ASTM E939-94(2012)
(Test Method)Standard Test Method of Field Testing Topical Applications of Compounds as Repellents for Medically Important and Pest Arthropods (Including Insects, Ticks, and Mites):I Mosquitoes (Withdrawn 2017)
Standard Test Method of Field Testing Topical Applications of Compounds as Repellents for Medically Important and Pest Arthropods (Including Insects, Ticks, and Mites):I Mosquitoes (Withdrawn 2017)
SIGNIFICANCE AND USE
This test method is an important part of the final phase of study in the development of mosquito repellents for personal use.
This test method is primarily designed to simulate a situation in which a person treated with a repellent is exposed to natural populations of attacking mosquitoes.
The simplicity of the test offers flexibility under a relatively wide range of circumstances and geographical locations. By following this test method, international testing with a variety of vector mosquito populations is no more difficult to accomplish than tests with various domestic species.
A number of people test topical applications of a repellent for the following reasons:
To determine how long the repellent is effective;
To establish the effective dosage range;
To establish the range of effectiveness on several mosquito genera and species in a number of geographical areas; and
To identify the material in terms of odor, staining capability, plasticizing effect, and oiliness or greasiness.
No repellent should be tested on humans without the written consent of the test volunteers (hereafter referred to as test subjects) and prior approval of competent authority, as designated in applicable laws and regulations governing experimentation on humans.
SCOPE
1.1 This test method is used to evaluate the repellency of promising compounds that have undergone primary laboratory studies and have been approved for skin application for secondary testing.
1.2 This test method is designed for the study of mosquito repellents, but with some modifications this test method can be used to determine the repellency of candidate compounds for other flying insects that attack humans.
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.
WITHDRAWN RATIONALE
This test method is used to evaluate the repellency of promising compounds that have undergone primary laboratory studies and have been approved for skin application for secondary testing.
Formerly under the jurisdiction of Committee E35 on Pesticides, Antimicrobials, and Alternative Control Agents, this test method was withdrawn in November 2017. This standard is being withdrawn without replacement due to its limited use by industry.
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Designation: E939 − 94 (Reapproved 2012)
Standard Test Method of
Field Testing Topical Applications of Compounds as
Repellents for Medically Important and Pest Arthropods
(Including Insects, Ticks, and Mites):I Mosquitoes
This standard is issued under the fixed designation E939; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope 4. Significance and Use
1.1 This test method is used to evaluate the repellency of
4.1 This test method is an important part of the final phase
promising compounds that have undergone primary laboratory
ofstudyinthedevelopmentofmosquitorepellentsforpersonal
studies and have been approved for skin application for
use.
secondary testing.
4.2 This test method is primarily designed to simulate a
1.2 This test method is designed for the study of mosquito
situation in which a person treated with a repellent is exposed
repellents, but with some modifications this test method can be
to natural populations of attacking mosquitoes.
used to determine the repellency of candidate compounds for
4.3 The simplicity of the test offers flexibility under a
other flying insects that attack humans.
relatively wide range of circumstances and geographical loca-
1.3 The values stated in SI units are to be regarded as
tions. By following this test method, international testing with
standard. No other units of measurement are included in this
a variety of vector mosquito populations is no more difficult to
standard.
accomplish than tests with various domestic species.
1.4 This standard does not purport to address all of the
4.4 A number of people test topical applications of a
safety concerns, if any, associated with its use. It is the
repellent for the following reasons:
responsibility of the user of this standard to establish appro-
4.4.1 To determine how long the repellent is effective;
priate safety and health practices and determine the applica-
4.4.2 To establish the effective dosage range;
bility of regulatory limitations prior to use.
4.4.3 To establish the range of effectiveness on several
mosquito genera and species in a number of geographical
2. Terminology
areas; and
2.1 Definitions of Terms Specific to This Standard:
4.4.4 To identify the material in terms of odor, staining
2.1.1 complete protection time (CPT)—the time from appli-
capability, plasticizing effect, and oiliness or greasiness.
cation of the repellent to the time of the first confirmed bite (a
secondbitebythesamespecieswithin30minofthefirst).This
4.5 No repellent should be tested on humans without the
permits any number of unconfirmed bites during the CPT.
written consent of the test volunteers (hereafter referred to as
test subjects) and prior approval of competent authority, as
3. Summary of Test Method
designated in applicable laws and regulations governing ex-
perimentation on humans.
3.1 Ameasured amount of the candidate material is applied
totheforearmorsometimesthelowerleg.Theseareasarethen
5. Apparatus
protected from rubbing and are continuously exposed to
mosquitoes in the field to determine the length of time the
5.1 Insect collection vials.
treatment provides either complete protection or a high level of
5.2 Aspirator.
protection.
5.3 Stereoscope (optional).
5.4 Standard References for Mosquito Identification, for
This test method is under the jurisdiction of ASTM Committee E35 on
determining species present in the field (optional).
Pesticides, Antimicrobials, and Alternative Control Agents and is the direct
responsibility of Subcommittee E35.12 on Insect Control Agents.
5.5 Temperature and Humidity Reading Equipment—
Current edition approved April 1, 2012. Published June 2012. Originally
Ideally, a continuous recording device such as a hygrothermo-
approved in 1983. Last previous edition approved in 2006 as E939 – 94 (2006).
DOI: 10.1520/E0939-94R12. graph should be used to record conditions during tests. If such
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
E939 − 94 (2012)
equipment is not available, readings should be made immedi- 8.2 For field tests, make the initial studies with a 25 %
ately before, midway during, and immediately after the tests ethanol solution of the candidate repellents (250 mgAI/mL).A
are made, using a sling psychrometer. compound or formulation may be retested at reduced or
increased concentrations, or at full strength, if warranted.
5.6 Air Speed Indicator and Light Meter, optional but
preferred if equipment is available.
NOTE 3—If the complete protection times (CPTs) of repellents are too
long for the available testing period or if the CPTs are too short for
5.7 Watch.
evaluation, adjust the concentrations accordingly. The termination of an
evaluation before a confirmed bite occurs is termed a “plus out” and
5.8 Headnets.
should be avoided.
5.9 Cotton Gloves.
8.3 Spread 1 mL of the repellent formulation or repellent
5.10 Battery-Operated Head Lamps,withredfiltersfortests
solution evenly over the forearm of the subject and compare
with nocturnally active species.
directlywithanotherrepellentofthesameconcentrationonthe
other arm (see Note 4). The behavior of some species neces-
5.11 Notebook, Test Sheets, and Pencils, for recording
sitates the use of the legs instead of the arms as treatment sites.
species, test data, date, and locality of the test.Asample work
This can be determined by observation before the tests begin.
sheet is attached with recorded results (see Annex A1.).
5.12 Water Supply, Mild Soap, and Paper Towels, for NOTE 4—The concentration of a compound being tested is not as
important as the assurance that it is paired with another compound of
washing treated skin.
equal concentration.
5.13 Clothing, should be appropriate to the season and
8.4 If legs are used as treatment sites, apply 1.5 mL on the
geographical area.
skin between the ankle and knee.
NOTE 1—There are differences of opinion as to whether the clothing
8.5 Determine the surface area of the limbs of each test
worn should be uniform in color and type, however, data available do not
subject so that treatment rates of candidate repellents and the
reject or confirm this contention.
standard are uniformly applied.Adjust the application rates for
5.14 List of Chemical Names, identifying the compounds or
differences in arm or leg size of different subjects.
mixtures, or both, to be tested.
8.6 Expose the treated arms or legs continuously to natural
NOTE 2—In the event of a medical emergency, the chemical list along
populations of the mosquito species being tested.
with pertinent toxicological data may be required.
8.7 The duration of effectiveness of the repellent is indi-
6. Reagents and Materials
cated by the CPT.
6.1 All test solutions are formulated on a weight to volume
8.8 Employ a balanced incomplete block (BIB) experimen-
basis (usually with 95 % ethanol).
tal design (round robin) when three to five chemicals are to be
6.2 Test chemicals in 25 % ethanol solutions. tested. With this design, test each repellent in the series on
opposite arms of a given number of subjects.
6.3 Test standard deet (N,N-diethyl-meta-toluamide) in a
25 % ethanol solution.
NOTE 5—Ideally, the number of subjects should equal the number of
candidate compounds, excluding the standard. Thus, if four compounds
6.4 For final development studies, formulations of lotion,
are to be tested, including the standard, three test subjects would be
liquids, creams, solids, or pressurized aerosols containing
required. To illustrate, the pairings would be: AB, AC, AD, BC, BD, and
ingredients for field evaluation are included.
CD. Subject 1 would testAB and CD; Subject 2,AC and BD, and Subject
3, AD and BC.
7. Sampling
8.9 Because of their variability in attractiveness, assign
repellentstotestsubjectsinarandomizedandbalancedfashion
7.1 Each candidate repellent is paired with each other
so that each subject does not wear the same combination of
repellent or a standard on the arms of a subject and exposed
materials more than once in a single BIB test series. For
simultaneously to the same insect population (See AnnexA1).
example, if compounds A, B, C, D, and E are tested with a
7.2 A round-robin or paired test is used in the experiments,
standard F, then Subject 1 will testAB, CD, and EF; Subject 2
usually based on the number of experimental materials being
will testAC, BE, and DF; Subject 3 will testAD, BF, and CE;
evaluated.
Subject 4 will testAE, BC, and CF; and Subject 5 will testAF,
7.3 Treatments are exposed to the mosquito population for
BC, and DE.
as long as the repellents are effective, and the biting activity
NOTE 6—If four or five compounds, excluding the standard, are tested
continues.
in a BIB series, one replication of the BIB is sufficient for a statistical
analysis. If three compounds are to be tested in this fashion, two
8. Procedure
replications of the round robin will be necessary.
8.1 Determine the identity of species of mosquito in the test 8.10 A direct comparison of the candidate versus the stan-
area prior to the test. Determine the time to begin and end tests dard repellent deet is used when fewer than three chemicals are
each day by the activity of the species to be tested. Diurnal to be tested. For these tests, four or more replications of tests
nocturnal or other patterns of mosquito behavior will govern with each chemical on at least three different subjects are
the scheduling of tests. necessary.
E939 − 94 (2012)
8.11 Express the results obtained by paired tests or BIB in a compartmented flat so that later they can be matched with
comparisons at CPT in minutes or hours. a record of the treatment and time of bite kept by each test
subject.
8.12 During the experiment, subjects shall avoid contact
8.22 The procedure makes possible the determination of
with the treated skin surfaces. Touching, rubbing, or abrasive
species biting, total numbers of bites during the exposure
action on the treated skin can affect the results. Avoid undue
period, percent protection provided by a repellent as compared
sweating or wetting of the treated skin except in special tests
with no treatment, and duration of protection against particular
designed to study the durability of repellen
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