CERTIFICATE OF ANALYSIS

Chain of Custody: 307491 Job Name: Task 3 ‐ Analysis of Official Samples Date Su

Client: US Food & Drug Adminitration Job Location: 2nd Group ‐ 10 Samples Date A

Address: Office of Cosmetics & Colors Job Number: CLIN 1 ‐ Task 3 (10 Samples) Rep

4300 River Road PO Number: HHSF223201810337P Date 

College Park, MD 20740 Person Su

Attention: John Gasper 

TEM LOD  TEM LOQ % Tremolite by TEM % Chrysotile by TEM % Total Tremolite &  Chrysotile by TEM

Using ASTM D5756 Mass 

Calculation

Using ASTM D5756 Mass 

Calculation

Using ASTM D5756 Mass 

Calculation

Using ASTM D5756 Mass 

Calculation

Using ASTM D5756 Mass 

Calculation

307491‐11 D‐51 0 00000144% 0.00000575% 0 00025% 0.00190% 0.00215% ND 17.8% 14.5% 

307491‐11A D‐51 0 00000133% 0.00000534% 0 00020% 0.00164% 0.00184% ND 17.9% 19 2% 

307491‐11B D‐51 0 00000151% 0.00000602% ND 0.00010% 0.00010% ND 18.0% 13.9% 

LOD = Limit of Detection LOQ = Limit of Quantification ND = Not Detected PLM = Polarized Light Microscopy TEM = Trans

Analytical Method(s):  PLM by Modified NY ELAP 198.6

TEM by Modified NY ELAP 198.4/ASTM D5756

Analyst(s): PLM

TEM

Technical Director: Andreas Sa

All results are to be consideredsigned by the Technical Directo

SUMMARY OF ANALYSIS

% Acid  Soluable 

%  Organics AMA  Sample ID Client  Sample ID %  Asbestos  by PLM

This report applies only to the sample, or samples, investigated and is not necessarily indicative of the quality or condition of apparently identical or similar products.  As a mutual protection to clients, the public, and these Laboratories, this report is submitted and acceaddressed and upon the condition that it is not to be used, in whole or in part, in any advertising or publicity matter nor shall it be reproduced, except in full, without prior written authorization from us.  Sample types, locations, and collection protocols are based upon 

and, unless collected by personnel of these Laboratories, we expressly disclaim any knowledge and liability for the accuracy and completeness of this information.  Residual sample material will be discarded in accordance with the appropriate regulatory guidelines, unle

accreditation applies only to polarized light microscopy of bulk samples and transmission electron microscopy of AHERA air samples.  This report must not be used to claim, and does not imply product certification, approval, or endorsement by NY ELAP,AIHA, NVLAP, N

reserved.  AMA Analytical Services, Inc.

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(b) (6)
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Sample Preparation 

Samples were prepared for PLM and TEM bulk analysis by Chon Simpha on May 22, 2019 through June 24, 2019. Sample 

preparation consisted of the following steps:

1) Label and weigh two 8mL glass vials for each sample in the set – one vial for the PLM preparation and one vial 

for the TEM preparation.

2) Weigh out 0.1 to 0.8 grams of material and place in corresponding 8mL glass vial. Record weight.

3) Burn samples at 480° C for at least 12 hours.

4) Record Post-Ash Weight.

5) Treat ashed sample with concentrated hydrochloric acid.

6) Filter acid reduced material onto a pre-weighed 47mm 0.4um PolyCarbonate filter.

7) Place filter into drying oven for 30 minutes and then record Post-Acid Reduced weight. 

8) Make four PLM slide preparations from the PLM residual ash for each sample in 1.550 dispersion oil. Make 

additional preparations in 1.605, 1.625, 1.680 and 1.700 dispersion oil as necessary for particle identification.

9) Weigh a portion of the residue from the TEM residual ash and place it into the corresponding pre-weighed 

100ml jar.

10) Fill the 100ml jar with deionized water

11) Sonicate the jars for approximate 5-minutes.
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12) Filter 0.2ml to 1ml of the solution onto a 47mm 0.22um MCE filter.

13) Dry the filter for 10 minutes then collapse, carbon coat, and place on a 3 TEM grids.

PLM Analysis

Analysis was performed in accordance with NY ELAP 198.6 protocols. The analysis was conducted using an Olympus BH2 polarized light microscope (PLM) equipped with a dispersion staining objective. All four slide preparations for each 

aliquot were examined. 400-point count was performed for those samples on which asbestos or a regulated amphibole 

was observed. If no asbestos was detected on any of the slides, the percentage of fibrous components was determined 

by visual estimation. The results of this analysis are detailed below in the Discussion and Interpretation of Analytical 

Findings section for each individual sample.

TEM Analysis

Analysis was performed in accordance with modified NY ELAP Method 198.4 protocols. The analysis was performed 

using a JEOL JEM-100CX II transmission electron microscope (TEM), equipped with a Thermo Fisher Quest Energy 

Dispersive X-Ray Analyzer (EDXA), at magnifications of 19,000x. Two grids for each aliquot were examined. Twenty (20)

grid openings per sample were examined.

Modifications to the NY ELAP 198.4 Method were:

1) The residue was not placed in alcohol and prepared using the quick drop method. To obtain a more uniform 

preparation, the residue was placed in a jar and filled with 100ml of deionized water. The jar was sonicated, and 

a portion of the solution was filtered onto a 47mm 0.22um MCE filter.

2) The tremolite and chrysotile were not visually estimated. The length and width of the observed particles were 

measured and the mass of each particle was calculated using the ASTM D5756 method. All particles identified as 

tremolite were included with the counts/concentrations, regardless of size and aspect ratio.

The results of this analysis are detailed below in the Discussion and Interpretation of Analytical Findings section for each 

individual sample.

Calculations 

ASTM D5756 Mass

M = π/4 L * W2 * D * 10-12

M = mass

L = length

W = width

D = density

Percent Calculation

EFA(mm2) * 100ml * MA(g) * RW(g)

 VF(ml) * IW(g) * AA(mm2) * RJ(g) 

The calculated value is then multiplied by 100 to convert it to percent.

EFA – Effective filter area

MA – Mass of asbestos

RW – Weight of residue

VF – Volume filtered

IW – Initial weight of the sample

AA – Area analyzed

RJ – Weight of residue placed into the jar
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Limit of Detection and Quantification

We used the mass of a 0.5 x 0.04-micron chrysotile fiber as the basis for our calculations. Limit of detection was defined 

as 1 fiber and limit of quantification was defined as 4 fibers.

Discussion and Interpretation of Analytical Findings:

PLM

All three aliquots of sample D-51 were analyzed by Peerawut Chaikeenee on June 26, 2019. No asbestos or nonasbestos amphibole variants were detected the samples. The results were calculated using the equations detailed in the 

calculations section. 

307491-11 NAD

307491-11A NAD

307491-11B NAD

 

TEM

Michael Greenberg analyzed sample 11 on June 20, 2019, 11A on June 24, 2019 and 11B on June 25, 2019. The sample 

consisted of talc particles and mica particles. Chrysotile was observed on all three aliquots. One tremolite particle of was 

observed on aliquot 11 and aliquot 11A. No tremolite was observed on aliquot 11B. The results were calculated using 

the equations detailed in the calculations section.

307491-11 0.00215%

307491-11A 0.00184%

307491-11B 0.00010%

The original preparation of 307491-11B did not have an even particulate distribution on the filter. A new preparation of 

11B was made. Out of precaution, additional preparations of 11 and 11A were also made. Analysis was performed on 

the original preparations of 11 and 11A. For 11B, the second preparation was analyzed.

Below are pictures, diffraction patterns, and chemistry from some of the observed particles. The unidentified peaks in 

chemistry spectra are copper, zinc, and carbon. Those peaks are from the TEM specimen holder and specimen grid.
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Talc particle from sample 307491-11

Diffraction pattern for the talc particle pictured above.
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Chemistry from the talc particle pictured above.

Mica particle from sample 307491-11
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Diffraction pattern from mica particle pictured above.

Chemistry from mica particle pictured above.
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Chrysotile particle from sample 11

Diffraction pattern from chrysotile pictured above.
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Chemistry from chrysotile pictured above.

Tremolite particle from sample 307491-11
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Diffraction pattern from tremolite particle pictured above.

Chemistry from tremolite pictured above.
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Tremolite particle from 307491-11A

Below are additional photos of some of the chrysotile matrices, bundles, and fibers counted:
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QC Discussion:

During preparation, one blank control sample and one reference control sample were prepared. These samples were 

prepared alongside the customer samples. The blank sample was prepared using Sigma-Aldrich Talc Powder, <10 
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micron. No asbestos was detected on the blank sample. The reference sample was made from the same Sigma-Aldrich 

talc powder spiked with 1% Chrysotile. The reference sample was analyzed and found to be within acceptable limits.

Our LIMS randomly selects samples for additional replicate and duplicate QC. 307491-11, 11A, and 11B/D-51 were not 

selected for any additional QC analysis.

Attachments:

The following items are attached to this case narrative for your reference:

1) Sample Log-In Sheet

2) Daily PLM Scope Calibration Log

3) Refractive Index Oil Calibration Log

4) Daily TEM Scope Calibration Log

5) QC Results Summary

6) Raw Data Sheets

a. Gravimetric Data

b. Filtration Worksheets

c. PLM Analysis

d. TEM Analysis

e. QC Samples

I certify that all information contained in this report pertaining to laboratory events, procedures, and protocols is true 

and accurately describes the handling of this project by AMA Analytical Services, Inc. and its personnel.

7/9/2019

Andreas Saldivar Date

Laboratory Director