s-DIANISIDINE 4,4'
METHYLENEBIS(2-CHLOROANILINE) (MOCA) s-TOLIDINE
Method no.: |
71 |
|
|
|
Matrix: |
Air |
|
Procedure: |
Samples are collected closed-face by drawing known
volumes of air through sampling devices consisting of
three-piece cassettes, each containing two sulfuric
acid-treated glass fiber filters separated by a spacer.
The sample filters are transferred to separate glass vials
containing 2 mL of deionized water within 10 h after sampling.
Quantitation is performed by analyzing the heptafluorobutyric acid
anhydride derivatives of the amines by gas chromatography using an
electron capture detector. |
|
Recommended air volume and sampling rate: |
100 L at 1 L/min |
|
|
|
|
s-Dianisidine |
MOCA |
s-Tolidine |
|
|
Target conc.: ppb
(µg/m3) |
1 (10) |
20 (218) |
1 (8.7) |
|
Reliable quantitation limits: ppt
(ng/m3) |
1.2 (12) |
40 (440) |
1.3 (11) |
|
Standard errors of estimate at the target
concentration: (Section 4.7.) |
7.8% |
5.8% |
8.0% |
|
|
|
Special requirements: |
Samples for s-dianisidine
must be shipped and stored at 0°C or colder to minimize loss of
analyte. These samples should be analyzed as soon as possible. |
|
Status of method: |
Evaluated method. This method has been subjected to
the established evaluation procedures of the Organic Methods
Evaluation Branch. |
|
Date: April 1988 Updated: July 1989 |
Chemist: Carl J.
Elskamp
|
Organic Methods Evaluation Branch OSHA Analytical
Laboratory Salt Lake City, Utah
1. General Discussion
1.1. Background
1.1.1. History
The previous OSHA-recommended procedures to determine airborne
concentrations of s-dianisidine, MOCA, and s-tolidine involved collection with an
untreated glass fiber filter, a bubbler containing 0.1 N HCl, and a
bubbler containing isopropyl alcohol, respectively (Ref. 5.1.). The
free amines were determined by high-performance liquid
chromatography using an ultraviolet detector. The procedures for
s-dianisidine and s-tolidine were OSHA laboratory
in-house methods which were never fully validated and
although the MOCA procedure had been validated, it is not certain
how efficient a bubbler is for collection of aerosols. Also bubblers
are an inconvenient means for taking personal air samples.
Methodology exists which has previously been validated for
benzidine, 3,3'-dichlorobenzidine,
2,4-toluenediamine, 2,6-toluenediamine
(Ref. 5.2.) and 4,4'-methylenedianiline (Ref. 5.3.).
The collection of air samples involves sampling on glass fiber
filters that had been treated with sulfuric acid. Thus the collected
amines are converted to the more stable and less volatile
corresponding amine salts on the filter surface. To further enhance
the stability of samples, the filters are transferred to small vials
containing 2 mL of deionized water within 10 h after sampling. The
analysis involves converting the amine salts to free amines by
addition of sodium hydroxide, extracting the amines into toluene,
removing a portion of the toluene extract, and derivatizing the free
amines in the extract with heptafluorobutyric acid anhydride (HFAA)
according to the reaction
RNH2 +
(C3F7CO)2O
->
RNHCOC3F7
+
C3F7COOH
The derivatives are analyzed by capillary gas chromatography
using an electron capture detector. This sampling and analytical
scheme was used in the validation of the following method for
s-dianisidine, MOCA and
s-tolidine.
Note: As a consequence of later evaluation tests done for
toluidine, this method has been updated. The sampling device now
consists of two acid-treated glass fiber filters assembled in a
three-piece cassette instead of a single
acid-treated filter with a support pad in a
two-piece cassette. Not only does this device offer
several advantages as listed in the toluidine method, it is now the
common sampler for several aromatic amines.
1.1.2. Toxic effects (This section is for information only and
should not be taken as the basis of OSHA policy.)
s-DIANISIDINE: s-Dianisidine has been shown to be carcinogenic
to rats and hamsters. There are no conclusive epidemiological
studies to show that s-dianisidine alone is carcinogenic to
humans. Most workers that are exposed to s-dianisidine are also exposed to other
related amines such as benzidine, which is widely believed to be a
human bladder carcinogen. (Ref. 5.4.) There are no OSHA or ACGIH
standards concerning occupational exposure to s-dianisidine.
MOCA: MOCA has been shown to be carcinogenic to rats and
mice. There are no conclusive epidemiological studies to show that
MOCA is carcinogenic to humans. (Ref. 5.5.) Exposure to MOCA results
in the same general toxic effects that are characteristic of
aromatic amines. These effects include cyanosis (a bluish or
purplish discoloration due to deficient oxygenation of the blood)
and methemoglobinemia (the presence of methemoglobin in the blood).
ACGIH has designated MOCA as a suspect human carcinogen and has
assigned it a TLV of 0.02 ppm with a "skin" notation. (Ref. 5.6.)
s-TOLIDINE: Rats that had been
administered s-tolidine
developed cancer in some tissues, but not in the bladder. In another
experiment, hamsters given s-tolidine orally did not develop cancer.
There is no strong evidence indicating that s-tolidine is or is not carcinogenic to
man. As with s-dianisidine, there is very
little information on the toxicity of occupational exposure to
s-tolidine alone. It is
usually used as a mixture with other aromatic amines, such as
benzidine. There have been cases where bladder cancers have been
found in workers making dyes and also in those exposed to a
combination of benzidine and s-tolidine. It is widely thought that
benzidine is a human bladder carcinogen. There is some evidence
indicating that s-tolidine
hastens the formation of cancerous tumors in rats that had been
exposed to benzidine. ACGIH has designated s-tolidine as being a suspected human
carcinogen. There is no TLV assigned to s-tolidine. (Ref. 5.7.)
1.1.3. Potential workplace exposure
s-DIANISIDINE: The principal use
for s-dianisidine is as a chemical
intermediate in the production of dyes. It has been reported that 89
dyes are produced with s-dianisidine being an intermediate.
probably the next most important use for s-dianisidine is as an intermediate in the
production of s-dianisidine
diisocyanate. It has also been used as a reagent for the detection
of a number of metals, thiocyanates, and nitrites. (Ref. 5.4.)
MOCA: MOCA is primarily used as a curing agent for
isocyanate containing polymers. It is also widely used for curing
liquid-castable polyurethane elastomers. Frequently it
is formulated with other aromatic diamines, such as
3,3'-dichlorobenzidine or
4,4'-methylenedianiline, to prepare special curing
agents. Small amounts of MOCA are used as curing agents for epoxy
and epoxy-urethane resin blends. (Ref. 5.5.)
s-TOLIDINE: The most important
use for s-tolidine is in the
manufacture of dyes. It is also used in some analytical chemistry
procedures. (Ref. 5.7.)
1.1.4. Physical properties and other descriptive information
s-DIANISIDINE (Ref.
5.4.) |
|
CAS no.: |
119-90-4 |
molecular weight: |
244.3 |
melting point: |
137-138°C |
description: |
Colorless crystals which turn violet on standing |
solubility: |
Almost insoluble in water, soluble in ethanol, ether,
acetone, benzene and chloroform; probably soluble in most
organic solvents and lipids |
chemical reactivity: |
A weak base; has the general characteristics of primary
aromatic amines |
synonyms: |
3,3'-Dimethoxybenzidine; bianisidine;
4,4'-diamino-3,3,dimethoxybiphenyl;
di-p-amino-di-m-methoxydiphenyl;
3,3'-dimethoxy-4,4'-diaminobiphenyl |
structural formula: |
|
|
MOCA (Ref. 5.5.) |
|
CAS no.: |
101-14-4 |
molecular weight: |
267.2 |
melting point: |
110°C |
description: |
Colorless crystals |
solubility: |
Almost insoluble in water; soluble in alcohol and ether
and probably in most organic solvents and lipids |
synonyms: |
di-(4-amino-3-chlorophenyl)methane;
3,3,-dichloro-4,4'-diaminodiphenylmethane;
bis amine; mboca;
methylenebis(ortho-chloroaniline); r,r'-methylenebis(ortho-chloroaniline) |
trade names: |
Curalin M; Curene 442; DACPM; MOCA; Cyanaset |
structural formula: |
|
|
s-TOLIDINE (Ref.
5.7.) |
|
CAS no.: |
119-93-7
| |
molecular weight: |
212.3 |
melting point: |
129-131°C |
description: |
White to reddish crystals or powder |
solubility: |
Slightly soluble in water, soluble in alcohol, ether or
dilute acids |
synonyms: |
3,3'-dimethylbenzidine |
structural formula: |
|
The analyte air concentrations throughout this method are
based on the recommended sampling and analytical parameters. Air
concentrations listed in ppb and ppt are referenced to 25°C and
760 mm Hg. Although the derivatives of the amines are analyzed,
the equivalent masses of the amines are listed throughout the
method. |
1.2. Limit defining parameters
1.2.1. Detection limit of the analytical procedure
The detection limits of the analytical procedure are 6.0, 5.5,
and 5.4 fg per injection for s-dianisidine, MOCA, and s-tolidine respectively. These are the
amounts of analytes which produce peaks with heights that are
approximately 5 times the baseline noise. (Section 4.1.)
1.2.2. Detection limit of the overall procedure
The detection limits of the overall procedure are 1.2, 44, and
1.1 ng per sample for s-dianisidine, MOCA, and s-tolidine respectively. These are the
amounts of analytes spiked on sample filters which allow recoveries
of amounts of analytes equivalent to the detection limits of the
analytical procedure. These detection limits correspond to air
concentrations of 1.2 ppt (12 ng/m3), 40
ppt (440 ng/m3), and 1.3 ppt (11
ng/m3) for s-dianisidine, MOCA, and s-tolidine respectively. (Section 4.2.)
1.2.3. Reliable quantitation limit
The reliable quantitation limits are 1.2, 44, and 1.1 ng per
sample for s-dianisidine,
MOCA, and s-tolidine
respectively. These are the smallest amounts of analytes which can
be quantitated within the requirements of a recovery of at least 75%
and a precision (±1.96 SD) of ±25% or better. These reliable
quantitation limits correspond to air concentrations of 1.2 ppt (12
ng/m3), 40 ppt (440
ng/m3), and 1.3 ppt (11
ng/m3) for s-dianisidine, MOCA, and s-tolidine respectively. (Section 4.3.)
The reliable quantitation limits and detection limits
reported in this method are based upon optimization of the
instrument for the smallest possible amount of analyte. When
the target concentration of an analyte is exceptionally higher
than these limits, they may not be attainable at the routine
operating parameters. |
1.2.4. Instrument response to the analyte
The instrument response over concentration ranges representing
one-half to wo times the target concentrations is
linear for all three analytes. (Section 4.4.)
1.2.5. Recovery
The recoveries of s-dianisidine, MOCA, and s-tolidine from samples used in a
15-day storage test remained above 79, 100, and 89%
respectively. The s-dianisidine samples were stored in a
refrigerator at 0°C and the samples for the other two analytes were
stored in a closed drawer at ambient temperature (about 21°C).
(Section 4.5.) The recovery of analyte from the collection medium
during storage must be 75% or greater.
1.2.6. Precision (analytical method only)
The pooled coefficients of variation obtained from replicate
injections of analytical standards at 0.5, 1, and 2 times the target
concentrations are 0.020, 0.034, and 0.025 for s-dianisidine, MOCA, and s-tolidine respectively. (Section 4.6.)
1.2.7. Precision (overall procedure)
The precisions at the 95% confidence level for the 15-day storage
tests are ±15.4, ±11.3, and ±15.7% for s-dianisidine, MOCA, and s-tolidine respectively. These include an
additional ±5% for sampling error. The s-dianisidine samples were stored in a
refrigerator at 0°C and the MOCA and s-tolidine samples were stored in a closed
drawer at ambient temperature (about 21°C). (Section 4.7.) The
overall procedure must provide results at the target concentration
that are ±25% or better at the 95% confidence level.
1.2.8. Reproducibility
Six samples for each analyte, spiked by liquid injection, and a
draft copy of this procedure were given to a chemist unassociated
with this evaluation. The s-dianisidine and s-tolidine samples were analyzed after 2
days of storage at 0°C and the MOCA samples were analyzed after 42
days of storage at -35°C. No individual sample result
deviated from its theoretical value by more than the precision of
the overall procedure as reported in Section 1.2.7. (Section 4.8.)
1.3. Advantages
1.3.1. The acid-treated filter provides a
convenient method of sampling for a number of aromatic amines.
1.3.2. The analysis is rapid, sensitive, and precise.
1.4. Disadvantages
1.4.1. Sample filters must be transferred to vials containing
water before being submitted to the laboratory for analysis.
1.4.2. Samples for s-dianisidine must be shipped and stored
under reduced temperatures and should be analyzed as soon as
possible.
2. Sampling Procedure
2.1. Apparatus
2.1.1. Samples are collected by use of a personal sampling pump
that can be calibrated within ±5% of the recommended flow rate with
the sampling filter in line.
2.1.2. Samples are collected closed-face using a sampling device
consisting of two sulfuric-acid treated
37-mm Gelman type A/E glass fiber filters contained in
a three-piece cassette. The filters are prepared by
soaking each filter with 0.5 mL of 0.26 N sulfuric acid. (0.26 N
sulfuric acid can be prepared by diluting 1.5 mL of 36 N sulfuric
acid to 200 mL with deionized water.) The filters are dried in an
oven at 100°C for 1 h and then assembled into
three-piece 37-mm polystyrene cassettes
without support pads. The front filter is separated from the back
filter by a polystyrene spacer. The cassettes are sealed with shrink
bands and the ends are plugged with plastic plugs.
2.1.3. Small sealable vials capable of holding at least 7 mL of
liquid are needed for sample shipment and storage. Glass
scintillation vials with caps containing Teflon liners are
recommended.
2.2. Reagents
Deionized water is needed for addition to the vials described in
Section 2.1.3.
2.3. Sampling technique
2.3.1. Immediately before sampling, remove the plastic plugs
from the filter cassettes.
2.3.2. Attach the cassette to the sampling pump with flexible
tubing and place the cassette in the employee's breathing zone.
2.3.3. After sampling, seal the cassettes with plastic plugs
until the filters are transferred to the vials containing deionized
water.
2.3.4. At some convenient time within 10 h of sampling, carefully
remove the filters from the cassettes and individually transfer them
to separate vials. Add approximately 2 mL of deionized water to each
vial. This can be done before or after the filters are transferred.
2.3.5. Seal the vials lengthwise with OSHA Form 21.
2.3.6. Ship and store samples for s-dianisidine at 0°C or colder.
2.3.7. Submit at least one blank filter with each sample set.
Handle the blank filters in the same manner as the air samples, but
draw no air through them.
2.3.8. Record air volumes (in liters) for each sample, along with
any potential interferences.
2.4. Retention efficiency
A retention efficiency study was performed by drawing 100 L of air
(76% relative humidity) at 1 L/min through six sample filters that had
been spiked with 1.00 µg of s-dianisidine. Instead of using backup pads,
blank acid-treated filters were used as backups in each
cassette. Upon analysis, the top filters were found to contain an
average of 90.1% (SD = 5.4) of the spiked amount. There was no
s-dianisidine found on the
bottom filters. Similar tests were done for 21.8 µg of MOCA and 0.868
µg of s-tolidine. Upon analysis,
the top filters were found to contain an average of 101.5% (SD = 4.5)
and 100.6% (SD = 4.2) of the spiked amounts of MOCA and s-tolidine respectively. There were no
detectable amounts of these two analytes found on the backup filters.
2.5. Extraction efficiency
2.5.1. The average extraction efficiencies from six filters for
each amine spiked at the target concentrations were 97.2, 95.7, and
99.2% for s-dianisidine, MOCA,
and s-tolidine respectively.
(Section 4.9.)
2.5.2. The stability of extracted and derivatized samples was
verified by reanalyzing the above samples 24 h later using fresh
standards. The average extraction efficiencies for the reanalyzed
samples were 98.9, 93.5, and 100.0% for s-dianisidine, MOCA, and s-tolidine respectively. (Section 4.9.)
2.6. Recommended air volume and sampling rate
2.6.1. The recommended air volume is 100 L.
2.6.2. The recommended sampling rate is 1 L/min.
2.6.3. If a smaller air volume is desired, the reliable
quantitation limits will be larger. For example, the reliable
quantitation limit for s-dianisidine for a 15-L air
sample would be 8.0 ppt.
2.7. Interferences (sampling)
2.7.1. Any compound in the sampled air that will react with the
sulfuric acid on the treated filters or with the collected analyte
is a potential sampling interference.
2.7.2. Suspected interferences should be reported to the
laboratory with submitted samples.
2.8. Safety precautions (sampling)
2.8.1. Attach the sampling equipment to the employees so that it
will not interfere with work performance or safety.
2.8.2. Follow all safety procedures that apply to the work area
being sampled.
3. Analytical Procedure
3.1. Apparatus
3.1.1. A GC equipped with an electron capture detector. For this
evaluation, a Hewlett-Packard 5890A Gas Chromatograph
equipped with a Nickel 63 electron capture detector and a 7673A
Automatic Sampler was used.
3.1.2. A GC column capable of separating the amine derivatives
from the solvent and interferences. A 15-m ×
0.32-mm i.d. (1.0-µm film)
SPB-5 fused silica column purchased from Supelco, Inc.
was used in this evaluation.
3.1.3. An electronic integrator or some other suitable means of
measuring peak areas or heights. A Hewlett-Packard
18652A A/D converter interfaced to a Hewlett-Packard
3357 Lab Automation Data System was used in this evaluation.
3.1.4. Small resealable vials with Teflon-lined caps capable of
holding 4 mL.
3.1.5. A dispenser or pipet for toluene capable of delivering 2.0
mL.
3.1.6. Pipets (or repetitive pipets with plastic or Teflon tips)
capable of delivering 1 mL, for dispensing the sodium hydroxide and
buffer solutions.
3.1.7. Repetitive pipets, one to deliver 25 µL of HFAA and one to
transfer 50-µL aliquots of MOCA samples and standards.
3.1.8. Disposable pipets to transfer the toluene layers after the
samples are extracted.
3.2. Reagents
3.2.1. Saturated and 0.5 N NaOH solutions, prepared from reagent
grade Na0H.
3.2.2. Toluene. American Burdick and Jackson "High Purity
Solvent" brand toluene was used.
3.2.3. Heptafluorobutyric acid anhydride (HFAA). HFAA from Pierce
Chemical Company was used.
3.2.4. Phosphate buffer, prepared from 136 g of reagent grade
potassium dihydrogen phosphate and 1 L deionized water. The pH is
adjusted to 7.0 with saturated sodium hydroxide solution.
3.2.5. s-Dianisidine, MOCA, s-tolidine, reagent grade. The s-dianisidine used in this evaluation was
purchased from Aldrich Chemical Company, Inc., Milwaukee WI. The
MOCA and s-tolidine were
purchased from CTC Organics, Atlanta, GA.
3.3. Standard preparation
3.3.1. CAUTION. THESE AROMATIC AMINES ARE OR SHOULD BE
CONSIDERED CARCINOGENIC TO HUMANS. Restrict use of pure compounds
and concentrated standards to regulated areas. Prepare concentrated
stock standards by diluting the pure amines with toluene. Prepare
analytical standards by injecting microliter amounts of diluted
stock standards into vials that contain 2.0 mL of toluene. In order
to keep the response of MOCA standards which are at or around the
target concentration (20 ppb for a 100-L air sample) in
the linear range of the electron capture detector used, a
furtherdilution was required. This was accomplished by adding
50-µL aliquots of the MOCA analytical standards to
vials containing 2.0 mL of toluene.
3.3.2. Add 25 µL of HFAA to each vial. Recap and shake the vials
for 10 s.
3.3.3. After allowing 10 min for the derivatives to form, add 1
mL of buffer to each vial to destroy the excess HFAA and to extract
the heptafluorobutyric acid that is formed.
3.3.4. Recap and shake the vials for 10 s.
3.3.5. After allowing the two layers to separate, analyze the
toluene (upper) layer of each standard by GC.
3.3.6. Bracket sample concentrations with analytical standard
concentrations. If sample concentrations are higher than the upper
range of prepared standards, prepare additional standards to
ascertain detector response or derivatize a smaller aliquot of the
toluene extract of the high samples using toluene as the diluent.
3.4. Sample preparation
3.4.1. The sample filters are received in vials containing
deionized water.
3.4.2. Add 1 mL of 0.5 N Na0H and 2.0 mL of toluene to each vial.
3.4.3. Recap and shake the vials for 10 min.
3.4.4. If the samples are to be analyzed for s-dianisidine or s-tolidine, allow the layers to separate
and transfer approximately 1 mL of the toluene (upper) layer of each
sample to separate vials with clean disposable pipets. For MOCA
samples, allow the layers to separate and transfer a
50-µL aliquot of the toluene layer of each sample to
separate vials, each containing 2.0 mL of toluene.
3.4.5. Add 25 µL of HFAA to each vial. Recap and shake the vials
for 10 s.
3.4.6. After allowing 10 min for the derivatives to form, add 1
mL of buffer to each vial to destroy the excess HFAA and to extract
the heptafluorobutyric acid that is formed.
3.4.7. Recap and shake the vials for 10 s.
3.4.8. After allowing the two layers to separate, analyze the
toluene (upper) layer of each sample by GC.
3.5. Analysis
3.5.1. GC conditions and information
zone temperatures: |
column, 250°C injector, 225°C
detector, 300°C |
|
gas flows: |
column |
2.3 mL/min hydrogen (35 kPa head pressure) |
|
make up |
45 mL/min nitrogen |
|
injection volume: |
1.0 µL |
|
split ratio: |
100:1 |
|
column: |
SPB-5, 1.0-µm film, 15-m × 0.32-mm i.d. fused
silica (Supelco, Inc.) |
|
retention times of derivatives:
|
s-Dianisidine, 5.5 min MOCA, 4.2
min s-tolidine, 3.9 min |
|
chromatograms: |
Section 4.10. |
3.5.2. Measure peak areas or heights by use of an integrator or
by other suitable means.
3.5.3. Construct a calibration curve by plotting response (peak
areas or heights) of standard injections versus micrograms of
analyte per sample. Bracket sample concentrations with standards.
3.6. Interferences (analytical)
3.6.1. Any compound that elutes in the same general time as the
HFAA derivative of the amine of interest is a potential
interference. Suspected interferences reported to the laboratory
with submitted samples by the industrial hygienist must be
considered before samples are derivatized.
3.6.2. GC parameters may be changed to possibly circumvent
interferences.
3.6.3. Retention time on a single column is not considered proof
of chemical identity. Analyte identity should be confirmed by GC/MS
if possible.
3.7. Calculations
The analyte concentration for samples is obtained from the
calibration curve in micrograms of analyte per sample. If any analyte
is found on the blank, that amount is subtracted from the sample
amounts. The air concentrations are calculated using the following
formulae.
µg/m3 = |
(micrograms of analyte per
sample) (1000)
(liters of air sampled) (extraction
efficiency) |
where extraction efficiencies are: |
97.2% (s-Dianisidine)
95.7% (MOCA) 99.2% (s-Tolidine) |
ppb = |
(µg/m3) (24.46)
(molecular weight of analyte) |
where |
24.46 is the molar volume (liters) at 25°C and 760
mm Hg molecular weights are: 244.3 (s-Dianisidine), 267.2
(MOCA), 212.3 (s-Tolidine) |
3.8. Safety precautions (analytical)
3.8.1. CAUTION. THESE AROMATIC AMINES ARE OR SHOULD BE
CONSIDERED CARCINOGENIC TO HUMANS. Restrict use of pure compounds
and concentrated standards to regulated areas.
3.8.2. Avoid skin contact and inhalation of all chemicals.
Restrict the use of chemicals to a fume hood if possible. Wear
safety glasses and a lab coat while in the lab area.
4. Backup Data
4.1. Detection limit of the analytical procedure
The injection volume listed in Section 3.5.1., 1.0 µL with a 1 to
100 split, was used in the determination of the detection limits of
the analytical procedure. The detection limits of 6.0 fg of
s-dianisidine, 5.5 fg of MOCA,
and 5.4 fg of s-tolidine were
determined by analyzing dilute standards equivalent to 1.20 ng of
s-dianisidine, 43.8 ng of MOCA,
and 1.09 ng of s-tolidine per
sample. (The samples are extracted into 2.0 mL of toluene. The MOCA
samples are further diluted by transferring a 50-µL aliquot to 2.0 mL
of toluene.) These amounts were judged to give peaks with heights
approximately 5 times the baseline noise. Chromatograms of such
injections are shown in Figures 4.1.1.
and 4.1.2.
4.2. Detection limit of the overall procedure
The detection limits of the overall procedure were determined by
analyzing filters spiked with loadings equivalent to the detection
limits of the analytical procedure. Samples were prepared by injecting
1.20 ng of s-dianisidine, 43.8
ng of MOCA, and 1.09 ng of s-tolidine onto acid-treated
filters. These amounts are equivalent to 1.2 ppt (12
ng/m3), 4O ppt (440
ng/m3), and 1.3 ppt (11
ng/m3) for s-dianisidine, MOCA, and s-tolidine respectively.
Table 4.2.1. Detection Limit of the Overall Procedure
for s-Dianisidine
|
sample no. |
ng spiked |
ng recovered |
|
1 2 3 4 5 6 |
1.20 1.20 1.20 1.20 1.20 1.20 |
1.07 1.27 1.13 1.01 1.25 1.31 |
|
Table 4.2.2. Detection Limit of the Overall Procedure
for MOCA
|
sample no. |
ng spiked |
ng recovered |
|
1 2 3 4 5 6 |
43.8 43.8 43.8 43.8 43.8 43.8 |
37.9 46.3 39.9 47.8 42.7 41.1 |
|
Table 4.2.3. Detection Limit of the Overall Procedure
for s-Tolidine
|
sample no. |
ng spiked |
ng recovered |
|
1 2 3 4 5 6 |
1.09 1.09 1.09 1.09 1.09 1.09 |
1.02 1.15 1.01 0.97 1.07 1.06 |
|
4.3. Reliable quantitation limit
The reliable quantitation limits were determined by analyzing
filters spiked with loadings equivalent to the detection limits of the
analytical procedure. Samples were prepared by injecting 1.20 ng of
s-dianisidine, 43.8 ng of MOCA,
and 1.09 ng of s-tolidine onto
acid-treated filters. These amounts are equivalent to 1.2
ppt (12 ng/m3), 40 ppt (440
ng/m3), and 1.3 ppt (11
ng/m3) for s-dianisidine, MOCA, and s-tolidine respectively.
Table 4.3.1. Reliable Quantitation Limit for
s-Dianisidine (Based on
samples and data of Table 4.2.1.)
|
sample |
% recovered |
statistics |
|
1 2 3 4 5 6 |
89.2 105.8 94.2 84.2 104.2
109.2 |
=
SD = Precision =
= |
97.8%
10.1% ±(1.96)(10.1%) ±19.8 |
|
Table 4.3.2. Reliable Quantitation Limit for MOCA
(Based on samples and data of Table 4.2.2.)
|
sample |
% recovered |
statistics |
|
1 2 3 4 5 6 |
86.5 105.7 91.1 109.1
97.5 93.8 |
=
SD = Precision =
= |
97.3%
8.69% ±(1.96)(8.69%) ±17.0 |
|
Table 4.3.3. Reliable Quantitation Limit for
s-Tolidine (Based on
samples and data of Table 4.2.3.)
|
sample |
% recovered |
statistics |
|
1 2 3 4 5 6 |
96.6 105.5
92.7 89.0 98.2 97.2 |
=
SD = Precision =
= |
96.0%
5.70% ±(1.96)(5.70%) ±11.2 |
|
4.4. Instrument response to the analyte
The instrument response to the analytes over the range of 0.5 to 2
times the target concentrations was determined from multiple
injections of analytical standards. These data are given in Tables
4.4.1. - 4.4.3. and Figures 4.4.1.
- 4.4.3.
The response is linear for the three analytes with slopes (in area
counts per micrograms of analyte per sample) of 150,400 for
s-dianisidine, 5080 for MOCA,
and 190,100 for s-tolidine.
Table 4.4.1. Instrument Response to s-Dianisidine
|
× target conc. µg/sample ppb |
0.5× 0.50 0.50 |
1× 1.00 1.00 |
2× 2.00 2.00 |
|
area counts
|
106412 103872 100769 106147 105799 103801
104467 |
195200 191643 189055 195327 188406 186838
191078 |
333377 337847 340122 335264 321881 325757
332375 |
|
Table 4.4.2. Instrument Response to MOCA
|
× target conc. µg/sample ppb |
0.5× 10.9 10.0 |
1× 21.8 20.0 |
2× 43.6 39.9 |
|
area counts
|
74749 75422 79894 75636 77790 75830
76554 |
127250 122572 133929 137936 132986 132634
131218 |
244357 255069 248678 232331 235188 239419
242507 |
|
Table 4.4.3. Instrument Response to s-Tolidine
|
× target conc. µg/sample ppb |
0.5× 0.434 0.50 |
1× 0.868 1.00 |
2× 1.736 2.00 |
|
area counts
|
115996 112884 110286 116518 115563 110950
113700 |
214903 209958 205280 212341 204580 201801
208144 |
367038 373575 371890 365314 349354 354703
363646 |
|
4.5. Storage test
Test atmospheres containing these potentially carcinogenic amines
could not be safely generated in our laboratory. Storage samples were
generated by spiking acid-treated filters with amounts of
analyte equal to the target concentrations (1.00 µg of s-dianisidine, 21.8 µg of MOCA, 0.868 µg of
s-tolidine).
Thirty-six samples were prepared for each analyte. One
hundred liters of air at 76% relative humidity were then drawn through
each filter. Within 1 h after the completion of drawing air through
the samplers, the filters were transferred to scintillation vials,
each containing 2 mL of deionized water. Six samples for each analyte
were analyzed immediately, fifteen were stored in a refrigerator at
0°C, and fifteen were stored in a closed drawer at ambient
temperature. Six samples for each analyte, three from refrigerated and
three from ambient storage, were analyzed at intervals over a period
of fifteen days. The results are given in Tables 4.5.1. -
4.5.3. and in Figures 4.5.1.1.,
4.5.1.2.,
4.5.2.1.,
4.5.2.2.,
4.5.3.1.
and 4.5.3.2.
The percent recovery versus days of storage was fit to the best
regression curve for each analyte under both storage conditions. The
standard errors of estimate are 7.8, 5.8, and 8.0% and the 95%
confidence limits (±1.96 SD) are ±15.4, ±11.3, and ±15.7% for
s-dianisidine, MOCA, and
s-tolidine respectively. These
values were obtained from Figures 4.5.1.1., 4.5.2.2. and 4.5.3.2.
Table 4.5.1. Storage Test for s-Dianisidine
|
days of |
|
% recovery |
storage |
|
refrigerated |
|
ambient |
|
0 0 3 5 8 12 15
|
|
94.0 102.8
87.5 77.0 86.4 73.2 85.6 |
97.6 91.2 92.4 83.3
88.6 72.8 89.5 |
90.6 89.4 92.5 84.2
85.8 77.0 84.6 |
|
94.0 102.8
74.5 81.2 62.6 69.4 53.7 |
97.6 91.2 70.2 49.9
68.8 34.4 70.0 |
90.6 89.4 72.0 63.9
54.6 45.9 67.8 |
|
Table 4.5.2. Storage Test for MOCA
|
days of |
|
% recovery |
storage |
|
refrigerated |
|
ambient |
|
0 0 3 5 8 12 15
|
|
98.2 104.9
98.6 101.4 101.8 102.0 101.3 |
95.0
97.0 103.4 108.4 100.6 99.0 102.3 |
95.4 100.4 104.7
99.7 102.3 100.9 104.6 |
|
98.2 104.9 102.5
98.4 102.6 99.7 103.2 |
95.0
97.0 100.8 102.0 104.8 97.2 99.2 |
95.4 100.4 101.9
99.4 104.3 98.1 100.6 |
|
Table 4.5.3. Storage Test for s-Tolidine
|
days of |
|
% recovery |
storage |
|
refrigerated |
|
ambient |
|
0 0 3 5 8 12 15
|
|
98.4 109.2 94.8 98.1 103.2
93.6 100.6 |
102.7 96.8 101.1 102.6
101.2 98.2 101.0 |
99.0 97.2 99.3 101.3 101.5
95.6 96.5 |
|
98.4 109.2
96.0 96.2 96.6 86.6 92.2
|
102.7
96.8 87.4 92.0 98.2 76.2 97.2 |
99.0 97.2 93.6 91.4 99.0 83.6 96.8 |
|
4.6. Precision (analytical method only)
The precision of the analytical method for each analyte is the
pooled coefficient of variation determined from replicate injections
of standards. The standards analyzed for these determinations are
described in Section 4.4. The precision of the analytical method for
each analyte is given in Tables 4.6.1. - 4.6.3.
Table 4.6.1. Precision of the Analytical Method for
s-Dianisidine (Based on the data
of Table 4.4.1.)
|
× target conc. µg/sample ppb |
0.5× 0.50 0.50 |
1× 1.00 1.00 |
2× 2.00 2.00 |
|
SD (area counts) |
2138 |
3594 |
7117 |
CV |
0.020 |
0.019 |
0.021 |
|
= 0.020 |
|
Table 4.6.2. Precision of the Analytical Method for
MOCA (Based on the data of Table 4.4.2.)
|
× target conc. µg/sample ppb |
0.5× 10.9 10.0 |
1× 21.8 20.0 |
2× 43.6 39.9 |
|
SD (area counts) |
1928 |
5443 |
8555 |
CV |
0.025 |
0.041 |
0.035 |
|
= 0.034 |
|
Table 4.6.3. Precision of the Analytical Method for
s-Tolidine (Based on the data of
Table 4.4.3.)
|
× target conc. µg/sample ppb |
0.5× 0.434 0.50 |
1× 0.868 1.00 |
2× 1.736 2.00 |
|
SD (area counts) |
2704 |
5054 |
9644 |
CV |
0.024 |
0.024 |
0.027 |
|
= 0.025 |
|
4.7.Precision (overall procedure)
The precision of the overall procedure is determined from the
storage data. The determination of the standard error of estimate
(SEE) for a regression line plotted through the graphed storage data
allows the inclusion of storage time as one of the factors affecting
overall precision. The SEE is similar to the standard deviation,
except it is a measure of dispersion of data about a regression line
instead of about a mean. It is determined with the following equation:
where |
n = k
= k = |
total no. of data points 2 for linear
regression 3 for quadratic regression |
Yobs
= |
observed % recovery at a given time |
Yest
= |
estimated % recovery from the regression line at the same
given time |
An additional 5% for pump error is added to the SEE by the addition
of variances. The precision at the 95% confidence level is obtained by
multiplying the SEE (with pump error included) by 1.96 (the
z-statistic from the standard normal distribution at the
95% confidence level). The 95% confidence intervals are drawn about
their respective regression lines in the storage graphs as shown in
Figures 4.5.1.1.,
4.5.1.2.,
4.5.2.1.,
4.5.2.2.,
4.5.3.1.
and 4.5.3.2.
The precisions of the overall procedure of ±15.4 for s-dianisidine, ±11.3 for MOCA, and ±15.7%
for s-tolidine were obtained
from Figures 4.5.1.1., 4.5.2.2. and 4.5.3.2. respectively.
4.8. Reproducibility
Six samples for each analyte were prepared by injecting microliter
quantities of standards onto acid-treated filters. The
s-dianisidine and s-tolidine samples were stored at 0°C for 2
days and the MOCA samples were stored at -35°C for 42
days. The samples were analyzed by a chemist unassociated with this
evaluation. The results are given in Tables 4.8.1. -
4.8.3. All of the amounts found were corrected for extraction
efficiency. The extraction efficiency is 97.2% for s-dianisidine, 95.7% for MOCA, and 99.2% for
s-tolidine. The precision of the
overall procedure is ±15.4% for s-dianisidine, ±11.3% for MOCA, and ±15.7%
for s-tolidine.
Table 4.8.1. Reproducibility Data for s-Dianisidine
|
sample no. |
µg found |
µg expected |
% found |
% deviation |
|
1 2 3 4 5 6 |
0.927 0.940 0.963 0.959 0.988 0.960 |
0.960 0.960 0.960 0.960 0.960 0.960 |
96.6 97.9 100.3 99.9 102.9
100.0 |
-3.4 -2.1 +0.3 -0.1 +2.9
0.0 |
|
Table 4.8.2. Reproducibility Data for MOCA
|
sample no. |
µg found |
µg expected |
% found |
% deviation |
|
1 2 3 4 5 6 |
20.80 20.05 20.42 19.82 21.90 21.59 |
22.07 22.07 22.07 22.07 22.07 22.07 |
94.2 90.8 92.5 89.8 99.2 97.8 |
-5.8 -9.2 -7.5 -10.2
-0.8 -2.2 |
|
Table 4.8.3. Reproducibility Data for s-Tolidine
|
sample no. |
µg found |
µg expected |
% found |
% deviation |
|
1 2 3 4 5 6 |
0.859 0.861 0.886 0.868 0.871 0.859 |
0.872 0.872 0.872 0.872 0.872 0.872 |
98.5 98.7 101.6
99.5 99.9 98.5 |
-1.5 -1.3 +1.6 -0.5 -0.1 -1.5 |
|
4.9. Extraction efficiency data
Six sample filters for each amine were spiked with the target
concentration amounts by liquid injection (1.00 µg of s-dianisidine, 21.8 µg of MOCA, and 0.868 µg
of s-tolidine). These samples
were analyzed to determine the extraction efficiencies. To determine
the stability of extracted and derivatized samples, these same samples
were reanalyzed after setting at room temperature for 24 h using fresh
standards. The results are given in Tables 4.9.1. -
4.9.3.
Table 4.9.1. Extraction Efficiency for s-Dianisidine,
|
sample no. |
% extracted |
reanalyzed after 24 h |
|
1 2 3 4 5 6
|
99.2 95.7 94.0 98.2 97.0 99.4
97.2 |
106.2
97.3 96.2 96.8 96.5 100.3
98.9 |
|
Table 4.9.2. Extraction Efficiency for MOCA
|
sample no. |
% extracted |
reanalyzed after 24 h |
|
1 2 3 4 5 6
|
95.4 92.7 99.5 94.0 92.2 100.5
95.7 |
94.4 94.1 96.6 92.9 90.3 92.8
93.5 |
|
Table 4.9.3. Extraction Efficiency for s-Tolidine
|
sample no. |
% extracted |
reanalyzed after 24 h |
|
1 2 3 4 5 6
|
101.7 97.1 96.5 100.5
97.7 101.8
99.2 |
107.5
95.9 98.9 98.2 95.8 103.7
100.0 |
|
4.10. Chromatogram
Chromatograms at the target concentrations are shown in Figures 4.10.1.
and 4.10.2.
The chromatograms are from 1.0-µL injections of
standards.
Figure 4.1.1.
Detection limit chromatogram for s-dianisidine and s-tolidine.
Figure 4.1.2. Detection
limit chromatogram for MOCA.
Figure 4.4.1. Instrument
response to s-dianisidine.
Figure 4.4.2. Instrument
response to MOCA.
Figure 4.4.3. Instrument
response to s-tolidine.
Figure 4.5.1.1.
s-Dianisidine refrigerated storage
samples.
Figure 4.5.1.2.
s-Dianisidine ambient storage
samples.
Figure 4.5.2.1. MOCA
refrigerated storage samples.
Figure 4.5.2.2. MOCA
refrigerated storage samples.
Figure 4.5.3.1.
s-Tolidine refrigerated storage
samples.
Figure 4.5.3.2.
s-Tolidine ambient storage
samples.
Figure 4.10.1.
s-Dianisidine and s-Tolidine
chromatogram.
Figure 4.10.2. MOCA
chromatogram.
5. References
5.1. "Chemical Information Manual", U.S. 1 Department of Labor,
Occupational Safety and Health Administration, OSHA Instruction CPL
2-2.43, Directorate of Technical Support; National
Technical Information Service: Springfield, VA, 22161, October 20,
1987.
5.2. Elskamp, Carl J. "OSHA Method No. 65; Benzidine,
3,3'-Dichloro-benzidine, 2,4-Toluenediamine,
2,6-Toluenediamine", OSHA Analytical Laboratory,
unpublished, Salt Lake City, UT, 84165, August 1987.
5.3. Elskamp, Carl J. "OSHA Method No. 57;
4,4'-Methylenedianiline", OSHA Analytical Laboratory,
unpublished, Salt Lake City, UT, 84165, January 1986.
5.4. "IARC Monographs on the Evaluation of the Carcinogenic Risk of
Chemicals to Man: Some aromatic amines, hydrazine and related
substances, N-nitroso compounds and miscellaneous
alkylating agents", International Agency for Research on Cancer: Lyon,
1974; Vol. 4, pp 41-47.
5.5. "IARC Monographs on the Evaluation of the Carcinogenic Risk of
Chemicals to Man: Some aromatic amines, hydrazine and related
substances, N-nitroso compounds and miscellaneous
alkylating agents", International Agency for Research on Cancer: Lyon,
1974; Vol. 4, pp 65-71.
5.6. "Documentation of the Threshold Limits Values and Biological
Exposure Indices", 5th ed.; American Conference of Governmental
Industrial Hygienists Inc.: Cincinnati, OH, 1986; p 392.4(86).
5.7. "Documentation of the Threshold Limits Values and Biological
Exposure Indices", 5th ed.; American Conference of Governmental
Industrial Hygienists Inc.: Cincinnati, OH, 1986; p 577.
|