1 

NIOSH Manual of Analytical Methods (NMAM) 5th Edition 

BACKUP DATA REPORT

NIOSH Method No. 8200

Title: Elements in tissue by ICP-AES

Analyte: Elements (Ca, Cu, Fe, K, Mg, Mn, Mo, Ni, P, Zn) 

 Author/developer: Ronnee N. Andrews, Ph.D., NIOSH

 Date: 8/30/2018

Disclaimer: Mention of any company or product does not constitute endorsement by the National Institute for Occupational 

Safety and Health, Centers for Disease Control and Prevention. In addition, citations to websites external to NIOSH do not 

constitute NIOSH endorsement of the sponsoring organizations or their programs or products. Furthermore, NIOSH is not 

responsible for the content of these websites. All web addresses referenced in this document were accessible as of the 

publication date. 
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NIOSH 8200: Elements in Tissue by ICP-AES

Back up Data Report

Method Validation for Rodent Nail Samples

Samples of rodent tissue have been analyzed previously using the acid digestion and sample analysis conditions 

listed in NIOSH 7300. NIOSH 7300 is an air sampling method and, as such, does not contain sampling details 

relevant to rodent tissue collection. A new method, NIOSH 8200, was developed to fully address sampling, 

preparation and analysis of tissue samples. After this method validation, all future rodent nail samples will be 

prepared and analyzed according to NIOSH 8200. 

Sample Preparation:

For the rat nails (NIOSH 8200), the following digestion procedure was followed:

1. Samples transferred to digestion vessels. In this case, the digestion vessels were either 125 mL Phillips 

beakers or Erlenmeyer flasks.

2. 10 mL 4:1 nitric acid: perchloric acid (concentrated and Optima grade) added to each sample and the 

samples covered with watchglasses. The amount of acid used initially varied based upon the amount of 

sample material to be digested. SRM 1577C used 15 mL of 4:1 nitric acid: perchloric acid (concentrated 

and Optima grade) for digestion.

3. Samples refluxed on hot plate initially at 120 – 130 °C. The temperature was then increased to 150 °C. 

Samples refluxed until digestion appeared complete (no visible solid). 

NOTE: Control rat nails refluxed for 5-6 days. SRM 1577C samples refluxed for 13 days. These times are 

highly variable. Recent studies with both nail samples and SRM bulks finished digestion (and sample 

analysis) in 1 week.

4. Watchglasses removed with a slight deionized water rinse and sample volume reduced to near-dryness 

by heating at 120 °C.

5. 0.5 mL 4:1 nitric acid: perchloric acid (concentrated and Optima grade) were added to each sample and, 

upon cooling to room temperature, sample digestates were transferred to dilution vessels and diluted to 

10 mL using deionized water. In this case, dilution vessels were 15 mL polystyrene centrifuge tubes.

All samples were analyzed on a Spectro Arcos ICP-AES using external calibration standards.

Determination of method limits of detection (LOD) and quantitation (LOQ)

In order to determine the method limits of detection and quantitation, seven reagent blank samples were 

digested and analyzed according to the method. In instances where the concentration detected by the ICP is less 

than the instrument’s internally calculated LOD, the result was noted in italics. The limits of detection (LOD) 

were calculated as 3.143 times the standard deviation of the average concentration and the limits of 

quantitation (LOQ) were calculated as 10 times the standard deviation of the average concentration. 

Please note: All analyzed elements are included below, even though the method is not validated for all of them. 

Also, the numbers listed next to the element pertain to the monitored wavelength (nm). 
3 

Table 1 Reported results from analysis of reagent blank samples used in the calculation of the limits of detection (LOD) and limits of 

quantitation (LOQ). Results reported in μg/sample.

Sample R Blk 1 R Blk 2 R Blk 3 R Blk 4 R Blk 5 R Blk 6 R Blk 7

Ag 328.068 0.02006 -0.024875 -0.022434 -0.019948 -0.017155 -0.028587 0.00639

Al 396.152 1.228 1.747 0.617259 0.894621 2.024 0.824497 1.324

As 193.759 0.092774 -0.050586 -0.050557 -0.041514 -0.034984 -0.066909 0.033453

Ba 455.404 -0.008365 -0.014292 -0.016756 -0.013449 -0.014373 -0.016224 -0.009579

Be 313.042 -0.005285 -0.006753 -0.00648 -0.006015 -0.007113 -0.007069 -0.00617

Ca 315.887 1.209 1.173 0.652289 0.974644 2.006 0.549532 1.424

Cd 226.502 0.031269 0.016114 0.024518 0.018098 0.019015 0.010278 0.026931

Co 228.616 0.006381 -0.015048 -0.013947 -0.012757 -0.014204 -0.014744 -0.00134

Cr 267.716 0.05222 -0.00916 -0.004945 -0.007282 -0.002837 -0.016587 0.019082

Cu 324.754 0.040663 -0.152997 -0.152636 -0.121204 -0.144777 -0.171575 -0.064351

Fe 259.941 0.304999 0.254413 0.087568 0.24097 0.556906 0.137938 0.241782

Fe 275.573 0.28761 0.189157 0.01678 0.173546 0.496848 0.065662 0.213135

In 230.606 -0.057503 -0.239714 -0.225797 -0.200741 -0.215013 -0.257486 0.157600

K 766.491 0.393265 -0.266618 -0.557722 -0.206051 0.428663 -0.427756 0.181406

La 408.672 -0.007089 -0.075737 -0.070929 -0.068543 -0.065945 -0.08678 -0.023689

Li 670.780 0.020077 0.00387 0.003184 0.003605 0.009343 0.002819 0.014114

Mg 279.079 0.137814 -0.327333 -0.294907 -0.303189 -0.178962 -0.351176 0.083316

Mn 257.611 0.019923 -0.014685 -0.023316 -0.020634 -0.005878 -0.022498 0.011659

Mo 202.095 0.018616 -0.01175 -0.012744 -0.009109 -0.002958 -0.013848 0.000473

Na 589.592 6.371 6.158 6.324 4.69 9.285 8.149 5.462

Ni 231.604 0.027576 -0.022321 -0.024231 -0.021527 -0.025661 -0.027086 0.003997

P 178.287 0.100522 0.037285 0.027322 0.034617 0.042917 0.033355 0.084851

Pb 220.353 0.250296 0.157012 0.184983 0.165768 0.182655 0.117439 0.238305

Sb 206.833 0.110342 -0.036767 -0.037742 -0.034879 0.021114 -0.041736 0.068063

Sc 361.384 -0.000616 -0.008108 -0.007714 -0.007329 -0.00742 -0.009515 -0.003061

Se 196.090 0.111126 -0.075861 -0.071874 -0.05863 -0.052823 -0.081248 0.056318

Sn 189.991 0.037058 -0.01963 0.007105 0.001925 0.000415 0.012423 0.011605

Sr 407.771 -0.006344 -0.008659 -0.010108 -0.008849 -0.009385 -0.009859 -0.005291

Te 214.281 0.104075 -0.064315 -0.074428 -0.061106 -0.053998 -0.07684 0.043781

Ti 334.941 0.011495 0.000584 -0.003835 -0.008018 0.023077 -0.009707 0.004449

Tl 190.864 0.062995 -0.015058 -0.014104 -0.00985 -0.003006 -0.012543 0.032369

V 311.071 0.020219 -0.008417 -0.007008 -0.006047 0.001233 -0.011068 0.012717

W 207.911 0.058878 -0.028473 -0.038108 -0.031176 0.175817 -0.042003 0.010435

Y 371.030 -0.008769 -0.021035 -0.019946 -0.020341 -0.020606 -0.02419 -0.013325

Zn 213.856 0.628706 0.534376 0.146918 0.425 0.772308 0.078812 0.279197

Zr 339.198 0.0135 -0.004539 -0.014445 -0.01153 0.033322 0.007769 0.009978
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Table 2 Calculated average reagent blank response, standard deviation, method LOD, and method LOQ. Results reported in μg/sample.

Sample Average R Blk Std. Dev. LOD LOQ

Ag 328.068 ND 0.0183 0.057 0.183

Al 396.152 ND 0.510 1.6 5.10

As 193.759 ND 0.0581 0.18 0.581

Ba 455.404 ND 0.00318 0.010 0.0318

Be 313.042 ND 0.000649 0.0020 0.00649

Ca 315.887 ND 0.492 1.5 4.92

Cd 226.502 ND 0.00712 0.022 0.0712

Co 228.616 ND 0.00846 0.027 0.0846

Cr 267.716 ND 0.0238 0.075 0.238

Cu 324.754 ND 0.0748 0.24 0.748

Fe 259.941 ND 0.150 0.47 1.50

Fe 275.573 ND 0.157 0.49 1.57

In 230.606 ND 0.0722 0.23 0.722

K 766.491 ND 0.419 1.3 4.19

La 408.672 ND 0.0281 0.088 0.281

Li 670.780 ND 0.00678 0.021 0.0678

Mg 279.079 ND 0.204 0.64 2.04

Mn 257.611 ND 0.0174 0.055 0.174

Mo 202.095 ND 0.0115 0.036 0.115

Na 589.592 [6.6] 1.57 4.9 15.7

Ni 231.604 ND 0.0207 0.065 0.207

P 178.287 ND 0.0288 0.091 0.288

Pb 220.353 [0.19] 0.0462 0.15 0.462

Sb 206.833 ND 0.0614 0.19 0.614

Sc 361.384 ND 0.00318 0.010 0.0318

Se 196.090 ND 0.0764 0.24 0.764

Sn 189.991 ND 0.0170 0.053 0.170

Sr 407.771 ND 0.00183 0.0058 0.0183

Te 214.281 ND 0.0709 0.22 0.709

Ti 334.941 ND 0.0116 0.037 0.116

Tl 190.864 ND 0.0302 0.095 0.302

V 311.071 ND 0.0119 0.037 0.119

W 207.911 ND 0.0795 0.25 0.795

Y 371.030 ND 0.00532 0.017 0.0532

Zn 213.856 ND 0.255 0.80 2.55

Zr 339.198 ND 0.0166 0.052 0.166

Variability of control rat nail samples

Control rat nail samples were supplied by research collaborators at HELD. Six samples were chosen at random 

for this experiment and the sample numbers are shown in the table. The average weight of the nail samples was 

39.3 mg. The results have been corrected for the average reagent blank. ND indicates the result was not 
5 

detected above the method LOD. Results in brackets are between the method LOD and LOQ. Results marked 

with * are outliers and were not included in the calculations.

Please note: All analyzed elements included below, even though the method is not validated for all of them. 

Also, the numbers listed next to the element pertain to the monitored wavelength (nm). 
6 

Table 3 Blank response for control rat nail samples, labelled A-F. Results reported in μg/g.

Sample A B C D E F

Ag 328.068 ND ND ND ND ND ND

Al 396.152 351 234 282 158 267 242

As 193.759 35.2 28.4 ND ND ND ND

Ba 455.404 0.926 0.846 1.06 1.13 0.862 1.07

Be 313.042 ND ND ND ND ND ND

Ca 315.887 15200 17900 21600 21600 18300 20300

Cd 226.502 [0.64] [0.96] [0.34] [0.97] [0.75] [0.31]

Co 228.616 ND ND ND ND ND ND

Cr 267.716 ND [2.6] ND ND ND ND

Cu 324.754 [16.] [14.] [16.] [16.] [18.] [16.]

Fe 259.941 [27.] [37.] [20.] [20.] [31.] [22.]

In 230.606 ND ND ND ND ND ND

K 766.491 1100 1230 1320 1330 1170 1380

La 408.672 ND ND ND ND ND ND

Li 670.780 [1.0] [0.94] ND [0.53] [1.0] [0.59]

Mg 279.079 366 422 495 483 423 478

Mn 257.611 ND ND ND ND ND ND

Mo 202.095 ND [1.0] ND ND ND ND

Na 589.592 2210 2390 3260 2590 2020 2990

Ni 231.604 ND ND ND ND ND ND

P 178.287 8040 9220 11100 11000 9390 10200

Pb 220.353 ND ND ND [5.6] ND ND

Sb 206.833 203.* 55.3 ND [6.9] 19.6 16.2

Sc 361.384 ND ND ND ND ND ND

Se 196.090 [8.0] [8.3] ND ND [8.6] ND

Sn 189.991 7.31 [3.9] ND [1.6] [4.3] [1.4]

Sr 407.771 3.90 4.51 5.48 5.42 4.38 4.99

Te 214.281 20.2 [15.] ND ND ND ND

Ti 334.941 9.06 4.84 [1.3] ND [1.6] [0.93]

Tl 190.864 ND [2.7] ND [2.5] [3.4] ND

V 311.071 ND ND ND ND ND ND

W 207.911 27.4 27.5 ND ND [11.] ND

Y 371.030 ND ND ND ND ND ND

Zn 213.856 102 107 97.9 115 109 116

Zr 339.198 ND ND 6.09* ND [1.6] ND
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Table 4 Average control rat nail sample response and method LOD and LOQs (based upon average sample weight of 39.3 mg). Results 

reported in μg/g.

Average Std. Dev. LOD LOQ

Ag 328.068 ND 0.361 1.5 4.65

Al 396.152 256 63.4 41 130

As 193.759 [12.] 15.7 4.6 14.8

Ba 455.404 0.983 0.120 0.25 0.810

Be 313.042 ND 0.0308 0.052 0.165

Ca 315.887 19200 2501 39 125

Cd 226.502 [0.66] 0.288 0.57 1.81

Co 228.616 ND 0.205 0.68 2.15

Cr 267.716 ND 0.871 1.9 6.06

Cu 324.754 [16.] 1.38 6.0 19.0

Fe 259.941 [26.] 7.00 12 38.2

In 230.606 ND 3.22 5.8 18.4

K 766.491 1260 107 34 107

La 408.672 ND 0.803 2.2 7.16

Li 670.780 [0.76] 0.246 0.54 1.72

Mg 279.079 444 49.7 16 51.9

Mn 257.611 ND 0.0632 1.4 4.43

Mo 202.095 ND 0.377 0.92 2.92

Na 589.592 2580 472 130 401

Ni 231.604 ND 0.354 1.7 5.28

P 178.287 9800 1153 2.3 7.34

Pb 220.353 ND 2.27 3.7 11.8

Sb 206.833 20.1 20.8 4.9 15.6

Sc 361.384 ND 0.0845 0.25 0.809

Se 196.090 ND 2.68 6.1 19.4

Sn 189.991 [3.3] 2.39 1.4 4.32

Sr 407.771 4.78 0.625 0.15 0.466

Te 214.281 ND 9.01 5.7 18.1

Ti 334.941 3.08 3.30 0.93 2.96

Tl 190.864 [2.5] 0.518 2.4 7.68

V 311.071 ND 0.254 0.95 3.03

W 207.911 [12.] 12.1 6.4 20.2

Y 371.030 ND 0.176 0.43 1.35

Zn 213.856 108 7.28 20 64.9

Zr 339.198 ND 0.513 1.3 4.22

Spiked control rat nail samples

Control rat nail samples were supplied by research collaborators at HELD. Samples were chosen at random for 

this study and the sample numbers are shown in the following results tables. Samples were spiked with 25.0, 
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110, or 200 μL of NIOSH-CAL-2A (Inorganic Ventures) and taken through the sample preparation and analysis 

procedures. Spiked concentrations are listed below. 

The individual results have been corrected for the average reagent blank from this sample set and the average 

control nail response. ND indicates the result was not detected above the method LOD. Results in brackets are 

between the method LOD and LOQ. In the summation tables for each level, the average recovery shown is the 

average of the recoveries calculated for each sample in that set and was not calculated based upon the average 

amount detected. This is due to varying weights of the samples.

The variability in the control nail samples (above section) is greater than the spiked amounts for Ca & P. Using 

the liquid spike, acceptable amounts of Ag, As, Ba, Be, Cd, Co, Cr, Cu, Fe, K, La, Mn, Ni, Pb, Se, Sr, Tl, V, Y, & Zn 

were recovered at each level and acceptable amounts of Al & Mg were recovered at the 110 and 200 μL levels. 

Please note: The numbers listed next to the element pertain to the monitored wavelength (nm).
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Table 5 Spiked concentrations (μg/sample) for the 3 levels: 25, 110, and 200 μL. 

25 μL 110 μL 200 μL

Ag 3.13 13.8 25

Al 6.25 27.5 50

As 12.5 55 100

Ba 1.25 5.5 10

Be 1.25 5.5 10

Ca 62.5 275 500

Cd 1.25 5.5 10

Co 1.25 5.5 10

Cr 1.25 5.51 10

Cu 6.25 27.5 50

Fe 6.25 27.5 50

K 93.8 413 750

La 3.13 13.8 25

Li 1.25 5.5 10

Mg 6.25 27.5 50

Mn 1.25 5.5 10

Ni 1.25 5.5 10

P 6.25 27.5 50

Pb 6.25 27.5 50

Se 9.38 41.3 75

Sr 1.25 5.5 10

Tl 6.25 27.5 50

V 3.13 13.8 25

Y 1.25 5.5 10

Zn 18.8 82.6 150
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Table 6 Response for control rat nail samples spiked at the 25 μL level, labelled G-L. Results reported in μg/g.

Sample G H I J K L

Ag 328.068 108 87.1 78.0 111 75.9 90.9

Al 396.152 [170] ND [93.] [130] [120] ND

As 193.759 433 341 410 522 296 352

Ba 455.404 45.9 36.5 32.5 46.7 32.5 38.7

Be 313.042 46.4 36.9 33.2 47.2 32.4 39.2

Ca 315.887 ND 1070 ND ND ND ND

Cd 226.502 44.1 34.4 30.7 45.0 30.2 36.3

Co 228.616 43.3 34.5 31.2 44.5 30.3 36.7

Cr 267.716 44.1 35.7 32.3 45.7 31.2 37.8

Cu 324.754 237 189 172 244 164 201

Fe 259.941 228 173 143 220 152 181

K 766.491 3900 3510 2793 4074 3010 3027

La 408.672 115 91.9 81.4 117 80.4 96.8

Li 670.780 60.0 47.4 43.8 58.7 43.6 50.4

Mg 279.079 205 186 60.4 158 132 [32.]

Mn 257.611 41.9 33.5 30.4 43.2 29.7 36.0

Ni 231.604 43.5 34.7 31.2 44.9 30.2 36.5

P 178.287 ND 19.0 ND ND ND ND

Pb 220.353 224 180 162 232 156 192

Se 196.090 352 278 269 372 244 290

Sr 407.771 46.0 36.4 32.3 47.5 33.0 38.2

Tl 190.864 216 172 154 222 151 182

V 311.071 115 93.1 84.2 120 81.4 97.7

Y 371.030 44.7 35.7 31.8 45.7 31.4 37.9

Zn 213.856 691 573 503 717 483 583
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Table 7 Summation table for control rat nail samples spiked at the 25 μL level. Average and standard deviation reported in μg/g. NOTE: % 

Recovery based upon average of recoveries for each sample and are not based upon the average amount detected (shown here).ND = not 

detected; N/A = value could not be calculated

Average Std. Dev. % Recovery

Ag 328.068 91.9 14.9 90.4

Al 396.152 [91.] 62.6 44.3

As 193.759 392 80.5 96.8

Ba 455.404 38.8 6.28 95.5

Be 313.042 39.2 6.38 96.6

Ca 315.887 ND - N/A

Cd 226.502 36.8 6.44 90.4

Co 228.616 36.7 6.02 90.4

Cr 267.716 37.8 5.99 93.2

Cu 324.754 201 32.9 99.1

Fe 259.941 183 34.6 89.8

K 766.491 3385 524 111.5

La 408.672 97.1 15.9 95.4

Li 670.780 50.7 7.20 125.1

Mg 279.079 129 69.4 62.8

Mn 257.611 35.8 5.70 88.2

Ni 231.604 36.9 6.16 90.7

P 178.287 ND - N/A

Pb 220.353 191 31.4 94.1

Se 196.090 301 50.3 98.7

Sr 407.771 38.9 6.46 95.8

Tl 190.864 183 30.3 90.1

V 311.071 98.6 16.0 97.0

Y 371.030 37.9 6.18 93.2

Zn 213.856 592 95.7 96.9
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Table 8 Response for control rat nail samples spiked at the 110 μL level, labelled M-R. Results reported in μg/g.

Sample M N O P Q R

Ag 328.068 338 308 355 329 386 257

Al 396.152 647 503 780 671 908 490

As 193.759 1380 1260 1470 1340 1610 1070

Ba 455.404 143 128 148 135 162 108

Be 313.042 140 129 152 137 165 109

Ca 315.887 4200 1990 1840 ND 2590 7830

Cd 226.502 140 128 147 136 159 106

Co 228.616 135 124 145 132 158 104

Cr 267.716 134 123 144 131 157 103

Cu 324.754 733 663 781 715 844 564

Fe 259.941 672 767 720 653 786 502

K 766.491 13800 12200 13600 12300 15000 10700

La 408.672 352 317 367 336 400 268

Li 670.780 209 188 223 203 241 166

Mg 279.079 643 536 670 518 761 562

Mn 257.611 130 119 140 126 150 99.1

Ni 231.604 132 121 142 130 154 102

P 178.287 ND ND ND ND ND 2400

Pb 220.353 673 615 722 659 783 515

Se 196.090 1040 950 1100 1010 1190 811

Sr 407.771 141 128 147 132 161 109

Tl 190.864 648 592 691 631 754 498

V 311.071 354 322 377 342 409 271

Y 371.030 141 127 147 134 160 107

Zn 213.856 2240 2050 2390 2190 2590 1710
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Table 9 Summation table for control rat nail samples spiked at the 110 μL level. Average and standard deviation reported in μg/g. NOTE: 

% Recovery based upon average of recoveries for each sample and are not based upon the average amount detected (shown here).ND =

not detected; N/A = value could not be calculated

Average Std. Dev. % Recovery

Ag 328.068 329 43.9 85.9

Al 396.152 667 161 86.4

As 193.759 1360 184 88.9

Ba 455.404 137 18.3 90.0

Be 313.042 139 19.2 90.8

Ca 315.887 2630 3424 38.7

Cd 226.502 136 18.1 89.2

Co 228.616 133 18.3 87.1

Cr 267.716 132 18.3 86.2

Cu 324.754 717 96.7 93.9

Fe 259.941 683 103 89.7

K 766.491 13000 1516 113.4

La 408.672 340 45.0 88.8

Li 670.780 205 26.3 134.5

Mg 279.079 615 93.2 81.0

Mn 257.611 127 17.7 83.4

Ni 231.604 130 18.0 85.2

P 178.287 ND - N/A

Pb 220.353 661 91.9 86.6

Se 196.090 1020 131 88.9

Sr 407.771 136 17.7 89.3

Tl 190.864 635 87.4 83.2

V 311.071 346 47.3 90.3

Y 371.030 136 18.1 89.2

Zn 213.856 2200 300 95.8
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Table 10 Response for control rat nail samples spiked at the 200 μL level, labelled S-X. Results reported in μg/g.

Sample S T U V W X

Ag 328.068 854 732 684 828 735 735

Al 396.152 1760 1630 1340 1860 1510 1520

As 193.759 3520 3060 2830 3400 3060 3050

Ba 455.404 363 306 290 350 311 310

Be 313.042 357 312 291 352 311 307

Ca 315.887 3070 10600 10100 7270 6980 13000

Cd 226.502 341 288 272 330 295 292

Co 228.616 341 292 271 326 295 291

Cr 267.716 344 294 271 327 294 293

Cu 324.754 1830 1570 1470 1770 1580 1570

Fe 259.941 1740 1460 1370 1650 1470 1460

K 766.491 36300 29500 29200 34400 30500 31000

La 408.672 908 761 722 873 774 770

Li 670.780 504 449 425 505 447 447

Mg 279.079 1480 1400 1310 1480 1310 1440

Mn 257.611 333 282 264 320 284 283

Ni 231.604 336 287 266 322 289 286

P 178.287 ND ND 275 ND ND 1520

Pb 220.353 1690 1450 1340 1620 1460 1450

Se 196.090 2520 2280 2140 2500 2340 2300

Sr 407.771 353 303 287 344 304 300

Tl 190.864 1640 1400 1310 1570 1420 1400

V 311.071 924 781 734 889 786 785

Y 371.030 362 304 287 348 309 307

Zn 213.856 5180 4500 4240 5100 4520 4470
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Table 11 Summation table for control rat nail samples spiked at the 200 μL level. Average and standard deviation reported in μg/g. NOTE: 

% Recovery based upon average of recoveries for each sample and are not based upon the average amount detected (shown here).ND = 

not detected; N/A = value could not be calculated

Average Std. Dev. % Recovery

Ag 328.068 761 65.3 89.0

Al 396.152 1600 186 93.6

As 193.759 3160 254 92.3

Ba 455.404 322 28.4 94.0

Be 313.042 322 26.6 94.0

Ca 315.887 8490 3465 50.8

Cd 226.502 303 26.7 88.6

Co 228.616 303 25.8 88.5

Cr 267.716 304 26.5 88.8

Cu 324.754 1630 137 95.3

Fe 259.941 1530 140 89.2

K 766.491 31800 2890 124.0

La 408.672 801 72.5 93.7

Li 670.780 463 33.4 135.4

Mg 279.079 1400 75.5 82.2

Mn 257.611 294 26.2 86.0

Ni 231.604 298 26.0 87.0

P 178.287 ND - N/A

Pb 220.353 1500 128 87.0

Se 196.090 2350 145 91.7

Sr 407.771 315 26.6 92.1

Tl 190.864 1460 124 85.2

V 311.071 816 73.3 95.4

Y 371.030 320 28.9 93.4

Zn 213.856 4670 382 91.0

SRM 1577C: Bovine Liver samples

SRM 1577C: Bovine Liver is the closest standard or certified reference material for animal tissue samples that we 

could obtain. It was prepared in 3 sets at 3 different target weights (levels): 100 (A-F), 150 (G-L), and 200 (M-R)

mg. SRM 1577C contains many elements: some that we do not typically report. Analyte concentrations in SRM 

1577C are found in Table 12. Results for the elements that we typically report are listed below.
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Table 12 Concentrations of elements in SRM 1577C

Element Concentration (μg/g) Designation

Ag 0.0059 Certified

As 0.0196 Certified

Ca 131 Certified

Cd 0.097 Certified

Co 0.3 Certified

Cr 0.053 Certified

Cu 275.2 Certified

Fe 197.94 Certified

K 10230 Certified

Mg 620 Certified

Mn 10.46 Certified

Mo 3.3 Certified

Na 2033 Certified

Ni 0.0445 Certified

Pb 0.0628 Certified

S 7490 Certified

Se 2.031 Certified

Sr 0.0953 Certified

V 0.00817 Certified

Zn 181.1 Certified

Cl 2870 Reference

Cs 0.0217 Reference

H 73500 Reference

Hg 0.00536 Reference

N 103000 Reference

P 11750 Reference

Rb 35.3 Reference

Sb 0.00313 Reference

Li 0.012 Information

Si 6 Information

The individual results have not been corrected for the average reagent blank response, but the average result 

for each concentration level has been corrected for it. ND indicates the result was not detected above the 

method LOD. Results in brackets are between the method LOD and LOQ. Standard deviations and % RSDs

calculated before average reagent blank correction. Results marked with * are outliers and were not included in 

the calculations. N/A indicates the value couldn’t be calculated.

The variability in the control nail samples is greater than the amounts present in the SRM for Ag, As, Ca, Cd, Cr, 

Ni, Pb, Sb, Sr, & V. The amounts of Co & Li detected in the samples were less than the LOQ for the method. 

Individual and summation data is displayed for As and Sb, but recovery was not calculated. This is due to 

artificially high amounts detected due to memory effects [1,2] coupled with amounts in SRM 1577C that are 

below the method LODs for these elements. Acceptable amounts of Ca, Cu, Fe, K, Mg, Mn, Mo, P, & Zn were 

recovered at each level using the bovine liver material.

Please note: The numbers listed next to the element pertain to the monitored wavelength (nm).
17

Table 13 Response for SRM 1577C at the low level, labelled A-F. Results reported in μg/g. Average sample weight is 0.1009 g 

Sample A B C D E F

Ag 328.068 ND ND ND ND ND ND

As 193.759 ND ND ND ND 1.94 ND

Ca 315.887 130 135 127 133 148 131

Cd 226.502 [0.43] ND ND ND ND ND

Co 228.616 ND ND ND ND ND ND

Cr 267.716 [0.71] ND ND ND ND ND

Cu 324.754 259 261 263 271 269 269

Fe 275.573 190 188 188 195 182 193

K 766.491 10400 10600 10800 11400 11100 10900

Li 670.780 [0.60] 1.39 [0.65] [0.59] 0.685 [0.58]

Mg 279.079 536 542 546 566 563 557

Mn 257.611 8.64 8.72 8.72 9.01 9.02 9.03

Mo 202.095 3.04 3.16 3.10 3.15 3.05 3.20

Na 589.592 2930 2910 2980 3050 3110 3000

Ni 231.604 ND ND ND ND ND ND

P 178.287 10700 10400 10800 11300 11200 11100

Pb 220.353 5.18 [2.0] [2.6] [2.7] [2.8] [3.1]

Sb 206.833 [5.0] ND ND [2.2] 41.4* ND

Se 196.090 [4.4] [3.6] [4.4] [4.0] [4.3] [4.3]

Sr 407.771 ND [0.080] ND ND ND ND

V 311.071 ND ND ND ND ND ND

Zn 213.856 165 166 164 179 183 176
18

Table 14 Summation table for SRM 1577C at the low level. Average and standard deviation reported in μg/g. % Recovery is based upon 

the average response. NOTE: ND = not detected; N/A = value could not be calculated

Average Std. Dev. % RSD % Recovery

Ag 328.068 ND - - N/A

As 193.759 ND - - N/A

Ca 315.887 134 7.63 5.7 102.3

Cd 226.502 ND - - N/A

Co 228.616 ND - - N/A

Cr 267.716 ND - - N/A

Cu 324.754 265 4.98 1.9 96.4

Fe 275.573 189 4.48 2.4 95.6

K 766.491 10800 370 3.4 105.9

Li 670.780 ND - - N/A

Mg 279.079 552 11.9 2.2 89.0

Mn 257.611 8.85 0.182 2.1 84.7

Mo 202.095 3.12 0.0632 2.0 94.5

Na 589.592 2940 74.3 2.5 144.5

Ni 231.604 ND - - N/A

P 178.287 10900 343 3.1 92.8

Pb 220.353 ND - - N/A

Sb 206.833 ND - - N/A

Se 196.090 [4.2] 0.314 7.5 205.2

Sr 407.771 ND - - N/A

V 311.071 ND - - N/A

Zn 213.856 172 8.16 4.7 94.9
19

Table 15 Response for SRM 1577c at the mid-level, labelled G-L. Results reported in μg/g. Average sample weight is 0.1510 g.

Sample G H I J K L

Ag 328.068 ND ND ND ND ND ND

As 193.759 [1.7] [1.6] [1.3] [1.6] [1.5] [1.5]

Ca 315.887 127 131 127 120 117 130

Cd 226.502 ND ND ND ND ND [0.14]

Co 228.616 [0.21] [0.22] [0.19] [0.21] [0.19] [0.23]

Cr 267.716 ND ND ND ND ND ND

Cu 324.754 262 271 251 257 249 265

Fe 275.573 188 193 176 183 176 186

K 766.491 10507 10897 10023 10245 10062 10709

Li 670.780 [0.43] [0.43] [0.39] [0.41] 0.453 [0.37]

Mg 279.079 535 550 512 520 506 539

Mn 257.611 8.75 9.09 8.31 8.63 8.33 8.81

Mo 202.095 3.14 3.30 2.96 3.12 3.02 3.22

Na 589.592 3180 3260 2920 3110 2990 3120

Ni 231.604 ND ND ND ND ND ND

P 178.287 10600 11100 10100 10400 10000 10800

Pb 220.353 [1.7] [1.8] [1.7] [1.5] [1.6] [1.5]

Sb 206.833 10.3 [1.7] ND 8.92 [1.6] [2.2]

Se 196.090 [4.0] [4.2] [3.6] [4.0] [3.8] [3.9]

Sr 407.771 [0.044] ND [0.040] ND ND ND

V 311.071 ND ND ND ND ND ND

Zn 213.856 164 176 165 163 157 170
20

Table 16 Summation table for SRM 1577C at the middle level. Average and standard deviation reported in μg/g. % Recovery is based upon 

the average response. NOTE: ND = not detected; N/A = value could not be calculated

Average Std. Dev. % RSD % Recovery

Ag 328.068 ND - - N/A

As 193.759 [1.5] 0.142 9.3 N/A

Ca 315.887 126 5.69 4.5 95.9

Cd 226.502 ND - - N/A

Co 228.616 [0.21] 0.0148 7.0 70.3

Cr 267.716 ND - - N/A

Cu 324.754 259 8.34 3.2 94.1

Fe 275.573 184 6.69 3.6 92.8

K 766.491 10400 356 3.4 101.5

Li 670.780 ND - - N/A

Mg 279.079 527 16.9 3.2 85.0

Mn 257.611 8.65 0.301 3.5 82.7

Mo 202.095 3.13 0.124 4.0 94.7

Na 589.592 3060 123 4.0 150.3

Ni 231.604 ND - - N/A

P 178.287 10500 408 3.9 89.4

Pb 220.353 ND - - N/A

Sb 206.833 4.34 4.14 95.3 N/A

Se 196.090 3.92 0.181 4.6 193.0

Sr 407.771 ND - - N/A

V 311.071 ND - - N/A

Zn 213.856 166 6.64 4.0 91.6
21

Table 17 Response for SRM 1577C at the high level, labelled M-R. Results reported in μg/g. Average sample weight is 0.2008 g.

Sample M N O P Q R

Ag 328.068 ND ND ND ND ND ND

As 193.759 [1.4] [1.3] [1.4] [1.4] [1.4] [1.5]

Ca 315.887 118 120 116 119 118 120

Cd 226.502 [0.13] [0.13] [0.12] [0.12] [0.12] [0.13]

Co 228.616 [0.24] [0.25] [0.23] [0.20] [0.22] [0.23]

Cr 267.716 ND ND ND ND ND ND

Cu 324.754 257 260 256 264 261 265

Fe 275.573 180 184 179 186 181 184

K 766.491 10100 10200 9950 10300 10000 10200

Li 670.780 0.398 [0.33] [0.30] 0.610 0.498 0.388

Mg 279.079 518 526 514 532 524 530

Mn 257.611 8.57 8.72 8.55 8.79 8.67 8.78

Mo 202.095 3.15 3.21 3.15 3.23 3.15 3.23

Na 589.592 3120 3210 3100 3230 3170 3230

Ni 231.604 ND ND ND ND ND ND

P 178.287 10400 10500 10300 10600 10400 10700

Pb 220.353 [1.3] [1.2] [1.2] [1.1] [1.2] [1.1]

Sb 206.833 [1.3] [1.5] [2.2] 3.49 9.28* [1.2]

Se 196.090 [3.7 [3.8] [3.7] [3.7] [3.7] 3.94

Sr 407.771 0.0419 0.0409 0.0336 0.0337 0.0325 0.0322

V 311.071 ND ND ND ND ND ND

Zn 213.856 163 168 163 169 167 169
22

Table 18 Summation table for SRM 1577C at the high level. Average and standard deviation reported in μg/g. % Recovery is based upon 

the average response. NOTE: ND = not detected; N/A = value could not be calculated

Average Std. Dev. % RSD % Recovery

Ag 328.068 ND - - N/A

As 193.759 [1.4] 0.0564 4.0 N/A

Ca 315.887 119 1.50 1.3 90.5

Cd 226.502 [0.12] 0.00564 4.6 127.4

Co 228.616 [0.23] 0.0155 6.7 76.6

Cr 267.716 ND - - N/A

Cu 324.754 261 3.60 1.4 94.7

Fe 275.573 182 2.67 1.5 92.2

K 766.491 10100 128 1.3 98.7

Li 670.780 [0.17] 0.116 27.6 1383.7

Mg 279.079 524 7.09 1.4 84.5

Mn 257.611 8.68 0.104 1.2 83.0

Mo 202.095 3.19 0.0402 1.3 96.5

Na 589.592 3150 55.2 1.7 154.8

Ni 231.604 ND - - N/A

P 178.287 10500 153 1.5 89.1

Pb 220.353 ND - - N/A

Sb 206.833 [1.9] 0.940 48.4 N/A

Se 196.090 [3.8] 0.102 2.7 185.0

Sr 407.771 0.0358 0.00440 12.3 37.6

V 311.071 ND - - N/A

Zn 213.856 166 2.63 1.6 91.9
23

Accuracy 

Table 19 Validation criteria, including range, bias, and accuracy calculated using SRM 1577C samples. 

Element nA Range, μg/sample Bias B SrT Accuracy (%)

Ca 18 13.0 – 26.6 -0.0956 0.00032 9.6

Cd 6 0.00964 – 0.0197 0.289 0.582 127.4

Co 12 0.0298 – 0.0610 -0.272 0.185 57.6

Cu 18 27.4 – 55.9 -0.0493 0.00008 4.9

Fe 18 19.7 – 40.2 -0.0649 0.00013 6.5

K 18 1020 – 2080 0.0622 0.00000 6.2

Mg 18 61.6 – 126 -0.155 0.00003 15.5

Mn 18 1.04 – 2.13 -0.165 0.00193 16.9

Mo 18 0.328 – 0.671 -0.0474 0.00774 6.0

Na 18 202 – 413 0.520 0.00002 52.0

P 18 1170 – 2380 -0.0950 0.00000 9.5

Se 18 0.202 – 0.413 0.943 0.0542 103.2

Sr 12 0.00947 – 0.0194 -0.592 0.460 134.9

Zn 18 18.0 – 36.8 -0.0712 0.00019 7.1

An = total number of results reported for each element, ND results not included

BSrT = precision [3,4] 

Storage

No storage studies were performed. The elements are expected to be stable in the nail structure.

Independent Laboratory User Check

An independent laboratory user check was performed (under contract #200-2016-M-89668) with Quality 

Chemical Laboratories (QCL) in Wilmington, NC. After determining that the laboratory could successfully prepare 

calibration curves and analyst spikes, control nail samples were submitted for preparation and analysis. A blind 

spiking solution (High-Purity Standards, Charleston, SC) was submitted and used to spike the control nails for the 

user check. The blind spiking solution contained: Mn and Ni at 5 μg/mL, Cu and Mo at 10 μg/mL, Fe at 15 μg/mL, 

Zn at 100 μg/mL, Mg at 300 μg/mL, K at 750 μg/mL, P at 5000 μg/mL, and Ca at 7000 μg/mL. Nails were spiked 

at 3 levels with 6 samples at each level. Spiked levels are shown in Table 19. 
24

Table 20 Spiked concentrations (μg/sample) for the 3 levels of the user check.

Element Level 1 (μg/sample) Level 2 (μg/sample) Level 3 (μg/sample)

Ca 700 1750 2800

Cu 1 2.5 4

Fe 1.5 3.75 6

K 75 187.5 300

Mg 30 75 120

Mn 0.5 1.25 2

Mo 1 2.5 4

Ni 0.5 1.25 2

P 500 1250 2000

Zn 10 25 40

The wavelengths monitored by QCL are listed in Table 20. Additionally, six control nail samples were also 

prepared and analyzed. The average blank control nail response was used to correct the results for latent 

analyte content. Average latent analyte content is shown in Table 20. 

Table 21 Wavelengths and reported detection limits (DL) and quantitation limits (QL) for the user check.

Element Wavelength (nm) DL (ppm)A QL (ppm)B

Ca 315.887 0.005 0.015

Cu 324.754 0.001 0.003

Fe 259.941 0.001 0.004

K 766.491 0.003 0.009

Mg 279.079 0.010 0.029

Mn 257.611 0.001 0.002

Mo 204.598 0.004 0.013

Ni 231.604 0.003 0.009

P 213.618 0.009 0.028

Zn 213.856 0.001 0.002

ADDDD = 3.3∗SSSS

SSSSSSSSSS , where SD is standard deviation of seven blank intensities and Slope is taken from the linearity of the curve

BQQ = 10∗SSSS

SSSSSSSSSS
25

Table 22 Average latent control nail response.

Element Average Concentration (μg/sample)

Ca 687.

Cu 0.635

Fe 0.258

K 42.1

Mg 12.9

Mn ND

Mo ND

Ni ND

P 336

Zn 3.30

Average percent recoveries of spiked nails, corrected for the latent analyte content are shown in Table 21. 

Table 23 Average percent recoveries of spiked control nails at three levels for the user check samples.

Element Level 1 (μg/sample) Level 2 (μg/sample) Level 3 (μg/sample)

Ca 97.7 105.4 107.2

Cu 107.6 99.3 102.6

Fe 87.4 88.1 87.5

K 95.2 94.3 101.5

Mg 112.3 103.2 106.6

Mn 93.1 93.6 91.4

Mo 95.2 95.5 91.9

Ni 88.2 88.2 86.3

P 94.4 100.0 103.0

Zn 101.1 99.6 106.1

The results show the successful analysis of blind spike control nail samples by an independent laboratory using 

the conditions described in this method.

Note: the laboratory contractor substituted a hot block digestor for the hot plate initially listed in the method. 

Before preparing the control nail samples with the hot block digestor, the contractor showed that the sample 

could adequately be digested using the sample preparation acids and heating temperatures. In fact, the sample 

preparation time was reduced as compared to that when using the hot plate due to its more controlled heating 

of the sample. Because of this change, the hot block was added to the method.

Conclusions

The digestion and analysis of rat nail samples using a NIOSH 8200 have been validated for Ca, Cu, Fe, K, Mg, Mn, 

Mo, P, & Zn. While acceptable amounts were recovered using SRM 1577C for Ca, the large variability (with 

respect to the amount present in SRM 1577C) in the control nail samples for Ca calls to question the applicability 

of SRM 1577C for the validation of rat nail samples for Ca. 

References

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Monviso Meta-ophiolite, Western Alps: Implications for fluid flow in Subduction Zones, Electronic appendix 2. 
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Additional note on LA-ICP-MS analytical procedures. J Petrology 52(6):1207-1236, 

https://dx.doi.org/10.1093/petrology/egr025. 

[2] EPA [1996]. Method 6010B, Inductively coupled plasma-atomic emission spectrometry. Washington, DC: U.S.

Department of the Interior, Environmental Protection Agency,

https://www.epa.gov/sites/production/files/documents/6010b.pdf.

[3] NIOSH [1995]. Guidelines for Air Sampling and Analytical Method Development and Evaluation. By Kennedy

ER, Fischbach TJ, Song R, Eller PM, Shulman SA. Cincinnati, OH: U.S. Department of Health and Human Services,

Centers for Disease Control and Prevention, National Institute for Occupational Safety and Health, DHHS

(NIOSH) Publication No. 95–117.

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PF, eds. NIOSH manual of analytical methods. 5th ed. Cincinnati, OH: U.S. Department of Health and Human

Services, Centers for Disease Control and Prevention, National Institute for Occupational Safety and Health,

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