p.1 

OAR Box 1846 

Prepped by Ollie Stewart 

Document Number: 

79) IV-A-1 

Docket Number: 

A-90-07 

Printed 11/18/2008 7:30:17 AM Sheet 79 of 93 
— P.2 

CRC Repdrf No. 551 

CHEMICAL METHODS FOR THE 

MEASUREMENT OF 

UNREGULATED DIESEL EMISSIONS 

Carbonyls/Aldehydes 

Particulate Characterization 

Sulfates 

PAH/N09PAH 

August 1987 

J'i'V FE3 IT1993 x ' 

I' " L _ . -—' 

{ EPA A1BJ30CICET 

COORDINATING RESEARCH COUNCIL, INC. 

219 PERIMETER CENTER PARKWAY, ATLANTA, GEORGIA 30348 
P.3 

»» COORDINATING RESEARCH COUNCIL 

INCORPORATED 

2 1 9 PERIMETER CENTER PARKWAY 

ATLANTA. GEORGIA 3034 6 

(404) 396-340 0 

CHEMICAL METHODS FOR THE MEASUREMENT OF 

UNREGULATED DIESEL EMISSIONS -

CARBONYLS/ALDEHYDES 

PARTICULATE CHARACTERIZATION 

SULFATES 

PAH/NO2PAH 

(CRC-APRAC PROJECT NO. CAPI-1-64) 

IN FORMULATING AND APPROVING REPORTS, THE 

APPROPRIATE COMMITTEE OF THE COORDINATING 

RESEARCH COUNCIL, INC. HAS NOT INVESTIGATED 

OR CONSIDERED PATENTS WHICH. MAY APPLY TO 

THE SUBJECT MATTER. PROSPECTIVE USERS OF THE 

REPORT ARE RESPONSIBLE FOR PROTECTING THEMSELVES AGAINST LIABILITY FOR INFRINGEMENT OF 

PATENTS. 

Prepared by the 

Chemical Characterization Panel 

of the 

CRC Program Group on Composition of Diesel Exhaust 

Edited By: 

J. M. Perez 

R. E. Jass 

D. G. Leddy 

August 1987 

Air Pollution Research Advisory Committee 

of the 

Coordinating Research Council, Inc. 
P.4 

TABLE OF CONTENTS 

Paige 

SECTION I - INTRODUCTION 3 

SECTION II - HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC (HPLC) 

DINITROPHENYLHYDRAZINE (DNPH) METHOD FOR CARBONYL COMPOUNDS 

A. Scope 7 

B. Definition 7 

C. Experimental 7 

D. Sample Collection 9 

E. Sample Analysis 9 

F. Precision and Accuracy 10 

G. Alternate Sampling Procedure 11 

SECTION III - DIESEL PARTICULATE CHARACTERIZATION 

A. Purpose 15 

B. Summary 15 

C. Definitions/Abbreviations 16 

D. Experimental 17 

SECTION 111.1 - TOTAL PARTICULATE MASS 

A. Procedures 21 

B. Measurement of Total Particulate Mass . 23 

SECTION III.2 - SOLVENT EXTRACTABLE FRACTIONS (SEF) 

A. Scope 27 

B. Extraction of SOF 27 

C. Extraction of TOE 28 

D. Determination of Sulfate 29 

E. Combined Water 30 

F. Repeatabi 1 ity 31 

SECTION IV - NITRATED POLYCYCLIC AROMATIC HYDROCARBONS 

A. Scope 33 

B. Background 33 

C. Definitions/Abbreviations 33 

D. Experimental Equipment 33 

E. Sol vents 34 

F. Procedure * 34 

G. Precision and Accuracy 35 

H. Alternate Column Configuration 35 

I. Alternate Method 35 
P.5 

SECTION V - POLYCYCLIC AROMATIC HYDROCARBONS 

A. Scope 39 

B. Background 39 

C. Definitions/Abbreviations 39 

D. Equipment 40 

E. Sol vents 40 

F. Procedures ^... 41 

REFERENCES 49 

TABLES 

Table I - Physical Properties of Carbonyl Compounds 55 

Table II - Storage Stability 56 

Table III - Comparison of SOF and Vacuum Removal of Organic 

Fraction 57 

Table IV - Elution of Polycyclic Aromatic Hydrocarbons on 

a Bondapak NH2 Column 58 

Table V - High Performance Liquid Chromatography of Polycyclic 

Aromatic Hydrocarbons: Fraction 1 59 

Table VI - High Performance Liquid Chromatography of Polycyclic 

Aromatic Hydrocarbons: Fraction 2 60 

Table VII - High Performance Liquid Chromatography of Polycyclic 

Aromatic Hydrocarbons: Fraction 3 61 

FIGURES 

Figure 1 - Sampling Cart (Schematic Diagram) 62 

Figure 2. - Carbonyl Analysis Method 63 

Figure 3 - Chromotogram of the Standard DNPH-Aldehyde Mixture 64 

Figure 4 - Formaldehyde Stability 65 

Figure 5 - Acrolein Stability 66 

Figure 6 - Total Aldehyde Stability 67 

Figure 7 - Soxhlet (Photograph) 68 

Figure 8 - Mass of Bound HoO Per Gram H2 S 04 in an Aqueous 

Solution Versus Relative Humidity (25°C) 69 

Figure 9 - Schematic of Nitropyrene Analysis System 70 

Figure 10 - HPLC PAH Method No. 1 71 

Figure 11 - HPLC Method Summary - Fractionation of the 

Diesel Extract 72 

Figure 12 - HPLC PAH Method No.. 2 73 

Figure 13 - Typical HPLC Traces, Method No. 2 74 

APPENDICES 

APPENDIX A - Membership: Chemical Characterization Panel of the 

CRC Diesel Exhaust Composition Group A-l 

• n -
P.6 

SECTIO N I 

INTRODUCTION 
P.7 

I. INTRODUCTION 

This Progress Report summarizes research conducted on unregulated 

diesel emissions methods by the Chemical Characterization Panel (CCP) 

of the Coordinating Research Council's (CRC) Diesel Composition Group 

over the past three years. (Membership of the Chemical Charactization 

Panel is liste d in Appendix A.) Methods are recommended for the 

measurement of aldehydes/carbonyls, sulfates, particulate characterization, polycyclic aromatic hydrocarbons (PAH) and nitrated PAH (N02-

PAH). Most of the methods proposed are the result of cooperative 

testing and are considered to be based on sound engineering practices. 

Two of the methods, PAH and N02PAH, are stil l being evaluated through 

cooperative round-robin study. 

The PAH method was developed under CRC contract at IITRI and previously evaluated in a round-robin by the group. Currently, a followup international round-robin is in progress. 

The N02PAH method has evolved primaril y as the result of work conducted at EPA-RTP. An alternate method was developed by A. D. Little , 

Inc. under contract for the CRC-APRAC CAPE-30 Project Group. 
P.8 

SECTIO N I I 

HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC (HPLC) 

DINITROPHENYLHYDRAZINE (DNPH) METHOD FOR CARBONYL COMPOUNDS 
P.9 

I I . HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC (HPLC) DINITROPHENYLHYDRAZINE (DNPH) METHOD FOR CARBONYL COMPOUNDS 

SCOPE 

The procedure incorporates an improved 2,4-dinitrophenylhydrazine 

method in which C^ to Cg carbonyl compounds are collected by 

bubbling diesel exhaust or diluted diesel exhaust through an 

acetonitrile solution of DNPH reagent and analyzed by HPLC using 

UV detection at 365 nm. Using an acetonitril e solution for 

sample collection allows direct injection of the sample into the 

HPLC, thereby eliminating many of the recovery steps and saving 

considerable sample preparation manhours. The method results in 

quantitative sample recoveries, improved accuracy and provides 

for a simple, rapid and low cost method for aldehyde analysis. 

The method was reported

 (1)*. 

B. DEFINITION 

Carbonyl Compounds - Compounds containing the C=0 functional 

chemical group. This procedure refers to Cj to Cg compounds 

containing this group, specifically the compounds shown in Table 

I. The method could also be applicable to higher molecular 

weight carbonyl compounds. 

EXPERIMENTAL 

1. Reagents - Reagents used in thi s procedure should be HPLC 

Grade (distilled in glass) or Equivalent Grade Solvents as 

indicated: 

Acetonitrile - Burdick and Jackson Labs, Inc., Fisher 

Scientific Co., J.T. Baker Chemical Co. or equivalent. 

2,4-Dinitrophenylhydrazine - Aldrich Chemical Co. or J.T. 

Baker, recrystallized twice from HPLC grade methanol or 

acetonitrile. 

Distilled, deionized water (HPLC Grade). 

2N HCl - Best Source. 

Ethanol - Best Source. 

Perchloric Acid - Best Source. 

* Numbers in ( ) are references at end of text. 
P.10 

Aldehyde Standards - Individual aldehydes for standard 

preparation should be best available grade. 

2,4-DNPH Derivatives - Method of preparation discussed in 

later'section. 

2. Equipment 

HPLC with UV detection (365 nm). Various units are available. 

5uZorbax ODS 4.6 mm x 25.0 cm HPLC column, DuPont Inst. Co. 

Midget Impingers (Light-Duty Diesels) or 100 mL Glass Impingers (Heavy-Duty Diesels). 

Flowmeter, pump and meter — required for sample collection 

(Figure 1). 

LC syringes as required. 

Melting Point Apparatus. 

Pretest Preparation 

a. Aldehyde-DNPH Derivatives - The 2,4-DNPH derivatives are 

prepared by reaction of the individual aldehydes with a 

saturated DNPH solution in 2N HCL. The derivatives are 

filtered, washed with 2N HCl and H2 0, air dried, and 

stored in sealed vials. Purity of the derivatives is 

' checked by melting point determinations, Table I. 

Purity can be improved by recrystallization from 

absolute ethanol or methanol, if required. Individual 

derivatives or a mixture of the standards should also be 

checked for purity by HPLC. 

b. Preparation of Solutions and Standards - A 6.25ymole/mL 

of 2,4-DNPH stock solution is prepared by addition of 

100 mL of acetonitrile to 120 mg of purified DNPH 

crystals. Dilution of the stock solution with acetonitrile is used to prepare a 1.6 mole/mL DNPH absorbing 

solution in the bubblers. Five drops of IN HCl0^ 

catalyst is added per each 20 mL of absorbing solution. 

The acid is added just prior to use. 

Standard solutions of the aldehydes are prepared by 

injecting 5.0 mL of the aldehyde into a septum sealed 

vial containing 5.0 mL of acetonitrile. The concentrations of the stock solutions are calculated from the 

densities of the aldehydes. The formaldehyde concentration is determined by the concentration of the 
P.11 

-9-. 

formalin solution by titration . Standard derivative 

solutions are prepared by injecting 2.0 to 20 uL of the 

aldehyde solution into a septum sealed vial containing 

the derivatizing reagent. Weighing solid DNPH hydrazones and dissolving them in acetonitrile can also be 

used to prepare standard derivatives. 

D. SAMPLE COLLECTION 

1. Sampling System - The sampling system can vary depending on 

the source of aldehydes. One system, Figure 1, was designed 

to sample diluted exhaust from various size dilution tunnels 

or heated Teflon

 K bags containing diluted exhaust. For 

sampling directl y from undiluted exhaust streams a short 

(approximately 2 meters X 0.635 cm O.D.) stainless steel 

line should be added to allow some cooling of the exhaust 

prior to entering the Teflon coated heated line. 

2. Sampling - A 1.6umole/mL DNPH solution is used for emission 

testing. This is obtained by diluting the stock solution as 

indicated above or the solution can be prepared by dis -

solving about 31 mg of purified DNPH in 100 mL of acetonitrile . 

For each light-dut y diesel (LDD) sample to be taken, the 

impingers are prepared by adding 10 mL of the DNPH solution 

and 2 drops (approximately 100 uL) of the IN HC104 to each 

impinger. Samplers are connected in series and placed in an 

ice bath. For sampling heavy-duty diesels (HDD), 20 ml of 

the DNPH solution with 4 drops IN HC104 are added to each 

impinger. Sample flow through the bubblers is maintained at 

1/2 L/min for LDD and 2-4 L/min HDD. Normally 10-30 L of 

exhaust (100-300 L at 9:1 dilutio n ratio) is adequate to 

obtain sufficient sample for a reliable analysis depending 

on the type of emission source. The lower the source concentration the more sample that is required for good quantitative results. As a guideline, aldehyde levels from 

LDD<:HDD<HD gasoline engines. 

Samples should be analyzed immediately or refrigerated until 

used. 

SAMPLE ANALYSIS 

1. Analysis Procedure - Samples should be analyzed as soon as 

possible, preferably within a few hours, to minimize the 

losses of certain species. The procedure for analysis is 

shown in Figure 2. Following an acceptable HPLC blank, the 

calibration standards are run. 
P.12 

•10 -

An example of an HPLC trace for the standards is shown in 

Figure 3. I t should be emphasized that the appearance may 

vary depending on the HPLC system utilized . However, the 

relative retention times of the peaks should closely agree 

with'those shown in Table I. I f not, retention times should 

be redetermined using a set of standards. The time required 

from injection to the appearance of the internal standard, 

benzaldehyde, is considered to be 1.0. The times for al l 

other peaks to elute are normalized to it . If an automated 

system is utilized, the samples can be sequenced running a 

standard check at the end of the series or every 4-6 samples 

depending on the number of samples to be analyzed. 

2. Sample Stability - The stability of samples indicates that 

losses of individual aldehydes, with the the exception of 

acrolein, are only about 10 percent in 24 hours. Experimentall y determined losses for formaldehyde, acrolein and total 

aldehydes are shown in Figures 4, 5, and 6. The acrolein 

losses as demonstrated on Figure 5 are concentration dependent. Higher concentrations approach 35% loss in 24 hours. 

Fortunately, this species is normally a minor component of 

the total aldehydes. Total aldehyde losses are 10 percent 

or less. Typically, sampling and analysis is completed 

within 24 hours. Storage stabilit y beyond 24 hours is shown 

on Table II . The data show that losses in total aldehydes 

within an eight week period were less than 5 percent. This 

is primarily due to acrolein and crotonaldehyde degradation. 

F. PRECISION AND ACCURACY 

The method precisio n and accuracy were determined in a 

cooperativ e round-robi n conducted by the Chemical 

Characterization Panel of the Diesel Exhaust Composition inhouse 

Group of the Coordinating Research Council, Inc., Air Pollution 

Research Advisory Committee (CRC-APRAC CAPI-1-64). The results 

are described in Reference 1. The concentration levels of 

aldehydes in the study are considered to be a worst case 

situation involving low levels of aldehydes. Reproducibility, 

i.e., agreement within a laboratory, was better than 20 percent 

relative standard deviation (r.s.d.). Agreement between laboratories , repeatability , was as high as 35 percent r.s.d. The 

precision and accuracy improved as the concentration of aldehydes 

in the samples increased. The concentration levels in the CRC 

study were of the order of 0.02y moles/ml and should be considered minimum levels required to achieve 20 percent r.s.d. 

reproducibility. With experience and analysis of samples within 

24 hours after collection, a precision and accuracy of 10 percent 

r.s.d. or better should be achievable. 
P.13 

-11-

G. ALTERNATE SAMPLING PROCEDURE 

Both General Motors Research and EPA-RTP have developed procedures utilizing DNPH-coated silica cartridges for sampling and 

analyzing carbonyl compounds in engine exhaust. S.B. Tejada's 

(EPA-RTP) method was presented at the CRC workshop in March 

1985(2). Basically, the procedure involves: 

1. Reagent purification. 

2. Preparation of DNPH-coated silica cartridges. 

3. Impinger solution. 

4. Cartridge sampling. 

5. HPLC conditions. 

One interesting aspect of thi s method is the presence of an 

unidentified "compound-x" that may be the decomposition product 

of acrolein in air. 

A second advantage of thi s procedure is the abilit y to readily 

collect field samples. Stabilit y of formaldehyde and acrolein 

appear improved on the cartridge. 

Tejada's procedure and the method for preparing cartridges have 

been published in the literature* 3'. 
P.14 

SECTIO N III. l 

TOTAL PARTICULATE MASS 
P.15 

•21-

III. 1 TOTAL PARTICULATE MASS 

A. PROCEDURES 

1. Operation of the Engine - Diesel particulate can be collected 1n a variety of tests. For steady-state particulate 

collection with or without a dilutio n tunnel, the engine 

operating procedures as described in SAE Procedure J1003(7) 

are guidelines for engine operation. For transient tests , 

the USEPA proceduresr8) can be used as a guideline. 

2. Dilution Ratio - The dilution ratio used to collect particulate 1s normally about 10-20 to 1, A1r to Exhaust. In 

practice, the actual dilution ratio 1s usually determined by 

the Incoming exhaust temperature and the amount of air 

required to achieve a 125°F (52°C) or lower filte r temperature. The time to collect a desired mass of particulate is 

also a factor 1n selecting the dilution ratio. References 9 

and 10 discuss these Interactions. Normally, the C02 levels 

of the raw exhaust and diluted exhaust, corrected for C02 in 

the Incoming air , are used to calculate dilutio n ratio in 

steady-state tests. Mass flow measurements are usually used 

in transient tests to determine the dilution. 

The dilutio n rati o can also be determined from the 

concentration of N0X in the raw and diluted exhaust. 

3. Tunnel Background - Tunnel background and conditioning can 

affect the levels of some specific hydrocarbon species such 

as PAH. The tunnel background 1s normally not a major 

factor In tota l particulate and SOF measurements I f some 

simple precautions are taken. Tunnel blanks should be 

periodically run by drawing dilution air through the filte r 

for an equivalent period of time as that used for sampling. 

Background levels should be well under IX of the total mass 

expected to be collected. On start-up of a test , engine 

- exhaust conditions (e.g., temperature, FID emissions) should 

be stabilized* 1 1 '

1 2 * before Introducing the exhaust Into the 

tunnel. Air only should be run through the tunnel during 

engine warm-up. A minimum of 20 minutes at the desired 

dilutio n rati o should be allowed for tunnel equilibration 

before collecting samples. I f the sampling point follows a 

high hydrocarbon emissions point (e.g., Low Idle or ligh t 

load conditions), a 15-minute high load burnout of the exhaust system may be desirable prior to setting the engine 

test condition. 
P.16 

•22-

4. Particulate Collection - Particulate collection methodology 

for transient tests is defined in the USEPA procedures'8'. 

For steady-state measurements, the methodology can be considerably different depending on the type of dilution tunnel 

used (13,14). The procedures used are usually those that 

have been developed by the researcher. The particulate 

characterization methods discussed in the following section 

do not depend on how the sample was collected. However, 

several precautionary items that wil l improve reproducibilit y in a laboratory are as follows: 

(a) Utiliz e Pallflex TX40HI20WW filters . (Filte r sizes 

from 47mm to 102mm are commonly used.) 

(b) Handle the filte r at al l times using tweezers or 

forceps. Avoid using fingers on the filter s or any 

device that may tear particles from the filter. 

(c) Condition the filte r by allowing i t to equilibrate in a 

constant temperature and humidity environment for 16-24 

hours prior to collection of the sample. After 

sampling, the filter s should be conditioned in the same 

environment prior to weighing. Room temperatures 

should not vary more than 20°+3°C during conditioning. 

The humidity, during conditioning, should be between 

30% and 70% and be controlled to a range of +10*-

(d) The filte r shall be weighed to the nearest 0.01 mg just 

prior to use. 

(e) Monitor the AP across the filte r during sampling. A 

AP greater than 13 cmHg is excessive and the termination of the test should be considered. In no case 

should the AP exceed 25 cmHg. 

(f) I t is preferable to collec t as large a sample in as 

short a time as possible. Sample times of 20 - 40 

minutes are common. The mass per filte r as listed are 

minimums desired: 

Filter Size, Minimum Mass 

mm mg 

47 1 

70 2 

90 3.5 

102 5 

At light particulate loads longer collection times may 

be required. 
•23-

(g) Filte r face velocities of 10-20 cm per second are 

suggested. 

(h) Two or three blank filter s from each lot of filter s 

should be chosen at random and then subjected to the 

SOF and sulfate analyses described in Section III . 

Normally the values for SOF on these blanks wil l be 

less than 0.1 mg and sulfates should be below detection 

limits. 

( i) Two blank filter s from each lot of filter s should be 

selected at random and stored in the constant temperature and humidity chamber or room. Each day the 

filter s can be weighed and recorded. Additional 

filter s loaded with standard particulate (NBS STD 

Reference Material 1650) or sulfuric add may be more 

sensitive to changes. The weight can be compared to 

previous day's values and can serve as an Indicator of 

the performance of the environmental chamber. 

B. MEASUREMENT OF TOTAL PARTICULATE MASS 

1. After the sample is obtained as per Section III.1A-4 , 

place the filte r in the constant temperature and humidity 

chamber or room and allow to equilibrate 1n the dark or 

under yellow light for 16-24 hours. As an option, see the 

following section (III.1B-2). 

2. Place the filte r 1n a vacuum desiccator saturated with NH40H 

for one hour. As an option, evacuate the desiccator with a 

water aspirator or vacuum pump (approximately 20 mm vacuum) 

and allow ammonia gas to leak into the desiccator until i t 

1s at atmospheric pressure. Let the filte r si t 1n the 

ammonia atmosphere for 15 minutes, and displace the ammonia 

from the desiccator with air. Place the filte r 1n the 

constant humidity chamber for 16-24 hours. 

If the NHJOH procedure 1s used, the combined water correction 1n Section III.2-E does not apply since water is 

removed during the equilibration. This method Is useful 1n 

handling samples containing high sulfate levels. It 

stabilizes the weight at about one mole of water per mole of 

sulfate*15*. 

3. Remove the filters from their containers with filter forceps 

and weigh to the nearest 0.01 mg. Record the weight. 
P.18 

SECTIO N III. 2 

SOLVENT EXTRACTABLE FRACTIONS (SEF) 
•27-

III.2 SOLVENT EXTRACTABLE FRACTIONS (SEF) 

A. SCOPE 

The methods of collection of a particulate sample from light duty 

diesel vehicles and heavy duty diesel engines are specified by 

USEPA procedures*8'. These samples are collected directly from 

diesel exhaust or from dilution tunnels using specific engine 

operating and sampling procedures and are composed of various 

subfractlons described 1n the next section. These fractions are 

of Interest 1n evaluating health effects Issues and effects of 

engineering or technology changes in the engine systems. Particulate samples can be obtained at transient or steady-state 

conditions depending on the purpose of the test. 

SEF fractions discussed include, SOF, TOE, Sulfate and Combined 

Water. 

B. EXTRACTION OF SOF - This fraction 1s obtained by extraction with 

dichloromethane (see note on page 17). 

1. Procedure 

(a) Carry out all operations under "yellow light" and minimize exposure to sunlight. Otherwise, keep the samples 

In the dark. 

(b) Fold the filter 1n halves, and finally quarters and/or 

eights with the particulate to the Inside. Bend over 

the top of the filte r before the final fold. Use the 

filter forceps and wear gloves and avoid tearing fibers 

from the filter. Carry out the filte r folding on a 

clean, plate glass surface. Place the filter s in the 

Soxhlet extractor with the point of the fold towards 

the base of the extractor. 

(c) Assemble the Soxhlet apparatus as Illustrated in 

Figure 7. For 47 mm filter s a micro Soxhlet unit 

equipped with a 30 ml flat bottomed flask 1s used. Put 

20 ml of dichloromethane In the flask. Wrap the flask 

and Soxhlet unit in aluminum foil . Start the water 

flowing through the condenser. Larger flasks and condensers may be required for large filters. 
P.20 

•29-

NOTE: The toluene:ethanol solvent may also remove some of the 

sulfates. Therefore, sulfate analysis should not be conducted on 

filters after a TOE extraction. 

D. DETERMINATION OF SULFATE 

The filte r from the SOF determination may be used to determine 

sulfates or a separate filte r may be used. If enough TPM has 

been collected for an accurate analysis, sufficient sulfate will 

have been collected. Basically, the procedure Involves extraction of the water soluble sulfates and analysis by ion chromatography. Instrument operating procedures are supplied by the 

manufacturer(17). 

1. Place the filte r 1n a 4 oz. Teflon capped bottle and add 20 

ml of an aqueous solution that 1s 0.003 molar 1n NaHCOo and 

0.0024 molar 1n Na2C03. Distilled, deionized water should 

be used. Some older units using older column technology may 

use a 60/40 Isopropyl alcohol/water solution for extraction. 

2. Shake bottles vigorously or place the bottles in a sonication bath with l-1nch of water in the bottom. Shake or 

sonicate until the filter breaks up or for 10 minutes. 

3. Filter the sample through a 0.45 vm filter on a Luer fitted 

syringe filter. 

4. Calibrate an 1on chromatograph with a standard potassium 

sulfate solution to obtain a calibration curve. 

5. Inject the filter sample directly Into the 1on chromatograph 

or dilute with the eluent if sulfate concentrations are too 

high. 

6.. Compare the response from the filter sample to the calibration curve. 

7. Calculate the mg of sulfate on the filte r and relate the 

results to the volume of exhaust sampled. 
P.21 

SECTIO N I V 

NITRATED POLYCYCLIC AROMATIC HYDROCARBONS 
P.22 

-33-

IV. NITRATED POLYCYCLIC AROMATIC HYDROCARBONS 

A. SCOPE 

This procedure defines the system and analytical method for the 

measurement of nitrated polycyclic aromatic hydrocarbons in 

diesel particulate samples by their conversion to amines and 

detection by fluorescence. 

B. BACKGROUND 

The determination of N02PAH compounds described in this procedure 

is based on an analytical method developed by S. Tejada at the 

U.S. EPA-RTP laboratory'21. The method was Initially developed 

for the analysis of 1-nitropyrene (1-NOP) and is currently undergoing cooperative round-robin testing. The method 1s an HPLC 

method and uses a Pt/Rh catalyst to convert the N02PAH to amines 

on column. The amines are then analyzed using fluorescence 

detection. 

C. DEFINITIONS/ABBREVIATIONS 

N02PAH — Nitrated Polycyclic Aromatic Hydrocarbons 

1-NOP — l-N1tropyrene 

HPLC — High Performance Liquid Chromatograph 

Pt/Rh — Platinum/Rhodium 

D. EXPERIMENTAL EQUIPMENT 

HPLC — Various units are available. 

Columns — 1. Pt/Rh catalyst coated on 5 inn Sperlsorb 5AY 

alumina.* (Available from Johnson-Mattey, Inc., 

Malvern, PA). Two columns (typical 5 cm x 4.6 mm) 

are required. 

* CAUTION - Methanol must be flushed from the column with water every 

time an end fitting 1s removed. (The catalyst has been observed to 

flame spontaneously when allowed to dry in air after use of an MeOH 

rich aqueous solution.) 
P.23 

-35-

4. The flow rate of the mobile phase is set at 1 ml/min with 

the first solvent (77% MeOH/23% water). 

5. Following the elution of the desired reduced N02PAH, e.g., 

1-NOP elutes at about 30 minutes after injection, the solvent flow is changed to pump #2 and 100% MeOH is used to 

clean the columns. After about 20 minutes, the column flow 

is switched back to pump #1 and the columns are equilibrated 

for 20 minutes prior to injection of the next sample. (See 

caution in Section IV-D.) 

6. Samples are quantified against standards obtained from the 

National Bureau of Standards(20). The NBS standards can 

also be incorporated as internal standards. 

G. PRECISION AND ACCURACY (Round-Robin in Progress) 

Estimated +20% or better. 

H. ALTERNATE COLUMN CONFIGURATION 

If Interference or peak separation problems do occur, the column 

system can be modified as described by Tejada(21 \ Tejada's 

column system 1s more complicated but does eliminate the overlap 

of some peaks. 

I. ALTERNATE METHOD 

A gas chromatography (GC) method utilizing an election capture 

detector (ECD) was developed by A.D. Little, Inc. under contract 

for CRC-APRAC CAPE-30. The method 1s reported in the 

I1terature(22). The method has been used by some CCP members. 

However, they did replace the splitless Injection of the sample 

with on-column Injection of the sample to obtain better results. 
P.24 

SECTIO N V 

POLYCYCLIC AROMATIC HYDROCARBONS 
P.25 

•39-

V. POLYCYCLIC AROMATIC HYDROCARBONS 

A. SCOPE 

This procedure defines the system and analytical method for the 

measurement of polycyclic aromatic hydrocarbons in diesel particulate samples by high performance liquid chromatography. 

B. BACKGROUND 

The analysis of PAH compounds 1n diesel exhaust and N02PAH compounds have received considerable attention relativ e to the 

health issues associated with diesel particulate. Two methods 

are discussed in thi s section. The firs t method evolved as a 

result of cooperative efforts of the CCP. However, the method is 

manpower intensive , requiring some 10-12 HPLC runs to analyze 

each sample. The method uses a normal phase chromatographic 

extrac t cleanup followed by an analysis using reverse phase 

columns and fluorescence detection for quantification. The compounds analyzed are found in Table IV. The method development 

was conducted under a CRC contract* 2 3 ' and a round-robin was 

conducted by the CCP(24). The second method requires only a 

silic a gel cleanup followed by an HPLC separation. I t is 

adequate for routine screening of samples but 1s not as comprehensive as Method No. 1. 

C. DEFINITIONS/ABBREVIATIONS 

CCP - Chemical Characterization Panel, CRC-APRAC CAPI-1-64 

In-House Group 

PAH - Polycyclic aromatic hydrocarbons-fused ring aromatic 

compounds 

N02PAH - Nitrated PAH 

ACN - Acetonitrile 

v/v - Volume/volume 

SOF - Dichloromethane extractable fraction 

TOE - Toluene:ethanol extractable fraction 

VOC - Vapor phase organic compounds 

HPLC - High performance liqui d chromatograph 
P.26 

•41-

PROCEDURES 

The analysi s of PAH compounds in diesel particulat e usuall y 

involves an extraction to remove the organic fraction containing 

the PAH's from the particulate or collection media. The extract 

undergoes a separation procedure to isolat e the PAH compounds. 

This i s followed by chromatographic analysi s of the PAH. The 

large number of compounds in diesel extracts make the precise and 

accurate analysi s of the PAH difficul t unless there i s prio r 

cleanup of the sample. 

Two methods are described in thi s report. Method 1 involves a 

more complicated preparation and analysi s of the PAH by rin g 

size. The fractions are analyzed by HPLC using specific conditions for each fraction. Considerable time and effort are 

required to analyze PAH by this method but excellent accuracy and 

precision are obtained. 

The second method, Method 2, requires less effort. I t involves 

cleanup of the sample using open column silica gel chromatography 

prior to analysis by reverse phase HPLC. 
P.27 

-43 -

METHOD 1 

This method was developed by the CRC(23) and evaluated in a roundrobin by the CCP(24)

. The method may be further modified as a result 

of a second round-robin currently in progress, but major changes are 

unlikely. A schematic summarizing the method is found in Figure 10. 

This method is applicable to SOF, TOE, and VOC samples. 

Step 1 - Extract Preparation 

§ Following the extraction of the SOF, TOE, or VOC, the extracts 

are reduced 1n volume to about 1 ml by evaporation of the solvent 

using a gentle stream of clean nitrogen in the Kuderna-Danish 

evaporator at 65°C. The solvent level is adjusted, i f required, 

to about 1 ml just prior to making a solvent exchange to heptane. 

(Isooctane can be used i f extract solubilit y problems are observed.) In the case of TOE fractions , a solvent exchange to 

dichloromethane must be conducted prior to the heptane exchange. 

• In the heptane exchange add 2 ml of heptane and evaporate the 

sample under a gentle stream of nitrogen to less than 0.5 ml. 

Add an additional 3 ml of heptane and concentrate the sample to a 

final volume of exactly 1 ml. The extract 1n heptane (50 pl) is 

fractionated using the normal phase Bondapak column. (Note: 

Small extract samples, less than 4 mg, or samples containing low 

PAH concentrations may require further concentrating of the heptane solution to obtain adequate samples from the fractionation 

in the next step. This can better be evaluated after conducting 

Step 2.) 

Step 2 - Extract Fractionation 

t The heptane solution (50 ul) 1s fractionated using normal phase 

HPLC on the 7.5 mm x 30 cm Bondapak amlnosllane column. The 

eluant 1s hexane at a flow rate of 3 ml/min. The eluting compounds are monitored using a UV detector at a wavelength of 254 

nm. Three fractions are collected as they elute from the column 

at approximately 0-11 minutes, 11-21 minutes and 21-40 minutes. 

A typical separation 1s found 1n Figure 11. Fraction 1 contains 

PAH with 3 rings or less, Fraction 2 contains 4-ring compounds, 

and Fraction 3 contains the PAH with 5 or more rings. The collection times may vary slightl y depending on experimental 

factors, e.g., column differences, sample composition, etc. I t 

1s Important, therefore, to periodically run a PAH standard. The 

actual collection times can be determined by analyzing a standard 

solution containing both Fluor and Chrysene. The collection time 

for Fraction 2 begins just prior to the elution of Fluor and ends 

jus t after the elution of Chrysene. The NBS standard125 ' contains both fluoranthene and chrysene, and can be used to establish actual collection times. 
P.28 

•44-

METHOD 1 - (Continued) 

The flow rates should be checked volumetrically to accurately 

establish the retention volume in the HPLC system from the UV 

detector to the collection point. 

Following the completion of the fractionation of the extract, the 

column should be backflushed. Use a linear gradient to 100% 

dichloromethane at a rate of 10% per minute. This is followed by 

a reverse gradient back to 100% hexane and equilibration of the 

column for 15 minutes at 100% hexane. 

Periodic regeneration of the column may be required if significant changes in the profile or retention times occur. Flushing 

the column for one hour with THF may be required*26'

27'. 

Step 3 - Fraction Analysis 

• The three fractions collected are concentrated to 5-8 ml at 

ambient temperature using a gentle stream of clean nitrogen. The 

samples are then transferred to 10 ml concentrator tubes and 

reduced in volume to 2 ml. A volume of 1 ml ACN is added along 

with sufficient dichloromethane to make a miscible solution. The 

solution is reduced in volume to about 0.5 ml under nitrogen. 

ACN is added to Fractions 1 and 2 to bring their volume to 

exactly 2.0 ml. Fraction 3 should be at exactly 0.5 ml. (Note: 

If samples are not used immediately, some readjustment of volumes 

with ACN may be required.) 

t The three fractions are analyzed using single column reverse 

phase HPLC and selective fluorescence detection. The specific 

conditions are described in Tables V, VI, and VII. 

• All three separations of the fractions are performed on the 4.6 

mm x 25 cm Vydac five-micron reverse-phase column using an 

acetonitrile/water gradient. Initial conditions are reestablished in each case by using a reverse phase gradient of 5% per 

minute until the initial conditions are attained. The system is 

allowed to reequilibrate for 15 minutes prior to the injection of 

the next sample. 

Additional details on the method can be found in References 23, 24, 

and 26. 
P.29 

-45-

METHOD 2 

This method was one utilized by two of the participants of the CCP and 

was submitted to the group for consideration in 1980(28). The method, 

which is summarized in Figure 12, was used in the round-robin and has 

been compared between laboratories. Typical traces are shown on 

Figure 13. Similar procedures have been used by panel members*5'

29'. 

Repeatability within the laboratory is normally around 10% r.s.d. I t 

is presented as a convenient method for analyzing large numbers of 

samples and can be automated for routine analysis. The separation of 

certain compounds such as BaA and BeP is not clean but the procedure 

is useful in screening samples. Method 1 is the preferred method when 

precision and accuracy of al l 11 compounds are of concern. 

Step 1 - Extract Preparation 

o The extract from the SOF or VOC are transferred to a KhudernaDanlsh evaporative concentrator flask along with rinsings of the 

flasks. The Kuderna-Danlsh units along with the M1cro-Snyder 

condensor are placed In an aluminum heater block at 40-45°C. The 

samples are blown down to a 1 ml volume with a gentle stream of 

clean, dry nitrogen. (TOE extracts have to undergo a solvent 

exchange to dichloromethane prior to undergoing Step 1.) The 

samples can be stored 1n 1-ml vials 1n the freezer until sufficient samples are obtained. (Six samples can be processed 

manually in an eight-hour day, or some two dozen can be run on an 

automatic processer overnite.) 

o The extracts are fractionated by open silica gel column chromatography to remove Interferin g oxidized material. The s11 lea 

gel columns can be purchased or prepared as follows: A small 

amount of Ottawa (white) sand Is placed In the bottom of an open 

1-cm column and 10 cc of chromatographic grade silica gel (60-230 

mesh) added. The gel should be activated overnite at 175°C. The 

column should be lightl y tapped to insure a reasonably tigh t 

column bed and a small amount of Ottawa sand 1s placed on top of 

the column. The column 1s rinsed with 25 ml of hexane. I f the 

columns are not going to be used Immediately, they should be kept 

wet by closing off the stopcock of the chromatographic column as 

the hexane level just reaches the top of the sand layer on top of 

the column. 

o To clean up the extract, each 1-ml sample Is placed on it s own 

column by pipette transfer and two rinsings of the Kuderna-Danlsh 

evaporator tube or vial are also added to the column. As the 

extract enters the column, i t is followed by 25 ml of hexane and 

then by 25 ml of dichloromethane. ALL eluting liquids are collected in one 100 ml beaker. When the last of the solvents has 

drained, about half of the mixture 1s poured into the evaporator 

tube and concentrated down to 2-3 ml. The remainder of the 

mixture along with two dichloromethane rinsings of the beaker are 
P.30 

-46-

METHOD 2 - (Continued) 

added and blown down under nitrogen at 40-45°C. If the original 

extract weight was greater than 4 mg, the samples are blown down 

to a final volume of 1 ml. If the extract weight was less than 4 

mg, the final volume is 0.5 ml. 

Next, transfer the sample to a 10-ml volumetric flask along with 

two washings of the sample container. Blow down with nitrogen to 

1 ml (>4 mg sample) or 0.2-0.3 ml (<4 mg sample) and bring the 

volume up to 10 ml with acetonitrile. Sample is transferred to a 

10 ml vial and stored in the freezer until ready for HPLC 

analysis. 

Step 2 - Sample Analysis 

• A Zorbax ODS column (4.6 mm x 25 cm) is used for the analysis. A 

short (4.6 mm x 5 cm) micro-Porasil C-18 column is used as a 

precolumn to prolong the lif e of the Zorbax column. 

t The NBS PAH standard' 2 1 ' is used to calibrate the system. One 

millilite r of the NBS standard is diluted to 10 mis with acetonitrile . This is further diluted 1:10 with acetonitrile. Ten 

microliter s of thi s solution is injected for the calibration. 

The HPLC operating conditions are: 

Flow = 1.0 ml/min. 

Solvent = ACN/Water. Initia l % ACN = 74% 

Final % ACN = 86% 

Time = 40 minutes 

(Waters 660 programer solvent Curve #7) 

Column = Zorbax ODS, 4.6 mm x 25 cm with C18 

u-Porasil precolumn 

Fluorescence Detector Conditions = Ex/Em 280/389 

t Solvents are degassed for at least one hour at approximately 150 

mm Hg vacuum. The solvents are maintained under a helium blanket 

during use by bubbling helium through the solvents at a slow 

rate. 

• Injection volumes of samples vary depending on the fina l total 

volume of sample and the milligrams of extract in the sample. A 

20-yl injection is a good starting point. 

• The solvent program is run in 40 minutes and then the fina l 

condition is held for 20 minutes. This is followed by a rapid 

return to the original conditions and a 10-minute equilibrium 

delay before injectin g the next sample. Total run time from 

injection to injection is 70 minutes. 
P.31 

REFERENCE S 
P.32 

-49-

REFERENCES 

1. Perez , J.M., Lipari, F. and Seizinger, D.E., "Cooperative Development of Analytical Methods for Diesel Emissions and Particulates, " SAE Paper No. 840413, Detroit, MI, March 1984. 

2. Tejada, S., CRC-APRAC CAPI-1-64 Chemical Characterizatio n of 

Diesel Exhaust Emissions Workshop II , Dearborn, MI, March 1985. 

3. Tejada, S.B., "Evaluation i f Silica Gel Cartridges Coated In Situ 

with Acidified 2,4-D1nitrophenylhydrazine for Sampling Aldehydes 

and Ketones 1n A1r," Intern . J. Environ., Anal. Chem., Vol. 26, 

pp. 167-185, 1986. 

4. Perez , J.M., "Chemical Characterizatio n of Diesel Exhaust - A 

Report of Cooperative Studies of Analytical Methods," Preprin t 

No. 83-48.5, 76th Annual APCA Meeting, Atlanta, GA, June 1983. 

5. Wall, J.C. and Hoekman, SX , "Fuel Composition Effects on HeavyDuty Diesel Particulat e Emissions," SAE Paper No. 841364, 

Baltimore, MD, October 1984. 

6. "Methylene Chloride Facing Regulation," Chemical and Engineering 

News, p. 17, March 17, 1986. 

7. SAE Recommended Practices, Society of Automotive Engineers, 400 

Commonwealth Drive, Warrendale, PA 15096. 

8. US EPA Heavy-Duty Diesel Transient Test, Code of Federal Regulations, Titl e 40, Part 86, Subpart N. 

9. Lipkea, W.H., Johnson, J.H., and Vuk, C.T., "The Physical and 

Chemical Character of Diesel Particulate Emissions - Measurement 

Techniques and Fundamental Considerations," SAE Paper No. 78018 

(SP-430), 1978. 

10. MacDonald, J.S., Plee, S.L., D'Arcy, J.B., and Schrek, R.M., 

"Experimental Measurements of the Independent Effects of Dilution 

Ratio and Filte r Temperature on Diesel Exhaust Particulat e 

Samples," SAE Paper No. 800185, Detroit, MI, February 1980. 

u 

11. Eisenberg, W.J., "Development of a Method for the Analysis of 

Polycyclic Aromatic Hydrocarbons 1n Diesel Exhaust Emissions, 

Final Report by I IT Research Institute to the Coordinating 

Research Council, Inc., Atlanta, GA, October 1983. 

12. Williams, R.L., Perez , J.M., and Griffing , M.E., "A Review of 

Sampling Condition Effects on Polynuclear Aromatic Hydrocarbons 

(PNA) from Heavy-Duty Diesel Engines," SAE Paper No. 852081, 

Tulsa, OK, October 1985. 
P.33 

-50-

REFERENCES - (Continued) 

13. Coordinating Research Council, Inc., "Informational Report on the 

Measurement of Diesel Particulate Emissions," CRC Report No. 522, 

June 1982. 

14. MacDonald, J.S., Barsic, N.J., Gross, G.P., Shahed, S.M., and 

Johnson, J.H., "Status of Diesel Particulate Measurement 

Methods," SAE Paper No. 840345, Detroit, MI, March 1984. 

15. Seizinger, D.E., Private Communication to the Chemical Characterization Panel of the CRC-APRAC Program Group on Composition of 

Diesel Exhaust, 1981. 

16. Shimpl, S.A. and Yu, M., "Determination of a Reliable and Efficient Diesel Particulate Hydrocarbon Extraction Process," SAE 

Paper No. 811183, Tulsa, OK, October 1981. 

17. DIONEX" Models 10/14 Ion Chromatograph Operation and Maintenance 

Manuel, DIONEX Corporation, Sunnyvale, CA, January 1979. 

18. Schuetzle, D. and Perez, J.M., "A CRC Cooperative Comparison of 

Extraction and HPLC Techniques for Diesel Particulate Emissions," 

Preprint No. 81-56.4, 74th Annual APCA Meeting, Philadelphia, PA, 

June 1982. 

19. Hare, C.T., "Supplemental Study of Fuel Property and Injection 

and Combustion System Type Effects on Emissions from Light-Duty 

Diesels," Final Report by Southwest Research Institute to CRCAPRAC CAPE-32-80 Project Group, Coordinating Research Council, 

Inc., Atlanta, GA, November 1985. 

20. Standard Materials, SRM #1650, National Bureau of Standards, 

Washington, D. C. 

21. Tejada, S.B., Zweidinger, R.B., and Sigsby, J.E., Jr., "Fluorescence Detector and Identification of N1tro-Der1vat1ves of Polynuclear Aromatic Hydrocarbons by On-Column Catalytic Reduction to 

Aromatic Amines," Anal. Chem., pp. 1827-1834, July 1986. 

22. Menzles, K.T., et al , "Development of a Routine Technique for 

Analysis of a Nitro-PAH In Diesel Particulate Extract," Final 

Report by A. D. Little , Inc. to CRC-APRAC CAPE-30-81 Project 

Group, Coordinating Research Council, Inc., Atlanta, Georgia, 

February 1985. 

23. Elsenberg, W.C, "Development of a Method for the Analysis of 

Polycyclic Aromatic Hydrocarbons In Diesel Exhaust Emissions," 

Final Report by IIT Research Institute to CRC-APRAC CAPE-30-81 

Project Group, Coordinating Research Council, Inc., Atlanta, GA, 

October 1983. 
P.34 

-51-

REFERENCES - (Continued) 

24. Eisenberg, W.C, Schuetzle, D., and Williams, R.L., "Cooperative 

Evaluation of Methods for the Analysis of PAH in Extracts from 

Diesel Particulate Emissions," SAE Paper No. 840414, Detroit, MI, 

March 1984. 

25. Standard Materials, Priority Pollutant PAH, SRM #1647, National 

Bureau of Standards, Washington, D. C. 

26. Eisenberg, W.C. and Cunningham, D.L.B., "Analysis of PAH in 

Diesel Emissions Using HPLC: A Method Development Study," 

Presented at the Eighth International Symposium on Polynuclear 

Aromatic Hydrocarbons, Columbus, OH, October 1983. 

27. Warner, I., Karlesky, D., and Skelly, D.C, "Reactivation of an 

Amino Bonded Phase Liquid Chromatographic Column," Anal. Chem., 

53:2146, 1981. 

28. Coordinating Research Council, Inc., "Informational Report on the 

Measurement and Characterization of Diesel Exhaust Emissions," 

CRC Report No. 517, December 1980. 

29. Swarin, S.J. and Williams, R.L., "Polynuclear Aromatic Hydrocarbons: Chemistry and Biological Effects," Bjorseth, A. and 

Dennis, A.J., editors, Battelle Press, Columbus, Ohio, 1980. 
P.35 

TABLE S 

AND 

FIGURE S 
TABLE I 

PHYSICAL PROPERTIES OF CARBONYL COMPOUNDS 

DNPH Derivative 

Aldehyde or Ketone 

Formaldehyde 

Acetaldehyde 

Acetone 

Propionaldehyde 

Acrolein 

n-Butyraldehyde 

Isobutyraldehyde 

Methylethyl ketone 

Crotonaldehyde 

n-Valeraldehyde 

I so-Valeraldehyde 

Caproaldehyde 

Enanthaldehyde 

Benzaldehyde 

p-Tolualdehyde 

IUC Name 

Methanal 

Ethanal 

2-Propanone 

Propanal 

Propenal 

Butanal 

2-Methylpropanal 

2-Butanone 

trans-2-Butenal 

Pentanal 

3-Methylbutanal 

Hexanai 

Heptanal 

Benzenecarbonal 

4-Methylbenzenecarbonal 

Chemical 

Formula 

CH20 

CH3CH0 

CH3COCH3 

CH3CH2CH0 

CH2:CHCH0 

CH3(CH2)2CHO 

CH3CH(CH3)CHO 

CH3C0CH2CH3 

CH3CH:CHCH0 

CH3(CH)3CH0 

(CH3)2CHCH2CH0 

CH3(CH2)4CH0 

CH3(CH2)5CH0 

C6H5CH0 

CH3C6H4CH0 

Molecular 

Weight 

30.03 

44.05 

58.08 

58.08 

56.07 

72.10 

72.11 

72.11 

70.09 

86.13 

86.13 

100.16 

114.18 

106.13 

120.14 

Melting 

Pt., °C 

-92 

-121 

-95 

-81 

-87.7 

-99 

-65.9 

-84 

-74 

-92 

-51 

-56 

-45 

-26 

— 

Mol. Wt. 

210.15 

224.18 

238.21 

238.21 

236.20 

252.23 

252.23 

252.23 

250.21 

266.26 

266.26 

279.28 

294.31 

286.25 

300.27 

Melting 

Pt., °C 

167 

167 

128 

156 

165 

122 

182 

— 

190 

106 

— 

104 

— 

237 

194 

Density 

0.815 

0.783 

0.792 

0.807 

0.841 

0.817 

0.794 

0.805 

0.858 

0.820 

0.805 

0.833 

0.850 

1.05 

1.02 

Rel Ret 

Time 

0.453 

0.574 

0.714 

0.795 

0.699 , 

Ol 

__ 1 

0.911 

— 

0.870 

— 

— 

1.23 

— 

1.00 

— 
TABLE II 

STORAGE STABILITY 

(ng/yL) 

SAMPLE NO. 

Formaldehyde 

Acetaldehyde 

Acrolein 

Acetone 

Propionald. 

Crotonald. 

Isobutryald. 

Benzald. 

Hexanai 

ORIG. 

23.0 

3.98 

1.03 

0.85 

0.86 

0.52 

0.58 

0.31 

0.19 

1 

A 

22.0 

3.82 

0.16 

0.83 

0.82 

0.42 

0.58 

0.29 

0.14 

A = 

B 

22.7 

3.97 

0.21 

0.89 

0.84 

0.47 

0.58 

0.31 

0.10 

6 weeks 

ORIG. 

23.6 

4.16 

0.94 

0.90 

0.91 

0.52 

0.61 

0.32 

0.15 

B = 

2 

A 

22.0 

3.86 

0.17 

0.89 

0.83 

0.32 

0.85 

0.27 

0.12 

2 months 

B 

22.3 

3.81 

0.20 

0.87 

0.84 

0.30 

0.74 

0.30 

0.10 

ND 

ORIG. 

14.1 

2.30 

0.77 

0.56 

0.47 

0.42 

0.34 

0.09 

0.07 

= Not 

3 

A 

13.5 

2.08 

O.ll 

0.56 

0.43 

0.14 

0.42 

ND 

ND 

Detected 

B 

13.8 

2.06 

0.14 

0.57 

0.47 

0.12 

0.35 

ND 

ND 

ORIG. 

14.1 

2.24 

0.67 

0.54 

0.47 

0.30 

0.32 

ND 

ND 

4 

A 

13.3 

2.01 

0.12 

0.53 

0.44 

0.14 

0.33 

ND 

ND 

B 

13.5 

1.96 

0.09 

0.52 

0.45 

0.12 

0.42 

ND 

ND 

I 

in 

i 
P.38 

•57-

TABLE III 

COMPARISON OF SOF AND VACUUM REMOVAL OF ORGANIC FRACTION 

Sample 

Size, mg 

9.90 

10.2 

7.77 

7.77 

SOF, 

W t. % 

36.4 

43.4 

— 

46.1 

Vacuum Temperatu 

200°F 275°F 

re* 

350°F 

Wt. Loss, % 

38.6 

--

22.0 

48.6 

41.4 

48.2 

39.5 

"" 

44.0 

49.2 

— 

51.0 

* All samples were under 15" vacuum (A15" Hg) for 6 hours. 
•58-

P.39 

TABLE IV 

HUJTUIU^ ^ 

P2tAJQNDAPAjU!H^ ^ 

Fluorene 

Anthracene 

Phenanthracene 

Fluoranthene 

Pyrene 

BenzU.anthracene 

Chrysene 

Benzo[k]fluoranthene 

Benz[a]pyrene 

Benzo[b]fluoranthene 

Benzo[g,h,i]perylene 

Elution 

Time, min. 

6.87 

9.36 

9.72 

12.45 

12.27 

17.61 

17.85 

21.45 

22.41 

23.04 

31.26 

Relative 

Elution 

TimeiJliOj 

0.55 

0.75 

0.78 

1.00 

0.98 

1.41 

1.43 

1.70 

1.80 

1.85 

2.51 

Fraction 

Collection 

Times 

Fraction 1 

0-11 min. 

Fraction 2 

11-21 min. 

Fraction 3 

21-40 min. 
P.40 

-59-

TABLE V 

HIGH PERFORMANCE LIQUID CHROMATOGRAPHY 

OF POLYCYCLIC AROMATIC HYDROCARBONS: FRACTION l 1 

PAH 

Fluorene 

Phenanthrene 

Anthracene 

Retention 

Time, min. 

16.50 

18.85 

20.92 

Fluorescence 

Wavelength, nm 

Excitation 

260 

260 

260 

Emission0 

370 

370 

370 

Detection 

Limit, ng2 

3.67 

0.93 

0.50 

1. HPLC elution conditions: 4.6 mm x 25.0 cm Vydac 5 micron 

reverse-phase column; linear gradient, acetonitrile/water (50/50, 

V/V) to acetonitrile/water (70/30, V/V) in 30 min. then to 100% 

acetonitrile In 10 min., followed by 10 min. at 100% acetonitrile . The system was returned to initia l conditions using a 

reverse gradient of 5%/min. for 10 min. and equilibrated at 

initia l conditions for 15 min.; flow: 1.0 ml/min., detector 

range: 1.0 (minimum range: 0.01); injection volume: 15 ul. 

2. The detection Umi t was defined as the on-column amount of 

compound that would give a peak that was 5 times the baseline 

noise under these instrumental conditions. 

3. A detector with a 370 nm emission cut-off filte r was used. 
P.41 

-60-

TABLE VI 

HIGH PERFORMANCE LIQUID CHROMATOGRAPHY 

OF POLYCYCLIC AROMATIC HYDROCARBONS: FRACTION 2 1 

Fluorescence 

Retention Wavelength, nm „ Detection, 

PAH 

Fluoranthene 

Pyrene 

Benz[a]anthracene 

Chrysene 

Time, min. 

10.50 

11.97 

17.40 

18.72 

Excitation 

260 

260 

280 

260 

EmissionJ 

370 

370 

389 

370 

Limit, ng 

1.22 

2.26 

0.51 

0.59 

1. HPLC elution conditions: 4.6 mm x 25.0 cm Vydac 5 micron 

reverse-phase column; isocratic elution, acetonitrile/water 

(70/30, V/V) for 15 min., then a linear gradient to 100% acetonitrile in 15 m1n., followed by 15 min. at 100% acetonitrile. The 

system Was returned to initia l conditions using a 10 min. 

gradient followed by an equilibration period of 15 min.; flow: 

1.0 ml/m1n.; detector range: 1.0 (minimum range: 0.01); injection volume: 30 yl. 

2. The'detection limit was defined as the on-column amount of 

compound that would give a peak that was 5 times the baseline 

noise under these instrumental conditions. 

3. A detector with a 370 and 389 nm emission cut-off filter was used 

in these analyses. 
P.42 

•61-

TABLE VII 

HIGH PERFORMANCE LIQUID CHROMATOGRAPHY 

OF POLYCYCLIC AROMATIC HYDROCARBONS: FRACTION 31 

Fluorescence 

Retention Wavelength, nm Detection, 

PAH 

Benzo[b]fluoranthene 

Benzo[k]fluoranthene 

Benzo[a]pyrene 

Benzo[g,h,i]perylene 

Time, min. 

32.34 

34.11 

35.73 

40.82 

Excitation 

280 

280 

280 

280 

Emission*1 

389 

389 

389 

389 

Limit, ng 

0.29 

0.23 

0.29 

1.71 

1. HPLC elution conditions: 4.6 mm x 25.0 cm Vydac 5 micron 

reverse-phase column; linear gradient, acetonitrile/water (50/50, 

V/V) to acetonitrile in 40 min., followed by 20 min. at 100% 

acetonitrile. The system was returned to initial conditions for 

15 m1n.; flow: 1.0 ml/min.; detector range: 1.0 (minimum range: 

0.01); injection volume: 30 pl. 

2. The detection limit was defined as the on-column amount of 

compound that would give a peak that was 5 times the baseline 

noise under these instrumental conditions. 

3. A detector with a 389 nm emission cut-off filter was used. 
P.43 

-62-

FIGURE 1 

SAMPLING CART 

(SCHEMATIC DIAGRAM) 

FILTERS 

(AS REQUIRED) 

.SOLENOIO 

ON-OFF 

DRY-GAS 

METER 

TEMP READO 

hBm-ji fflbr*-c GAS VOLUME 

EMERGENCY POWER 
P.44 

•63 -

FIGURE 2 

CARBONYL ANALYSIS METHOO 

PREPARATION* 

I 

SAMPUNG 

HPLC ANALYSIS" 

TIMS (MIN) 

START 

0 

a 

9 

15 

20 

30 

STOP 

8 

9 

15 

20 

30 

35 

I 

I 

FLOW RATE 

(MUMIN) 

0.7 

1.0 

1.0 

1.0 

1.0 

2.0 

1 

SOLVENT — 

GRADIENT CONC 

(VOL %) 

87ACN/33 H,0 

67ACNJ33 H,0 

G RAO IENT TO 85 ACN 

GRADIENT TO 100 ACN 

100 ACN 

REVERSE GRA0IENT 

TO 67 ACN/23 H,0 

•ONPH SOLUTIONS 

STANOAROS 

••HPLC — 4.8 mm x 25 cm column — 

5M Zorfiax OOS; 600 PSI (initial): 

UV-365 nm; sensitivity 0.2 AUFS 
P.45 

-64-

1X1 

LU££ 

cu 

T3 

CU >> 

T3 CO -C 

>,T3 OJ 

OJ 

-o £ <U 

CO 01 "O 

LL-O 

0) 0> -C' >>"0 0» T3 "O >> 0) 

"O "O Jl£i-T3 r >>-_: OJ "O OJ 

>,> > -oa><0>><U.--0J"O>," O j=_= n--o_._c-.oj-o>,.::>, i. a> c i.r- >, . 4)"Di-r ai r 

"O "O •<- QJ C « +•> "O i— r— (0 QJ-O OJ 

r— i— 0J__OC3i — <-<03"Or-" O 

(_ <_r— O •<- OJ Q <0 > i . r— r— (O r— 

X E -M O -M --.-M I N I O) O <_ +- (O 

Q. E QJ J. OJ O O O C Oi- H X Q.-M zouuui-i.intiiA a i <u a; o 

Q-.<<<Q.(J)HC.M > Q..C _c o 

1 

O 

(A 

ii 

9 

es 

a 
FIGUR

E 4 

250 FORMALDEHYD

E STABILITY 

_g> o . 

E at 

<a 

• Ui 

Q >-

I UJ 

SAMPLE 

1 

200 

150 

i c\ tn 

i 

100 

tc 

o 

50 

1 

16 

24 

HOURS 

32 

40 

48 
-fi 

a. 

E 

I 

-i 

o 

UU 

80 

eo 

40 

20 

- \ 

^ 

^ _ 

1 

FIGURE 5 

ACROLEIN STABILITY 

1 1 

^ 

1 1 

SAK 

1 

I 

cn 

• 

16 24 32 

ELAPSED TIME — HOURS 

40 48 

i-
500, 

FIGURE 6 

TOTAL ALDEHYDE STABIUTY 

Engine Operation 

Condition 

No. 1 

•2 400| 

o. 

E 

S3 

UJ o 

>-

i 

UJ 

3001 

• 

i 

2 2001 

No. 2 

100« 

16 

ELAPSED TIME 

24 

HOURS 

32 40 

fc 
P.49 

-68-

FIGURE 7 - SOXHLET 
-

* * • 

o 

CO 

C\J 

X 

o 

CVJ 

a: 

o> 

-69-

FIGURE 8 

MASS OF BOUND H-0 PER GRAM H2S04 

IN AN AQUEOUS SOLUTION VERSUS RELATIVE HUMIDITY (25°C) 

7 

6 

5 

4 

3 

2 

1 

0 

- j 

~ / 

I 

-jiT^ i . 9 

J J 

50 

1 1 1 

0 20 40 60 80 100 

Relative Humidity, % 

P.50 

-
P.51 

-70 -

FIGURE 9 

SCHEMATIC OF NITROPYRENE ANALYSIS SYSTEM 

Solvent 

Reservoir 

77: Methanol 

23Z Water .i----> 

I Flow 

1.1 mi/min 

I 

[Pump J 

Solvent 

Reservoir 

1002 Methanol 

Reduction Catalyst 

3 inch x 4.6 mn 

packed with ground 

up 3-way catalyst 

70 mesh 

For removal of oxygen 

rD 

r*. Septum-less 

N' Injector 

Analytical Column 

Zorbax ODS Column 

23 cm * 4.6 mm 

i 

Analytical Coluan 

Zorbax OOS Column 

15 cm * 4.6 mm 

Reduction Catalyst 

3 inch

 x 4,6 _a 

packed with ground-up 

3-way catalyst, 70 mesh 

Waste 

Fluorescence Detector 

360 nm excitation 

430 nm emission for 

1-nitropyrene 
P.52 

-71-

FIGURE 10 

HPLC PAH METHOD NO. 1 

FRACTION 1 

(3 OR LESS RINGS) 

DIESEL PARTICULATE 

EXTRACTION 

KUDERNA-DANISH 

(SOLVENT EXCHANGE) 

EXTRACT FRACTIONATION 

(BONDAPAK NH,)* 

FRACTION 2 

(4 RINGS) 

SOLVENT EXCHANGE AND 

SAMPLE CONCENTRATION 

FRACTION 3 

(5 OR MORE RINGS) 

VYDAC 5 pm COLUMN SEPARATION 

SEE TABLE V SEE TABLE VI SEE TABLE VII 

* See Figure 11. 
FIGURE 11 

HPLC METHOD SUMMARY - FRACTIONATION OF THE DIESEL EXTRACT 

STEP1 

FRACTION -

FRACTIONATION 

UV DETECTION 

BONDAPACK NH2 

i 

STEP 2 VYDAC 5 COLUMN — FLUORESCENCE DETECTION 

FRACTION I FLUORENE 

ANTRACENE 

PHENANTHENE 

FRACTION II FLUORANTHENE 

PYRENE 

BENZ(A)ANTHRACENE 

CHRYSENE 

FRACTION III BENZO(B)FLUORANTHENE 

BENZO(K)FLUORANTHENE 

BENZO(A)PYRENE 

BENZO(GHI)PERYLENE 

SEPARATION CHECKED 

USING NBS SRM #1647 

(16 PAH STANDARDS) 
P.54 

•73-

FIGURE 12 

HPLC PAH METHOD NO. 2 

DIESEL PARTICULATE 

EXTRACTION - MECL2 

SILICA GEL SEPARATION 

HEXANE/MECL2 

SAMPLE CONCENTRATED 

TO 1-ml (KUDERNA-DANISH 

-

) 

SOLVENT EXCHANGE (ACN) 

-

HPLC-FLUORESCENCE 

EX/EM = 280/389 

ZORBAX ODS 
P.55 

-74-

FIGURE 13 

TYPICAL HPLC TRACES - METHOO 2 

A. PAH STANDARD .01'N 

r-mr 

9. JIESEL ENGINE 

PARTICULATE 

EXTRACT <SOP> 

•~^h—^ 

W: