Technology Transition Workshop

DART Mass Spectrometry of

Writing Inks
Roger Jones
Midwest Forensics Resource Center and
Ames Laboratory—USDOE,
Iowa State University

Midwest Forensics Resource Center

 Technology Transition Workshop
DART MS of Writing Inks
• Comparison to Conventional Ink Analysis
• Spectrum Acquisition
– Presentation of Sample
– Instrumental Conditions and Data Acquisition
– NIST Mass Spectral Search Program
• Interference from Paper
• Effects of Writing Age
– Identification of Ink Components
• Ink Identification Using NIST Software
 Technology Transition Workshop
Comparison to Conventional
Methods
• Optical Inspection
– Side-by-side comparison under various
illuminations through various filters
– Strengths: Quick, cheap, non-destructive, often
sufficient
– Weaknesses: Requires physical reference,
provides minimal information, no library
possible, cannot identify ink
 Technology Transition Workshop
Comparison to Conventional
Methods
• Thin Layer Chromatography
– Side-by-side comparison, separates and
detects dyes and fluorescent components
– Strengths: Cheap, usually sufficient, more
information than optical inspection, can build
library, may identify ink
– Weaknesses: Slow, destructive, requires
physical reference, sensitive to environment,
less information than DART, requires physical
sample library
 Technology Transition Workshop
Comparison to Conventional
Methods
• DART MS
– Elemental formula determination of all volatile
and semi-volatile components
– Strengths: Quick, non-destructive, no physical
reference, provides lots of information,
database library, can identify ink,
complementary to dye-sensitive methods
– Weaknesses: Expensive equipment, sensitive
to vehicle, paper occasionally interferes
 Technology Transition Workshop
MS of Extracts vs. DART MS in Situ
DART MS of Ink on Paper ESI-MS of Extracts
139
5 Black Bic
100 Bic Wide Body 100
183
367 Ballpoints Bic Wide Body 372

50 351 50 358
374
269
0 0
100 Bic Velocity 100
Bic Velocity

Relative Intensity (m/z 372 = 100)

Relative Intensity (m/z 139 = 100)

50 50

0 0
100 Bic Round Stic 100
Bic Round Stic

50 50

0 0
100 Bic Cristal Grip 100
Bic Cristal Grip

50 50

0 0
100 Bic Atlantis 100
Bic Atlantis

50 50

0 0
100 200 300 400 500 100 200 300 400 500
m/z m/z
 Technology Transition Workshop
Presentation of Sample

Modified Wikipedia image, with acknowledgement to JEOL USA, Inc., and author R. B. Cody
Technology Transition Workshop
Full Page in Open Sample Holder
Technology Transition Workshop
Full Page in Open Sample Holder
Technology Transition Workshop
Sample Held by Cover-Plate Hook
1/8-inch hole in cover plate gives access to sample.
Technology Transition Workshop
Page in Closed Sample Holder
Technology Transition Workshop
Sample in Position for Analysis
 Technology Transition Workshop
Instrumental Conditions
Mount wired to
spectrometer
inlet

–3500 V +150 V

3.25
L/min
He

255 °C +650 V +20 V, 80 °C

Modified Wikipedia image, with acknowledgement to JEOL USA, Inc., and author R. B. Cody
 Technology Transition Workshop
Raw Spectra from Chromatogram
Zebra Rubber 80 Ballpoint on Hammermill Fore DP Paper
50000 50000
Intensity (counts)

Intensity (counts)
269
40000 40000
315
30000 30000
359
20000 279 20000 279

10000 10000

0 0
100 200 300 400 500 600 100 200 300 400 500 600

m/z m/z
3.0e+7
Total Ion Count

2.5e+7 PEG 600

2.0e+7 30 second averages

1.5e+7
1.0e+7 Ink on Paper

5.0e+6 Blank Paper

0.0
0 1 2 3 4 5

Time (min)
 Technology Transition Workshop
Final Spectrum from Raw Spectra
Zebra Rubber 80 Ballpoint on Hammermill Fore DP Paper
50000 50000
Intensity (counts)

Intensity (counts)
269
40000 40000
315
30000 30000
359
20000 279 20000 279

10000 10000

0 0
100 200 300 400 500 600 100 200 300 400 500 600

m/z m/z
50000
Intensity (counts)

40000 269
315
30000
359
20000
279
10000

0
100 200 300 400 500 600

m/z
Technology Transition Workshop
DART MS of 3 Main Ink Types
269
100 Zebra Rubber 80
Ballpoint
75
50
255
25 315 359 403
100 139
0
Relative Intensity
150
100 Bic Cristal Gel Roller
75
50
25
106 168 239
0
239
100 Mont Blanc
75
Le Grand Rollerball

50 195
121 283
403
25
0
100 200 300 400

m/z
 Technology Transition Workshop
NIST Spectral Search Software
• NIST Mass Spectral Search Program
– Demo Version 2.0d is a free download.
– Fully functional software with a small library
• Search Types
– Spectral match; reverse/forward, with m/z limits,
identity/similarity, match ring number
– Search formula, name, CAS number, NIST
registry number, peaks
– Constraints; molecular weight, elemental
abundance, peak presence
 Technology Transition Workshop
NIST Spectrum Search Results
Distribution of matches Comparing unknown and highlighted match

Unknown Spectrum of unknown

List of matches Spectrum of highlighted match

 Technology Transition Workshop
Calculating Match Quality
Match Quality, Q, combines two factors, F1 and F2,
that range from 0 to 1.
F1 is a mass-weighted dot product of the two
spectra:

F1 
 M ( A A
L U )1/ 2

 MAL  MAU  1/ 2

Sums are over M; M = m/z

AL = relative intensity of library peak
AU = relative intensity of unknown peak
From Stein, J. Am. Soc. Mass Spec., 1994, 5, 316.
 Technology Transition Workshop
Calculating Match Quality
F2 compares ratios of successive peaks:
n n
 1  NU & L
 AL ,i   AU ,i 
F2        
A  A 
 NU & L  i  2  L ,i 1   U ,i 1 
NU&L = number of peaks common to both unknown
and library spectra. Sum is over those peaks.
n = 1 if AL ratio < AU ratio, n = -1 if AU ratio < AL ratio.

Q
999
NU F1  NU & L F2 
NU  NU & L
Q is 999 for a perfect match.
 Technology Transition Workshop
Test of Paper Effects
• Lines written freehand on 16 different types of
paper with three pens—1 ballpoint, 1 gel, 1
fluid ink.
• Stored in closed file in drawer for 8–9 months.
• Three spectra acquired for each ink on each
paper. Samples handled with gloves.
• All spectra for one ink placed in a NIST library.
• All spectra searched against the library and
match qualities determined for each spectrum.
Reverse matching with no m/z limit.
 Technology Transition Workshop
Papers Used in Test
White Office Paper Envelopes
Champion Benefit Writing Brown Manila
Crane’s Resume White Ames Lab Letterhead
Great White Multipurpose 20
Hammermill Color Copy Miscellaneous Types
Hammermill Color Laser Ampad Evidence (ruled legal
Gloss pad)
Hammermill Fore DP Colored paper, goldenrod
Hammermill Great White Copy Post-It Note, yellow
HP Color Inkjet Whatman #1 filter paper
Staples Copy Notepaper with printing under
writing
 Technology Transition Workshop
Ballpoint Spectrum Match Qualities
Distribution of Match Qualities of One Spectrum with
45 Spectra of the Same Ink on Different Paper:
Black Zebra Rubber 80 Ballpoint Ink on Hammermill Fore
Percent of Ballpoint Samples

30% Median Match Quality = 880

20%

10% Hammermill Color Laser Gloss

0%
1000 900 800 700 600 500

Match Quality Index

 Technology Transition Workshop
100 Unknown Being Matched
Hammermill Fore DP #1
80

Spectra Matching 60
40
20

a Ballpoint 0
100 Best Match on Different Paper
Manila Envelope #1
80

Spectrum 60
Match Quality = 925

Relative Intensity
40
20
Spectral Matching for a sample of 0
Zebra Rubber 80 Black Ballpoint on 100 Worst Match Other Than
Hammermill Laser Gloss
80
Hammermill Fore DP Paper 60
Post-It Note #2
Match Quality = 816
40
20
0
100 Worst Match
Hammermill Laser Gloss #1
80
Match Quality = 548
60
40
20
0
100 200 300 400 500 600

m/z
 Technology Transition Workshop
Median Match Qualities for Ballpoint
Distribution of Median Match Qualities with 45 Spectra
of the Same Ink on Different Paper:
Black Zebra Rubber 80 Ballpoint Ink
Percent of Ballpoint Samples

30%

20%

10% Hammermill Color Laser Gloss

0%
1000 900 800 700 600 500

Median Match Quality Index

 Technology Transition Workshop
Gel Ink Spectrum Match Qualities
Distribution of Match Qualities of One Spectrum with
45 Spectra of the Same Ink on Different Paper:
Black Bic Cristal Gel Roller Ink on Hammermill Fore DP
Percent of Gel-Ink Samples

30% Median Match Quality = 854

20%

10%

0%
1000 900 800 700 600 500

Match Quality Index

 Technology Transition Workshop
Median Match Qualities for Gel Ink
Distribution of Median Match Qualities with 45 Spectra
of the Same Ink on Different Paper:
Black Bic Cristal Gel Roller Ink
Percent of Gel-Ink Samples

30%

20%

10%

0%
1000 900 800 700 600 500

Median Match Quality Index

 Technology Transition Workshop
Fluid Ink Spectrum Match Qualities
Distribution of Match Qualities of One Spectrum with
45 Spectra of the Same Ink on Different Paper:
Blue Mont Blanc Le Grand Roller Ink on Hammermill Fore
Percent of Fluid-Ink Samples

30% Median Match Quality = 777

20%

10%

0%
1000 900 800 700 600 500

Match Quality Index

 Technology Transition Workshop
Median Match Qualities for Fluid Ink
Distribution of Median Match Qualities with 45 Spectra
of the Same Ink on Different Paper:
Blue Mont Blanc Le Grand Rollerball Ink
Percent of Fluid-Ink Samples

30%

20%

H. Color Laser Gloss

Crane's Resume
10%

0%
1000 900 800 700 600 500

Median Match Quality Index

 Technology Transition Workshop
Effects of Writing Age
• Lines written freehand periodically on white
copy paper (Hammermill Fore DP).
• Stored in closed folder in desk drawer with
blank pages surrounding each written page.
• Samples handled with gloves during analysis.
• Three spectra at each age averaged for time
plots.
 Technology Transition Workshop
200 332 Days Old

150

Relative Intensity (m/z 367 = 100)

183 367
100
139
Black Bic 50 165
351
374
100 269
Round Stic 0
200

199
Ballpoint at 200

100
139
4 Days Old

Two Ages 150

183 367
100

351
50 269
374
165
0
100 200 300 400

m/z
 Technology Transition Workshop
Components of Bic Ballpoint
Observed Chemical Actual Most Probable
m/z Formula m/z Identity
100.076 [C5H9NO+H]+ 100.076 N-methyl-2-pyrrolidone
139.076 [C8H10O2+H]+139.076 Phenoxyethanol
165.094 [C10H12O2+H]+165.092 Various fragrant liquids
183.102 [C10H14O3+H]+ 183.102 Diethylene glycol monophenyl ether

199.142 [C11H18O3+H]+ 199.133 Various fragrant liquids

269.166 [C17H20N2O+H]+ 269.166 Michler’s ketone
351.215 uncertain ---- ----
367.210 [C20H30O6+H]+ 367.212 Dibutylglycol phthalate
374.264 [C25H30N3+2H]+ 374.260 Crystal violet
Technology Transition Workshop
Crystal Violet Cationic Dye
Technology Transition Workshop
Crystal Violet DART Spectrum
Relavtive Intensity (m/z 372 = 100) 120
[Crystal Violet] +
372.242
100

[Crystal Violet + 2H] +

80 374.259

60

[Methyl Violet] + [Crystal Violet + H2O] +

40
358.232 390.260

345.205 [Crystal Violet + O2] +

20 404.237

0
300 320 340 360 380 400 420 440

m/z
 Technology Transition Workshop
300

250 m/z 100

m/z 139
m/z 199

Relative Intensity (m/z 367 = 100)

200

Aging of 150

Black Bic 100

50
Round Stic
0
Ballpoint 150
m/z 165
m/z 183
m/z 269
m/z 351
100

50

0
0 100 200 300

Age (days)
 Technology Transition Workshop
150
100 336 Days Old

80

Relative Intensity (m/z 150 = 100)

60

Black Bic 40 151

20 148
Cristal Gel 0
106 168
195 239

150
Roller Ink at 100 9 Days Old

80
Two Ages 60 151 301

40
195
20 239
148 168
106
0
100 150 200 250 300 350 400

m/z
 Technology Transition Workshop
Components of Bic Gel Ink
Observed Chemical Actual Most Probable
m/z Formula m/z Identity
106.088 [C4H11NO2+H]+ 106.087 Diethanolamine
148.100 C7H16O3+ 148.110 Dipropylene glycol monomethyl ether
150.114 [C6H15NO3+H]+ 150.113 Triethanolamine
151.102 [C6H14O4+H]+ 151.097 Triethylene glycol
168.127 C10H16O2+ 168.115 Dipentene diepoxide
195.124 [C8H18O5+H]+ 195.123 Tetraethylene glycol
239.151 [C10H22O6+H]+ 239.149 Pentaethylene glycol
301.186 [C18H24N2O2+H]+ 301.192 ----
or [C19H24O3+H]+ 301.180
 Technology Transition Workshop
m/z 148
70
m/z 151
m/z 239
60 m/z 301

Relative Intensity (m/z 150 = 100)

50

Aging of 40

Black Bic 30

20
Cristal Gel
10

Roller Ink 30
0
m/z 106
m/z 168
20 m/z 195

10

0
0 100 200 300

Age (days)
 Technology Transition Workshop
239
100 340 Days Old

80

Relative Intensity (m/z 239 = 100)

Blue 60

Mont Blanc 40 121 195

256
283
300 403
20 299
Le Grand 0
151 212 327

239
Rollerball 100 4 Days Old 283

80
Fluid Ink at 60 195 299
343
327

Two Ages 40 121

256 300
255 344
212 387
211 388
20 431 432 476
150 151
371 475
415
0
100 200 300 400 500

m/z
 Technology Transition Workshop
239
100 340 Days Old

80

Relative Intensity (m/z 239 = 100)

Blue 60

Mont Blanc 40 121 195

256
283
300 403
20 299
Le Grand 0
151 212 327

239
Rollerball 100 4 Days Old 283

80
Fluid Ink at 60 195 299
343
327

Two Ages 40 121

256 300
255 344
212 387
211 388
20 431 432 476
150 151
371 475
415
0
100 200 300 400 500

m/z
 Technology Transition Workshop
Components of Mont Blanc Ink
Observed m/z Chemical Formula Probable Identity
121.075 [C8H8O+H]+ Acetophenone
403.193 Not determined Not in ink
151.101, 195.126, 239.152, [HO(C2H4O)nH+H]+ Polyethylene
283.178, 327.207, 371.233, n = 3 to 9 glycols
415,254
211.132, 255.159, 299.185, [C8H10O(C2H4O)n+H] Polyethylene
343.214, 387.243, 431.267, + n = 3 to 9 glycol ethyl phenyl
475.292 diethers
212.154, 256.179, 300.206, C16H20(C2H4O)n+ or Polyethylene
344.231, 388.257, 432.288, C7H12N2(C2H4O)m+ glycol derivatives
476.313 n = 0 to 6, m = 2 to 8
 Technology Transition Workshop
100
m/z 121
m/z 255
m/z 256
80 m/z 283
m/z 299

60

Relative Intensity (m/z 239 = 100)

Aging of Blue 40

Mont Blanc 20

Le Grand 0
m/z 195

Rollerball Fluid 80 m/z 300

m/z 327
m/z 343
m/z 344

Ink
60 m/z 403

40

20

0
0 100 200 300

Age (days)
 Technology Transition Workshop
Ink Identification
• Library of 166 inks: 76 ballpoints, 50 fluid inks
and 40 gel inks.
• Written 10 to 35 months before testing.
• 2 spectra of each ink placed in NIST library for
the correct ink type. Handled with gloves.
• Every spectrum tested as an unknown.
– Perfect match with self is ignored, so Hit #1 is
next best match
• Ink correctly identified if Hit #1 is the other
spectrum for the same ink.
 Technology Transition Workshop
Ink Identification Test Results

Black Inks Blue Inks

Ink
Type No. of Inks Correct No. of Inks Correct
in Library Identifications in Library Identifications
Ballpoint 48 99% 28 100%

Fluid 26 81% 24 77%

Gel 21 93% 19 97%

All (black 166 92% -- --

& blue)
 Technology Transition Workshop
Unknowns that Matched Incorrectly
• Black Ballpoints – 1 error
– PaperMate Flexgrip Elite matched Papermate
Flexgrip Ultra
• Blue Ballpoints – 0 errors
• Black Gels – 3 errors
– 2 Pentel Sunburst Gels matched a Parker Gel
Refill and an Integra Roller Gel
– Zebra Jell 3 matched Parker Gel Refill
• Blue Gels – 1 error
– Pentel Hybrid H2 matched Pentel Sunburst Gel
 Technology Transition Workshop
Unknowns that Matched Incorrectly
• Black Fluids – 10 errors
2 PaperMate Liquid Expresso Staedtler Silver Ball 404
Pilot Precise Rolling Ball Uni-ball Deluxe Roller
Sheaffer Slim Rollerball Refill Uni-ball Vision Roller
Staedtler Lumocolor 2 Waterman RollerBall Refill

• Blue Fluids – 11 errors

Bic Z4 Parker Roller Ball Refill
2 Cross Porous Tip Refill 2 Sheaffer Slim Rollerball Refill
2 PaperMate Flair 2 Zebra Zeb-Roller 2000
PaperMate Liquid Expresso
 Technology Transition Workshop
30% Correct Match

Match Quality 20%

of Correct

Percentage of Matches
10%
and Closest
Incorrect 0%
Closest Incorrect Match
12%
Matches for 9%

Black 6%

Ballpoints 3%

0%
1000 900 800 700 600 500
Forward-Matching Match Qualities
 Technology Transition Workshop
Correct Match
20%

Match Quality 15%

of Correct 10%

Percentage of Matches
and Closest 5%

Incorrect 0%
Closest Incorrect Match
12%
Matches for 9%

Blue 6%

Ballpoints 3%

0%
under
1000 900 800 700 600 500 400
400
Forward-Matching Match Qualities
 Technology Transition Workshop
20% Correct Match

Match Quality 15%

of Correct 10%

Percentage of Matches
Pentel Sunburst
and Closest 5%

Incorrect 0%
12% Closest Incorrect Match
Matches for 9%

Black Gel 6%

Inks 3%

0%
under
1000 800 600 400
300
Forward-Matching Match Qualities
 Technology Transition Workshop
30% Correct Match

Match Quality 20%

of Correct

Percentage of Matches
10%
and Closest
Incorrect 0%
Closest Incorrect Match
12%
Matches for 9%

Black Fluid 6%

Inks 3%

0%
under
1000 900 800 700 600 500 400
400
Forward-Matching Match Qualities
 Technology Transition Workshop
Match Qualities in Ink Identification
Closest Incorrect Difference Between
Correct Match Correct & Incorrect
Ink Type Match
Average Median Average Median Average Median
Black 882.3 888.5 749.7 750.0 138.4 119.0
Ballpoint
Blue 878.0 884.0 626.0 620.5 252.0 249.0
Ballpoint
Black 773.2 792.5 647.7 695.0 125.5 83.0
Fluid
Blue Fluid 702.3 743.5 563.7 592.0 138.6 130.0

Black Gel 793.9 832.0 609.0 602.5 188.3 155.5

Blue Gel 752.2 719.0 590.3 560.5 152.7 125.0

 Technology Transition Workshop
Acknowledgements
• National Institute of Justice
– This project was supported by Award No. 2006-DN-
BX-K017 awarded by the National Institute of
Justice, Office of Justice Programs, US Department
of Justice. The opinions, findings, and conclusions
or recommendations expressed in this exhibition
are those of the author and do not necessarily
reflect the views of the Department of Justice.
• John McClelland, principal investigator
• Susan Lorge, worked out optimum conditions
and acquired library data