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DETERMINATION OF HEAVY METALS
(ARSENIC, CADMIUM, LEAD AND MERCURY) IN 2 20/11/12 ACM 005
COSMETIC PRODUCTS

1. SCOPE AND FIELD OF APPLICATION

The method describes the determination of heavy metals (arsenic, cadmium, lead and mercury) in
cosmetic products.

2. PRINCIPLE
Organic matter in sample is digested by high pressure microwave digestion and determine the amount
of heavy metals, i.e. arsenic (As), cadmium (Cd), lead (Pb) and mercury (Hg) by using graphite furnace-
atomic absorption spectrophotometer (GF-AAS), cold vapour-atomic absorption spectrophotometer (CV-
AAS) and hydride generation-atomic absorption spectrophotometer (HG-AAS).

3. REAGENTS
All reagents must be of analytical grade.
3.1 Nitric acid
3.2 Hydrochloric acid
3.3 Hydrogen peroxide 30 % v/v
3.4 Reductant
3.4.1 For Hg, prepare fresh solution of either
3.4.1.1 1.1 % w/v stannous chloride in 3 % v/v hydrochloric acid or
3.4.1.2 0.2 % w/v sodium borohydride in 0.05 % w/v sodium hydroxide
3.5 Deionised water, resistivity  18.2 megaohm-cm
3.6 Standard calibration solutions
As, Cd, Pb and Hg standard stock solutions concentration of 1,000 g/mL
3.6.1 As :
3.6.1.1 For GF-AAS
Prepare As standard calibration solutions concentration of 10, 20, 30, 40 and
50 g/L in 0.5% v/v nitric acid.
3.6.1.2 For HG-AAS
3.6.1.2.1 Prepare As intermediate standard solution concentration of 1 µg/mL.
3.6.1.2.2 Pipette 200, 400, 600, 800 and 1000 µL from 3.6.1.2.1 into five
separate 100 mL volumetric flasks and continue under 5.3.
3.6.2 Cd :
Prepare Cd standard calibration solutions concentration of 1, 2, 3, 4 and 5 g/L
in 0.5 % v/v nitric acid.
3.6.3 Pb :
Prepare Pb standard calibration solutions concentration of 10, 20, 30, 40 and
50 g/L in 0.5 % v/v nitric acid.
3.6.4 Hg :
Prepare Hg standard calibration solutions concentration of 1, 2, 3, 4 and 5 g/L
in 3 % v/v hydrochloric acid.

Note : It is suggested that for different system e.g. different instrument, the manufacturer’s recommendation
for concentration of standard calibration solutions should be followed.

3.7 Modifier for GF-AAS

3.7.1 For As :

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DETERMINATION OF HEAVY METALS
(ARSENIC, CADMIUM, LEAD AND MERCURY) IN 2 20/11/12 ACM 005
COSMETIC PRODUCTS

Palladium modifier solution (Pd-modifier) concentration of 10 mg/L. Prepare

Pd-modifier concentration 1,000 g/mL in 0.5 % v/v nitric acid.
3.7.2 For Pb and Cd :
Mix 1:1 of 0.2 % w/v Mg(NO3)2.6H2O in 0.5 % v/v nitric acid and 0.2 % w/v
NH4H2PO4 in 0.5 % v/v nitric acid.

Note : It is suggested that for different system e.g. different instrument, the manufacturer’s recommendation
for concentration of reductant and modifier should be followed.

3.8 Reagent for pretreatment of As

Mix 1:1 of 10 % w/v potassium iodide and 10 % w/v ascorbic acid.

4. APPARATUS
Normal laboratory equipment, and:
4.1 Whatman paper No. 1
4.2 Microwave digestion :
4.2.1 Condition :

sample type max. power (W) max. temp. max. pressure time
(oC) (bar) (mins)
cream 800 200 75 55
powder 1000 200 75 55
lipstick 900 200 75 55

4.2.2 Example of the microwave digestion condition

For Anton Paar microware digester, set the condition as follows :

Sample type : cream

Phase Power Time (mins) Final Power Max. Temp Hold time Fan
(W) Ramp (W) (oC) (mins) speed
1 250 5 400 200 0 1
2 400 0 400 200 20 1
3 400 10 800 200 0 1
4 800 0 800 200 5 1
5 0 0 0 - 15 3

Sample type : powder

Phase Power Time (mins) Final Power Max. Temp Hold time Fan
(W) Ramp (W) (oC) (mins) speed
1 250 5 450 200 0 1
2 450 0 450 200 20 1
3 450 10 1,000 200 0 1
4 1,000 0 1,000 200 5 1
5 0 0 0 - 15 3

Sample type : lipstick

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DETERMINATION OF HEAVY METALS
(ARSENIC, CADMIUM, LEAD AND MERCURY) IN 2 20/11/12 ACM 005
COSMETIC PRODUCTS

Phase Power Time (mins) Final Power Max. Temp Hold time Fan
(W) Ramp (W) (oC) (mins) speed
1 250 5 400 200 0 1
2 400 0 400 200 20 1
3 400 10 900 200 0 1
4 900 0 900 200 5 1
5 0 0 0 - 15 3

Note: It is suggested that for other microwave digester equipment, the manufactuer’s
recommendations should be followed.

4.3 Quartz or Tetrafluoromethane (TFM) vessel 50 mL

4.4 Graphite Furnace-Atomic Absorption Spectrophotometer (As, Cd and Pb)

4.4.1 Condition :
Element wavelength pyrolysis atomized temp. Modifier (µL) injection
(nm) (oC) (oC) volume (µL)
As 193.7 1250 2100 5 20
Cd 228.8 650 1650 5 20
Pb 283.3 550 1550 5 20

4.4.2 Example of the GF-AAS condition

For Perkin Elmer GF-AAS, set the condition as follows :

Element : As
Step Temperature Time (mins) Hold Time Internal Flow
(oC) Ramp (mins) mL/min
1 110 11 30 250
2 130 15 30 250
3 1250 10 20 250
4 2100 0 5 0
5 2450 1 4 250
6 - - - 250

Element : Cd
Step Temperature Time (mins) Hold Time Internal Flow
(oC) Ramp (mins) mL/min
1 110 1 30 250
2 130 15 30 250
3 650 10 20 250
4 1650 0 5 0
5 2350 1 3 250
6 - - - 250
Element : Pb
Step Temperature Time (mins) Hold Time Internal Flow

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DETERMINATION OF HEAVY METALS
(ARSENIC, CADMIUM, LEAD AND MERCURY) IN 2 20/11/12 ACM 005
COSMETIC PRODUCTS

(oC) Ramp (mins) mL/min

1 110 1 30 250
2 130 15 30 250
3 550 10 20 250
4 1550 0 5 0
5 2350 1 3 250
6 - - - 250

Note: It is suggested that for GF-AAS instument, the manufactmer’s recommendation should be
followed.

4.5 Hydride Generation-Atomic Absorption Spectrophotometer

4.5.1 Example of the HG-AAS condition
For Perkin Elmer HG-AAS with Flow Injection Analysis System (FIAS), set the
condition as follows :
Element wavelength reducing agent carrier atomization injection volume
(nm) temp. (oC) (µL)
As 193.7 0.2 % w/v NaBH4 10 % v/v 900 500
HCl

Note: It is suggested that for other HG-AAS instument, the manufacturer’s recommendation should
be followed.

4.6 Cold vapour-Atomic Absorption Spectrophotometer

4.6.1 Example of the CV-AAS condition
For Perkin Elmer CV-AAS with Flow Injection Analysis System (FIAS), set the
condition as follows :
Element wavelength reducing agent carrier atomization injection
(nm) temp. ( C)
o volume (µL)
Hg 253.7 1.1% w/v SnCl2 or 3% v/v HCl 300 500
0.2 % w/v NaBH4

4.6.2 For GBC CV-AAS, set the condition as follows :

Element wavelength (nm) reducing agent carrier
Hg 253.7 1.1 %w/w SnCl2 or 3 %v/v HCl
0.2 % w/v NaBH4

Note : It is suggested that for other CV-AAS intrument, the manufacturer’s recommendation should
be followed.

4.7 Electrodeless Discharge Lamp or Hollow Cathode Lamp : As, Cd, Pb and Hg

5. PROCEDURE
5.1 Sample Preparation :
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COSMETIC PRODUCTS

Prepare the Method Blank as in sample preparation but without adding the sample. Sample
preparation can be carried out as follows :
5.1.1 Microwave digestion (for As, Cd, Pb and Hg)
5.1.1.1 Accurately weigh, to the nearest 0.1 mg in duplicate, 0.15 - 0.20 g of sample
into a high pressure resistance 50 mL quartz or TFM vessel. Avoid contact with
the side of the vessel.
5.1.1.2 Add 3 mL conc. nitric acid and 30 % v/v hydrogen peroxide 1 mL by using
graduate pipette. If sample contain talcum or pigment, add conc. hydrochloric
acid 1 mL.
5.1.1.3 Close the vessel lid. Leave for about 15 minutes to ensure complete reaction.
Digest in microwave digestion system at the specified program.
5.1.1.4 After cooling to room temperature, add deionised water 20 mL to the digested
solution, rinse the inner wall and lid thoroughly. Filter through Whatman paper
no.1 into 50 mL volumetric flask and dilute to volume with deionised water.
Note : If the sample is not completely digested, further re-digest the uncomplete digested
sample by putting the original vessel into microwave under the condition as specified
in 4.2.
5.2 Spiked sample preparation for determination of percent recovery
Percent recovery of elements in sample should be carried out at least one sample in a set of
analysis.
5.2.1 Accurately weigh, to the nearest 0.1 mg in duplicate, 0.15 – 0.20 g of sample into a 50 mL
quartz or TFM vessel.

5.2.2 Pipette standard solutions into the vessel as follows :

Element Conc. (µg/mL) Volume (µL) Final conc.(ug/L)
As 5 300 30
Cd 0.5 300 3.0
Pb 5 600 300 30
Hg 0.5 300 3.0
Note : It is suggested that for different system e.g. different instrument, the manufacturer’s recommendation
for concentration of standard calibration solutions should be followed.

5.2.3 Perform as in 5.1.1.1 to 5.1.1.4.

5.3 Pretreatment for As (for analyses by HG-AAS)

5.3.1 Pipette 10 mL each of deionised water (as standard blank), the reagent blank and the
sample solution into separate 100 mL volumetric flasks.
5.3.2 Add 10 mL of conc. hydrochloric acid and 10 mL of reagent for pretreatment of As (3.8) to
each of the solutions in 5.3.1 and five levels of stand solutions from 3.6.1.2.2. Allow them
to stand for 45 minutes at ambient temperature. Dilute to volume with water. The final
concentrations of the standard solutions are 2, 4, 6, 8 and 10 g/L, respectively.
5.3.3 These solutions are used for analyses by HG-AAS.

5.4 Running sequence

Sequentially inject or aspirate reagent blank, standard and sample solutions into the GF-AAS or
CV-AAS or HG-AAS at the specified condition (4.4 to 4.6) as follows :

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COSMETIC PRODUCTS

- 0.5 % v/v nitric acid or 3 % v/v hydrochloric acid

- Standard calibration solutions (5 levels)
- Standard check (Standard calibration solution ; 1 level)
(i.e. As 20 or 6 g/L, Cd 2 g/L, Pb 20 g/L and Hg 2 g/L)
- 0.5 % v/v nitric acid for GF-AAS or 3 % v/v hydrochloric acid for CV-AAS or 10% v/v
hydrochloric acid for CV-AAS
- Method blank, replicates 1 and 2
- Sample solution, replicates 1 and 2
- Spiked sample, replicates 1 and 2
- Standard check
Note : the difference of response of standard solution for standard check should not differ
from calibration curve by not more than 10 %
5.5 Response (absorbance; peak height or peak area) of standard calibration solutions are plotted
versus the concentration of each standard solution by the instrument. The concentration of As, Cd,
Pb and Hg in sample solutions are calculated and recorded. Then calculate g/g of As, Cd, Pb and
Hg in sample.

6. CALCULATION
n
conc. of As, Cd,Pb, Hg in sample sol ( g/L) x sample solution volume (mL)
As, Cd, Pb, Hg = g/g
sample weight (g) x 1,000

% recovery = conc. of As,Cd,Pb,Hg in sample ( g/g) - conc. of As, Cd, Pb, Hg in unspiked sample ( g/g) x 100

conc. of As, Cd, Pb, Hg added in sample ( g/g)

7. REMARKS
7.1 Method validation information
7.1.1 Precision
7.1.1.1 Within day
Element Content in cream (µg/g) % Relative standard deviation
As 1.7 2
Cd 1 3.8
Pb 40 2.9
Hg 0.1 4.4

7.1.1.2 Different days

Element Content in cream (µg/g) p-value (n=5,  = 0.05)
As 1.7 0.94
Cd 1 0.86
Pb 40 0.96
Hg 0.1 0.62
7.1.2 Recovery
Percent recovery of As, Cd, Pb and Hg from spiked cream are 97-115 %, 86-105 %, 88-
110 % and 88-111 %, respectively.

7.1.3 Linearity and Range

Linearity of response over the range of concentration are as follows :

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(ARSENIC, CADMIUM, LEAD AND MERCURY) IN 2 20/11/12 ACM 005
COSMETIC PRODUCTS

Element Range (g/L) Correlation coefficient, r

As 5.0-50 0.99921
Cd 0.5-5.0 0.99997
Pb 5.0-50 0.99925
Hg 0.5-5.0 0.99815

7.1.4 Limit of quantitation (LOQ) and limit of detection (LOD)

Element Limit of quantitation, LOQ Limit of Detection, LOD (g/g)
(g/g)
As 2.5 0.5
Cd 0.5 0.1
Pb 5.0 1.0
Hg 0.5 0.1

7.2 The requirement of As, Cd, Pb and Hg in cosmetic products according to the ASEAN Cosmetic
Decretive (ACD) are as follows :
1) ACD : September, 2007 Annex II - Part 1 List of substances which must not form part of the
composition cosmetic products, As, Cd, Pb and Hg are prohibited substances.
2) On 13 December 2007, the ASEAN Cosmetic Committee (ACC) accepted the maximum
contaminated level of As, Pb, and Hg of 5, 20 and 1 g/g respectively.
3) Unavoidable level for cadmium is not available in ACD.
7.3 Results report.
It is recommended 2 decimal places of test results should be reported.
7.3.1 If the result is higher than the LOD, but below the LOQ listed above, it shall be reported as
“Concentration of As, Cd, Pb or Hg is below 2.5, 0.5, 5.0 or 0.5 g/g” as stated in the table
7.1.4.
7.3.2 If the result is higher than the LOQ listed above, the concentration of As, Cd, Pb or Hg in
sample will be reported.
7.3.3 If the result is below the LOD i.e. 0.5, 0.1, 1.0 and 0.1 g/g for As, Cd, Pb and Hg,
respectively, it shall be reported as “As, Cd, Pb or Hg is not detected”
7.4 When concentration is high level, the sample extract (5.1 and 5.2) should be diluted to
concentration within the calibration range.

7.5 Reductant for Hg

7.5.1 Using sodium borohydride as a reductant may cause some abnormal phenomena i.e. false
positive peak of mercury could be observed in particular for eye shadow sample.
7.5.2 Using stannous chloride as a reductant may cause some abnormal phenomena i.e. dilution
of high concentration of mercury in sample solution could not recover the correspond
concentration (e.g. face cream or mask sample).

Appendix A
Informative
Wet digestion for determination of mercury

Sample solution for determination of Hg may be prepared by wet digestion. However, this technique
involves a low recovery of Hg as compared to the microwave digestion technique.
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COSMETIC PRODUCTS

A.1 REAGENTS
All reagents must be of analytical grade
A.1.1 Nitric acid
A.1.2 Deionised water, resistivity > 18.2 megaohm-cm

A.2 APPARATUS
Normal laboratory equipment, and
A.2.1 Water bath
A.2.2 Block heater
A.2.3 Digestion tube 50 mL
A.2.4 Refrigerator
A.2.5 Whatman paper No.40

A.3 PROCEDURE
A.3.1 Accurately weigh 0.5 g sample into a digestion tube with screw cap and add 7 mL of conc.
nitric acid.
A.3.2 Heat the sample solution in a block heater at 60C maximum for at least 3 hours.
A.3.3 Cool and dilute to volume (50 mL) with water. Stand for 24 hours in the refrigerator for
cream and lipstick samples. Filter the solution through Whatman paper No. 40.
A.3.4 The digested solutions are used for analyses by CV-AAS.

Appendix B
Informative
Sample preparation by dry ashing

Sample solution for determination of As, Cd and Pb may be prepared by dry ashing.

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COSMETIC PRODUCTS

B.1 REAGENTS
All reagents must be of analytical grade
B.1.1 50 % w/v Magnesium nitrate
B.1.2 Hydrochloric acid , 6 M
B.1.3 Deionised water, resistivity > 18.2 megaohm-cm

B.2 APPARATUS
Normal laboratory equipment, and
B.2.1 Silica dish
B.2.2 Muffle furnace
B.2.3 Water bath
B.2.4 Heating mantle
A.2.5 Whatman paper No.1

B.3 PROCEDURE
B.3.1 Accurately weigh 2.5 g sample into a silica dish and add 3 mL of 50 % w/v magnesium
nitrate.
B.3.2 Dry on the water bath and ash the residue first in the heating mantle until no more fume
and then in the muffle furnace at 500 o C for 3 hours.
B.3.3 Cool, add 25 mL 6 M hydrochloric acid, filter into a 50 mL volumetric flask and dilute to
volume with water. For As, continue under 5.2 to 5.3

Harmonised method:
 Issued by the chemical analysis group at the harmonization workshop in Kuala-Lumpur, on
September 13th to 17th, 2004
 Approved by the harmonization workshop delegates workshop in Kuala-Lumpur, on September 13th
to 17th, 2004,
 Modified after the Bangkok training, Nov 29th to Dec 3rd, 2004

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DETERMINATION OF HEAVY METALS
(ARSENIC, CADMIUM, LEAD AND MERCURY) IN 2 20/11/12 ACM 005
COSMETIC PRODUCTS

 Modified and approved after the Brunei workshop, Aug 30th to 31st, 2005
 Modified and approved after the final review in Singapore, Nov 30th to Dec 2nd, 2005
 Modified and approved after the Malaysia workshop, Jul 10th to 12th , 2006
 Modified and approved after the ACTLC meeting in Laos, May 20th to 24th , 2013

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