J Forensic Sci, January 2016, Vol. 61, No.

1
doi: 10.1111/1556-4029.12897
TECHNICAL NOTE Available online at: onlinelibrary.wiley.com

CRIMINALISTICS

Chun-Chieh Chen,1,2 M.S.; Yung-Chien Yu,3 M.S.; Henry C. Lee,4 Ph.D.; Yun-Seng Giang,2 Ph.D.; and
Sheng-Meng Wang,2 Ph.D.

Latent Fingerprint Development on Thermal

Paper Using Traditional Ninhydrin and
1,2-indanedione

ABSTRACT: Thermal paper poses a significant challenge to latent print development as it tends to change color when traditional fingerprint
development formulations are applied to it. In this study, the optimal components of ninhydrin, 1,2-indanedione, 1,8-diazafluoren-9-one (DFO),
and 5-methylthioninhydrin (5-MTN) for yielding clear fingerprints on thermal paper were determined by systematically adjusting the relative
amounts of the reagents, polar solvents, and the nonpolar diluent petroleum ether, followed by validation on text-printed thermal paper. Specifi-
cally, 3.0% ethyl acetate as the polar solvent in petroleum ether was found to be the optimal combination; the optimal dilution ratios of
ninhydrin, DFO, and 5-MTN original solutions with petroleum ether were 1 to 2, 11, and 7, respectively. The optimal concentration of 1,2-in-
danedione in petroleum ether was 0.125 g/L, with a string of 0.5% ethyl acetate in petroleum ether.

KEYWORDS: forensic science, fingerprint, latent print, thermal paper, ninhydrin, 1,2-indanedione, 1,8-diazafluoren-9-one, 5-methylthionin-
hydrin

Thermal paper is a heatsensitive chemical-coated paper that
changes color when exposed to heat. It is commonly used for
thermal printers and lightweight printing devices such as cash
registers and credit card terminals. Equipment utilizing thermal
paper has grown smaller in size, and the size of the thermal
paper can also be adjusted accordingly. The heat-sensitive layer
of the thermal paper has also improved tremendously and has
become ubiquitous in financial, commercial, and retail usage.
Consequently, thermal paper evidence is commonly encountered
in criminal investigations. Important investigative leads can be
obtained from the surface of the thermal paper, including not
only the information contained therein such as date and time of
transaction(s) but also physical evidence such as latent finger-
prints.
The detection of fingerprints is a crucial piece of evidence that
can point to the identity of the possible suspect or potential
witnesses and can lead to the eventual solving of the case. The
retrieval, development, and subsequent identification of latent
fingerprints are important steps in the investigative process.
However, the surface layer of the thermal paper poses a signifi-
cant challenge for latent print development. Thermal paper tends
1 (8,9). Another method is to integrate polyvinylpyrrolidones
Forensic Science Section, Changhua County Police Bureau, 778, Sec. 2,
Jhong-jheng Rd., Changhua City 50004, Taiwan.
2
Department of Forensic Science, Central Police University, 56, Shu-Jen
Road, Kwei-San, Taoyuan 33304, Taiwan.
3
Forensic Science Center, Criminal Investigation Bureau, 5, Lane 553,
Chung Hsiao East Road Section 4, Xin Yi area, Taipei 10675, Taiwan.
4
Henry C. Lee Institute of Forensic Science, University of New Haven,
300 Boston Post Road, West Haven, CT 06516.
Received 2 May 2014; and in revised form 10 Dec. 2014; accepted 30
Dec. 2014.
to change color when traditional fingerprint development formu-
lations such as ninhydrin and 1,2-indanedione are applied to it.
This not only causes the disappearance or masking of the infor-
mation printed on the thermal paper but also impedes the devel-
opment of latent fingerprints for subsequent comparison.
It has been reported in the literature that preventing discol-
oration of thermal paper is possible by avoiding direct application
of polar solvents. Researchers also reported that chemicals like
DMAC (1) and ninhydrin can detect latent fingerprints when
applied to the thermal paper using vapor-phase fuming under low
pressure and high temperature (2). It has also been reported that
the application of a ninhydrin solution with 3MTM NovectTM
Hydrofluoroether products (HFE-71IPA in HFE-7100) to detect
latent fingerprints can prevent thermal paper from changing color
(3). The use of 1,2-indanedione solution with HFE-7100 to detect
latent fingerprints will also not cause thermal paper to change
color (4). Other investigators suggest that using specialty equip-
ment such as vacuum metal deposition (5) or fuming polar sol-
vents such as hydrochloric acid (6), acetic acid, and acetone will
develop latent fingerprints on thermal paper (7). Decolorizing the
thermosensitive layer of the thermal paper after ninhydrin treat-
ment using acetone or G3 solution, or using hemiketals such as
ThermaNin or isononyl ninhydrin (INON), has also been reported
(PVPs) into a ninhydrin (10) or 1,8-diazafluoren-9-one (DFO)
(11) solution to prevent black staining of the thermal paper.
While working on real case samples at Changhua County
Police Bureau, Taiwan (ROC), we found that there are many
obstacles in using amino acid-sensitive reagents like ninhydrin
and 1,2-indandione to recover latent fingerprints on thermal
paper. Most of the forensic laboratories in smaller jurisdictions
have often failed to develop latent fingerprints using traditional

© 2015 American Academy of Forensic Sciences 219

220 JOURNAL OF FORENSIC SCIENCES
formulae, presumably because they do not have vacuum equip-
ment and/or access to specialized chemicals such as ThermaNin
or INON. Thus, the problem stems from the use of polar sol-
vents in the formulations; however, that cannot be avoided
because ninhydrin, 1,2-indandione, and DFO are all insoluble in
nonpolar solvents. We wish to report the most convenient and
effective method for the development of latent prints on thermal
paper without discoloring stains by modifying the traditional
formulations, that is, adjusting the concentration of the polar
solvent in the standard solutions.
Materials and Methods
The objective of this study was to refine the traditional
fingerprint development formulations, that is, ninhydrin and
1,2-indanedione, and determine their respective optimal composi-
tions for developing latent prints on thermal paper. Ninhydrin,
1,2-indanedione, 5-methylthioninhydrin (5-MTN), and DFO
were of reagent grade and were purchased from BVDA Interna-
tional (Haarlem, Netherland). Methyl-tert-butyl ether (MTBE)
was of reagent grade and was obtained from Riedel-de Haen
(Seelze, Germany). Ethyl acetate, glacial acetic acid, methanol,
and ethanol were obtained from Katayama chemicals (Osaka,
Japan), and petroleum ether was from Nihon Shiyaku Reagent
(Osaka, Japan). All aforementioned reagents were of extra pure
grade. Samples were photographed in both absorbance (visible
light) and fluorescent modes using a Canon EOS 20D or 30D
digital camera with operation conditions set as “exposure, auto-
matic white balance, and ISO 400.” Observation and documenta-
tion in the fluorescent mode were performed using an alternative
light source (Omnichromeâ SpectrumTM 9000 [Chino, California,
USA]) at 505 nm and a Canon EOS 20D digital camera with a
100-mm AF Micro-Nikkor lens and Kenko YA3 orange filter.
Experiment 1: Optimization of Polar Solvent Concentration for
the Prevention of Thermal Paper Discoloration
Five different brands of thermal paper were acquired exter-
nally: PARLIKING, HONWEN, FUJI, AURORA, and DAIWA.
Four polar solvents (acetic acid, ethyl acetate, methanol, and
ethanol) were each diluted with petroleum ether to the final con-
centrations of 0.5%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%,
4.0%, 4.5%, 5.0%, 5.5%, and 6.0%. These solutions were
applied to the five different thermal papers to determine the
maximum concentration limit that will not unfavorably change
the color of the blank thermal paper. There were a total of 240
(5 brands 9 4 solvents 9 12 concentrations) applications of the
solutions in this experiment.
Experiment 2: Optimization of Developing Reagent
Concentration for Quality Fingerprinting
Five donors’ latent fingerprints were investigated in this
experiment. Each donor was instructed to deposit his fingerprints
on ten strips of thermal paper to form one set. The fingerprinted
thermal paper strips were stored overnight in a closed drawer at
room temperature.
The test solutions of ninhydrin, DFO, and 5-MTN were pre-
pared as follows:
 Ninhydrin test solutions
The ninhydrin original solution was prepared by dissolving
500 mg of ninhydrin in 4.5 mL of ethanol and 0.5 mL of
acetic acid while adding 0.2 mL of ethyl acetate and made
up to 100 mL with petroleum ether. The original solution
was then topped off with varying amounts of petroleum ether
(100, 200, 300, 400, 500, 600, 700, 800, and 1400 mL).
These intermediate solutions were then further diluted with
2, 3, 4, 5, 6, 7, 8, 9, and 15 volumes of petroleum ether to
achieve the ninhydrin test solutions. An appropriate ninhy-
drin test solution was applied to each set of fingerprinted
thermal paper in the fume hood and then placed in a closed
drawer for 24 h. There were a total of 50 (10 concentra-
tions 9 5 strips) applications of the test solutions in this
experiment.
 5-MTN test solutions
The 5-MTN original solution was prepared by dissolving
300 mg of 5-MTN in 10 mL of MTBE, 2.5 mL of ethanol,
and 1.5 mL of ethyl acetate, while adding 1 mL of acetic
acid. The dilutions of the original solution and the applica-
tions of the test solutions were performed in the same man-
ner as those for the ninhydrin test solutions. There were a
total of 50 (10 concentrations 9 5 strips) applications of the
test solutions in this experiment.
 DFO test solutions
The DFO original solution was prepared by dissolving 50 mg
of DFO in 10 mL of methanol and 10 mL of ethyl acetate,
while adding 12 mL of acetic acid. The dilutions of the origi-
nal solution and the applications of the test solutions were
performed in the same manner as those for the ninhydrin test
solutions. There were a total of 50 (10 concentrations 9 5
strips) applications of the test solutions in this experiment.
Experiment 3: Optimization of 1,2-indanedione Solution for
Quality Fingerprinting
FUJI thermal paper was selected as the substrate to deposit
fingerprints. The five fingerprint donors were instructed to per-
form heavy exercises for perspiration and then vertically depos-
ited their fingerprints onto the thermal paper in a single column.
Each strip of the thermal paper was cut into two parts, dividing
each fingerprint into halves that were treated separately with dif-
ferent developing reagents. The two treated halves were recom-
bined, and a direct visual comparison of the fingerprint ridges
was made between the two halves. For the sake of validity, each
print–reagent pair was carried out twice. The treated prints were
first air-dried in the fume hood and then placed in a closed
drawer for at least 48 h. Note that the development of the finger-
prints must be performed at room temperatures. Heat must be
avoided because the heat-sensitive layers of the thermal paper
would change color immediately if exposed to high tempera-
tures. Observations and comparisons of the developed finger-
prints were performed using an alternative light source (ALS) at
505 nm with an orange filter.
 Concentration effect of acetic acid
The effect of different concentrations of acetic acid on the
development of latent fingerprints was investigated. A stan-
dard development formula comprised of 0.125 g/L 1,2-in-
danedione and 0.5% ethyl acetate but containing no acetic
acid was used. As a variation, we added acetic acid to the
standard development formula at different concentrations of
0.5%, 1.0%, 1.5%, 2.0%, 2.5%, and 3.0%. One half of the
fingerprints were treated with the standard development for-
mula and the other half with the development formula con-
taining the appropriate concentration of acetic acid. The
appropriate FUJI thermal papers were then recombined, and
 CHEN ET AL. . LATENT FINGERPRINT DEVELOPMENT ON THERMAL PAPER 221

a direct visual comparison of the quality of the fingerprints

Results and Discussion
was made. There were a total of 30 (6 concentrations 9 5
strips) comparisons in this subexperiment. Experiment 1: Optimization of Polar Solvent Concentration for
 Concentration effect of ethyl acetate the Prevention of Thermal Paper Discoloration
The effect of different concentrations of ethyl acetate on the
While the low-polarity petroleum ether would not render any
development of fingerprints was studied. A standard devel-
damage or discoloration to the thermal paper, even if directly
opment formula, comprised of 0.125 g/L 1,2-indanedione
applied to it, all four polar solvents (acetic acid, ethyl acetate,
and 0.5% ethyl acetate but no acetic acid, was used. As a
methanol, and ethanol) could cause thermal paper to change
variation, we added ethyl acetate to the standard develop-
color (i.e., turning gray or black) immediately once applied. The
ment formula at different concentrations of 1.0%, 1.5%,
rate at which thermal paper discolored and the degree of discol-
2.0%, 2.5%, 3.0%, and 3.5%. One half of the fingerprints
oration on the heat-sensitive surface was dependent on the con-
were treated with the standard development formula and the
centration of the relevant polar solvent in petroleum ether.
other half with the development formula containing the
Taking acetic acid in petroleum ether, for example, Fig. 1
appropriate concentration of ethyl acetate. The appropriate
shows the actual discoloring effects of the polar solvent at var-
FUJI thermal papers were then recombined and a direct
ious concentrations on the five brands of (blank) thermal
visual comparison of the quality of the fingerprints was
paper. The maximum concentration limit of acetic acid in pet-
made. There were a total of 30 (6 concentrations 9 5 strips)
roleum ether previously determined for not discoloring PAR-
comparisons in this subexperiment.
LIKING, HONWEN, FUJI, AURORA, and DAIWA thermal
 Concentration effect of 1,2-indanedione
paper too significantly to interfere with the fingerprinting
The effect of different concentrations of 1,2-indanedione in the
development was found to be 1.0%, 1.0%, 2.0%, 1.5%, and
development of latent fingerprints was also studied. We used a
2.0%. Similarly, Table 1 summarizes all the corresponding
standard development formula, comprised of 0.125 g/L 1,2-in-
maximum concentration limits of acetic acid, ethyl acetate,
danedione and 0.5% ethyl acetate but containing no acetic
methanol, and ethanol in petroleum ether for not discoloring
acid. As a variation, an appropriate amount (0.008, 0.016,
the five thermal papers significantly. Referring to the actual sit-
0.031, 0.063, 0.25, 0.375, and 0.50 g/L) of 1,2-indanedione
uations of the five thermal papers, the respective polar sol-
was further added to the standard development formula. One
vent’s optimal concentration that would not discolor the
half of the fingerprints were treated with the standard develop-
thermal paper too significantly to interfere with the fingerprint-
ment formula and the other half with the development formula
ing development was determined to be 1.0%, 3.0%, 1.0%, and
containing variable concentration of 1,2-indanedione. The
1.0% for acetic acid, ethyl acetate, methanol, and ethanol in
appropriate FUJI thermal papers were then recombined, and a
petroleum ether, respectively. The best overall polar solvent for
direct visual comparison of the quality of the fingerprints was
all five thermal papers was determined to be 3.0% ethyl acet-
made. There were a total of 35 (7 concentrations 9 5 strips)
ate in petroleum ether.
comparisons in this subexperiment.

FIG. 1––The discoloring effects of various concentrations of acetic acid in petroleum ether on five brands of (blank) thermal paper. The thickly framed
entries correspond to the previously determined maximum concentration limit for not significantly discoloring PARLIKING, HONWEN, FUJI, AURORA, and
DAIWA thermal paper as to interfere with fingerprint development.
222 JOURNAL OF FORENSIC SCIENCES

TABLE 1––Maximum concentration limits of four polar solvents that do not prints were found on the thermal paper. Thus, the optimal ninhy-
significantly discolor the five thermal papers so as to interfere with finger- drin–petroleum ether ratio was determined to be 1:2 (Fig. 3a).
print development.†
The above dilution study was not only applicable to the tradi-
tional ninhydrin solution but also to other amino acid reagents such
Concentration of Brands of Thermal Paper as DFO and 5-MTN. DFO and 5-MTN, when diluted with 11 and
Polar Solvent in 7 volumes of petroleum ether, respectively, would also develop
PARLIKING HONWEN FUJI AURORA DAIWA
Petroleum Ether distinct prints without causing interfering discoloration (Fig. 4).
Acetic Acid 1.0 % 1.0 % 2.0 % 1.5 % 2.0 %
Experiment 3: Optimization of 1,2-indanedione Solution for
Ethyl Acetate 3.0 % 3.0 % 4.5 % 3.5 % 4.5 %
Quality Fingerprinting
Methanol 1.0 % 1.0 % 1.0 % 1.5 % 1.5 %
• Concentration Effect of Acetic Acid
Ethanol 1.0 % 1.0 % 1.5 % 2.0 % 2.5 % A visual comparison indicated that the addition of the rela-
tively more polar and hence, more destructive acetic acid to
the standard development formula, 1,2-indanedione did not
†
The thickly framed entries correspond to the respective optimal con- improve but significantly worsened the fluorescence intensity
centration of the polar solvents, where the most thickly framed entry, i.e., of the developed fingerprint ridges (Fig. 5).
3.0% ethyl acetate in petroleum ether, represents the best polar solvent overall.
• Concentration Effect of Ethyl Acetate
A visual comparison indicated that increasing the concentration
Experiment 2: Optimization of Ninhydrin Concentration for of the relatively more polar and hence, more destructive ethyl
Quality Fingerprinting acetate in the development formula, 1,2-indanedione from 1.0%
to 3.5% did not improve but significantly worsened the fluores-
As soon as the original ninhydrin formula containing 0.5 g nin- cence intensity of the developed fingerprint ridges (Fig. 6).
hydrin, 4.5 mL ethanol, 0.5 mL acetic acid, 0.2 mL ethyl acetate, • Concentration effect of 1,2-indanedione
and 100 mL petroleum ether was applied, not only did the surface A visual comparison indicated that there was no significant
of the thermal paper turn dark but also the fingerprint ridges difference in the fluorescence intensity between the finger-
became blurry due to the discoloring effects of the polar solvents prints on thermal paper developed with 0.125 and 0.25 g/L
(Fig. 2a). At between 1:1 and 1:4, different clarities of the finger- 1,2-indanedione (Fig. 7). However, when the concentration of
prints on the thermal paper were readily developed (Fig. 2a,b). 1,2-indandione fell below 0.063 g/L, the fluorescence inten-
However, at a ninhydrin–petroleum ether ratio of 1:4, the ninhydrin sity of the fingerprint ridges worsened significantly. Thus, the
started to be insufficient, and the developed fingerprints became minimum concentration of 1,2-indanedione in petroleum ether
less distinct (Fig. 3b). At more dilute ratios than 1:5, no finger- should be 0.125 g/L.

FIG. 2––(a) Except for the leftmost strip of fingerprinted thermal paper totally discolored by undiluted ninhydrin formula containing highly concentrated
polar solvents, the other four entries on the right show different clarities of fingerprints on thermal paper developed with 1:1, 1:2, 1:3, and 1:4 ninhydrin in
petroleum ether, respectively; (b) A closer look of the foregoing original entry where both the thermal paper and the fingerprint ridges are discolored.
 CHEN ET AL. . LATENT FINGERPRINT DEVELOPMENT ON THERMAL PAPER 223

FIG. 3––(a) Distinct fingerprint on thermal paper developed with 1:2 ninhydrin in petroleum ether; (b) The 1:4 ninhydrin solution in petroleum ether is a lit-
tle too weak to develop a distinct fingerprint.

FIG. 4––Distinct fingerprints developed on thermal paper: (a) DFO in petroleum ether (1:11) under ALS; (b) 5-MTN in petroleum ether (1:7) under visible
light.

the development of latent prints on thermal paper, highly con-

Optimal Formulae of 1,2-indanedione and Ninhydrin for Latent
Fingerprint Development on Thermal Paper
Because both 1,2-indanedione and ninhydrin were insoluble in
petroleum ether, polar solvents such as acetic acid, ethanol, or
ethyl acetate were used to help dissolve 1,2-indanedione or nin-
hydrin prior to the addition of petroleum ether. Unfortunately, in
centrated polar solvent(s) will not only discolor the thermal
paper but also blur the fluorescence of the ridges. On the basis
of our experimental results, for the 1,2-indanedione formula, the
selected polar solvent was ethyl acetate, and its amount should
be <3.0%. For the ninhydrin formula, the selected polar solvent
was ethanol, and its amount should be <1.0%. Specifically, the
224 JOURNAL OF FORENSIC SCIENCES
FIG. 5––The addition of 3% acetic acid (top) worsens the intensity of the developed fingerprints on thermal paper in both the absorbance mode (left) or the
fluorescence mode (right).
FIG. 6––Increasing ethyl acetate in the 1,2-indanedione formula from
0.5% (right) to 3.0% (left) worsens the fluorescent intensity of the developed
fingerprint ridges.
optimal 1,2-indanedione working solution was prepared by dis-
solving 0.125 g/L of 1,2-indanedione in 0.5% ethyl acetate (but
without acetic acid) and adding 1 L of petroleum ether.
Diluting the original ninhydrin solution is the most practical
method for smaller jurisdictions. The optimal ninhydrin–petro-
leum ether ratio was determined to be 1:2. In fact, the proposed
optimal 1,2-indanedione and ninhydrin formulae were mutually
validated by developing two latent fingerprints of the same
source on text-printed thermal paper of the same kind. The
development process would take a minimum of 48 h inside a
drawer at room temperature, and this would have an effect on
timely processing. Care must also be taken while using petro-
leum ether given its highly flammable nature. The fingerprint
FIG. 7––There is no significant difference in fluorescent intensity of the
fingerprints on thermal paper developed with 0.125 g/L (left) and the ones
developed with 0.25 g/L (right) 1,2-indanedione.
development results were photographed under ALS at 505 nm
for 1,2-indanedione and under visible light for ninhydrin
(Fig. 8).
Conclusions
For criminal investigations in practice, most small laboratories
do not have special equipment and/or may lack access to new
reagents such as ThermaNin or INON. Our research suggests
that the most convenient and effective approach to develop latent
fingerprints on thermal paper without causing thermal paper dis-
coloration is to modify traditional amino acid sensitive reagents.
The results obtained in this study demonstrated that the proposed
 CHEN ET AL.
FIG. 8––The proposed optimal 1,2-indanedione and ninhydrin formulae are mutually validated by developing two latent fingerprints of the same source on
text-printed thermal paper of the same kind.
optimal solutions for 1,2-indanedione, ninhydrin, DFO, and 5-
MTN can all yield clear fingerprints on thermal paper. Com-
pared to those reported in the literature, all of our methods
involved no heating, and therefore, 1,2-indanedione took a bit
longer to develop clear prints. As far as the resulting fluores-
cence intensity is concerned, our optimized ninhydrin formula
yielded prints comparable to ones in the literature within a com-
parable time frame, while 1,2-indanedione, 5-MTN, and DFO
gave slightly weak yet usable fluorescence.
For the sake of safety, a corner of the thermal paper should
always be tested first for the suitability of the chemical solution.
For situations where there are two opportunities to process the
same latent fingerprint on the same thermal paper, it is suggested
that a systematic 2-stage process be employed (12,13): (i) apply
the foregoing optimal 1,2-indanedione formula and (ii) apply the
foregoing optimal ninhydrin formula. This dual application will
significantly increase the success rate of developing latent prints
on thermal paper.
Acknowledgments
Scientific advice from Professors Eugene Lee and Stella Tan
of the National University of Singapore was highly appreciated.
Financial support provided by Ministry of Interior, Taiwan
(ROC) (Grant No. 101-0805-05-05-02) and Wintek Corporation.
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Additional information and reprint requests:
Sheng-Meng Wang, Ph.D.
Department of Forensic Science
Central Police University
56 Shu-Jen Road, Kwei-San
Taoyuan 33304
Taiwan
E-mail: wang531088@mail.cpu.edu.tw