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Forensic Science International 280 (2017) 130138

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Forensic Science International


journal homepage: www.elsevier.com/locate/forsciint

Differentiation of red lipsticks using the attenuated total reection


technique supported by two chemometric methods
Marta Gadysz, Magorzata Krl* , Pawe Koscielniak
Laboratory for Forensic Chemistry, Department of Analytical Chemistry, Faculty of Chemistry, Jagiellonian University in Krakw, Gronostajowa 2, 30-387
Krakw, Poland

A R T I C L E I N F O A B S T R A C T

Article history:
Received 19 June 2017 The main challenge for the identication and differentiation of lipsticks for forensic purposes is the fact
Received in revised form 2 September 2017 that they have a similar chemical composition in particular, samples of the same hue. The objective of
Accepted 25 September 2017 the presented research was to solve this problem using a nondestructive analytical technique
Available online 6 October 2017 attenuated total reection spectroscopy (ATR). 38 red lipsticks produced by 20 different manufacturers
were examined in optimized experimental conditions. To facilitate discrimination and provide more
Keywords: extensive analyses of the obtained data, two chemometric techniques: principal component analysis and
Red lipsticks cluster analysis were used. Ultimately, nine groups of investigated lipstick samples with comparable
Differentiation
chemical compositions were differentiated. Moreover, lipstick smears on six different interfering
Attenuated total reection
surfaces were analyzed and spectral subtraction was performed in order to identify individual samples.
Principal component analysis
Cluster analysis The established approach succeeded in identifying the index number and the manufacturer of the
samples by using an in-lab built ATR spectra library. The developed method demonstrates a great
potential for the differentiation and identication of red lipsticks with a very similar hue. It also seems to
have good prospects for future application in forensic science investigations.
2017 Elsevier B.V. All rights reserved.

1. Introduction occurring in smaller quantities are antioxidant materials and


perfumes [3,4].
Lipstick residues and smears may be signicant forensic There have been many publications regarding lipstick exami-
evidence in investigations, in particular in cases of rape, sexual nation, reporting results from simple optical [5] to modern
assault, murder and burglary. They may be found left on a variety of analytical methods (i.e. CE [6], HPLC [7,8], UHPLCMS [9],
surfaces for instance, tissue paper, cigarette butts, glasses, GCMS [10], ICP-OES [11], ICPMS [12], AAS [11,13], LIBS [14,15],
drinking cups, clothing etc., and constitute indirect evidence XRF [16]). However, most of them have focused on (aspects of) the
linking a suspect with the victim or crime scene [14]. Neverthe- negative impact of the lipstick components on humans and the
less, identication of lipstick samples and differentiation between environment. Among other things, the content of azo [6], xanthene
them can be a major challenge, because the majority of lipsticks [7,17] or other organic colorants [8,9], as well as hazardous
have a similar composition. They are composed of three main nitrosamines [10] and heavy metals (e.g. Pb) [1116,1820] in
ingredients: oils (typically 4070%) a mixture of castor, lipsticks have been studied. In the case of forensic lipstick
vegetable, mineral or lanolin oil; waxes (approximately 15%) examination, the most common technique is nondestructive
especially beeswax or carnauba; and colouring agents (roughly 8%) Raman spectroscopy [2,3,21,22]. It should be pointed out here,
organic dyes such as erythrosine, amaranth, rhodamine, however, that in every mentioned article, Raman spectroscopy has
tartrazine, derivatives of uorescein, and inorganic pigments, been accompanied by problems associated with uorescence. An
particularly titanium dioxide and iron oxides. Other constituents in-situ method of surface enhanced resonance Raman scattering
(SERRS) detection of the colorants in lipstick smears on both glass
and cotton surfaces was reported [23], but a surfactant was
required to obtain SERRS spectra of the dyes and pigments in such
waxy samples. Moreover, the technique of neutron activation
* Corresponding author at: Laboratory for Forensic Chemistry, Department of analysis (NAA) a nondestructive and highly sensitive but also
Analytical Chemistry, Faculty of Chemistry, Jagiellonian University in Krakw, very time-consuming and expensive method was employed to
Gronostajowa 2, 30-387 Krakw, Poland.
study the presence of trace elements in samples of lipsticks [24].
E-mail address: krolm@chemia.uj.edu.pl (M. Krl).

https://doi.org/10.1016/j.forsciint.2017.09.019
0379-0738/ 2017 Elsevier B.V. All rights reserved.
M. Gadysz et al. / Forensic Science International 280 (2017) 130138 131

Another technique described in the literature used to differentiate combined. This procedure is accomplished repeatedly until all
lipstick is TLC [1,4,25]. However, although the information samples are arranged into one cluster. It is worth pointing out
obtained by the TLC analytical methods was usually sufcient that several methods for measuring similarity and combining
for lipstick differentiation from the forensic point of view, these clusters are available. Their choice depends on the investigated
methods have the important drawback of destructive sample samples and this should be evaluated experimentally [31]. In this
preparation. study, Euclidean distances were calculated and Wards method
One of the techniques, which to the best of our knowledge was employed. Furthermore, in the literature, over 20 different
has not yet been applied to lipstick examination is attenuated total rules for stopping an obtained dendrogram are known. In this
reection spectroscopy (ATR). Two signicant advantages of ATR, article, Mojenas stopping rule was considered to be the most
namely that the sample is not destroyed and no sample efcient.
preparation is required, have led to this technique being widely According to Mojenas stopping rule, one should select the
employed in forensic science investigations [2629]. Consequent- number of groups where the following inequality (Eq. (1)) is
ly, the aim of the presented research was to develop the rst ATR- fullled
based procedure enabling differentiation of red lipsticks. Addi-
tionally, chemometric methods such as principal component di1 > d ksd 1
analysis (PCA) and cluster analysis (CA) were applied in order to where d0,dn, dn1 are distances corresponding to n, n  1, . . .
facilitate discrimination and provide more extensive analyses of
1 clusters. The terms d, sd, and k are the mean, the standard
the obtained data. Lastly, the in-lab built ATR spectra library, deviation of the d values, and a constant, respectively [32]. As
enabling identication of all investigated samples, was tested by
stated by Milligan and Cooper [33], the value of k should be 1.25.
analyzing traces of random lipsticks on six different, potentially
interfering surfaces.
3.3. Correlation method
2. Experimental
Correlation coefcients (CC) were computed as Pearson
The solutions (isopropanol and methanol) used throughout the correlation coefcients by means of Thermo Scientic OMNIC
experiments were supplied by Sigma-Aldrich (Germany). 38 red software. It seems important to note that the used algorithm
lipsticks (for 5 randomly selected 5 items of the same index additionally included elimination of the effects of baseline
number were bought) of a very similar hue representing variation.
20 different manufacturers were either purchased in local shops
or donated by Inglot Sp. z o.o. (cosmetics company). Detailed 4. Results and discussion
information about all of them is presented in Table 1.
The experiments were carried out using a Thermo Nicolet Firstly, the parameters of the experiments were optimized. It is
iS50 FTIR (Thermo Fisher Scientic Co, Waltham, MA, USA) with a known that increasing the number of scans reduces the signal-to-
Smart Orbit micro-ATR accessory. All spectra were collected from noise level of data, and enables one to distinguish small peaks from
650 to 4000 cm1 by putting small amounts of a sample noise, and that lower resolution can have an effect on the
(approximately 10 mg) on the ZnSe crystal and carefully pressing differentiation of peaks. Therefore, 16, 20 and 60 scans, and
it with an ATR pressure tower. After each measurement, the surface resolutions of 4 and 2 cm1 were tested. Despite the fact that only
of the crystal was cleaned with 50% v/v isopropanol and then with slight differences were observed between registered spectra,
methanol in order to avoid contamination. A new background was 20 scans per second with 4 cm1 spectral resolution for both
collected prior to the analysis of each new sample. background and samples were considered to be optimal.
Processing of the obtained spectra was carried out using The homogeneity (intra-variability) of lipsticks was investigat-
Thermo Electrons OMNIC 9 software. All statistical analyses were ed by analyzing spectra obtained from three different parts of
performed using Statistica 12.5 PL software (StatSoft, Tulsa, OK, several randomly selected lipstick samples (L3, L10, L14, L21, L31,
USA) and OriginPro 2017 software (OriginLab Corporation, North- L32, L34 and L36) during one day. As demonstrated in exemplary
ampton, MA, USA). Fig. 1a presenting the results obtained for three lipsticks, no visual
differences were found on the spectra. Consequently, it was
3. Applied chemometric/computational methods considered that the samples were homogeneous and their
composition was stable during one day.
3.1. Principal component analysis Additionally, during three days the inter-variability of lipstick
was examined using 5 of the previously selected lipsticks. The
PCA is a well-known multivariate statistical method enabling similarity of the spectra was calculated as correlation coefcient
reduction of the dimensionality of the original dataset by creating a (L10: 99.90  0.04; L31: 99.91  0.04; L32: 99.94  0.01; L34:
linear combination of variables called principal components (PCs). 99.83  0.06; L36: 99.92  006). The presented results provided
However, the determination of the optimal number of components the basis for claiming that the analyzed samples of the lipstick
that should be chosen is a crucial step. Inappropriate estimation of exhibited almost unchanged chemical composition also within a
PCs could lead to loss of information. The Scree test, Kaisers few days. Moreover, both above-mentioned experiments proved
stopping rule and the percentage of cumulative variance are the the high intra- and inter-repeatability of the proposed ATR-FTIR
most frequently used strategies [30], and these were applied in this method.
study. Furthermore, PCs are a very useful tool for demonstrating Finally, the variation of chemical composition of lipstick
the relationships between analyzed samples. samples within the same series was also investigated during
one day by analyzing ve items (ae) of ve samples (L10, L31, L32,
3.2. Cluster analysis L34 and L36) from the same manufacturer with the same index
number. As it can be seen in exemplary Fig. 1b the obtained
In CA, samples are assembled in high dimensional space. At spectra for samples L36 ae almost completely overlapped.
the beginning, each sample constitutes its own separate cluster, Therefore, it was ascertained that the chemical composition of
and then two objects, which are the closest to each other, are analyzed lipsticks that were both produced by the same
132 M. Gadysz et al. / Forensic Science International 280 (2017) 130138

Table 1
List of lipsticks examined during the present study.

Code of lipstick Manufacture Index numbera Seriesb Comment


L1 INGRID 289 Wonder shine
L2 INGRID 288 Wonder shine
L3 MISSLYN M20.94 Rouge a levres High passion
L4 MISSLYN M20.87 Rouge a levres Cayun pepper
L5 GOSH 005 Velvet touch lipstick Matt, classic red
L6 p2 030 Full color lipstick Challenge authority
L7 BourjoisParis 13 Rouge Edition Rouge jet set
L8 e 02 Longlasting lipstick All you need is red
L9 REVLON 720 Super lustrous lipstick Fire & Ice
L10a LOREAL Paris 330 Color riche Matte, cocorico
L10b LOREAL Paris 330 Color riche Matte, cocorico
L10c LOREAL Paris 330 Color riche Matte, cocorico
L10d LOREAL Paris 330 Color riche Matte, cocorico
L10e LOREAL Paris 330 Color riche Matte, cocorico
L11 MAYBELLINE New York 955 Color sensational Craving coral, matte
L12 SEPHORA B06 Rouge Baume SPF20 Soft red
L13 SEPHORA R05 Rouge Cream Hot Tango
L14 SEPHORA N 62 Rounge Brilliance Stay Together
L15 SEPHORA N 19 Color lip last Pure red-matt
L16 CATRICE 430 Ultimate color Hotn Spice
L17 CATRICE 440 Ultimate color Hungs-and Hibis-Kisses
L18 CATRICE 140 Ultimate stay Behind the red curtain
L19 LOVELY 8 Creamy color
L20 WIBO 1 Matte Intense
L21 MISS SPORTY 301 My BFF My cheeky cherry
L22 EVELINE COSMETICS 486 Aqua Platinum
L23 EVELINE COSMETICS 710 Color edition red in love
L24 RIMMEL 510 Moisture renew Mayfair red lady
L25 ASTOR 203 Perfect stay fabulous
L26 MAX FACTOR 715 Color elixir Ruby tuesday
L27 MANHATTAN 550 Soft rouge lipstick Watermelon
L28 INGLOT 176 Lipstick
L29 INGLOT 180 Lipstick
L30 INGLOT 429 Lipstick matte
L31a INGLOT 44 Slim gel lipstick
L31b INGLOT 44 Slim gel lipstick
L31c INGLOT 44 Slim gel lipstick
L31d INGLOT 44 Slim gel lipstick
L31e INGLOT 44 Slim gel lipstick
L32a INGLOT 21 AMC lip pencil matte
L32b INGLOT 21 AMC lip pencil matte
L32c INGLOT 21 AMC lip pencil matte
L32d INGLOT 21 AMC lip pencil matte
L32e INGLOT 21 AMC lip pencil matte
L33 INGLOT 64 AMC lip paint
L34a INGLOT 12 HD lip tint matte
L34b INGLOT 12 HD lip tint matte
L34c INGLOT 12 HD lip tint matte
L34d INGLOT 12 HD lip tint matte
L34e INGLOT 12 HD lip tint matte
L35 MANHATTAN 45 M Xtreme last & shine
L36a MANHATTAN 45 N Perfect creamy & care
L36b MANHATTAN 45 N Perfect creamy & care
L36c MANHATTAN 45 N Perfect creamy & care
L36d MANHATTAN 45 N Perfect creamy & care
L36e MANHATTAN 45 N Perfect creamy & care
L37 MANHATTAN 45 F Perfect creamy & care
L38 MANHATTAN 90 A Perfect creamy & care
a
The index number is a number assigned to each lipstick in a given series.
b
Series name refers to cosmetics collection. The same series name indicates the same collection of lipsticks.

manufacturer and had the same index number were characterized asymmetric (2916 cm1) C H stretching vibrations and asym-
by very good repeatability. metric CH3 stretching vibrations (2954 cm1). The last range
(below 1800 cm1) contains many peaks. CO stretching vibra-
4.1. Visual inspection of spectra tions (1730 cm1), CH2 scissor deformation (1465 cm1), CO
stretching and CH bending vibrations (1172 cm1), and CH2
As seen in Fig. 2, a typical spectrum of a lipstick sample can be rocking mode (712 cm1) are just some of them. All of the
divided into three main ranges. Generally, the rst range (between mentioned peaks could be attributed to various oils and waxes,
3740 and 3100 cm1) includes one broad band, which is attributed especially castor oil and beeswax two common ingredients in
to water and hydroxyl groups. The second range (from 3050 to lipsticks [3]. Therefore, it can be supposed that the most
2775 cm1) takes account of symmetric (2850 cm1) and informative and distinguishing range is below 1800 cm1.
M. Gadysz et al. / Forensic Science International 280 (2017) 130138 133

Fig. 2. Typical lipstick spectrum, represented by L13.

are distinctive and consequently they can be effortlessly distin-


guished.
Concisely, according to PCA analysis, red lipsticks can be divided
into the following six groups: G1 {L34}, G2 {L32}, G3 {L31, L33}, G4
{L10}, G5 {L15, L25} and G6, which includes all the remaining
samples.
In an attempt to nd the most efcient grouping of lipsticks,
cluster analysis was performed in order to classify lipstick samples
in the G6, based on similarities of their ATR spectra in the range
from 1800 to 650 cm1 (the ngerprint region). Accordingly, the
dendrogram presented in Fig. 4 was created, whereas the results
obtained from Mojenas stopping rule are shown in Table I in the
ESM. They indicated that the value of 1.19 should be used as the
stopping distance.
As a consequence, the dendrogram was divided into the
Fig. 1. Investigation of a) homogeneity of lipsticks L21, L14 and L3; b) variability of following four groups of lipstick samples: G6a {L1, L8, L20, L27,
lipsticks from the same manufacturer and with the same index number: L36 ae. L28, L29, L30, L35, L36a, L37, L38,}, G6b {L4, L5, L13, L17, L21, L22,

4.2. Differentiation of red lipsticks

4.2.1. Chemometric methods


A primary discrimination of lipstick samples was performed
using the PCA technique, taking into account all 38 of the collected
ATR spectra registered in the range from 4000 to 650 cm1. Fig. I in
the Electronic Supplementary Material (ESM) shows a plot for
deciding on the number of variables a scree test. Interpretation
of this plot indicates that only the rst three components should be
retained in the present study (the line drops precipitously),
because sufcient useful chemical information is provided by
these three components. They also complied with the Kaiser
criterion, which only recommends components with eigenvalues
greater than unity. Furthermore, approximately 90% of the total
variance in the spectra was explained by them: 70.17%, 10.38%,
8.14% for the rst, second, and third PC, respectively. Thus, it is
reasonable to base inferences solely on these PCs.
The PCA score plot of the rst three PCs, demonstrating the
separation of samples, is presented in Fig. 3. Additionally, in Fig. II
in the ESM, two-dimensional plots are presented to make the
visual analysis easier. As seen, most of the lipsticks, except L10, L15,
L25, L31, L32, L33 and L34, were classied into one multi-element
class. Additionally, it was noted that lipstick L15 is very similar to
L25 and lipstick L31 is comparable to L33. Moreover, lipsticks L10,
L32 and L34 are each completely separated from all other lipsticks. Fig. 3. The PCA score plot of the rst three PCs for all investigated lipsticks, taking
This nding suggests that some of the analyzed lipstick samples into account ATR spectra registered in the range from 4000 cm1 to 650 cm1.
134 M. Gadysz et al. / Forensic Science International 280 (2017) 130138

Fig. 4. Cluster analysis of lipsticks from G6, taking into account ATR spectra registered in the range from 1800 cm1 to 650 cm1.

L26}, G6c {L2, L3, L6, L11, L14, L16, L18, L23, L24} and G6d {L7, L9, G6 b. Besides, strong absorption in the range from 1100 to
L12, L19}. Ultimately, as a result of chemometric analysis, nine 1000 cm1 is characteristic for G6 a, b and, as was mentioned
groups of red lipstick samples with comparable chemical before, these bands are connected with phosphates and ether/
compositions were created: G1G5 and G6 ad. alcoholethers [34]. It is noteworthy that there are also three
It is worth noting that according to chemometric classication, unique bands for G6 d at approximately 1387, 1376 and
all the investigated lipstick samples manufactured by Manhattan 1363 cm1.
were classied into one group (G6 a). On the other hand, the Summarizing, visual comparison of the obtained spectra led to
lipstick samples from Sephora were categorized into various conrmation of the classication ascertained according to the
groups (G6 b, c, G5), while some of lipstick samples manufactured chemometric results.
by Inglot were placed in group G6 a and some in G1G3.
4.2.3. Correlation method
4.2.2. Visual classication In order to make the analysis more objective as well as to verify
A visual comparison of the analyzed spectra was also the existence of an unequivocal dependence between the
performed. In the rst step of differentiation between lipstick composition of the lipstick samples and the manufacturer (or
samples, one should look at both the relative intensities and the series), the correlation method was applied.
degree of resolution of bands from 3050 to 2775 cm1. As can be At rst, the variability of the same lipstick sample, s, was
seen in Fig. 5, this region enables differentiation between all ascertained as the mean CC (correlation coefcient) value
groups, except between G2 and G6, and between G4 and G5. calculated for ve items (a-e) of the ve samples (L10, L31, L32,
Further distinction between groups was based on the bands L34 and L36) of lipsticks of the same index number, the same series
occurring below 1800 cm1. As is evident in Fig. 6a, the most and the same manufacturer. The mentioned samples were
characteristic feature for G5 is a lack of band at 1745 cm1 representatives of different PCA groups so they characterized by
corresponding to the carbonyl group. Moreover, in G4 one can see different chemical composition. It is worth pointing out that the
a weak doublet at 1375 and 1363 cm1, whereas in G5 there is CCs value were evaluated by analyzing samples on three successive
only a band at 1363 cm1. Both of these bands are due to CH3 days and they ranged between 0.04% and 0.33%. On this basis the
deformation [34]. following assumptions related to ATR spectra of each examined
The differentiation of G2 from G6 is presented in Fig. 6b. On the pair of lipstick samples have been made:
face of it, spectra obtained for these groups are completely unlike
each other in the range from 1800 to 650 cm1. The most  if CC  100% 3s, it is highly possible that the spectra
noteworthy difference is strong absorption near 1000 and correspond to the same lipsticks,
1100 cm1 in G2. It is highly possible that these peaks correspond  if CC < 100% 3s, the spectra are considered as coming from
to phosphates and ethers or alcoholethers, respectively, which different lipsticks.
are found in many plasticizers [34].
Moreover, the differences between G6 ad are selected in Fig. 6c In short, the similarity limit in the case of the present study is a
only representative spectra of lipstick samples from the groups value of 99.01%.
have been presented. As far as one can see, the region where Comparing the spectra of lipsticks produced by different
differences are most clearly visible is from 1400 to 800 cm1. In this manufacturers, it was ascertained that they are usually characterized
region, a little broad band (especially in G6 ac) at 1150 cm1 by CCs considerably less than 99.00% (see Table II in ESM), e.g., for L15
characteristic for CO stretch in nonaromatic ethers is observable. (Sephora) and L3 (Misslyn), the CC = 55.81% or for L6 (p2) and L20
Moreover, the existence of a siloxane group is most likely due to a (Wibo), the CC = 92.58%. There were a few cases where the
band occurring at 1108 cm1, which is most visible in spectra from compositions of two lipsticks originating from different
M. Gadysz et al. / Forensic Science International 280 (2017) 130138 135

Fig. 5. Differentiation between groups based on the range from 3050 cm1 to 2775 cm1.

manufacturers were very similar, e.g., for L3 (Misslyn) and L22 they may differ to a very limited extent, e.g. only slightly in the
(Eveline Cosmetics) CC = 99.45%, for L29 (Inglot) and L4 (Misslyn) concentration of colour additives. However, further data are
CC = 99.55%. needed in order to conrm this assumption. It has not escaped our
If the investigated samples were from the same manufacturer notice that there were also lipsticks produced by the same
but from different series, most of them could also be easily manufacturer in the same series with an index number differing
distinguished. It can be seen in Table 2 that CCs between spectra of only by one, e.g., L1 Ingrid no. 289 and L2 Ingrid no. 288, which
lipstick samples produced by Inglot, Manhattan and Sephora were revealed, without any doubt, two different spectra (CC = 98.52%).
not higher than 98.50%. Moreover, it is evident that lipsticks It seems important to reiterate that samples from the same
manufactured by Inglot are characterized by the most variable manufacturer, the same series and having the same index number
composition (CC from 96% to 24%). (for instance L36 ae), as was mentioned before, were considered
The obtained results conrmed the theoretical assumption that to be indistinguishable. The CC between spectra was much higher
lipstick samples from the same manufacturer and from the same than 99.01%.
series are more similar to each other than lipstick samples from
different series (see CC for L28 and L29 from Inglot, or L36 and 4.3. Analysis of lipsticks traces on various surfaces preliminary
L37 from Manhattan). On the other hand, some exceptions can be examination
observed: sample L35 from the Xtreme last & shine series is
undistinguished (CC between 99.11% and 99.49%) from the samples Due to the fact that lipstick samples can be found left on various
from the Perfect creamy & care series. Moreover, the data suggest surfaces, it is justied to investigate the possibility of identifying
that a comparison of samples with a similar index number (from lipstick traces in the form of such smears.
the same manufacturer and from the same series) can cause Firstly, one of the samples (L28) was left as a lipstick mark on
problems. In these cases, the CCs between samples were calculated various surfaces in order to assess which surfaces caused
to be greater than 99.01%, and in consequence, they were not interfering bands. The surfaces studied included: envelope, paper
differentiable with the use of the presented method. The (80 g/m2), tissue (100% cellulose), cigarette butts, and a transparent
discrimination of such lipsticks samples can be problematic and plastic cup, as well as the white collar of a shirt. As can be seen in
136 M. Gadysz et al. / Forensic Science International 280 (2017) 130138

Table 2
Spectra correlation coefcients between samples from the same manufacturer: a)
Inglot; b) Manhattan; c) Sephora. Lipsticks samples from the same series are bolded.

a)
L28 100.0
L29 99.57 100.0
L30 96.44 96.70 100.0
L31 91.67 91.50 87.83 100.0
L32 90.24 90.90 95.44 83.29 100.0
L33 79.95 79.48 74.51 88.50 65.70 100.0
L34 27.11 25.90 24.67 40.22 17.65 53.76 100.0
L28 L29 L30 L31 L32 L33 L34

b)
L27 100.0
L35 98.46 100.0
L36a 97.79 99.11 100.0
L37 98.24 99.49 99.65 100.0
L38 97.93 99.26 99.83 99.78 100.0
L27 L35 L36a L37 L38

c)
L12 100.0
L13 94.87 100.0
L14 95.95 96.69 100.0
L15 62.50 57.46 55.17 100.0
L12 L13 L14 L15

In striving to nd the most efcient approach to identication of


lipstick marks, two spectral subtractions were performed: (1) in
the whole range of registered spectra, (2) in the ngerprint region.
The effectiveness of the proposed approach to the identication of
lipsticks marks was veried by analyzing six different traces of red
lipstick (L7, L12, L18, L23, L28 and L35) smudges on all the above-
mentioned surfaces.
Overall, the procedure was carried out in a few steps by:
measuring the surface spectrum and the spectrum of the lipstick
on the same surface, then subtracting one from the other in the
whole range of the rejected spectrum and in the ngerprint region
(from 1800 to 650 cm1), and subsequently identifying the
spectrum by using an in-lab built ATR spectra library. The obtained
results indicate that the second approach solely enables one to
connect lipstick marks with the manufacturer and index number
(see Table 3). It should be highlighted that the described
subtraction approach did not yield pure spectra of lipstick samples
additional or negative bands appeared in some cases.
Nevertheless, a comparison of the spectral subtraction result with
the spectra collected in the in-lab built library enabled in every
case without exception the successful identication of lipstick
samples.

5. Conclusions

ATR spectroscopy was successfully used for the differentiation


of 38 red lipsticks of a very similar hue. Although no unambiguous
trends could be identied within manufacturers or series and the
differentiation of lipsticks is a complicated issue, combining the
two chemometric methods (PCA and CA) enabled the distinguish-
Fig. 6. Differentiation of lipsticks according to ATR spectra registered in the range
from 1800 cm1 to 650 cm1: a) G4 represented by L25, G5 represented by L10; b) ing of nine groups from investigated lipsticks of similar ATR
G2 represented by L32, G6 represented by L12; c) G6a represented by L35, G6b spectra. Similarities cannot be assumed, in advance, on the basis
represented by L26, G6c represented by L3, and G6d represented by L19. of manufacturer, series or index number compatibility. However,
visual analysis revealed bands that were characteristic for
Fig. III in the ESM, all of the analyzed surfaces, except the appropriate groups and conrmed the classication obtained
transparent plastic cup, provided additional bands to the lipstick by chemometric techniques. Furthermore, the approach sup-
spectra. ported by the correlation coefcient enabled deeper differentia-
In order to isolate lipstick spectra from surface spectra, Thermo tion of all studied lipstick spectra. And, importantly, the
Electrons OMNIC software was used. This is one of the most differentiation could be achieved non-destructively in a matter
difcult issues in the identication of samples in forensic science. of minutes.
M. Gadysz et al. / Forensic Science International 280 (2017) 130138 137

Table 3
A comparison of the two approaches for the identication of lipsticks in the form of lipstick marks smudged on interfering surfaces. Correct identication of lipsticks is
indicated by +.

Lipstick Code The range of spectral subtraction

4000650 cm1 1800650 cm1

Cigarette butts Envelope Paper Tissue White collar of shirt Cigarette butts Envelope Paper Tissue White collar of shirt
L7      + + + + +
L12      + + + + +
L18      + + + + +
L23      + + + + +
L28      + + + + +
L35      + + + + +

Table 4
Discriminating power of PCA, CA and CC analysis. [7] S. Scalia, S. Simeoni, Assay of xanthene dyes in lipsticks by inverse supercritical
uid extraction and HPLC, Chromatographia 53 (2001) 490494.
Method Discriminating power (DP)
[8] S.C. Rastogi, V.J. Barwick, S.V. Carter, Identication of organic colourants in
PCA 0.29 cosmetics by HPLC-diode array detection, Chromatographia 45 (1997) 215
CA 0.51 228.
Correlation method 0.93 [9] Y. Xian, Y. Wu, X. Guo, Y. Lu, H. Luo, D. Luo, Y. Chen, Simultaneous
determination of 11 restricted dyes in cosmetics by ultra-high-performance
liquid chromatography/tandem mass spectrometry, Anal. Methods 5 (2013)
19651974.
[10] Q. Ma, H.-W. Xi, C. Wang, H. Bai, G.-C. Xi, N. Su, L.-Y. Xu, J.-B. Wang,
Determination of ten volatile nitrosamines in cosmetics by gas chromatogra-
To sum up, on the basis of the performed PCA, CA and CC phy tandem mass spectrometry, Chinese J. Anal. Chem. 39 (2011)
12011207.
differentiations, the discriminating power (DP) of the ATR analysis
[11] . Ferreira Batista, A. dos Santos Augusto, E.R. Pereira-Filho, Chemometric
of lipstick samples has been calculated according to the following evaluation of Cd, Co Cr, Cu, Ni (inductively coupled plasma optical emission
equation: DP = 1  2 M/(N(N  1)) (where M is number of matching spectrometry) and Pb (graphite furnace atomic absorption spectrometry)
pairs; N is total number of analysed lipsticks). The obtained DP concentrations in lipstick samples intended to be used by adults and children,
Talanta 150 (2016) 206212.
values were listed in Table 4. [12] N.M. Hepp, Determination of total lead in 400 lipsticks on the U.S. market
These results conrmed the above-mentioned conclusion that using a validated microwave-assisted digestion, inductively coupled plasma-
PCA and CA enables some preliminary classication whereas the mass spectrometric method, J. Cosmet. Sci. 63 (2012) 159176.
[13] S. Gunduz, S. Akman, Investigation of lead contents in lipsticks by solid
correlation coefcient is the best choice for the lipsticks sampling high resolution continuum source electrothermal atomic absorption
discrimination. spectrometry, Regul. Toxicol. Pharm. 65 (2013) 3437.
Ultimately, by using the developed method and in-lab built ATR [14] M.A. Gondal, Z.S. Seddigi, M.M. Nasr, B. Gondal, Spectroscopic detection of
health hazardous contaminants in lipstick using Laser Induced Breakdown
spectra library, it was possible to detect and connect some lipsticks Spectroscopy, J. Hazard. Mater. 175 (2010) 726732.
left as lipstick marks with their manufacturer and index number, [15] A. dos Santos Augusto, E. Ferreira Batista, E.R. Pereira-Filho, Direct chemical
even on interfering surfaces. This nding conrms that the ATR inspection of eye shadow and lipstick solid samples using laser-induced
breakdown spectroscopy (LIBS) and chemometrics: proposition of classica-
technique is a useful tool in lipstick differentiation and identica-
tion models, Anal. Methods 8 (2016) 58515860.
tion. All things considered, it seems reasonable to assume that the [16] E. Da Silva, A. Matthews David, A. Pejovi c-Mili
c, The quantication of total lead
proposed method shows promise for future application in forensic in lipstick specimens by total reection X-ray uorescence spectrometry, X-
Ray Spectrom. 44 (2015) 451457.
science investigations as a good alternative to routinely used
[17] Ch.Ch. Wang, A.N. Masi, L. Fernandez, On-line micellar-enhanced spectro-
methods. uorimetric determination of rhodamine dye in cosmetics, Talanta 75 (2008)
135140.
[18] I. Al-Saleh, S. Al-Enazi, Trace metals in lipsticks, Toxicol. Environ. Chem. 93
Appendix A. Supplementary data (2011) 11491165.
[19] I. Al-Saleh, S. Al-Enazi, N. Shinwari, Assessment of lead in cosmetic products,
Regul. Toxicol. Pharm. 54 (2009) 105113.
Supplementary data associated with this article can be found, [20] P. Piccinini, M. Piecha, S.F. Torrent, European survey on the content of lead in lip
in the online version, at https://doi.org/10.1016/j.forsciint.2017. products, J. Pharm. Biomed. Anal. 76 (2013) 225233.
09.019. [21] P. Gardner, M.F. Bertino, R. Weimer, E. Hazelrigg, Analysis of lipsticks using
Raman spectroscopy, Forensic Sci. Int. 232 (2013) 6772.
[22] F. Salahioglu, M.J. Went, S.J. Gibson, Application of Raman spectroscopy for the
diff ; ;erentiation of lipstick traces, Anal. Methods 5 (2013) 53925401.
References [23] C. Rodger, V. Rutherford, D. Broughton, P.C. White, W.E. Smith, The in-situ
analysis of lipsticks by surface enhanced resonance Raman scattering, Analyst
[1] S. Singh, V. Saran, M. Mishra, A.K. Gupta, Forensic examination of lipstick by 123 (1998) 18231826.
the various physio-chemical and instrumental method, Int. J. Soc. Relev. [24] G. Misra, V.K. Mittal, Neutron activation analysis of lipsticks using g-ray
Concern 3 (2015) 17. spectrometry, J. Appl. Spectrosc. 71 (2004) 270274.
[2] M. Lpez-Lpez, N. zbek, C. Garca-Ruiz, Confocal Raman spectroscopy to [25] A.F.L. bin Abdullah, Y. Marimuthu, Ch. Kah Haw, N.F. binti Mohamad Said, N.Z.
trace lipstick with their smudges on different surfaces, Talanta 123 (2014) M. Muslim, N.F.N. Hassan, M.H. Yaacob, Y. Chong Hooi, Forensic discrimination
135139. of lipsticks by thin layer chromatography and gas chromatographymass
[3] F. Salahioglu, M.J. Went, Differentiation of lipsticks by Raman Spectroscopy, spectrometry, Malays. J. Forensic Sci. 2 (2011) 2228.
Forensic Sci. Int. 223 (2012) 148152. [26] J. Manheim, K.C. Doty, G. McLaughlin, I.K. Lednev, Forensic hair differentiation
[4] B. Joshi, K. Verma, J. Singh, A comparison of red pigments in different lipsticks using attenuated total reection fourier transform infrared (ATR FT-IR)
using thin layer chromatography (TLC), J. Anal. Bioanal. Tech. 4 (2013) 14. spectroscopy, Appl. Spectrosc. 70 (2016) 11091117.
[5] Y. Ehara, Y. Marumo, Identication of lipstick smears by uorescence [27] M.C.A. Marcelo, K.C. Mariotti, M.F. Ferrao, R.S. Ortiz, Proling cocaine by ATR-
observation and purge-and-trap gas chromatography, Forensic Sci. Int. 96 FTIR, Forensic Sci. Int. 246 (2015) 6571.
(1998) 110. [28] M. Szafarska, M. Wo zniakiewicz, M. Pilch, J. Zieba-Palus, P. Koscielniak,
[6] D. Li, Z. Wang, L. Wang, X. Xu, H. Zhang, Ultrasonic extraction coupled with Computer analysis of ATR-FTIR spectra of paint samples for forensic purpose, J.
capillary electrophoresis for the determination of azo dyes in lipsticks using Mol. Struct. 924926 (924) (2009) 504.
ionic liquid as dynamic coating and background electrolyte, Chin. J. Chem. 29 [29] C.M. Orphanou, The detection and discrimination of human body uids using
(2011) 147152. ATR FT-IR spectroscopy, Forensic Sci. Int. 252 (2015) e10e16.
138 M. Gadysz et al. / Forensic Science International 280 (2017) 130138

[30] B.R. Kowalski (Ed.), Chemometrics: Mathematics and Statistics in Chemistry, [33] G.W. Milligan, M.C. Cooper, An examination of procedures for determining the
Reidel, Dordrecht, 1984. number of clusters in a data set, Psychometrika 50 (1985) 159179.
[31] D.L. Massart, L. Kaufman, The Interpretation of Analytical Chemical Data by the [34] G. Socrates, Infrared and Raman characteristic group frequencies, Tables and
Use of Cluster Analysis, John Wiley & Sons, New York, 1983. Charts, John Wiley and Sons, Ltd., Chichester, 2001.
[32] R. Mojena, Hierarchical grouping methods and stopping rules: an evaluation,
Comput. J. 20 (1977) 359363.