, method
ofadministra- tion, volume and type of suspension medium, and durationrE servation have all
been shown to influence adverse responses to toxic substances. These and other factors have
been discussed in
detail in earler editions of this textbook (Doull, 1980). Because of this inherent variability in
LDso estimates, it is now recognized that for most purposes it is only neessary to characterize
the LD50 within an order of magnirude range such as 5 to 50 mgfkg, 50 to 500 mg/kg, and so
on.
There are several traditional approaches to determining th LD50 and its 95 percent confidence
limit as well as the slope of the probit line. The reader is referred to the classic works of Litch.
field and Wilcoxon (1949), Bliss (1957), and Finney (1971) for a description of the mechanics of
these procedures. A computer pro- gram in BASIC for determining probit and Iog-probit or logit
cor- relations has been published (Abou-Setta et al., 1986). These tra- ditional methods for
determining LDsos require a relativelyje ni.imber of animals (4() to 50). Other statistical
techniques that re quire fewer animals, such as the moving averages method of Thompson and
Weill (WeiI, 1952), are available but do not providt confidence limits for the LD50 and the slope
of the probit line, Finney (1985) has succinctly summarized the advantages and de- ficiencies of
many of the traditional methods. For most circum- stances, an adequate estimate of the LD50
and an approximation of the 95 percent confidence intervals can be obtained wirh as few as 6
to 9 animals, using the up-and-down method as modified by Bruce (1985). When this method
was compared with traditional methods that typically utilize 40 to 50 animals, excellent agree.
ment was obtained for all 10 compounds tested (Bruce, 1987). In mice and rats the LD50 is
usually determined as described above, but in the Iarger species only an approximation of the
LD50 is ob- tained by increasing thedose in the same animal until serious loxic effects are
evident.
If there is a reasonable Iikelihood of substantial exposure to the material by dermal or
inha]atiort exposure, acute dermal and acute inhalation studies are performed. Wher anirnals
are exposed acutely to chemicals in the air they breathe or the water they (fish) live in, the dose
the animals receive is usually not known. For these situations, the lethal concentration 50
(LC50) is usually determined; that is, the concentration ofchemical in the air or water that
causes death to 50 percent of the animals. In reporting an LC50, it is im- perative that the time
of exposure be indicated. Theaci dermal rabbits. The site of application is shaved. The test
substance is kept in contact with the skin for 24 h by wrapping the skin with an impervious
plastic material. At the end of the exposure period, the wrapping is removed and the skin is
wiped to remove any test substance still remaining. Ani- mals are observed at various intervals
for 14 days, and the LD5() is calculated. If no toxicity is evident at 2 gfkg, further acute dermal
toxicity testing is usually not performed. Acute inhalation studies are performed that are similar
to other acute toxicity studies ex- cept that the route of exposure is inhalation. Most often, the
length of exposure is 4 h.
Although by themselves LD50 and LC50 values are of Iimited significance, acute Iethality studies
are essential for characterizing the toxic effects ofchemicals and their hazard to humans. The
most meingfu1 scientific information derived from acute lethality tests comes from clinical
observations and postmortem examination of animals rather than from the specific LD50 value.