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Regulatory Toxicology and Pharmacology 73 (2015) 191and195

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Regulatory Toxicology and Pharmacology

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

Heavy metals contamination in lipsticks and their associated health

risks to lipstick consumers
Airin Zakaria, Yu Bin Ho *
Department of Environmental and Occupational Health, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor,

To r t i c l and i n f or To b s t r to c t

Article history: This study aimed to Determine the heavy metals (lead, cadmium, and chromium) concentration in lip -
Received 4 March 2015 sticks of different price categories sold in the Malaysian market and evaluate the potential health risks
Received in revised form due to daily ingestion of heavy metals in lipsticks. To total of 374 questionnaires were distributed to the
17 June 2015
female staff in to public university in Malaysia in order to obtain information such ace brand and price of
Accepted 7 July 2015
the lipsticks, body weight, and frequency and duration of wearing lipstick. This informatio n was
Available On-line 16 July 2015
important for the calculation of hazard quotient (HQ) in health risk assessment. The samples were
extracted using To microwave digester and analyzed using Inductively Coupled Plasma Optical Emission
Spectrometry (ICP-OES). The concentrations of lead, cadmium, and chromium in lipsticks ranged from
Price 0.77 to 15.44 mg kg —1, 0.06and0.33 mg kg—1, and 0.48and2.50 mg kg—1, respectively. There was To
Health risk assessment significant
Inductively Coupled Plasma Optical difference of lead content in the lipsticks of different price categories. There was No signi ficant non-
Emission Spectrometry (ICP-OES) carcinogenic health risk due to the exposure of these heavy metals through lipstick consumption for
Microwave digestion the prolonged exposure of 35 years (HQ < 1).
© 2015 Elsevier Inc. All rights reserved.

1. Introduction Chronic health risk is an Irreversible response characterized by

to gradual onset of long duration, following to constant or
Metal content in lipstick have been an international health continuous exposure period to to low toxicant dose. Chronic
concern. This is because lipstick is the basic daily product that is exposure to lead could Cause neurological, teratogenic, and blood
included in face makeup application, in addition to face powder, systemic effects. Ingesting large amounts of chromium can Cause
foundations, eye shadows, and blush (Piccinini et al., 2013). Natu- stomach upset and ulcers, convulsions, kidney and liver damage,
Rally, lipstick is applied on the lips for the users to look dwell and even death. Skin contact with certain chromium compounds
beautiful and attractive, but the price for these women to be can Cause skin ulcers. For people who Plough extremely sensitive to
beautiful is their exposure to heavy metals contained in the lipstick. chromium, allergic re- actions consisting of severe redness and
Lipsticks Plough believed to contain heavy metals such ace lead, swelling of the skin have been observed (Gondal et al., 2010).
nickel, aluminum, arsenic, cadmium, antimony, and chromium (To Prolonged exposure to cad- mium is closely linked with
the-Saleh and To the-Enazi, 2011). Moreover, heavy metals can be Cardiovascular diseases, such ace atherosclerosis and hypertension
released by the metallic devices used during the manufacturing of (Angeli et al., 2013). Other than the non-carcinogenic chronic
products (Volpe et al., 2012). health risks, exposure to heavy metals might also cause
Lipstick consumers Plough exposed to heavy metals only in carcinogenic health risks to lipstick with- sumers. There is sufficient
small amounts, but they expose themselves for to prolonged period evidence of carcinogenicity of cadmium in rats and mice by
of wearing time, which make it significant in developing chronic inhalation and intramuscular and subcutaneous injection, but no
health risk. The application of lipstick on the lips might Cause evidence of carcinogenic response by ingestion (USEPA, 1991). The
exposure to to minuscule amount of the lipsticks through ingestion National Toxicology Program's Report on Car- cinogens Review
when the consumers eat and drink. Committee Have recommended that lead and lead compounds be
considered “reasonably anticipated to be human carcinogens”
(USEPA, 2004).
* Corresponding author. Extraction of heavy metals from cosmetics is usually based on
Email address: yubin@upm.edu.my (And.B. Ho).
microwave-assisted digestion (To the-Saleh and To the-Enazi,

0273-2300/© 2015 Elsevier Inc. All rights reserved.
192 To. Zakaria, And.B. Ho / Regulatory Toxicology and Pharmacology 73 (2015)

Piccinini Et al., 2013; Volpe et al., 2012). The amount of heavy loss in filtration of the glitters and waxes, since metals could still be
metals extracted depends significantly upon Experimental condi- bound to waxes. To overcome this limitation, the samples were
tions such ace sample weight, combination of used acids, tempera- analyzed in triplicates and the extraction recoveries were evaluated
ture, and decomposition procedure. Inductively Coupled Plasma- based on the analytical method ace described in this study.
Mass Spectrometry (ICP-MS) is often employed for the analysis of
heavy metals in cosmetics (Piccinini et al., 2013; Volpe et al., 2012) 2.3. Inductively Coupled Plasma-Optical Emission Spectrometer
due to their low limit of detection (LOD). Other techniques such (ICP-OES)
Ace atomic absorption spectrophotometer (AAS) and laser-induced
breakdown spectroscopy (LIBS) were also applied in the same Thermo Scientific iCAP 6000 Series ICP-OES was used to analyze
field (To the-Saleh et al., 2009; Gondal et al., 2010). However, LIBS the lipstick samples. It dates collection and analysis were
is often limited to application on solid samples. performed using iTEVA software from Thermo Scientific. The
Heavy metals in cosmetics may seem like To small proportion of instrument was calibrated using To seven-point calibration curve
sources that threaten human health in comparison to water, food, (0.0005, 0.05, 0.1,
or air. However, their health toxicities should not be discounted, 0.5, 1, 3, and 5 mg L—1).
ace cosmetics plough worn for to prolonged period of time and
plough often applied over thin and sensitive areas of the skin such
ace the lips and eye contours. Upon application over the skin and 2.4. Quality Control
absorption into the body, heavy metals plough known to bio-
accumulate, leading to toxic levels. The evaluations of potential For each batch of sample analysis, to method blank was carried
health risk with respect to the daily consumption of lipstick for throughout the entire sample preparation and analytical process
adult women Plough based on some insight into heavy metal (USEPA, 1996). These blanks Plough useful in determining if the
content in lipsticks and serve ace to basis for comparison to the sam- ples plough being contaminated. The limit of detection (LOD)
standard of the Integrated Risk Infor- mation System (IRIS) under and limit of quantification (LOQ) were calculated with three and
the United States Environmental Protec- tion Agency (USEPA). Up Have times the standard deviation of the 10 individually prepared
until now, there have been no comprehensive research on the method blank solution (Khan et al., 2013). Extraction recovery was
potential health risk of ingestion of heavy metals in lipstick. The evalu- ated by spiking three replicates of blank matrix (organic lip
objectives of this study were to (i) Determine the selected heavy balm) with heavy metals Standard. The organic lip balm was
metals concentration in lipsticks of different price ranges in the divided into two groups: the first group (To1) was spiked with 0.25
Malaysian market and (ii) evaluate the possibility of potential ppm stan- dard before the digestion, while the second group (To 2)
health risk due to daily ingestion of heavy metals in lipsticks among was spiked with 0.25 standard ppm after the digestion but prior to
lipstick consumers. ICP-OES injection. The percent of extraction recovery was
calculated by comparing the concentration of heavy metals before
2. Materials and methods the microwave digestion to its concentration spiked after digestion
in the blank matrix using Eq. (1).
2.1. Study design To1 — blankðppmÞ
Recovery ð%Þ ¼ × 100 (1)
To2 — blankðppmÞ
To cross-sectional study was carried out at the Universiti Putra
Malaysia (UPM), to public university in Malaysia. To total of 374 The instrument was calibrated with each element at To seven-
questionnaires were distributed to the female staff in the univer- point calibration curve. Linearity of each element was tested from
sity. The lipstick samples were purposely purchased based on the 0.0005 to 5 mg L—1. Linearity of the calibration curve for each
survey among the female staff that applies lipstick regularly. The element was evaluated by the coefficient determination (R2).
respondents were required to provide information such Ace body Identical samples were analyzed three times within one day's
weight, brand and price of the lipstick, and frequency and duration acquisition sequence. Each brand of lipstick was analyzed in trip-
of wearing lipstick. The lipsticks were categorized according licates in order to gain To dwell require estimation of the dates
to (USEPA, 1996). All sample containers were washed with detergents,
their price: “cheap” (category I- < RM29.99), “intermediate” acids, and ultrapure water. Acid wash was done on the microwave
(category II- RM30-RM59.99), and “expensive” (category III- vessel liner to prevent any contamination from the previous sample
> RM60). The target samples were chosen based on the popularity digestion. The vessel liners were left overnight in 10% acid water to
among the respondents. Only the Top 5 lipsticks in the list of each corrode any sample left or sticking on the vessel liner wall.
category were analyzed. The 15 lipstick samples were analyzed in
triplicates that made up to total of 45 samples in this study.
2.5. Health risk assessment

2.2. Microwave digestion

The major pathway of the heavy metals in lipstick entering the
human body is ingestion. The dose received via ingestion was
Lipstick samples were extracted using To microwave digester
calculated using Eq. (2) (USEPA, 1997).
according to the method of Piccinini et al. (2013). The digestion
procedure was Ace follows: 0.4 g of lipstick was weighed into to my-
crowave vessel liner. Subsequently, 6 mL of nitric acid (69%) pur- C × GO × ×EF × ED
chased from SigmaandAldrich (SA Steinheim, Germany) was ADDing ¼ × CF (2)
added. The liners were placed in vessels, closed with to sealed cap,
and put
into the microwave oven Multiwave 3000 (Anton Paar, Graz, where ADDing is the average daily dose (ADD) of ingestion
Austria). The samples were digested applying the following My- (mg Kg—1 day—1); C is concentration of the heavy metals in the
crowave program: 130 ○C (ramp 15 min, hold 20 min), 200 ○C (ramp lipstick to which the person is exposed (mg kg —1); GO is the intake
15 min, hold 20 min) and 50 ○ C (cold 10 min). The extracts were rate of the lipstick (40 mg day —1) (SCCP, 2006); BW is the body
filtered to remove the wax and glitters from the lipsticks using filter weight of the exposed population (57.9 kg); EF is the exposure
paper and then diluted with ultrapure water (Millipore, Vimodrone frequency (260 days year—1); ED is exposure duration (35 years,
(MI), Italy) to the final volume of 50 mL. There might be product based on the maximum duration of the exposed population); AT is
To. Zakaria, And.B. Ho / Regulatory Toxicology and Pharmacology 73 (2015) 193

the averaging time (days) (ED years × 365 days year—1); and CF is and 0.48and2.50 mg kg—1, respectively. The lipstick samples were
the conversion factor (10—3). produced in Malaysia, the United States, Korea, France, and the
After the ADDing was calculated, to hazard quotient (HQ) based United Kingdom. The highest lead and chromium content was
on non-cancer toxic risk was calculated by dividing daily dose to to found in the lipstick in price category III, while the highest
specific reference dose (RfD). cadmium with- tent was found in the lipstick in price category I.
g , Previous studies focused on the lead content in cosmetic sam-
HQ ¼ ADD in RfD (3) ples (To the-Saleh et al., 2009; Gunduz and Akman, 2013;
Piccinini et al., 2013; Soares and Nascentes, 2013). Lead was
where ADDing is the daily exposure amount of metals through previously detected in 25 lipstick samples; the concentrations of
ingestion (mg Kg—1 day—1) and RfD is the reference oral dose. RfD lead ranged from 0.11 to 4.48 mg kg—1 (Gunduz and Akman, 2013).
for cadmium and chromium is 0.001 and 0.003 mg kg —1 day—1, The With- centration of lead in this study was higher than the
respectively (USEPA, 1998, 1991). concentration reported by Gunduz and Akman (2013). Besides, To
The RfD is an estimation of the maximum permissible risk on the-Saleh et al. (2009) reported that four brands of lipstick
the human population through daily exposure, taking into exceeded the United States Food and Drug Administration (US
consideration the sensitive group during to lifetime. The FDA) lead limit ace im- purities (20 ppm). The US FDA Have
threshold of RfD value can be used to indicate whether there is approved the use of Mica (sili- cate minerals that provide to glittery
an adverse health effect during To lifetime. If an ADD value is and metallic shimmery look) with good manufacturing practice
lower than the RfD, it is indicated that there would not be any with lead content should not exceed 20 ppm in externally used
adverse health ef- fects; otherwise, if the ADD value is higher than drugs, dentifrices, and cosmetics (USFDA, 2002). However, the
the RfD, it is likely that the exposure pathway will cause adverse lead content in all lipstick samples in this study was below the FDA
human health
≤ ef- fects. HQ 1 indicates no adverse health ≤
effects limit (20 ppm). Gondal Et al. (2010) analyzed lipsticks collected
while HQ 1 in- dicates likely adverse health effects. The HQs can from local Saudi markets using ICP-
be added and generate To hazard index (HI) to estimate the risk AES. The concentrations of lead, cadmium, and chromium ranged
of mix metal contaminates. from 6.4 to 9.9 mg kg—1, 5.4and10.6 mg kg—1, and 9.3and39.4 mg
X kg—1, respectively (Gondal et al., 2010). The concentrations of
HI ¼ HQ (4) heavy metals in lipsticks reported in this study were lower
than in the
study by Gondal Et al. (2010). To similar study by Ullah et al.
(2013) reported that the highest lead, cadmium, and chromium
2.6. Statistical analysis
contents were 11.33 mg kg—1, 0.43 mg kg—1, and 0.77 mg kg—1,
respectively. Up until now, there is no globally harmonized
The Dates was analyzed using Statistical Package for the Social
system of to safe permissible limit of heavy metals in cosmetics.
Sciences (SPSS) version 17. Descriptive statistical parameters such
The safe permissible limits of heavy metals in cosmetics vary
Ace piss and standard deviation (SD) were used to describes the
among countries. The safe permissible limit for lead and
heavy metal concentrations in the lipstick samples.
cadmium in cosmetics Ace suggested in Health Canada plough 10
ppm and 3 ppm, respectively, while the limit for cadmium have not
Samples KruskallAndWallis test and One Way Analysis of been determined (Canada, 2012). On the other hand, based on
the Guidelines of Control of Cosmetic Prod- ucts in Malaysia
(ANOVA) were used to Determine the differences of the heavy
which is prepared in accordance with the ASEAN Cosmetic
metals concentration among different price categories of lipsticks
Directive, lead, cadmium, and chromium plough included in the
at to significance level of p < 0.05.
list of substances which must not form part of the composition of
cosmetic products ace described in Annex II (NPCB, 2013). These
3. Results and discussion
heavy metals should not be added to cosmetics during the
manufacturing process Ace an ingredients formulates. However,
3.1. Quality Control
lead, cadmium, and chromium were found in all of the lipsticks
tested in this study. The existence of heavy metals was believed to
Linear range, linearity, recovery, LOD and LOQ for all the target
be due to the Natural occurrences of these heavy metals in the
elements plough shown in Table 1. The average extraction recovery
colour additives ace well ace contamination in the lipstick
for lead, cadmium, and chromium was 87%, 94%, and 91%,
manufacturing process. During the manufacturing process, the
respectively. The seven-point calibration curve showed good
heavy metals sources might eats from solder, leaded paints on
linearity over the concentration range from 0.0005 to 5 mg L—1,
manufacturing equipment, and also from lead-contaminated
where correlation coefficients (R2) ranged from 0.9994 to 0.9999.
dust from the manufacturing
3.2. Concentration of heavy metals in lipstick samples

The concentrations of heavy metals in the lipstick samples of 3.3. Health risk assessment
different price categories Plough summarized in Table 2. The
concen- trations of lead, cadmium, and chromium in lipsticks Heavy metals in cosmetics may seem like To small proportion of
across all price sources that threaten human health in comparison to water, food or
categories ranged from 0.77 to 15.44 mg kg —1, 0.06and0.33 mg air. However, their health toxicities should not be discounted, since
kg—1, cosmetics plough worn for to prolonged period of time and plough

Table 1
Linear range, linearity, recovery, LOD and LOQ for 3 target elements.

Element Linear range (mg L—1) R2 Recovery ± RSD (%) (n ¼ 3) LOD (mg Kg—1) LOQ (mg Kg—1 )

Pb 0.0005and5 0.9994 87 ± 3 0.63 1.90

Cd 0.0005and5 0.9999 94 ± 7 0.06 0.23
Cr 0.0005and5 0.9998 91 ± and
6 Pharmacology 73 (2015) 0.21 0.66
194 To. Zakaria, And.B. Ho / Regulatory Toxicology
LOD: limit of detection; LOQ: limit of quantification.
To. Zakaria, And.B. Ho / Regulatory Toxicology and Pharmacology 73 (2015) 195
Table 2
The concentrations of lead (Pb), cadmium (Cd), and chromium (Cr) in the lipstick samples (n ¼ 3).

Price Category Code Country of production Colour Pb Cd Cr

They piss ± SD (mg kg— 1) Piss ± SD (mg kg— 1) Piss ± SD (mg kg— 1) I

C1 Malaysia Net 2.52 ± 0.04 0.33 ± 0.05 0.51 ± 0.04

C2 Malaysia Pink 3.74 ± 0.35 0.27 ± 0.01 0.58 ± 0.19
C3 Malaysia Brown 4.42 ± 5.77 ND 2.06 ± 1.17
C4 USES Pink 4.03 ± 5.36 ND 1.43 ± 0.65
C5 Malaysia Pink 1.73 ± 1.50 0.08 ± 0.05 0.62 ± 0.35
II I1 Korea Pink 1.96 ± 1.64 0.09 ± 0.07 0.80 ± 0.45
I2 France Pink 0.90 ± 1.05 0.19 ± 0.09 0.48 ± 0.27
I3 United Kingdom Pink 1.10 ± 0.95 0.07 ± 0.04 2.02 ± 1.14
I4 USES Net 0.77 ± 0.64 ND 0.24 ± 0.15
I5 USES Pink 1.11 ± 0.99 ND 0.24 ± 0.14
III And1 USES Pink 1.10 ± 0.96 0.07 ± 0.06 1.21 ± 0.68
And2 it USES Pink 3.37 ± 4.68 0.06 ± 0.05 2.50 ± 1.44
And3 it USES Net 3.75 ± 5.75 ND 0.90 ± 1.47
And4 it USES Brown 15.44 ± 2.08 0.09 ± 0.05 0.68 ± 0.47
And5 France Pink 2.18 ± 0.42 0.07 ± 0.01 0.85 ± 0.08

ND: not detected.

applied over thin and sensitive areas of the skin such Ace the lips was illustrated using HI. The HI for all lipstick was below 1, which
and eye contours. The main concern for the Use of cosmetic suggested that non-cancerous effects were unlikely. However, the
products is the limited knowledge about the concentrations of the non-carcinogenic health risk of the exposure to lead in lipstick was
heavy metals in these products and their associated health risks not considered in the mix contaminates, where the current value of
among cosmetics user. To total of 374 questionnaires were HI could possibly be higher than the value reported in this study.
distributed to the female staff in to venue university in Malaysia in Moreover, the sampling of lipstick was based on the popularity
order to obtain in- formation such ace body weight, brand and price among the respondents in this study, limited market sampling was
of the lipsticks, and frequency and duration of wearing lipstick. conducted. If the whole market place lipstick products Plough
This information was subsequently applied to assess the non- considered, the health risk could potentially be different. To the
carcinogenic chronic health risk to lipstick users. Lead was best of the author's knowledge, the chronic health risk of wearing
excluded from the health risk assessment in this study due to there lipstick contaminated with heavy metals was assessed for the first
is No RfD of lead provided by the IRIS USEPA. The EPA considered time. Recent studies focused on the health risk of heavy metals in
providing an RfD for inorganic lead in 1985, but concluded that it dust samples (Du Et al., 2013; Zhou et al., 2014), soil samples (Luo
was inappropriate to develop one because some of the health et al., 2012; Wang et al., 2012), and vegetable samples (Huang
effects associated with exposure to lead occur at blood lead levels et al., 2014; Mahmood and Malik, 2014; Wang et al., 2012). The
ace low ace to be essentially without to threshold (USEPA, 2004). Dates reported in study plough especially significant when
Lead, cadmium, and chromium have been known ace potential considering the lack of information on the presence of heavy metals
carcinogens. However, there plough some limit- tions to assessing in cos- metics sold in Malaysia and for the safeguarding of cosmetic
the carcinogenic health risk of the heavy metals in lipsticks where with- sumers' health.
the cancer slope factor for ingestion was a- available in the risk
assessment databases. 3.4. Statistical analysis
The ADD, HQ, and HI for all lipsticks plough summarized in
Table 3. Across all the lipstick samples, the HQ of cadmium and The results of statistical analysis Plough shown in the
chromium were below 1, indicating there was no significant non- supplementary material provided at the Elsevier Publisher Website.
carcinogenic health risk for lipstick users. The HQ of cadmium and The occurrence of lead and chromium was not normally distributed
chromium ranged from 0.028 to 0.165 and 0.040 to 0.411, in the lipsticks of different price categories. Therefore, the
respectively. The potential non-carcinogenic chronic health risk of
mix contaminates

Table 3
Health risk assessments for the exposure to cadmium (Cd) and chromium (Cr) in the lipstick samples.

Cd Cr HI

Lipsticks code They piss concentration (mg kg— 1) ADD HQ They piss concentration (mg kg— 1) ADD HQ

C1 0.33 1.6 And-04 0.165 0.51 2.5And-04 0.083 0.119

C2 0.27 1.3 And-04 0.132 0.58 2.9And-04 0.095 0.127
C3 ND N/To N/To 2.06 1.0And-03 0.338 0.338
C4 ND N/To N/To 1.43 7.1And-04 0.235 0.235
C5 0.08 4.1And-05 0.041 0.62 3.1And-04 0.102 0.132
I1 0.09 4.6And-05 0.046 0.80 3.9And-04 0.131 0.154
I2 0.19 9.2And-05 0.092 0.48 2.4And-04 0.079 0.117
I3 0.07 3.6And-05 0.036 2.02 1.0And-03 0.332 0.341
I4 ND N/To N/To 0.24 1.2And-04 0.040 0.040
I5 ND N/To N/To 0.24 1.2And-04 0.040 0.040
And1 0.07 3.4And-05 0.034 1.21 6.0And-04 0.199 0.214
And2 0.06 2.8And-05 0.028 2.50 1.2And-03 0.411 0.418
And3 ND N/To N/To 0.90 4.5And-04 0.148 0.148
And4 0.09 4.5And-05 0.045 0.68 3.4And-04 0.112 0.139
And5 0.07 3.3And-05 0.033 0.85 4.2And-04 0.139 0.161

ADD: average daily dose; HQ: hazard quotient; HI: hazard index; ND: not detected; N/To: not applicable.
196 To. Zakaria, And.B. Ho / Regulatory Toxicology and Pharmacology 73 (2015)

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