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In toxicology, the median lethal dose, LD50 (abbreviation for "lethal dose, 50%"), LC50 (lethal concentration, 50%) or LCt50 is a toxic unit that measures the lethal dose of a given substance. The value of LD50 for a substance is the dose required to kill half the members of a tested population after a specified test duration. LD50 figures are frequently used as a general indicator of a substance's acute toxicity. A lower LD50 is indicative of higher toxicity.

The term LD50 is generally attributed to John William Trevan. The test was created by J. W. Trevan in 1927. The term semilethal dose is occasionally used in the same sense, in particular with translations of foreign language text, but can also refer to a sublethal dose. LD50 is usually determined by tests on animals such as laboratory mice. In 2011, the U.S. Food and Drug Administration approved alternative methods to LD50 for testing the cosmetic drug Botox without animal tests.

Conventions

The LD50 is usually expressed as the mass of substance administered per unit mass of test subject, typically as milligrams of substance per kilogram of body mass, sometimes also stated as nanograms (suitable for botulinum), micrograms, or grams (suitable for paracetamol) per kilogram. Stating it this way allows the relative toxicity of different substances to be compared and normalizes for the variation in the size of the animals exposed (although toxicity does not always scale simply with body mass). For substances in the environment, such as poisonous vapors or substances in water that are toxic to fish, the concentration in the environment (per cubic metre or per litre) is used, giving a value of LC50. But in this case, the exposure time is important (see below).

The choice of 50% lethality as a benchmark avoids the potential for ambiguity of making measurements in the extremes and reduces the amount of testing required. However, this also means that LD50 is not the lethal dose for all subjects; some may be killed by much less, while others survive doses far higher than the LD50. Measures such as "LD1" and "LD99" (dosage required to kill 1% or 99%, respectively, of the test population) are occasionally used for specific purposes.

Lethal dosage often varies depending on the method of administration; for instance, many substances are less toxic when administered orally than when intravenously administered. For this reason, LD50 figures are often qualified with the mode of administration, e.g., "LD50 i.v."

The related quantities LD50/30 or LD50/60 are used to refer to a dose that without treatment will be lethal to 50% of the population within (respectively) 30 or 60 days. These measures are used more commonly within radiation health physics, for ionizing radiation, as survival beyond 60 days usually results in recovery.

A comparable measurement is LCt50, which relates to lethal dosage from exposure, where C is concentration and t is time. It is often expressed in terms of mg-min/m. ICt50 is the dose that will cause incapacitation rather than death. These measures are commonly used to indicate the comparative efficacy of chemical warfare agents, and dosages are typically qualified by rates of breathing (e.g., resting = 10 L/min) for inhalation, or degree of clothing for skin penetration. The concept of Ct was first proposed by Fritz Haber and is sometimes referred to as Haber's law, which assumes that exposure to 1 minute of 100 mg/m is equivalent to 10 minutes of 10 mg/m (1 × 100 = 100, as does 10 × 10 = 100).

Some chemicals, such as hydrogen cyanide, are rapidly detoxified by the human body, and do not follow Haber's law. In these cases, the lethal concentration may be given simply as LC50 and qualified by a duration of exposure (e.g., 10 minutes). The material safety data sheets for toxic substances frequently use this form of the term even if the substance does follow Haber's law.

For disease-causing organisms, there is also a measure known as the median infective dose and dosage. The median infective dose (ID50) is the number of organisms received by a person or test animal qualified by the route of administration (e.g., 1,200 org/man per oral). Because of the difficulties in counting actual organisms in a dose, infective doses may be expressed in terms of biological assay, such as the number of LD50s to some test animal. In biological warfare infective dosage is the number of infective doses per cubic metre of air times the number of minutes of exposure (e.g., ICt50 is 100 medium doses - min/m).

Limitation

As a measure of toxicity, LD50 is somewhat unreliable and results may vary greatly between testing facilities due to factors such as the genetic characteristics of the sample population, animal species tested, environmental factors and mode of administration.

There can be wide variability between species as well; what is relatively safe for rats may very well be extremely toxic for humans (cf. paracetamol toxicity), and vice versa. For example, chocolate, comparatively harmless to humans, is known to be toxic to many animals. When used to test venom from venomous creatures, such as snakes, LD50 results may be misleading due to the physiological differences between mice, rats, and humans. Many venomous snakes are specialized predators on mice, and their venom may be adapted specifically to incapacitate mice; and mongooses may be exceptionally resistant. While most mammals have a very similar physiology, LD50 results may or may not have equal bearing upon every mammal species, such as humans, etc.

Examples

Note: Comparing substances (especially drugs) to each other by LD50 can be misleading in many cases due (in part) to differences in effective dose (ED50). Therefore, it is more useful to compare such substances by therapeutic index, which is simply the ratio of LD50 to ED50.

The following examples are listed in reference to LD50 values, in descending order, and accompanied by LC50 values, {bracketed}, when appropriate.

Substance Animal, route LD50
{LC50}
LD50 : g/kg
{LC50 : g/L}
standardised 		Reference
Water (H2O) rat, oral >90,000 mg/kg >90
Sucrose (table sugar) rat, oral 29,700 mg/kg 29.7
Corn syrup rat, oral 25,800 mg/kg 25.8
Glucose (blood sugar) rat, oral 25,800 mg/kg 25.8
Monosodium glutamate (MSG) rat, oral 16,600 mg/kg 16.6
Stevioside (from stevia) mice and rats, oral 15,000 mg/kg 15
Gasoline (petrol) rat 14,063 mg/kg 14.0
Vitamin C (ascorbic acid) rat, oral 11,900 mg/kg 11.9
Glyphosate (isopropylamine salt of) rat, oral 10,537 mg/kg 10.537
Lactose (milk sugar) rat, oral 10,000 mg/kg 10
Aspartame mice, oral 10,000 mg/kg 10
Urea (OC(NH2)2) rat, oral 8,471 mg/kg 8.471
Cyanuric acid rat, oral 7,700 mg/kg 7.7
Cadmium sulfide (CdS) rat, oral 7,080 mg/kg 7.08
Ethanol (CH3CH2OH) rat, oral 7,060 mg/kg 7.06
Sodium isopropyl methylphosphonic acid (IMPA, metabolite of sarin) rat, oral 6,860 mg/kg 6.86
Melamine rat, oral 6,000 mg/kg 6
Taurine rat, oral 5,000 mg/kg 5
Melamine cyanurate rat, oral 4,100 mg/kg 4.1
Fructose (fruit sugar) rat, oral 4,000 mg/kg 4
Sodium molybdate (Na2MoO4) rat, oral 4,000 mg/kg 4
Sodium chloride (table salt) rat, oral 3,000 mg/kg 3
Aspirin (acetylsalicylic acid) rat, oral 1,944 mg/kg 1.944
Delta-9-tetrahydrocannabinol (THC) rat, oral 1,270 mg/kg 1.27
Cannabidiol (CBD) rat, oral 980 mg/kg 0.98
Methanol (CH3OH) human, oral 810 mg/kg 0.81
Trinitrotoluene (TNT) rat, oral 790 mg/kg 0.790
Arsenic (As) rat, oral 763 mg/kg 0.763
Ibuprofen rat, oral 636 mg/kg 0.636
Formaldehyde (CH2O) rat, oral 600–800 mg/kg 0.6
Solanine (main alkaloid in the several plants in Solanaceae amongst them Solanum tuberosum) rat, oral (2.8 mg/kg human, oral) 590 mg/kg 0.590
Alkyl dimethyl benzalkonium chloride (ADBAC) rat, oral
fish, immersion
aquatic invertebrates, immersion 		304.5 mg/kg
{0.28 mg/L}
{0.059 mg/L} 		0.3045
{0.00028}
{0.000059}
Coumarin (benzopyrone, from Cinnamomum aromaticum and other plants) rat, oral 293 mg/kg 0.293
Psilocybin (from magic mushrooms) mouse, oral 280 mg/kg 0.280
Hydrochloric acid (HCl) rat, oral 238–277 mg/kg 0.238
Ketamine rat, intraperitoneal 229 mg/kg 0.229
Paracetamol (acetaminophen) rat, oral 200 mg/kg 0.2
Caffeine rat, oral 192 mg/kg 0.192
Arsenic trisulfide (As2S3) rat, oral 185–6,400 mg/kg 0.185–6.4
Sodium nitrite (NaNO2) rat, oral 180 mg/kg 0.18
Methylenedioxymethamphetamine (MDMA, ecstasy) rat, oral 160 mg/kg 0.18
Uranyl acetate dihydrate (UO2(CH3COO)2) mouse, oral 136 mg/kg 0.136
Dichlorodiphenyltrichloroethane (DDT) mouse, oral 135 mg/kg 0.135
Uranium (U) mice, oral 114 mg/kg (estimated) 0.114
Bisoprolol mouse, oral 100 mg/kg 0.1
Cocaine mouse, oral 96 mg/kg 0.096
Cobalt(II) chloride (CoCl2) rat, oral 80 mg/kg 0.08
Cadmium oxide (CdO) rat, oral 72 mg/kg 0.072
Thiopental sodium (used in lethal injection) rat, oral 64 mg/kg 0.064
Demeton-S-methyl rat, oral 60 mg/kg 0.060
Methamphetamine rat, intraperitoneal 57 mg/kg 0.057
Sodium fluoride (NaF) rat, oral 52 mg/kg 0.052
Nicotine mouse and rat, oral

human, smoking

50 mg/kg 0.05
Pentaborane human, oral 50 mg/kg 0.05
Capsaicin mouse, oral 47.2 mg/kg 0.0472
Vitamin D3 (cholecalciferol) rat, oral 37 mg/kg 0.037
Piperidine (from black pepper) rat, oral 30 mg/kg 0.030
Heroin (diamorphine) mouse, intravenous 21.8 mg/kg 0.0218
Lysergic acid diethylamide (LSD) rat, intravenous 16.5 mg/kg 0.0165
Arsenic trioxide (As2O3) rat, oral 14 mg/kg 0.014
Metallic arsenic (As) rat, intraperitoneal 13 mg/kg 0.013
Sodium cyanide (NaCN) rat, oral 6.4 mg/kg 0.0064
Chlorotoxin (CTX, from scorpions) mice 4.3 mg/kg 0.0043
Hydrogen cyanide (HCN) mouse, oral 3.7 mg/kg 0.0037
Carfentanil rat, intravenous 3.39 mg/kg 0.00339
Nicotine (from various Solanaceae genera) mice, oral 3.3 mg/kg 0.0033
White phosphorus (P) rat, oral 3.03 mg/kg 0.00303
Strychnine (from Strychnos nux-vomica) human, oral 1–2 mg/kg (estimated) 0.001–0.002
Aconitine (from Aconitum napellus and related species) human, oral 1–2 mg/kg 0.001–0.002
Mercury(II) chloride (HgCl2) rat, oral 1 mg/kg 0.001
Cantharidin (from blister beetles) human, oral 500 μg/kg 0.0005
Aflatoxin B1 (from Aspergillus flavus mold) rat, oral 480 μg/kg 0.00048
Plutonium (Pu) dog, intravenous 320 μg/kg 0.00032
Bufotoxin (from Bufo toads) cat, intravenous 300 μg/kg 0.0003
Brodifacoum rat, oral 270 μg/kg 0.00027
Caesium-137 (
Cs) 		mouse, parenteral 21.5 μCi/g 0.000245
Sodium fluoroacetate (CH2FCOONa) rat, oral 220 μg/kg 0.00022
Chlorine trifluoride (ClF3) mouse, absorption through skin 178 μg/kg 0.000178
Sarin mouse, subcutaneous injection 172 μg/kg 0.000172
Robustoxin (from Sydney funnel-web spider) mice 150 μg/kg 0.000150
VX human, oral, inhalation, absorption through skin/eyes 140 μg/kg (estimated) 0.00014
Venom of the Brazilian wandering spider rat, subcutaneous 134 μg/kg 0.000134
Amatoxin (from Amanita phalloides mushrooms) human, oral 100 μg/kg 0.0001
Dimethylmercury (Hg(CH3)2) human, transdermal 50 μg/kg 0.000050
TBPO (t-Butyl-bicyclophosphate) mouse, intravenous 36 μg/kg 0.000036
Fentanyl monkey 30 μg/kg 0.00003
Venom of the Inland Taipan (Australian snake) rat, subcutaneous 25 μg/kg 0.000025
Ricin (from castor oil plant) rat, intraperitoneal
rat, oral 		22 μg/kg
20–30 mg/kg 		0.000022
0.02
2,3,7,8-Tetrachlorodibenzodioxin (TCDD, in Agent Orange) rat, oral 20 μg/kg 0.00002
Tetrodotoxin from the blue-ringed octopus intravenous 8.2 μg/kg 0.0000082
CrTX-A (from Carybdea rastonii box jellyfish venom) crayfish, intraperitoneal 5 μg/kg 0.000005
Latrotoxin (from widow spider venom) mice 4.3 μg/kg 0.0000043
Epibatidine (from Epipedobates anthonyi poison dart frog) mouse, intravenous 1.46-13.98 μg/kg 0.00000146
Batrachotoxin (from poison dart frog) human, sub-cutaneous injection 2–7 μg/kg (estimated) 0.000002
Abrin (from rosary pea) mice, intravenously

human, inhalation

human, oral

0.7 μg/kg

3.3 μg/kg

10–1000 μg/kg

0.0000007

0.0000033

0.00001–0.001

Saxitoxin (from certain marine dinoflagellates) human, intravenously

human, oral

0.6 μg/kg

5.7 μg/kg

0.0000006

0.0000057

Pacific Ciguatoxin-1 (from ciguateric fish) mice, intraperitoneal 250 ng/kg 0.00000025
Palytoxin (from Palythoa coral) mouse, intravenous 45 ng/kg

2.3–31.5 μg/kg

0.000000045

0.0000023

Maitotoxin (from ciguateric fish) mouse, intraperitoneal 50 ng/kg 0.00000005
Polonium-210 (
Po) 		human, inhalation 10 ng/kg (estimated) 0.00000001
Diphtheria toxin (from Corynebacterium) mice 10 ng/kg 0.00000001
Shiga toxin (from Shigella bacteria) mice 2 ng/kg 0.000000002
Tetanospasmin (from Clostridium tetani) mice 2 ng/kg 0.000000002
Botulinum toxin (from Clostridium botulinum) human, oral, injection, inhalation 1 ng/kg (estimated) 0.000000001
Ionizing radiation human, irradiation 3–5 Gy (Gray) —

Poison scale

Negative values of the decimal logarithm of the median lethal dose LD50 (−log10(LD50)) on a linearized toxicity scale encompassing 11 orders of magnitude. Water occupies the lowest toxicity position (1) while the toxicity scale is dominated by the botulinum toxin (12).

The LD50 values have a very wide range. The botulinum toxin as the most toxic substance known has an LD50 value of 1 ng/kg, while the most non-toxic substance water has an LD50 value of more than 90 g/kg; a difference of about 1 in 100 billion, or 11 orders of magnitude. As with all measured values that differ by many orders of magnitude, a logarithmic view is advisable. Well-known examples are the indication of the earthquake strength using the Richter scale, the pH value, as a measure for the acidic or basic character of an aqueous solution or of loudness in decibels. In this case, the negative decimal logarithm of the LD50 values, which is standardized in kg per kg body weight, is considered −log10(LD50).

The dimensionless value found can be entered in a toxin scale. Water as the baseline substance is neatly 1 in the negative logarithmic toxin scale.

Animal rights concerns

Animal-rights and animal-welfare groups, such as Animal Rights International, have campaigned against LD50 testing on animals. Several countries, including the UK, have taken steps to ban the oral LD50, and the Organisation for Economic Co-operation and Development (OECD) abolished the requirement for the oral test in 2001 (see Test Guideline 401, Trends in Pharmacological Sciences Vol 22, February 22, 2001).

Procedures

A number of procedures have been defined to derive the LD50. The earliest was the 1927 "conventional" procedure by Trevan, which requires 40 or more animals. The fixed-dose procedure, proposed in 1984, estimates a level of toxicity by feeding at defined doses and looking for signs of toxicity (without requiring death). The up-and-down procedure, proposed in 1985, yields an LD50 value while dosing only one animal at a time.

See also

Other measures of toxicity

Related measures

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Further reading

  • Lipnick RL, Cotruvo JA, Hill RN, Bruce RD, Stitzel KA, Walker AP, et al. (March 1995). "Comparison of the up-and-down, conventional LD50, and fixed-dose acute toxicity procedures". Food and Chemical Toxicology. 33 (3): 223–231. doi:10.1016/0278-6915(94)00136-C. PMID 7896233.

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