Talanta 266 (2024) 124954

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Talanta
journal homepage: www.elsevier.com/locate/talanta

Multi-class analysis of 100 drug residues in cosmetics using

high-performance liquid chromatography-quadrupole time-of-flight
high-resolution mass spectrometry
Dan Chen a, 1, Ying Chen a, 1, Yuan Zhang a, 1, Juan Du b, Han Xiao b, Zong Yang c, Jia Xu b, *
a
Guangdong Institute of Sport Science, Guangzhou, 510663, PR China
b
Institute of Maternal and Child Health, Wuhan Children’s Hospital (Wuhan Maternal and Child Healthcare Hospital), Tongji Medical College, Huazhong University of
Science and Technology, Wuhan, 430016, PR China
c
Asia Pacific Technical Support Center of SCIEX, Shanghai, 200050, PR China

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

Keywords: Cosmetics are an important aspect of the lives of many people. With an increasing demand for cosmetics, con­
Prohibited substances sumers pay more attention to their efficacy and composition. To improve their efficacy, prohibited substances,
HPLC–Q–TOF–HRMS such as hormones, glucocorticoids, antibiotics, antifungals and antihistamines, may be added to cosmetics. We
Toner
developed a rapid method for the multi-class analysis of drug residues in toner and lotion cosmetic samples using
Lotion
Cosmetic
high-performance liquid chromatography coupled with quadrupole time-of-flight high-resolution mass spec­
trometry (HPLC–Q–TOF–HRMS). The primary variables in the extraction and purification steps were studied to
minimize the interference of the sample matrix. The non-information-dependent sequential window acquisition
of all theoretical fragment ion spectra (SWATH®) mode was used to improve the data acquisition efficiency. The
secondary product ion peak areas were used for quantification to obtain a satisfactory matrix effects. The vali­
dation experiments confirmed that the developed method exhibited good linearity (5–200 ng/L) with correlation
coefficients (R) ≥ 0.9902. Our developed method was then successfully applied to 92 real cosmetic samples. The
calibration curve established by this method can be used for retrospective quantitative analysis over long du­
rations without re-calibration. This method is efficient and suitable for screening and controlling multi-class
prohibited substances in the cosmetics industry to reduce potential risks.

1. Introduction challenges in the detection of these compounds in cosmetics. According

With the improvement in living standards, the cosmetics industry has
experienced considerable growth owing to the increasing attention paid
to personal image. Cosmetics and personal care products have become
an essential part of our daily lives [1,2]. Functional cosmetics for
repairing damaged skin, delaying skin aging, and whitening or rejuve­
nating skin are gaining popularity [3,4]. Most products are safe and
reliable owing to strict market regulations. However, to artificially in­
crease the efficacies of their products, manufacturers may add illegal
additives, such as hormones, glucocorticoids, antibiotics, antifungals
and antihistamines [5–8]. These additives can adversely affect the safe
use of cosmetics, as well as human health [9–11].
The wide variety and diverse structures of prohibited additives pose
to the Chinese “Hygienic Standard for Cosmetics” legislation, more than
1000 substances are prohibited from use in cosmetics, of which only 4%
can be detected using the established detection methods. Multiple pro­
hibited additives cannot be analyzed using these detection methods, and
it is not feasible to determine the entire composition of each cosmetic
product [12]. Prohibited additives originate from various sources, and
existing detection methods fail to detect multiple unknown prohibited
substances, such as drug derivatives that are present in cosmetics [13].
In addition, cosmetics may contain multiple prohibited additives at low
concentrations, making it challenging to detect all of these substances
using a single detection method; this necessitates the development of
techniques that are more sensitive than those currently in use [14,15].
Cosmetic products contain complex ingredients that cause considerable

* Corresponding author.
E-mail address: xujia0113@hust.edu.cn (J. Xu).
1
These authors contributed equally to this work.

https://doi.org/10.1016/j.talanta.2023.124954
Received 20 March 2023; Received in revised form 25 June 2023; Accepted 14 July 2023
Available online 17 July 2023
0039-9140/© 2023 Elsevier B.V. All rights reserved.
D. Chen et al. Talanta 266 (2024) 124954

Table 1
HPLC-Q-TOF HRMS, parameters for the target compounds.
No. Compound Diagnostic ion Chemical Productions (m/z) Theoreticalmass (m/z)
Formula
ion 1 ion 2 ion 3

1 Oxymetholone [M+H]+ C21H32O3 99.045 107.085 215.181 333.242

2 Boldenone [M+H]+ C19H26O2 121.063 135.116 173.096 287.201
3 Trenbolone [M+H]+ C18H22O2 199.112 253.158 227.143 271.169
4 Stanozolol [M+H]+ C21H32N2O 81.044 121.101 203.155 329.259
5 Metandienone [M+H]+ C20H28O2 121.064 149.132 283.205 301.216
6 Nandrolone phenylpropionate [M+H]+ C27H34O3 105.068 257.189 239.179 407.258
7 Methyltestosterone [M+H]+ C20H30O2 109.064 97.064 285.221 303.232
8 Nandrolone [M+H]+ C18H26O2 109.064 257.190 145.101 275.201
9 Methenolone enanthate [M+H]+ C27H42O3 187.148 83.049 303.232 415.321
10 Testosterone [M+H]+ C19H28O2 97.064 253.195 123.080 289.216
11 Danazol [M+H]+ C22H27NO2 148.076 310.216 120.081 338.211
12 Testosterone cypionate [M+H]+ C27H40O3 97.064 271.206 107.085 413.305
13 Testosterone isocaproate [M+H]+ C25H38O3 97.064 109.064 175.148 387.289
14 Methenolone acetate [M+H]+ C22H32O3 187.147 83.048 303.232 345.242
15 Androstenedione [M+H]+ C19H26O2 97.064 109.064 123.081 287.201
16 Fluticasone propionate [M+H]+ C25H31F3O5S 293.153 205.066 313.159 501.192
17 clobetasol propionate [M+H]+ C25H32ClFO5 278.166 147.080 355.146 467.200
18 Fluocinonide [M+H]+ C26H32F2O7 121.065 337.144 319.133 495.219
19 Prednisolone acetate [M+H]+ C23H30O6 147.080 307.170 385.201 403.212
20 Triamcinolone acetonide acetate [M+H]+ C26H33FO7 339.160 439.212 147.081 477.228
21 Beclomethasone dipropionate [M+H]+ C28H37ClO7 319.169 57.033 291.174 521.230
22 Betamethasone dipropionate [M+H]+ C28H37FO7 279.173 337.179 319.169 505.260
23 Flumethasone [M+H]+ C22H28F2O5 253.122 121.065 235.112 411.198
24 Fludrocortisone acetate [M+H]+ C23H31FO6 239.144 325.180 143.086 423.218
25 Betamethasone valerate [M+H]+ C27H37FO6 279.173 147.080 337.179 477.265
26 Triamcinolone acetonide [M+H]+ C24H31FO6 213.127 147.080 339.159 435.218
27 Triamcinolone [M+H]+ C21H27FO6 225.128 147.081 357.170 395.186
28 Chlormadinone acetate [M+H]+ C23H29ClO4 301.135 267.174 133.101 405.183
29 Medroxyprogesterone [M+H]+ C24H34O4 123.080 327.230 97.064 387.253
30 Lincomycin [M+H]+ C18H34N2O6S 126.128 359.218 287.195 407.221
31 Roxithromycin [M+H]+ C41H76N2O15 679.434 158.116 116.107 837.532
32 Enrofloxacin [M+H]+ C19H22FN3O3 342.161 245.109 316.182 360.172
33 Norfloxacin [M+H]+ C16H18FN3O3 302.130 233.108 205.077 320.140
34 Pefloxacin [M+H]+ C17H20FN3O3 316.145 290.109 233.109 334.156
35 Ciprofloxacin [M+H]+ C17H18FN3O3 314.130 231.057 205.077 332.140
36 Ofloxacin [M+H]+ C18H20FN3O4 318.162 261.104 344.114 362.151
37 Sarafloxacin [M+H]+ C20H17F2N3O3 299.099 368.121 368.121 386.131
38 Enoxacin [M+H]+ C15H17FN4O3 303.125 206.073 234.104 321.136
39 Lomefloxacin [M+H]+ C17H19F2N3O3 265.114 237.084 223.068 352.147
40 Nalidixic acid [M+H]+ C12H12N2O3 187.048 215.080 159.054 233.092
41 Oxolinic acid [M+H]+ C13H11NO5 244.058 216.029 160.039 262.071
42 Flumequine [M+H]+ C14H12FNO3 244.075 202.028 174.035 262.087
43 Danofloxacin [M+H]+ C19H20FN3O3 340.145 255.057 82.065 358.156
44 Difloxacin hydrochloride [M+H]+ C21H19F2N3O3 382.135 299.099 285.084 400.147
45 Orbifloxacin [M+H]+ C19H20F3N3O3 295.106 267.038 254.067 396.153
46 Sparfloxacin [M+H]+ C19H22F2N4O3 292.126 251.087 230.073 393.173
47 Fleroxacin [M+H]+ C17H18F3N3O3 326.147 269.090 222.059 370.137
48 Sulfamerazine [M+H]+ C11H12N4O2S 92.049 156.011 199.098 265.075
49 Sulfapyridine [M+H]+ C11H11N3O2S 156.011 184.087 108.044 250.064
50 Sulfamethoxypyridazine [M+H]+ C11H12N4O3S 92.049 156.011 215.093 281.070
51 Sulfamethoxazole [M+H]+ C10H11N3O3S 92.049 156.011 160.087 254.059
52 Sulfadoxine [M+H]+ C12H14N4O4S 156.076 92.049 245.103 311.081
53 Sulfathiazole [M+H]+ C9H9N3O2S2 156.012 92.050 190.043 256.021
54 Sulfamethizole [M+H]+ C9H10N4O2S2 156.011 92.049 108.044 271.032
55 Trimethoprim [M+H]+ C14H18N4O3 230.116 123.066 81.045 291.145
56 Sulfisoxazole [M+H]+ C11H13N3O3S 156.011 92.049 108.044 268.075
57 Sulfamoxole [M+H]+ C11H13N3O3S 156.011 155.988 108.044 268.075
58 Sulfabenzamide [M+H]+ C13H12N2O3S 156.011 92.049 108.044 277.064
59 Sulfaphenazole [M+H]+ C15H14N4O2S 158.071 92.049 222.034 315.091
60 Sulfamethazine [M+H]+ C12H14N4O2S 124.087 186.033 92.049 279.091
61 Sulfadiazine [M+H]+ C10H10N4O2S 156.011 92.048 158.002 251.060
62 Sulfaquinoxaline [M+H]+ C14H12N4O2S 92.049 156.011 235.099 301.075
63 Sulfachloropyridazine [M+H]+ C10H9ClN4O2S 156.011 130.017 92.049 285.021
64 Sulfameter [M+H]+ C11H12N4O3S 92.049 156.011 215.093 281.070
65 Sulfisomidine [M+H]+ C12H14N4O2S 124.087 186.033 92.049 279.091
66 Sulfamonomethoxine [M+H]+ C11H12N4O3S 215.093 156.011 92.049 281.070
67 Sulfadimethoxine [M+H]+ C12H14N4O4S 156.076 245.103 92.049 311.081
68 Sulfaguanidine [M+H]+ C7H10N4O2S 156.012 163.036 92.050 215.060
69 Sulfanitran [M+H]+ C14H13N3O5S 134.060 198.023 294.055 336.065
70 Sulfapyrazole [M+H]+ C16H16N4O2S 172.086 236.050 156.011 329.107
71 Ipronidazole [M+H]+ C7H11N3O2 109.075 123.091 81.045 170.092
72 Metronidazole [M+H]+ C6H9N3O3 128.045 82.053 111.042 172.072
73 Ornidazole [M+H]+ C7H10ClN3O3 128.045 82.052 128.066 220.048
(continued on next page)

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D. Chen et al. Talanta 266 (2024) 124954

Table 1 (continued )
No. Compound Diagnostic ion Chemical Productions (m/z) Theoreticalmass (m/z)
Formula
ion 1 ion 2 ion 3

74 Desloratadine [M+H]+ C19H19ClN2 259.134 73.065 282.104 311.131

75 Chlorpheniramine maleate [M+H]+ C16H19ClN2 230.071 167.072 118.065 275.131
76 Astemizole [M+H]+ C28H31FN4O 135.080 218.153 308.156 459.255
77 Tripelennamine [M+H]+ C16H21N3 211.121 120.068 119.059 256.181
78 Bromopheniramine [M+H]+ C16H19BrN2 274.021 167.072 245.992 319.080
79 Diphenhydramine [M+H]+ C17H21NO 167.083 166.077 151.054 256.170
80 Methylprednisolone [M+H]+ C22H30O5 161.096 253.159 280.183 375.217
81 Prednisolone [M+H]+ C21H28O5 147.080 173.096 223.112 361.201
82 Dexamethasone [M+H]+ C22H29FO5 237.127 147.080 279.174 393.207
83 Altrenogest [M+H]+ C21H26O2 227.142 251.143 159.080 311.201
84 Progesterone [M+H]+ C21H30O2 97.064 123.081 145.101 315.232
85 Prednisone [M+H]+ C21H26O5 147.080 237.127 265.159 359.185
86 Thiamphenicol [M+H]+ C12H15Cl2NO5S 198.059 307.991 118.064 356.012
87 Betamethasone [M+H]+ C22H29FO5 237.127 147.080 279.174 393.207
88 Beclomethasone [M+H]+ C22H29ClO5 147.081 391.166 211.112 409.178
89 Hydrocortisone [M+H]+ C21H30O5 121.065 327.195 145.101 363.217
90 Cortisone [M+H]+ C21H28O5 163.112 121.066 145.102 361.201
91 Hydroxyprogesterone [M+H]+ C21H30O3 97.064 123.081 313.215 331.227
92 Dexamethasone acetate [M+H]+ C24H31FO6 237.128 147.080 291.175 435.218
93 Griseofulvin [M+H]+ C17H17ClO6 165.053 285.051 69.033 353.079
94 Clotrimazole [M+H]+ C22H17ClN2 165.070 277.078 241.102 345.115
95 Econazole [M+H]+ C18H15Cl3N2O 125.014 193.053 282.985 381.032
96 Fluconazole [M+H]+ C13H12F2N6O 220.069 238.080 169.046 307.111
97 Ketoconazole [M+H]+ C26H28Cl2N4O4 489.146 244.006 82.053 531.156
98 Miconazole [M+H]+ C18H14Cl4N2O 158.975 227.014 123.000 414.993
99 Bifonazole [M+H]+ C22H18N2 243.114 228.092 165.069 311.154
100 Ciclopirox olamine [M+H]+ C21H21N 117.069 141.069 91.054 288.175

matrix interference during the detection of prohibited additives [16,17].
Therefore, it is imperative to establish a rapid and sensitive analytical
technique for screening and confirming multiple known prohibited ad­
ditives and unknown drug derivatives in cosmetic and to establish a
comprehensive integrated analytical strategy for accurate
quantification.
Different analytical techniques such as high performance liquid
chromatography coupled with ultraviolet detector (HPLC–UV) [18,19],
high performance liquid chromatography coupled with diode array
detection (HPLC–DAD) [20,21], gas chromatography tandem mass
spectrometry (GC–MS) [22–24], and high performance liquid chroma­
tography tandem mass spectrometry (HPLC–MS) [25,26] have been
reported for the determination of compounds in cosmetics.
High performance liquid chromatography coupled with high-
resolution mass spectrometry (HPLC–HRMS) is well established for
screening and identifying contaminants in food and the environment. It
can be employed for the accurate qualitative analysis and non-
directional screening of unknown substances owing to its highly accu­
rate resolution, ability to determine isotopic peak distributions, and full-
scan capabilities [27–30]. In HRMS methods, the MS parameters need
not be optimized for each analyte using reference materials. Addition­
ally, HRMS record multi-stage scans of screened target compounds,
which can be further confirmed by matching the obtained fragments
with fragmentation patterns from a mass spectral library [31,32]. It can
effectively differentiate between co-eluting homologous- and
heterologous-compounds, reducing the requirements for chromato­
graphic separation and extensive sample pretreatment. Further, it can
analyze an unrestricted number of compounds, and data on target and
non-target compounds can be collected and retrospectively analyzed
without the need tore-inject samples using a re-extractor [33]. Consid­
ering that unknown prohibited substances in cosmetics are primarily
known drug derivatives, HRMS can be used to effectively screen for the
congeners of prohibited substances owing to its unique, robust data
acquisition and processing capabilities [34–36].
In this study, based on the diversity, concealment, variability, and
unpredictability of prohibited substances in cosmetics, we established a
novel integrated research strategy to screen and confirm known pro­
hibited substances, identify unknown prohibited substances, and
accurately quantify the substances in cosmetics using HRMS combined
with a mass spectral tree similarity-filter technique. A method based on
high-performance liquid chromatography coupled to quadrupole time-
of-flight high-resolution mass spectrometry (HPLC–Q–TOF–HRMS)
was established for rapidly screening and quantitative detection of 100
prohibited substances in toner and lotion samples.
2. Materials and methods
2.1. Chemicals and reagents
The 100 selected analytes were purchased from Anpel (Shanghai,
China). Oasis PRiME HLB solid phase extraction (SPE) purification
cartridge were acquired from Waters (Milford, MA, USA). LC–C18 SPE
purification cartridge was acquired from Merck (Darmstadt, Germany).
HPLC-grade methanol and acetonitrile were supplied by Merck (Darm­
stadt, Germany). HPLC-grade formic acid was purchased from Sigma-
Aldrich (St. Louis, MO, USA). Ultrapure water (18.2 MΩ) was pre­
pared using a Merck Milli-Q system (Lane End, UK). All other solvents
were of analytical grade unless stated otherwise and were used without
further purification.
The standard stock solutions (100 μg/mL for each compound) were
prepared in methanol and stored in a refrigerator to prevent degrada­
tion. Working standard solutions containing all the compounds were
freshly prepared by diluting the stock solutions with ultrapure water to
the desired concentrations.
2.2. Instrumentation
The HPLC–Q–TOF–HRMS system used for analysis consisted of a
Shimadzu Nexera LC-30A UHPLC system (Tokyo, Japan) with a degas­
ser, programmable pump, autosampler and thermostated column oven,
coupled with a Q–TOF high-resolution mass spectrometer (X500R
Q–TOF, AB Sciex, Darmstadt, Germany) equipped with an electrospray
ionization source (ESI). A Vortex-Genie2 mixer obtained from Scientific
Industries (Bohemia, USA) was used for sample extraction.

3
D. Chen et al.
Fig. 1. The effects of the extraction solvent on the target compound recoveries in toner (a) and lotion (b) samples.
2.3. Sample preparation
The cosmetics samples (0.5 g) were accurately weighed into 10 mL
centrifuge tubes and then mixed with 4 mL of acetonitrile. Then, the
mixture ultrasonicated for 10 min and centrifuged at 10000 rpm for 10
min. Finally, the supernatants of the toner samples were filtered through
0.22 μm nylon filters. The supernatants of the lotion samples were
extracted using PRiME HLB SPE cartridges and filtered through 0.22 μm
nylon filters.
2.4. HPLC–Q–TOF analysis
Reversed-phase gradient liquid chromatographic separation was
performed using a Waters Xbridge C18 column (2.1 × 150 mm, 5 μm).
The column temperature was maintained at 40 ◦ C. The mobile phase
comprised ultrapure water containing 0.01% formic acid (mobile phase
A) and acetonitrile (mobile phase B). The linear gradient program was as
follows: 0.0–1.0 min, 3% B; 1.0–1.5 min, linear gradient to 15% B;
1.5–7.0 min, linear gradient to 45% B; 7.0–19.0 min, linear gradient to
4
Talanta 266 (2024) 124954
98% B; 19.0–22.0 min, 98% B. The total run time was 22 min. The in­
jection volume was 10 μL. The gradient flow rate was set at a constant
0.5 mL/min.
The positive mode ESI (ESI+) parameters were set as follows: ion
spray voltage floating , 5.5 kV; source temperature , 500 ◦ C; curtain gas ,
55 psi; declustering potential , 65 V; nebulising gas , 55 psi; and heater
gas , 55 psi. The SCIEX OS1.5 software (SCIEX) was used for instrument
control and data processing.
3. Results and discussion
3.1. Establishment of the standard spectral library
A mixed standard solution containing 100 target compounds
(Table 1) with a mass concentration of approximately 50 μg/L per
compound was prepared and loaded into the instrument for testing. Data
were acquired in the information-dependent acquisition (IDA) mode to
obtain accurate relative molecular masses, retention times, isotopic
peaks, and characteristic fragment ion (MS/MS) mass spectra of the 100
D. Chen et al.
Fig. 2. The effects of the purification cartridges on the target compound recoveries in toner (a) and lotion (b) samples.
compounds and thereby establish a standard spectral library of the
compounds.
The experimental samples were extracted and analyzed, collected
data were imported into the data analysis module of SCIEX OS1.5, and
sample mass spectra were compared with those in the standard spectral
library. According to European Union guidelines (SANTE/11945/2015),
two ions with a mass accuracy of no greater than 5 ppm and an anion
abundance ratio within 30% of the standards are required for compound
identification [37]. Therefore, the following identification criteria were
considered for the compounds: the retention time of the extracted ion
chromatographic peak in the test sample must correspond to that of the
standard; the exact mass deviation must not exceed 5 ppm; the differ­
ence in the isotopic abundance ratio must not exceed 10%; the matching
score between the MS/MS spectrum of the compound and that of the
mass spectral library must be more than 70 points; and the difference in
the abundance ratio between the two qualitative product ions must be
less than 20%. The concentrations of test compounds in the samples
were calculated from the corresponding calibration curves of qualitative
ion peak areas.
5
Talanta 266 (2024) 124954
3.2. Optimization of the HPLC–Q–TOF–HRMS method
3.2.1. Chromatographic method
Preliminary positive and negative ion scans of the 100 compounds
indicated that the ESI + mode produced the strongest molecular ion
peaks [M+H]+. Therefore, the ESI + mode was applied in subsequent
experiments. Formic acid was added to the organic mobile phase to
improve the ionization efficiency. Different mobile phase systems (0.1%
formic acid in acetonitrile–water and acetonitrile–water) were evalu­
ated and the results were compared to determine the optimal peak
separation and response intensity. Upon addition of formic acid, the
precursor ion peak intensities and resolutions of most compounds
improved [38]. Therefore, 0.1% formic acid in acetonitrile was selected
as the optimal mobile phase. The elution gradient was adjusted based on
the presence of representing the compounds.
3.2.2. Mass spectrometric method
The non-information-dependent sequential window acquisition of all
theoretical fragment ion spectra (SWATH®) mode was used to improve
the data acquisition efficiency [39]. Unlike the IIDA mode, in which a
D. Chen et al. Talanta 266 (2024) 124954

Fig. 3. The matrix effect in toner samples quantified by the precursor ion or secondary product ion peak areas.

Fig. 4. The matrix effect in lotion samples quantified by the precursor ion or secondary product ion peak areas.

secondary scan is triggered only when the determination criteria are mass, and isotopic peak ratios of the precursor ions. The molecular
met, the SWATH® mode acquires continuous, panoramic, compositions and fragmentation pathways of the compounds were
high-throughput, non-dependent mass spectral data [40]. In addition, it determined from the secondary ion fragment spectra.
acquires secondary fragments of all the precursor ions within each
window via software deconvolution and intelligent window partitioning
3.3. Optimization of sample preparation procedures
of the precursor ion mass ranges [41]. First, a blank matrix sample was
analyzed in the SWATH® mode to obtain the total ion current. Next, all
3.3.1. Extraction optimization
compounds detected within 1–25 min were sorted according to their
Cosmetic samples are complex matrices comprising functional in­
mass numbers and subjected to continuous uniform segmentation using
gredients (e.g., preservatives, colorants, UV–light filters, antioxidants,
software according to their relative molecular masses. SWATH® scan
fragrances, and hair-coloring materials) and essential raw materials (e.
parameters were subsequently set based on these segments. The mo­
g., waxes, oils, polymers, and fatty materials) [42]. The primary chal­
lecular structures of the target compounds were determined using
lenges in extracting cosmetic samples for analysis include the efficient
chromatographic separation and MS, and their elemental compositions
extraction of target analytes from complex matrices and the subsequent
were determined based on the parent ion peak retention time, exact
cleanup steps to eliminate matrix interference and facilitate

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D. Chen et al. Talanta 266 (2024) 124954

Table 2
Analytical performance of HPLC-Q-TOF HRMS method for the target compounds.
No. Linear equation R Toner Lotion

RT (min) LOD (μg/kg) Linear range (ng/mL) RT (min) LOD (μg/kg) Linear range (ng/mL)

1 y= 27.08189 x - 62.35367 0.9996 14.68 9.91 10–200 14.67 39.64 10–200

2 y= 584.89610 x + 423.47451 0.9987 9.49 9.43 10–200 9.51 37.70 10–200
3 y= 238.96157 x + 1439.44725 0.9920 9.16 9.80 10–200 9.17 39.20 10–200
4 y= 168.90498 x + 226.06286 0.9956 9.82 11.57 10–200 9.83 11.57 5–100
5 y= 996.20172 x + 1353.10030 0.9941 10.08 10.73 10–200 10.10 21.45 5–100
6 y= 420.13998 x + 288.31401 0.9956 17.84 10.34 10–200 17.84 10.34 10–200
7 y= 312.39464 x + 5188.01097 0.9907 11.09 10.43 10–200 11.11 20.85 10–200
8 y= 287.25172 x + 182.06039 0.9978 9.76 10.62 10–200 9.76 21.23 10–200
9 y= 265.45929 x + 320.65294 0.9977 20.98 10.33 10–200 20.98 41.30 10–200
10 y= 212.98169 x + 147.39801 0.9982 10.40 11.75 10–200 10.39 23.50 5–100
11 y= 471.73041 x + 720.31552 0.9984 13.62 3.88 10–200 13.64 7.77 10–200
12 y= 597.89087 x + 1170.56753 0.9983 20.75 20.22 10–200 20.76 40.44 10–200
13 y= 341.70871 x + 239.62626 0.9976 19.40 10.61 10–200 19.41 21.22 10–200
14 y= 135.97375 x + 368.57177 0.9932 15.64 9.64 10–200 15.65 19.29 10–200
15 y= 700.60294 x + 708.65550 0.9961 9.49 10.12 10–200 11.18 20.24 10–200
16 y= 518.06903 x + 584.40579 0.9911 13.34 3.93 10–200 13.34 9.83 10–200
17 y= 88.43076 x - 13.40204 0.9972 13.25 4.17 10–200 13.24 20.85 10–200
18 y= 284.81700 x + 335.96514 0.9950 11.81 3.88 10–200 11.82 7.76 10–200
19 y= 253.67900 x + 247.93056 0.9967 9.38 10.75 10–200 9.39 21.50 10–200
20 y= 433.85693 x - 72.67058 0.9996 11.72 10.75 10–200 11.72 10.75 10–200
21 y= 801.21859 x + 1242.09872 0.9903 14.28 9.85 10–200 14.28 9.85 10–200
22 y= 430.95400 x + 147.02846 0.9965 13.72 9.98 10–200 13.74 19.96 10–200
23 y= 82.07114 x + 588.15390 0.9902 8.49 10.83 10–200 8.50 21.65 10–200
24 y= 428.98558 x + 553.31485 0.9909 9.59 11.29 10–200 9.61 22.58 10–200
25 y= 504.70956 x + 453.58872 0.9963 12.48 10.69 10–200 12.48 21.39 10–200
26 y= 58.93621 x - 118.25372 0.9984 9.01 10.48 10–200 9.02 10.48 10–200
27 y= 324.91205 x + 3676.66699 0.9950 5.61 40.74 10–200 5.60 40.74 10–200
28 y= 735.76826 x + 341.47555 0.9975 13.77 12.43 10–200 n.d. n.d. n.d.
29 y= 89.30334 x + 58.93598 0.9993 13.91 11.30 10–200 13.91 22.59 10–200
30 y= 457.92317 x + 425.63554 0.9975 3.83 10.12 10–200 n.d. n.d. n.d.
31 y= 136.43012 x + 251.54468 0.9920 8.26 9.66 10–200 8.28 19.32 10–200
32 y= 750.61986 x + 219.56528 0.9959 4.71 10.00 10–200 4.69 20.00 10–200
33 y= 136.34611 x + 250.94138 0.9921 4.28 20.00 10–200 4.28 40.00 10–200
34 y= 16.07825 x + 280.28072 0.9961 4.35 40.00 5–100 3.76 40.00 5–100
35 y= 535.04842 x + 20.01385 0.9993 4.40 10.00 10–200 4.40 40.00 5–100
36 y= 94.89931 x + 135.40996 0.9981 4.31 10.00 10–200 3.71 40.00 5–100
37 y= 57.00847 x + 185.30096 0.9931 5.07 10.00 10–200 5.11 40.00 5–100
38 y= 146.87265 x + 480.43644 0.9929 4.07 40.00 10–200 3.47 40.00 10–200
39 y= 54.16358 x + 207.72666 0.9996 4.57 10.00 10–200 4.57 40.00 10–200
40 y= 265.30109 x - 627.41024 0.9980 8.41 10.00 10–200 8.43 10.00 10–200
41 y= 86.94341 x + 129.64599 0.9986 6.93 10.00 10–200 6.95 10.00 10–200
42 y= 93.84070 x + 380.62933 0.9964 8.66 10.00 10–200 8.65 10.00 10–200
43 y= 367.52257 x + 210.12331 0.9989 4.55 10.00 10–200 4.55 40.00 10–200
44 y= 183.10928 x + 772.52048 0.9959 5.17 10.00 10–200 5.19 40.00 10–200
45 y= 872.81500 x + 751.72005 0.9968 4.79 10.00 10–200 4.84 40.00 10–200
46 y= 552.87023 x + 478.04182 0.9952 5.15 10.00 10–200 5.21 40.00 10–200
47 y= 164.54894 x + 164.52326 0.9914 4.33 10.00 10–200 3.76 40.00 10–200
48 y= 28.56419 x + 158.65058 0.9991 4.84 10.00 10–200 4.84 20.00 10–200
49 y= 2738.84112 x + 1618.11487 0.9943 4.58 10.00 10–200 4.60 20.00 10–200
50 y= 392.46666 x + 562.87826 0.9961 5.35 10.00 10–200 5.33 20.00 10–200
51 y= 825.57969 x + 48.88933 0.9930 6.42 10.00 10–200 6.44 20.00 10–200
52 y= 825.57969 x + 48.88933 0.9930 7.40 4.00 10–200 7.40 10.00 10–200
53 y= 1014.52335 x + 987.27027 0.9970 4.38 10.00 10–200 4.39 40.00 10–200
54 y= 326.90522 x + 272.19147 0.9917 5.24 10.00 10–200 5.24 20.00 10–200
55 y= 1892.83624 x + 1215.19537 0.9978 4.24 10.00 10–200 4.23 10.00 10–200
56 y= 2100.58624 x + 724.06847 0.9920 6.74 10.00 10–200 6.73 20.00 10–200
57 y= 48.44243 x + 228.20084 0.9956 6.74 10.00 10–200 6.73 20.00 10–200
58 y= 961.41649 x + 482.78613 0.9985 7.26 10.00 10–200 7.27 20.00 10–200
59 y= 552.87023 x + 478.04182 0.9952 7.56 10.00 10–200 7.57 20.00 10–200
60 y= 1892.83624 x + 1215.19537 0.9978 5.33 40.00 10–200 5.33 40.00 10–200
61 y= 552.87023 x + 478.04182 0.9952 4.27 10.00 10–200 4.28 20.00 10–200
62 y= 10.56147 x - 507.70370 1.0000 7.40 10.00 10–200 7.42 20.00 10–200
63 y= 75.73024 x + 75.86250 0.9973 6.09 10.00 10–200 6.10 20.00 10–200
64 y= 245.47364 x + 381.05625 0.9961 5.35 10.00 10–200 5.33 20.00 10–200
65 y= 47.77515 x + 140.68735 0.9998 5.33 40.00 10–200 5.33 40.00 10–200
66 y= 131.89681 x + 89.57149 0.9958 5.35 10.00 10–200 5.33 20.00 10–200
67 y= 344.87787 x + 120.10910 0.9948 7.40 4.00 10–200 7.40 10.00 10–200
68 y= 277.75810 x + 793.50489 0.9920 1.50 40.00 10–200 1.48 40.00 10–200
69 y= 204.45954 x + 54.98299 0.9978 8.66 4.00 10–200 8.65 20.00 10–200
70 y= 3725.04890 x + 4006.84843 0.9913 7.98 10.00 10–200 7.99 10.00 10–200
71 y= 222.95072 x + 848.03048 0.9995 6.97 10.00 10–200 6.98 10.00 10–200
72 y= 420.65997 x + 272.83098 0.9921 3.79 10.00 10–200 3.81 40.00 10–200
73 y= 1668.36890 x + 983.50105 0.9996 5.58 10.00 10–200 5.58 20.00 10–200
(continued on next page)

7
D. Chen et al. Talanta 266 (2024) 124954

Table 2 (continued )
No. Linear equation R Toner Lotion

RT (min) LOD (μg/kg) Linear range (ng/mL) RT (min) LOD (μg/kg) Linear range (ng/mL)

74 y= 184.31934 x + 126.19590 0.9995 5.28 40.00 10–200 5.28 40.00 10–200

75 y= 1011.56745 x + 96.53339 0.9956 5.71 20.00 10–200 5.73 10.00 10–200
76 y= 224.66415 x + 578.48735 0.9941 5.81 40.00 10–200 5.82 40.00 10–200
77 y= 723.73774 x + 318.46557 0.9944 5.57 10.00 10–200 6.45 10.00 5–100
78 y= 51.46289 x + 235.84566 0.9995 5.97 10.00 10–200 5.97 10.00 10–200
79 y= 1011.56745 x + 96.53339 0.9956 6.85 10.00 10–200 6.86 20.00 5–100
80 y= 16.99505 x + 31.04191 0.9914 8.27 10.73 10–200 8.28 21.46 10–200
81 y= 27.08189 x - 62.35367 0.9996 7.64 10.78 10–200 7.44 10.78 10–200
82 y= 584.89610 x + 423.47451 0.9987 8.48 11.36 10–200 8.50 45.44 10–200
83 y= 238.96157 x + 1439.44725 0.9920 11.75 11.96 10–200 11.77 23.91 5–100
84 y= 168.90498 x + 226.06286 0.9956 13.66 11.81 10–200 13.66 11.81 10–200
85 y= 996.20172 x + 1353.10030 0.9941 7.47 11.03 10–200 7.47 11.03 10–200
86 y= 420.13998 x + 288.31401 0.9956 5.00 11.20 10–200 n.d. n.d. n.d.
87 y= 312.39464 x + 5188.01097 0.9907 8.48 10.49 10–200 8.50 20.98 10–200
88 y= 287.25172 x + 182.06039 0.9978 8.76 9.90 10–200 8.77 19.80 10–200
89 y= 265.45929 x + 320.65294 0.9977 7.55 10.30 10–200 7.57 41.20 10–200
90 y= 212.98169 x + 147.39801 0.9982 7.64 10.10 10–200 7.65 20.20 10–200
91 y= 471.73041 x + 720.31552 0.9984 11.3 11.94 10–200 11.30 23.87 10–200
92 y= 597.89087 x + 1170.56753 0.9983 9.01 10.20 10–200 9.02 10.20 10–200
93 y= 341.70871 x + 239.62626 0.9976 9.81 10.00 10–200 9.83 10.00 10–200
94 y= 135.97375 x + 368.57177 0.9932 8.49 10.00 10–100 8.50 40.00 10–200
95 y= 700.60294 x + 708.65550 0.9961 9.36 4.00 10–200 9.39 20.00 10–200
96 y= 518.06903 x + 584.40579 0.9911 5.47 10.00 10–200 5.48 20.00 10–200
97 y= 88.43076 x - 13.40204 0.9972 7.63 10.00 10–100 7.64 40.00 2–50
98 y= 284.81700 x + 335.96514 0.9950 10.05 10.00 10–200 10.07 10.00 2–50
99 y= 253.67900 x + 247.93056 0.9967 8.58 10.00 10–200 8.58 40.00 2–50
100 y= 433.85693 x - 72.67058 0.9996 7.96 10.00 10–200 7.67 10.00 2–50

n.d.: Not detected.

instrument-based testing. Solvent extraction has been employed exten­
sively as an effective technique for extracting cosmetic samples [43,44].
According to the Chinese “Hygienic Standard for Cosmetics”, there are
more than 1000 substances are banned in cosmetics, and we examined
100 prohibited substances with diverse chemical properties in this
study. Therefore, a broad-spectrum extraction solvent (acetonitrile or
methanol) was selected for the extraction of multiple compounds. To
simplify the experimental steps and maximize the extraction rate, 27
representative compounds were selected from the 100 target compounds
for method development and optimization. The effects of the extraction
solvent on the recovery of the target compound from the toner and
lotion samples were compared experimentally (Fig. 1). An equal volume
of acetonitrile or methanol was added to the samples before vortexing,
and they were allowed to rest. Particle precipitation was observed at the
bottom of the centrifuge tubes in the acetonitrile group, possibly
because the toner and lotion samples contained glycerol, low-carbon
fatty acids, moisture, oligosaccharides, amino acids, and plant ex­
tracts. Acetonitrile can alter the solubility of oligosaccharides, poly­
saccharides, and amino acids, causing them to precipitate out of
solution, which can facilitate the partial removal of matrix components
in cosmetics. In the toner samples, the extraction efficiencies of the
target compounds were similar when acetonitrile or methanol was used
as the extraction solvent, whereas in the lotion samples, acetonitrile had
a higher extraction efficiency for the target compounds than did meth­
anol. Therefore, acetonitrile was used as the extraction solvent.
3.3.2. Optimization of purification conditions
Acetonitrile can be used to extract salt and lipid components from
samples as well as the target compounds. Therefore, the extract was
purified using a solid-phase extraction (SPE) cartridge to reduce the
effects of extracted compounds on the results. Impurities (e.g., lipids)
were retained on the cartridge, while the target compounds and
extraction solvent passed through the cartridge. The extract was then
filtered through a membrane filter and analyzed. This protocol elimi­
nates the need for additional steps, such as elution and solvent exchange,
and achieves a simple and rapid purification. The purification effects of
the C18 and PRiME HLB SPE cartridges were investigated. As shown in
Fig. 2, after purifying the toner samples using these two cartridges, the
peak areas of the target compounds were significantly reduced. Because
that the impurities in the toner samples could be removed using only
acetonitrile and centrifugal freezing, the toner samples were not further
purified using SPE. Perphenazine was not recovered from the lotion
samples purified using the C18 SPE cartridge. However, after treatment
with the PRiME HLB SPE cartridge, peaks corresponding to all the target
compounds were detected, with improved response intensities for most
of the compounds. In addition, the chromatograms of the HLB-purified
PRiME extracts contained fewer interfering peaks adjacent to the
target compounds than did those of the unpurified extracts (Fig. 2).
Therefore, the PRiME HLB SPE cartridge was further used to purify the
lotion extracts.
3.4. Method validation
3.4.1. Matrix effects (MEs)
After HPLC–Q–TOF–HRMS data acquisition, quantification was
performed based on the peak area of the precursor or secondary product
ion. Precursor ions are conventionally used for quantification because of
their convenience; however, they can be adversely affected by MEs or
interference from adjacent peaks. By contrast, quantification using
secondary product ions is based on the highly selective SWATH® mode.
The standards prepared in the blank matrix solution and pure solvent
were quantitatively analyzed using the aforementioned quantification
methods, and the respective MEs were determined and compared. ME
changes within 20% are acceptable according to the SANTE/11945/
2015 regulation [45]. There were more compounds with a satisfactory
ME (|ME|≤20%) in the toner and lotion samples when the
high-precision secondary fragment quantification method was used than
when the precursor ion peak area was used, although a few compounds
with moderate MEs (20% < |ME|<50%) were observed (Figs. 3 and 4).
Therefore, the secondary product ion peak areas were selected for
quantification. In addition, because cosmetic matrices can vary notably,
pure solvent calibration curves were used for quantification.

8
D. Chen et al. Talanta 266 (2024) 124954

Table 3
Accuracy and recovery of HPLC-Q-TOF HRMS method for the target compounds.
No. Toner Lotion

Low concentration Medium concentration High concentration Low concentration Medium concentration High concentration

Recovery RSD Recovery RSD Recovery RSD Recovery RSD Recovery RSD Recovery RSD
(%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%)

1 80.64 4.89 76.31 7.94 69.40 6.58 49.81 13.37 34.97 24.56 35.25 31.04
2 80.10 2.61 81.85 2.61 84.43 0.49 59.44 23.74 59.42 11.93 80.62 13.47
3 87.62 3.38 92.19 3.58 92.54 3.78 55.63 34.00 63.55 15.83 96.41 11.48
4 67.44 5.09 74.74 5.08 75.88 8.22 36.24 26.71 43.44 26.90 56.71 16.17
5 76.95 4.29 89.95 6.18 88.39 5.95 42.45 28.08 51.44 12.61 78.74 14.32
6 67.56 9.72 64.84 14.50 60.52 0.54 33.60 6.01 37.75 16.25 32.93 10.50
7 75.66 2.83 84.11 6.73 89.83 5.17 34.13 34.68 45.11 16.99 81.26 10.43
8 86.42 7.91 85.02 4.84 82.13 2.83 53.89 12.29 50.32 8.29 63.74 10.81
9 69.23 4.93 59.87 16.10 54.91 4.63 52.77 35.84 40.66 5.65 26.38 8.61
10 80.11 12.11 91.28 2.37 95.10 5.27 45.01 24.95 47.91 10.17 69.17 10.01
11 84.39 2.53 79.76 7.24 76.14 3.26 34.45 27.87 42.32 26.75 50.56 15.29
12 55.04 3.74 57.72 17.07 50.24 5.68 53.79 23.36 39.65 22.00 33.12 19.84
13 83.60 7.89 64.99 22.38 49.73 0.74 50.00 16.78 36.49 9.27 23.88 19.33
14 81.95 11.26 75.41 9.56 71.97 3.44 52.51 13.19 43.04 24.71 39.49 1.25
15 80.11 2.62 81.85 2.61 84.43 0.49 27.68 17.76 39.55 11.52 60.83 3.19
16 81.58 7.32 80.91 9.94 75.64 2.79 58.51 25.19 53.03 27.90 58.68 26.69
17 74.05 6.43 84.83 6.64 89.13 7.57 48.59 23.74 51.63 20.96 58.98 18.71
18 81.97 5.16 82.94 2.35 80.43 0.70 39.63 21.42 47.62 22.72 65.92 24.83
19 83.14 0.82 91.27 7.48 96.07 2.83 56.48 26.80 62.89 18.63 92.65 26.18
20 78.31 6.56 86.19 3.03 83.93 4.63 33.57 33.91 46.56 19.46 64.97 15.85
21 83.80 7.01 81.49 7.90 74.87 4.16 55.25 24.29 50.57 31.37 55.20 24.46
22 85.60 8.43 91.73 6.08 88.55 2.99 46.45 35.85 50.55 36.03 57.87 23.34
23 89.35 8.33 95.07 3.59 98.01 2.91 83.60 25.27 76.73 20.43 101.79 18.83
24 83.57 3.48 88.89 4.60 93.35 3.78 52.80 18.74 53.80 16.01 73.12 18.36
25 78.05 1.55 85.96 7.16 86.20 6.14 55.04 24.70 50.62 20.41 67.65 21.00
26 77.50 3.27 85.86 7.32 88.44 4.16 56.80 35.55 69.82 21.28 91.99 18.24
27 95.95 6.82 108.97 7.41 90.05 1.43 48.43 28.86 70.85 19.95 90.29 27.88
28 83.35 12.06 85.30 7.87 79.66 2.05 n.d. n.d. n.d. n.d. n.d. n.d.
29 75.66 7.99 81.81 8.98 84.74 2.46 47.97 18.90 49.63 13.92 55.44 12.76
30 75.92 5.46 64.03 16.89 65.95 8.38 n.d. n.d. n.d. n.d. n.d. n.d.
31 51.04 13.22 69.97 6.90 73.00 2.74 50.56 19.32 69.52 12.87 90.82 18.51
32 62.78 3.36 54.17 15.54 53.43 7.05 56.91 26.93 35.45 64.07 55.57 16.39
33 51.45 28.18 54.00 16.73 54.20 7.95 56.61 23.97 48.71 31.82 62.56 25.25
34 25.52 15.52 36.49 18.29 44.48 14.23 37.92 28.75 50.52 13.04 52.25 11.03
35 49.81 14.75 48.73 9.86 55.32 11.58 34.16 29.44 41.05 28.69 57.55 21.15
36 61.89 8.20 55.87 11.86 57.89 8.70 44.48 10.81 49.10 9.12 55.96 12.34
37 49.91 8.79 56.48 3.94 60.82 5.99 63.04 20.88 58.67 35.11 73.16 13.88
38 34.72 0.51 42.24 15.31 58.40 8.84 51.63 15.90 48.22 37.12 63.57 13.77
39 72.83 7.82 65.60 15.45 63.96 3.33 58.27 20.32 59.80 12.35 71.19 17.78
40 64.72 7.27 94.87 13.73 96.79 3.62 53.84 13.57 83.72 12.31 100.96 14.10
41 60.72 14.34 68.99 45.58 61.77 4.50 72.51 11.64 77.28 11.24 82.93 10.45
42 66.70 1.64 76.51 11.30 76.83 1.00 59.33 27.24 73.48 11.15 97.55 12.60
43 59.97 7.65 45.53 12.70 39.94 7.35 48.64 30.62 41.94 20.73 46.89 26.33
44 111.22 0.55 72.02 4.08 45.97 4.42 70.48 13.71 70.98 17.66 85.84 18.69
45 72.56 4.53 66.97 9.47 68.04 7.55 70.96 22.88 64.86 20.66 82.65 15.39
46 55.80 4.63 65.25 12.03 68.01 3.75 62.96 24.69 71.08 14.68 72.08 9.63
47 56.55 6.99 58.17 15.65 61.25 13.96 46.64 35.65 56.76 13.00 62.36 7.21
48 59.63 15.11 67.27 17.76 70.98 5.17 32.80 21.85 51.49 33.08 68.44 6.07
49 62.77 10.53 69.17 17.91 72.02 6.92 41.41 43.78 57.66 38.46 73.53 31.05
50 61.74 8.11 74.12 13.28 74.31 3.22 41.09 39.27 68.35 17.13 84.81 6.77
51 75.69 14.48 85.07 12.96 81.74 2.84 85.47 21.97 77.94 13.71 82.80 12.96
52 73.02 4.08 89.48 23.26 89.51 3.37 61.95 28.11 71.49 12.73 89.48 3.93
53 69.86 9.29 80.11 3.99 75.81 2.58 19.84 19.07 60.72 30.19 75.91 27.00
54 70.21 5.04 81.70 6.50 83.47 2.73 62.67 19.33 75.19 19.88 95.55 12.13
55 64.37 8.95 62.67 16.04 62.75 5.10 69.04 29.92 76.75 16.78 77.46 15.81
56 85.43 8.63 86.37 10.27 78.24 2.22 90.21 19.59 87.50 13.85 98.61 12.94
57 85.43 8.63 86.37 10.27 78.24 2.22 90.21 19.59 87.50 13.85 98.61 12.94
58 88.49 3.45 93.00 12.67 85.15 1.56 91.63 20.93 92.62 13.07 97.33 6.02
59 79.31 3.31 90.42 7.65 88.72 4.49 79.81 29.00 81.66 32.87 99.81 29.38
60 66.23 7.71 73.26 12.01 71.14 3.43 60.91 30.93 77.25 20.18 85.23 9.31
61 48.76 6.43 61.38 1.99 86.31 6.83 31.81 16.53 59.38 45.08 82.76 41.93
62 79.57 5.42 92.87 11.04 90.41 0.56 59.20 33.40 71.74 20.71 95.64 16.23
63 83.62 1.92 88.21 7.12 80.76 3.73 98.99 22.66 93.81 6.40 98.96 4.06
64 61.74 8.11 74.12 13.28 74.31 3.22 50.83 31.83 73.37 16.01 86.69 6.65
65 66.23 7.71 73.26 12.01 71.14 3.43 61.39 30.82 77.57 20.14 85.44 9.31
66 61.74 8.11 74.12 13.28 74.31 3.22 45.52 35.99 71.33 16.66 86.76 6.72
67 73.02 4.08 89.48 23.26 89.51 3.37 61.95 28.11 71.49 12.73 89.48 3.93
68 103.41 11.84 85.35 7.83 66.31 11.02 92.59 21.37 76.74 8.75 87.31 23.75
69 97.92 7.22 97.33 5.30 88.46 4.99 62.88 16.83 62.08 6.92 69.05 4.87
70 93.03 13.45 120.27 21.61 111.21 7.44 75.23 68.95 94.33 34.13 107.97 24.58
(continued on next page)

9
D. Chen et al. Talanta 266 (2024) 124954

Table 3 (continued )
No. Toner Lotion

Low concentration Medium concentration High concentration Low concentration Medium concentration High concentration

Recovery RSD Recovery RSD Recovery RSD Recovery RSD Recovery RSD Recovery RSD
(%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%)

71 101.41 3.34 97.41 13.54 89.52 1.04 91.25 31.54 83.69 20.02 88.31 32.23
72 88.57 8.79 101.43 0.67 88.24 6.87 100.05 6.26 100.69 36.62 115.46 38.04
73 89.17 2.55 92.11 4.33 89.57 3.34 94.85 32.58 87.29 23.33 96.09 27.96
74 29.56 13.09 43.85 20.77 61.56 4.01 68.19 11.78 44.90 11.42 62.16 17.42
75 88.24 5.48 94.42 9.46 104.16 2.91 70.13 11.20 84.81 7.68 81.32 15.55
76 58.19 5.50 60.48 8.29 56.62 9.76 39.20 7.45 49.55 13.40 91.68 8.91
77 46.11 7.97 56.22 18.73 61.64 2.77 47.23 19.57 68.86 8.17 70.34 9.88
78 60.24 13.20 66.76 12.66 72.68 1.30 27.87 14.07 61.18 6.85 86.06 7.70
79 74.32 11.63 67.58 25.88 54.06 2.28 48.21 23.17 68.88 6.63 85.23 3.81
80 85.00 3.20 88.51 6.20 86.87 4.20 75.36 21.20 75.71 22.26 88.41 13.32
81 83.39 5.13 91.37 3.07 92.32 2.87 86.45 10.64 83.22 14.30 94.12 8.59
82 83.99 4.27 96.97 3.31 98.52 0.53 69.31 16.83 71.71 10.80 93.80 11.04
83 66.23 9.05 92.60 8.72 86.22 4.00 36.75 28.47 47.69 15.73 64.64 9.50
84 79.10 7.34 78.20 6.88 73.87 6.77 35.76 9.34 41.62 18.82 52.92 17.37
85 83.21 5.14 94.73 10.38 96.11 3.01 84.77 17.65 81.55 14.55 87.76 13.64
86 90.98 11.36 60.02 15.74 74.78 10.33 n.d. n.d. n.d. n.d. n.d. n.d.
87 83.98 4.27 96.97 3.31 98.52 0.53 70.72 18.44 75.01 10.90 92.07 8.05
88 88.04 2.46 88.96 6.10 89.92 0.82 65.09 27.22 70.61 18.69 98.72 18.27
89 90.10 4.23 88.86 7.20 89.38 3.51 87.23 21.84 75.87 11.94 87.62 8.17
90 83.39 5.13 91.37 3.07 92.32 2.87 70.99 19.29 73.38 9.11 92.04 12.02
91 79.92 2.20 83.82 7.01 88.84 1.39 21.52 31.56 41.76 17.91 75.86 9.00
92 77.50 3.27 85.86 7.32 88.44 4.16 56.80 35.55 69.82 21.28 87.55 10.41
93 79.36 6.45 84.48 8.46 81.69 5.00 42.45 18.95 52.37 5.71 73.01 11.56
94 77.22 20.74 71.31 13.28 72.14 8.56 27.04 29.21 53.27 25.25 63.78 7.75
95 55.82 4.98 76.05 14.33 82.48 5.29 34.32 34.46 50.01 23.50 55.02 14.45
96 91.44 1.38 89.18 3.24 89.56 5.06 65.39 11.67 74.70 5.99 82.93 4.96
97 69.51 4.01 62.77 7.93 51.66 2.37 16.75 12.50 29.07 29.06 49.40 20.44
98 40.49 17.77 71.83 9.78 83.21 1.91 43.79 7.65 49.87 11.07 46.20 6.50
99 53.73 10.63 74.59 13.51 83.36 2.73 52.00 9.33 56.73 4.33 54.95 2.22
100 57.22 11.67 78.28 9.20 89.10 2.74 49.79 17.83 53.24 9.40 72.99 9.22

n.d.: Not detected.

3.4.2. Calibration curves and limits of detection (LODs)
For the toner samples, the calibration curves were linear within the
range of 10–200 ng/L for 97 prohibited substances, 10–100 ng/L for two
prohibited substances, and 5–100 ng/L for pefloxacin. For the lotion
samples, the curves were linear within the ranges of 10–200, 5–100 and
2–50 ng/L for 83, 10 and four prohibited substances, respectively. Good
linearity was obtained for all target substances with correlation coeffi­
cient (R) of 0.9902–1.0000. Compounds 28, 30 and 86 were not detected
in the lotion samples using the proposed method. These compounds may
have been completely adsorbed by the PRiME HLB SPE cartridges.
The LODs were determined by the standard addition method and
calculated from the MS data for each substance at a signal-to-noise ratio
of 3 (Table 2). The LODs of the 100 prohibited substances in the toner
and lotion samples were 3.88–40.74 and 7.76–45.44 mg/kg, respec­
tively. In the toner samples, the LODs of the 88 target substances were
lower than 12 mg/kg. In the lotion samples, the LODs of 20 target
substances were below 20 mg/kg, and those of 45 target substances were
20–40 mg/kg.
and recoveries of less than 50% for 38 compounds. The recoveries of the
medium-concentration spikes ranged from 29.07% to 100.69%, with
RSDs of 4.33%–64.07% and recoveries of less than 50% for 26 com­
pounds. The recoveries of the high-concentration spikes ranged from
23.88% to 115.46% with RSDs of 1.25%–41.93% and recoveries of less
than 50% for eight compounds. The complexity of the sample matrix
affected the recovery; specifically, a large effect on low-concentration
spikes and a small effect on high-concentration spikes were observed.
3.5. Application of our optimized method to cosmetic samples
In total, 92 commercially available toner and lotion samples were
analyzed for 100 prohibited substances. No positive samples were
detected, possibly because the selected samples were well-known
branded products manufactured through strict quality control processes.
3.6. Retrospective analysis

3.4.3. Accuracy and recovery
Blank cosmetic samples were spiked with a mixed reference solution
to investigate the recoveries; the added concentrations were 0.10, 0.20,
and 0.60 mg/kg (Table 3). For the toner samples, the recoveries of the
low-concentration spikes ranged from 25.52% to 111.22%, with relative
standard deviations (RSDs) of 0.51%–28.18% and recoveries of less than
50% for seven compounds. The recoveries of the medium-concentration
spikes ranged from 36.49% to 120.27% with RSDs of 0.67%–45.58%
and recoveries of less than 50% for nine compounds. The recoveries of
the high-concentration spikes ranged from 39.94% to 111.21%, with
RSDs of 0.49%–14.23% and recoveries of less than 50% for three com­
pounds. For the lotion samples, the recoveries of the low-concentration
spikes ranged from 16.75% to 100.05%, with RSDs of 6.01%–68.95%
The stability of the calibration curve was established by injecting the
same standard solution for one year. No significant changes were
observed in the quantitative results (RSD <20.0%), indicating that the
calibration curve can be applied without re-calibration for an extended
period and for retrospective quantitative analysis.
4. Conclusions
HRMS is an important tool for screening and quantifying target and
non-target compounds in complex samples and allows for confident
characterization of organic compounds. Combining HRMS with HPLC
significantly expands the chemical space that can be sampled in complex
sample matrices. However, the method established in this study ex­
cludes ultra-trace compounds, compounds present in only a few

10
D. Chen et al.
samples, as well as degradation products not currently available in the
spectral library. Considering the limitations of the instrumentation,
inorganic and metal impurities are outside the scope of this study.
In this study, 100 prohibited substances were selected as target
compounds to establish a standard spectral library for quantitative
identification and detection in toner and lotion samples. Combining
SWATH® mode acquisition with fragment ion peak area quantification
ensured the accuracy of the qualitative and quantitative results and
reduced MEs. The toner and lotion samples do not require complex
pretreatment and can be directly injected for detection by a “one-step”
purification process. The 100 prohibited substances in the established
standard spectral library cannot include all prohibited substances. If
unidentified compounds are found in cosmetic samples, they will be
confirmed by mass spectrometry. Based on accurately measured mass
and isotope patterns, the molecular composition of unidentified com­
pounds is assigned to the detected m/z features, and if the molecular
composition matches at least one mass listed in the mass spectrometry
database, the compound can be referenced and finally confirmed by the
standards. Therefore, this study established a screening method for
HPLC–Q–TOF–HRMS that can be employed for the rapid screening and
accurate quantification of prohibited substances in toner and lotion
samples.
CRediT authorship contribution statement
Dan Chen: Methodology, Validation, Formal analysis. Ying Chen:
Methodology, Validation, Investigation. Yuan Zhang: Formal analysis,
Data curation, Investigation, Resources. Juan Du: Investigation. Han
Xiao: Conceptualization, Resources. Zong Yang: Conceptualization,
Data curation, Resources. Jia Xu: Conceptualization, Project adminis­
tration, Supervision, Writing – review & editing.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
Data will be made available on request.
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