NISTIR 7880-22

NIST Micronutrients Measurement Quality Assurance Program

Summer 2001 Comparability Studies

Results for Round Robin L Fat-Soluble Vitamins and Carotenoids in Human Serum

and Round Robin 15 Ascorbic Acid in Human Serum

David L. Duewer Sam A. Margolis (Retired)

Katherine E. Sharpless Jeanice B. Thomas

karenwTypewritten Texthttp://dx.doi.org/10.6028/NIST.IR.7880-22

karenwTypewritten Text

NISTIR 7880-22

NIST Micronutrients Measurement Quality Assurance Program

Summer 2001 Comparability Studies

Results for Round Robin L Fat-Soluble Vitamins and Carotenoids in Human Serum

and Round Robin 15 Ascorbic Acid in Human Serum

David L. Duewer Sam A. Margolis (Retired)

Katherine E. Sharpless Jeanice B. Thomas

Chemical Sciences Division Materials Measurement Laboratory

2013

U.S. Department of Commerce , Acting Secretary

National Institute of Standards and Technology Patrick D. Gallagher, Under Secretary of Commerce for Standards and Technology and Director

karenwTypewritten Text

karenwTypewritten Texthttp://dx.doi.org/10.6028/NIST.IR.7880-22

karenwTypewritten Text

(This page intentionally blank)

ii

Abstract

The National Institute of Standards and Technology coordinates the Micronutrients Measurement Quality Assurance Program (MMQAP) for laboratories that measure fat- and water-soluble vitamins and carotenoids in human serum and plasma. This report describes the design of and results for the Summer 2001 MMQAP measurement comparability improvement studies: 1) Round Robin L Fat-Soluble Vitamins and Carotenoids in Human Serum and 2) Round Robin 15 Total Ascorbic Acid in Human Serum. The materials for both studies were shipped to participants in June 2001; participants were requested to provide their measurement results by mid-September, 2001.

Keywords

Human Serum Retinol, α-Tocopherol, γ-Tocopherol, Total and Trans-β-Carotene

Total Ascorbic Acid

iii

Table of Contents

Abstract............................................................................................................................................... iii Keywords ............................................................................................................................................ iii Table of Contents ............................................................................................................................... iv Introduction......................................................................................................................................... 1 Round Robin L: Fat-Soluble Vitamins and Carotenoids in Human Serum ................................ 1 Round Robin 15: Vitamin C in Human Serum............................................................................... 2 References ............................................................................................................................................ 3

Appendix A. Shipping Package Inserts for RR15........................................................................ A1 Appendix B. Final Report for RR15 ..............................................................................................B1 Appendix C. “All-Lab Report” for RR15 ......................................................................................C1 Appendix D. Representative “Individualized Report” for RR15 ............................................... D1 Appendix E. Shipping Package Inserts for RR15 ......................................................................... E1 Appendix F. Final Report for RR15............................................................................................... F1 Appendix G. “All-Lab Report” for RR15..................................................................................... G1 Appendix H. Representative “Individualized Report” for RR15............................................... H1

iv

Introduction

Beginning in 1988, the National Institute of Standards and Technology (NIST) has coordinated the Micronutrients Measurement Quality Assurance Program (MMQAP) for laboratories that measure fat- and water-soluble vitamins and carotenoids in human serum and plasma. The MMQAP provides participants with measurement comparability assessment through use of interlaboratory studies, Standard Reference Materials (SRMs) and control materials, and methods development and validation. Serum-based samples with assigned values for the target analytes (retinol, alpha-tocopherol, gamma/beta-tocopherol, trans- and total beta-carotene, and total ascorbic acid) and performance-evaluation standards are distributed by NIST to laboratories for analysis.

Participants use the methodology of their choice to determine analyte content in the control and study materials. Participants provide their data to NIST, where it is compiled and evaluated for trueness relative to the NIST value, within-laboratory precision, and concordance within the participant community. NIST provides the participants with a technical summary report concerning their performance for each exercise and suggestions for methods development and refinement. Participants who have concerns regarding their laboratory’s performance are encouraged to consult with the MMQAP coordinators.

All MMQAP interlaboratory studies consist of individual units of batch-prepared samples that are distributed to each participant. For historical reasons these studies are referred to as “Round Robins”. The MMQAP program and the nature of its studies are described elsewhere. [1,2]

Round Robin L: Fat-Soluble Vitamins and Carotenoids in Human Serum

Participants in the MMQAP Fat-Soluble Vitamins and Carotenoids in Human Serum Round Robin L comparability study (hereafter referred to as RR15) received three lyophilized and two liquid-frozen human serum test samples for analysis. Unless multiple vials were previously requested, participants received one vial of each serum. These sera were shipped on dry ice to participants in June 2001. The communication materials included in the sample shipment are provided in Appendix A.

Participants are requested to report values for all fat-soluble vitamin-related analytes that are of interest to their organizations. Not all participants report values for the target analytes, and many participants report values for non-target analytes.

The final report delivered to every participant in RR15 consists of three documents:

• A cover letter for the current study, a brief description of the other two documents, and a discussion of our analysis of the overall results that may be of broad interest. This cover letter is reproduced as Appendix B.

• The “All-Lab Report” that lists all of the reported measurement results, a number of consensus statistics for analytes reported by more than one participant, and the mean median and pooled SD from any prior distributions of the serum. This report also provides a numerical “score card” for each participant’s measurement comparability for the more commonly reported analytes. This report is reproduced as Appendix C.

1

• An “Individualized Report” that graphically analyzes each participant’s results for all analytes reported by at least five participants. This report also provides a graphical summary of their measurement comparability. The graphical tools used in this report are described in detail elsewhere [3]. An example “Individualized Report” is reproduced as Appendix D.

Round Robin 15: Vitamin C in Human Serum

Participants in the MMQAP Vitamin C in Human Serum Round Robin 15 comparability study (hereafter referred to as RR15) received three frozen serum test samples and a solid ascorbic acid control material for analysis. Unless multiple vials were previously requested, participants received one vial of each material. These sample materials were shipped on dry ice to participants in June 2001. The communication materials included in the sample shipment are provided in Appendix E.

The test serum materials were prepared by adding equal volumes of 10 % metaphosphoric acid (MPA) to human serum that had been spiked with ascorbic acid. While these samples contain some dehydroascorbic acid, its content is variable. Therefore, the participants report only total ascorbic acid (TAA, ascorbic acid plus dehydroascorbic acid). Participants are also encouraged to prepare calibration solutions from the supplied solid control to enable calibrating their serum measurements to the same reference standard.

The final report delivered to every participant in RR15 consists of three documents:

• A cover letter for the current study, a brief description of the other two documents, and a discussion of our analysis of overall results that may be of broad interest. This cover letter is reproduced as Appendix F.

• The “All-Lab Report” that summarizes all of the reported measurement results and provides several consensus statistics. This report is reproduced as Appendix G.

• An “Individualized Report” that graphically analyzes each participant’s results for TAA, including a graphical summary of their measurement comparability. The graphical tools used in this report are described in detail elsewhere [3]. An example “Individualized Report” is reproduced as Appendix H.

2

References

1 Duewer DL, Brown Thomas J, Kline MC, MacCrehan WA, Schaffer R, Sharpless KE, May WE, Crowell JA. NIST/NCI Micronutrients Measurement Quality Assurance Program: Measurement Repeatabilities and Reproducibilities for Fat-Soluble Vitamin-Related Compounds in Human Sera. Anal Chem 1997;69(7):1406-1413.

2 Margolis SA, Duewer DL. Measurement Of Ascorbic Acid in Human Plasma and Serum: Stability, Intralaboratory Repeatability, and Interlaboratory Reproducibility. Clin Chem 1996;42(8):1257-1262.

3 Duewer DL, Kline MC, Sharpless KE, Brown Thomas J, Gary KT, Sowell AL. Micronutrients Measurement Quality Assurance Program: Helping Participants Use Interlaboratory Comparison Exercise Results to Improve Their Long-Term Measurement Performance. Anal Chem 1999;71(9):1870-1878.

3

Appendix A. Shipping Package Inserts for RR15

The following three items were included in each package shipped to an RR15 participant: • Cover letter • Datasheet • Packing List and Shipment Receipt Confirmation Form

The cover letter and datasheet were enclosed in a sealed waterproof bag along with the samples themselves. The packing list was placed at the top of the shipping box, between the cardboard covering and the foam insulation.

A1

June 8, 2001

Dear Colleague:

UNITED STATES DEPARTMENT OF COMMERCE National Institute of Standards and Technology Gaithersburg, Maryland 20899-0001

Enclosed is the set of samples for the second quality assurance round robin exercise (Round Robin L) for 2001. You will find one vial of each of three lyophilized and two liquid-frozen serum samples for analysis along with a form for reporting your results. When reporting your results, please submit one value for each analyte for a given serum sample. If an obtained value is below your limit of quantitation, please indicate this result on the form by using NQ (Not Quantitated). For" analytes not measured, please leave a blank. Results are due to NIST by September 14, 2001. Results received two weeks after the due date will not be included in the summary report for this round robin study. The feedback report concerning the study will be provided around mid-October.

Lyophilized samples should be reconstituted with 1.0 mL of HPLC-grade water or equivalent. We recommend that dissolution be facilitated with 3 to 5 min agitation in an ultrasonic bath or at least 30 min at room temperature with intermittent swirling. (CAUTION: Vigorous shaking will cause foaming and possibly interfere with accurate measurement.· The rubber stopper contains phthalate esters that will leach into the sample upon intermittent contact of the liquid sample with the stopper. These esters absorb strongly in the UV region and elute near retinal in most LC systems creating analytical problems.) Pipette a known volume of serum from the vial for analysis. (The final volume of the reconstituted sample is greater than 1.0 mL). Liquid-frozen samples 274 and 275 should not be reconstituted.

For consistency, we request that laboratories use the following absorptivities (E 1 % cm) in ethanol: retinal, 18~3 at 325 nm; retinyl palmitate, 975 at 325 nm; a-tocopherol, 75.8 at 292 nm; y-tocopherol, 91.4 at 298 nm; a-carotene, 2800 at 444 nm (in hexane); P-carotene, 2560 at 450 nm (in ethanol), 2592 at 452 nm (in hexane); lycopene, 3450 at 472 nm (in hexane).

Please mail or fax your results for Round Robin L to:

Micronutrients Measurement Quality Assurance Program NIST 100 Bureau Drive Stop 8392 Gaithersburg, MD 20899-8392 Fax: (301) 977-0685

If you have questions regarding this round robin exercise, please call me at (301) 975-3120;e-mail me at jbthomas@nist.gov; or mail/fax queries to the above address.

e~13~ lmtn ce Brown Thoma'.s arch Chemist

Analytical Chemistry Division Chemical Science and Technology Laboratory

Enclosures

NISI A2

Participant #: ________ Date: _____________

Round Robin L NIST Micronutrients Measurement Quality Assurance Program

Analyte 274 275 276 277 278 Units* total retinol

trans-retinol retinyl palmitate

α-tocopherol γ/β-tocopherol δ-tocopherol

total β-carotene trans-β-carotene

total cis-β-carotene total α-carotene

trans-α-carotene

total lycopene trans-lycopene

total β-cryptoxanthin total α-cryptoxanthin

total lutein total zeaxanthin

total lutein&zeaxanthin ubiquinone-10 (Q10)

phylloquinone (K1)

25-hydroxyvitamin D

Other analytes?

* we prefer µg/mL Was sera 274 & 275 frozen when received? Yes | No

Comments:

Mail: M2QAP NIST, Stop 8392 Please return results on-or-before Fax: 301-977-0685 Gaithersburg, MD 20899-8392 14-Sep-2001 Email: David.Duewer@NIST.gov

A3

Participant #: ________ Date: _____________

Round Robin L NIST Micronutrients Measurement Quality Assurance Program

Packing List and Shipment Receipt Confirmation Form

The enclosed bubble-pack (should) contain one vial each of the following five sera:

Serum Form Reconstitute? #274 Liquid frozen No #275 #276 #277 #278

Liquid frozen Lyophilized Lyophilized Lyophilized

No Yes (1 ml H2O) Yes (1 ml H2O) Yes (1 ml H2O)

Please 1) Open the pack immediately 2) Check that it contains one vial each of the above samples 3) Check if sera #274 and #275 arrived frozen 4) Store the samples upright at -20 ˚C or below until analysis 5) Complete the following information 6) Fax the completed form to us at 301-977-0685

(or email requested information to david.duewer@nist.gov)

1) Date this shipment arrived: ___________________________ 2) Are all five vials intact? Yes | No

If "No", which one(s) were damaged?

3) Was there any dry-ice left in cooler? Yes | No 4) Did sera #274 and #275 arrive frozen? Yes | No 5) At what temperature are you storing the samples? _______ ˚C 6) When do you anticipate analyzing these samples? ________________________

Your prompt return of this information will help control M2QAP expenses.

The M2QAP Gang

Mail: M2QAP NIST, Stop 8392 Fax: 301-977-0685 Gaithersburg, MD 20899-8392 Email: David.Duewer@NIST.gov

A4

mailto:David.Duewer@NIST.govmailto:david.duewer@nist.gov

Appendix B. Final Report for RR15

The following three pages are the final report as provided to all participants: • Cover letter. • An information sheet that:

o describes the contents of the “All-Lab” report, o describes the content of the “Individualized” report, o describes the nature of the test samples and details their previous distributions,

if any, and o summarizes aspects of the study that we believe may be of interest to the

participants.

B1

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NIST CENTENNIALC

October 31, 2001

Dear Colleague:

UNITED STATES DEPARTMENT OF COMMERCE National Institute of Standards and Technology Gaithersburg, Maryland 20899-

Enclosed is the summary report of the results for Round Robin L (RR 50) for fat-soluble vitamins and carotenoids. Included in this report are: a summary of data for all laboratories; the measurement comparability summary for evaluating laboratory performance; lyophilized vs. fresh-frozen commutability data, a summary of individual laboratory performance and interlaboratory accuracy and precision; and a summary of the NIST assigned value (NA V) vs. your laboratory value for the analytes that you measured. As in previous reports, the NIST-assigned values are equally weighted means of the medians from this interlaboratory comparison exercise and the means from the analyses performed by NIST.

Data for evaluating laboratory performance in RR 50 are provided in the comparability summary (Score Card) on page 6 of the "All Lab Report." Laboratory comparability is summarized as follows: results rated 1 to 3 are within 1 to 3 standard deviation(s) of the assigned value, respectively; those rated 4 are >3 standard deviations from the assigned value.

If you have concerns regarding your laboratory's performance, we suggest that you obtain and analyze a unit of SRM 968c, Fat-Soluble Vitamins, Carotenoids, and Cholesterol in Human Serum. If your measured values do not agree with the certified values, we suggest that you contact us for consultation.

Intent-to-participate forms for the 2002 QA program were mailed about two weeks ago. This form will provide us with formal notification of your intent to participate in the program for the upcoming year. The program will consist of two round robin studies for the fat-soluble vitamins and carotenoids and one study for vitamin C in serum. To participate in the fat-soluble vitamins and carotenoids in serum studies, the participation fee is $1600 for U.S. laboratories and $2000 for non-U.S. laboratories. To participate in the vitamin C in serum study, the participation fee is $800 for U.S. laboratories and $1000 for non-U.S. laboratories. We ask that you return the form to us by no later than November 30, 2001.

B2

The following publication has been released. Please contact the corresponding author for reprints.

"Stability of Ascorbic Acid in Solutions Stored in Autosampler Vials," Margolis, S. A. and Park, E., Clin. Chem., 47, 1463-4, 2001.

If you have any questions regarding this report, please contact David Duewer at 301/975-3935; e-mail: david.duewer@nist.gov, or me at 301/975-3120; e-mail: jbthomas@nist.gov; fax: 301/977-0685.

· ce Brown Thomas Research Chemist Analytical Chemistry Division Chemical Science and Technology Laboratory

cc: L. C. Sander S. A. Wise

Enclosures

B3

The NIST M2QAP Round Robin XLIX (RR50) report consists of:

Page 1 Your values, the number of labs reporting values, and our assigned values.

2 to “Four Plot” summaries of your current and past measurement performance, one page for n each analyte you report that is also reported by at least 10 other participants.

n+1 The “target” plot version of your “Comparability Summary” scores.

Page “All Lab” Report 1-4 5a 5b 6

Samples.

Serum

274

275

276

277

278

“Individualized” Report

A listing of all results and statistics for analytes reported by at least two laboratories. A list of results for the four analytes reported by only one laboratory.

A legend for the above two lists. The “Comparability Summary” (or “Score Card”).

The five sera below were distributed in RR50.

Description

liquid-frozen material prepared from a native serum; partner to lyophilized serum 276

liquid-frozen material prepared from a native serum; partner to lyophilized serum 277

lyophilized material prepared from a native serum; partner to liquid-frozen serum 274

lyophilized material prepared from a native serum; partner to liquid-frozen serum 275

lyophilized material prepared from a native serum

Prior Distribution

as serum 267 in RR48

as serum 271 in RR49

as serum 270 in RR49

as serum 266 in RR48

as serum 192 in RR30 (3/94); serum 199 in RR32 (9/94); serum 218 in RR36 (3/96); and serum 250 in RR44 (9/98)

Qualitative Observations Several participants noted the presence an “insoluble stringy clot” in Serum 275 and/or 277. We had similar reports from the earlier distributions of these sera. However, since the solids do not appear to affect measurements, we chose to redistribute this {liquid-frozen, lyophilized} pair to help complete our commutability study.

All participants received their samples still frozen. We thank all of you who promptly confirmed receipt of the samples. Such prompt confirmation greatly simplifies our package delivery tracking and thus helps contain M2QAP costs. We will be requesting similar confirmation in all future studies.

Quantitative Results There has been no significant change in the median level or in the variability of any measurand in serum 278 over eight years storage at –80 °C.

While the liquid frozen and lyophilized sera of the commutability pairs were prepared from the same serum pool, the measurand concentrations in the lyophilized sera are expected and generally observed to be about 0.96x that of their liquid-frozen analogues. This is a simple dilution effect, since the lyophilized sera are reconstituted with 1 mL of water rather than to their original 1 mL total volume.

B4

Appendix C. “All-Lab Report” for RR15

The following six pages are the “All-Lab Report” as provided to all participants, with two exceptions:

• the participant identifiers (Lab) have been altered. • the order in which the participant results are listed has been altered.

The data summary in the “All-Lab Report” has been altered to ensure confidentiality of identification codes assigned to laboratories. The only attributed results are those reported by NIST. The NIST results are not used in the assessment of the consensus summary results of the study.

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1.5

9 1.

84 1

.56

1.72

FS

V-C

I 0.

800

0.54

0 0.

750

0.52

0 1.

060

0.03

0 0.

050

0.02

0 0.

040

0.04

0 6.

92

7.51

6.4

9 7.

13 8

.56

2.38

2.1

0 2.

27 1

.97

2.61

FS

V-C

K 0.

670

0.47

0 0.

700

0.51

0 1.

190

6.03

7.

12 6

.58

7.52

8.0

4 2.

63 2

.20

2.67

2.2

0 2.

95

FSV-

CL

0.44

4 0.

330

0.43

7 0.

316

0.66

8 8.

75 1

0.08

8.3

1 9.

14 8

.68

2.67

2.2

1 3.

02 2

.06

2.58

FS

V-C

R

0.72

0 0.

580

0.70

0 0.

500

1.16

0 5.

70

7.80

5.6

0 7.

10 6

.60

FSV-

CV

0.51

0 !0

.88

0.46

7 0.

217

1.46

3 0.

050

0.05

0 nd

0.

020

0.03

0 7.

18

7.97

6.5

0 7.

93 8

.05

2.27

1.8

8 2.

10 1

.91

2.34

0.

101

0.33

8 0.

090

0.11

4 0.

127

FSV-

CW

>0.

587

>0.3

97 >

0.55

4 >0

.387

>0.

945

0.58

7 0.

397

0.55

4 0.

387

0.94

5 0.

016

0.04

8 0.

015

0.04

5 0.

020

7.35

8.

32 6

.83

7.98

8.2

1 2.

47 2

.06

2.31

1.9

9 2.

42

0.09

0 0.

110

0.09

0 0.

100

0.12

0 FS

V-C

X >0

.59

>0.4

0 >0

.56

>0.3

8 >0

.89

0.59

0.

40

0.56

0.

38

0.89

6.

55

7.36

6.2

4 6.

91 7

.21

FSV-

CZ

0.70

0 0.

470

0.65

0 0.

460

1.21

0 2.

40

2.70

2.2

0 2.

60 2

.80

FSV-

DB

0.64

0 0.

470

0.62

0 0.

460

1.11

0 6.

74

7.98

6.2

5 7.

71 7

.15

FSV-

DD

>0.

715

>0.4

86 >

0.63

9 >0

.322

>1.

072

0.71

5 0.

486

0.63

9 0.

322

1.07

2 FS

V-D

F 0.

660

0.47

0 0.

620

0.44

0 1.

096

FSV-

DI

0.68

8 0.

469

0.66

0 0.

452

1.09

1 0.

015

0.03

8 0.

010

0.04

8 0.

020

7.30

8.

30 7

.19

8.16

8.5

9 2.

30 1

.89

2.25

1.8

4 2.

38

0.12

9 0.

156

0.12

5 0.

149

0.18

0 FS

V-D

Q

6.58

7.

15 5

.88

6.42

6.6

2 2.

66 1

.96

2.24

1.8

1 2.

32

FSV-

DR

0.

610

0.42

0 0.

560

0.40

0 1.

090

5.96

6.

67 5

.59

6.65

6.4

7 FS

V-D

U

!0.9

3 0.

510

0.69

0 0.

470

1.12

0 !6

.78

6.44

5.5

0 6.

24 6

.24

FSV-

EH

0.58

5 0.

415

0.56

8 0.

400

0.94

4 0.

035

0.05

8 0.

033

0.05

6 0.

032

6.26

7.

03 5

.79

6.36

9.3

4 1.

96 1

.83

2.10

1.6

5 2.

86

0.13

3 0.

121

0.12

8 0.

157

0.23

0 FS

V-EQ

0.

619

0.44

2 0.

578

0.38

6 1.

036

5.72

6.

86 5

.42

5.91

7.0

0 FS

V-FB

0.

680

0.49

0 0.

690

0.48

0 1.

250

7.33

7.

65 6

.73

7.39

8.0

2 FS

V-FJ

0.

700

0.50

0 0.

670

0.48

0 1.

190

7.33

8.

38 6

.92

8.51

8.5

5 2.

31 2

.00

2.22

1.9

2 2.

36

N

38

38

39

39

39

8 8

8 8

8 11

12

10

12

11

44

45

45

45

45

25

25

25

25

25

6

6 6

6 6

Min

0.

444

0.33

0 0.

437

0.21

7 0.

668

0.58

7 0.

397

0.55

4 0.

322

0.89

0 0.

012

0.03

7 0.

003

0.02

0 0.

020

2.40

2.

70 2

.20

2.60

2.8

0 1.

96 1

.55

1.65

1.4

9 1.

72

0.09

0 0.

110

0.09

0 0.

100

0.12

0 M

edia

n 0.

668

0.47

0 0.

622

0.45

0 1.

096

0.64

2 0.

457

0.63

3 0.

435

1.07

4 0.

024

0.05

2 0.

021

0.05

1 0.

032

6.28

7.

08 5

.88

6.70

7.1

2 2.

33 1

.95

2.21

1.8

4 2.

36

0.13

1 0.

155

0.11

5 0.

137

0.15

9 M

ax

0.80

0 0.

580

0.75

0 0.

522

1.46

3 0.

715

0.50

0 0.

700

0.47

0 1.

140

0.05

0 0.

098

0.04

9 0.

091

0.05

3 8.

75 1

0.08

8.3

1 9.

14 9

.34

2.67

2.2

1 3.

02 2

.20

2.95

0.

283

0.34

0 0.

248

0.26

0 0.

364

SD

0.05

5 0.

037

0.06

9 0.

047

0.08

0 0.

045

0.02

4 0.

034

0.04

3 0.

054

0.01

4 0.

013

0.00

9 0.

008

0.00

9 0.

74

0.65

0.5

5 0.

64 0

.81

0.12

0.1

1 0.

13 0

.09

0.12

0.

022

0.12

1 0.

025

0.02

9 0.

065

CV

8 8

11

11

7 7

5 5

10

5 57

25

42

15

28

12

9

9 10

11

5

6 6

5 5

17

79

22

21

41

Npa

st

44

35

35

44

45

11

12

12

11

0 11

14

12

15

8

52

44

44

52

45

29

25

25

29

21

6 7

7 6

9 M

edia

npas

t 0.

663

0.47

0 0.

650

0.44

6 1.

079

0.66

8 0.

445

0.60

3 0.

440

0.02

8 0.

057

0.02

7 0.

053

0.05

3 6.

32

7.12

6.0

0 6.

81 7

.14

2.34

2.0

1 2.

30 1

.85

2.45

0.

121

0.11

8 0.

111

0.15

2 0.

110

SDpa

st

0.08

1 0.

051

0.05

0 0.

045

0.10

6 0.

033

0.02

3 0.

031

0.03

0 0.

014

0.01

4 0.

013

0.01

4 0.

035

0.64

0.

58 0

.54

0.68

0.5

5 0.

27 0

.16

0.19

0.1

5 0.

27

0.20

1 0.

065

0.06

6 0.

180

0.02

0

NIS

Ta

NIS

Tb

NN

IST

4 4

4 4

4 4

4 4

4 4

4 4

4 4

4 4

4 4

4 4

1 1

1 1

Mea

n 0.

624

0.44

1 0.

599

0.43

5 1.

016

0.62

4 0.

441

0.59

9 0.

435

0.95

9 5.

83

6.88

5.7

5 6.

41 6

.88

2.22

1.9

1 2.

10 1

.71

2.25

0.

214

0.10

9 0.

139

0.13

6

0.64

1 0.

448

0.63

1 0.

460

1.06

8 0.

641

0.44

8 0.

631

0.46

0 1.

019

0.60

7 0.

435

0.56

9 0.

411

0.97

0 0.

607

0.43

5 0.

569

0.41

1 0.

909

5.49

6.

55 5

.49

6.65

6.8

7 2.

14 1

.76

1.92

1.6

3 2.

12

6.25

7.

21 6

.06

6.17

6.8

9 2.

31 2

.06

2.27

1.8

0 2.

37

nq

0.21

4 0.

109

0.13

9 0.

136

Srep

0.

005

0.01

2 0.

007

0.01

9 0.

026

0.00

5 0.

012

0.00

7 0.

019

0.02

7 0.

16

0.16

0.1

4 0.

12 0

.28

0.14

0.0

6 0.

07 0

.06

0.13

0.

032

0.00

5 0.

021

0.01

6 Sh

et

0.00

6 0.

017

0.00

6 0.

022

0.01

9 0.

006

0.01

7 0.

006

0.02

2 0.

033

0.25

0.

13 0

.16

0.48

0.1

5 0.

05 0

.10

0.04

0.0

7 0.

09

Sanl

0.

025

0.00

9 0.

042

0.03

5 0.

073

0.02

5 0.

009

0.04

2 0.

035

0.08

6 0.

48

0.47

0.4

5 0.

34 0

.02

0.11

0.2

1 0.

24 0

.11

0.18

SN

IST

0.02

6 0.

023

0.04

3 0.

046

0.08

0 0.

026

0.02

3 0.

043

0.04

6 0.

096

0.56

0.

51 0

.50

0.60

0.3

2 0.

18 0

.24

0.25

0.1

5 0.

24

NAV

N

AU

0.64

6 0.

456

0.61

0 0.

443

1.05

6 0.

063

0.04

3 0.

071

0.04

9 0.

101

0.63

1 0.

447

0.61

5 0.

436

1.01

8 0.

052

0.03

7 0.

055

0.04

6 0.

127

0.02

4 0.

052

0.02

1 0.

051

0.03

2 0.

017

0.02

1 0.

017

0.02

1 0.

018

6.05

6.

98 5

.81

6.55

7.0

0 0.

81

0.66

0.5

5 0.

67 0

.83

2.27

1.9

3 2.

15 1

.78

2.30

0.

25 0

.25

0.26

0.2

1 0.

25

0.12

6 0.

184

0.11

2 0.

138

0.14

7 0.

101

0.12

8 0.

026

0.02

9 0.

067

All L

ab R

epor

t Pa

ge 1

/ 6

Rou

nd R

obin

L L

abor

ator

y R

esul

ts

All R

esul

ts in

µg/

mL

Total β-Carotene

trans-β-Carotene

Total cis-β-Carotene

Total α-Carotene

trans-α-Carotene

C3

Lab

274

275

276

277

278

274

275

276

277

278

274

275

276

277

278

274

275

276

277

278

274

275

276

277

278

FSV-

BA

0.49

8 0.

313

0.44

9 0.

304

0.49

2 0.

294

0.42

5 0.

286

0.45

8 0.

476

0.02

2 0.

019

0.02

4 0.

018

0.03

3 0.

024

0.02

0 0.

024

0.01

6 0.

019

FSV-

BB

0.50

0 0.

331

0.45

2 0.

318

0.53

4 0.

314

0.43

5 0.

301

0.50

9 0.

481

0.01

9 0.

017

0.01

7 0.

017

0.02

5 0.

030

0.01

7 0.

030

0.00

9 0.

024

FSV-

BD

FSV-

BE

0.47

8 0.

349

0.47

1 0.

327

0.53

0 FS

V-BF

0.

583

0.40

5 0.

632

0.40

7 0.

790

0.02

4 0.

013

0.02

7 0.

009

0.01

1 FS

V-BG

0.

513

0.36

2 0.

474

0.31

3 0.

518

0.03

8 0.

022

0.02

9 0.

024

0.02

2 FS

V-BH

0.

619

0.45

0 0.

589

0.41

1 0.

683

0.41

0 0.

547

0.37

4 0.

624

0.58

1 0.

038

0.04

0 0.

042

0.03

7 0.

059

0.04

0 0.

016

0.02

5 0.

012

0.

017

FSV-

BI

0.52

5 0.

348

0.49

3 0.

331

0.55

7 0.

035

0.02

0 0.

033

0.01

6

0.02

2 FS

V-BJ

0.

573

0.35

7 0.

546

0.32

3 0.

578

0.03

8 0.

025

0.03

8 0.

016

0.02

5 FS

V-BK

FS

V-BL

FS

V-BM

FS

V-BN

0.

560

0.36

4 0.

461

0.32

5 0.

588

0.33

7 0.

435

0.30

4 0.

544

0.52

8 0.

032

0.02

7 0.

026

0.02

1 0.

044

0.02

8 0.

014

0.02

4 0.

015

0.01

9 FS

V-BO

0.

509

0.36

9 0.

472

0.35

3 0.

641

0.03

7 0.

024

0.03

9 0.

021

0.03

2 FS

V-BP

0.

549

0.38

3 0.

495

0.34

6 0.

565

0.02

5 0.

015

0.02

0 0.

023

0.01

7 FS

V-BQ

FS

V-BR

FS

V-BU

0.

576

0.35

9 0.

502

0.40

0 0.

596

0.03

2 0.

025

0.03

3 0.

022

0.02

6 FS

V-BV

0.

542

0.34

9 0.

498

0.34

0 0.

552

0.02

4 0.

015

0.02

3 0.

012

0.01

7 FS

V-BW

0.

540

0.36

0 0.

510

0.33

0 0.

560

0.04

1 0.

023

0.03

8 0.

030

0.02

4 FS

V-BX

>0.

498

>0.3

36 >

0.48

3 >0

.313

>0.

522

0.33

6 0.

483

0.31

3 0.

522

0.49

8 0.

033

0.02

3 0.

033

0.02

1 0.

023

FSV-

CB

0.50

8 0.

351

0.47

3 0.

340

0.53

5 0.

024

0.01

4 0.

023

0.01

1 0.

015

FSV-

CC

FS

V-C

D

0.52

2 0.

331

0.50

9 0.

310

0.56

6 0.

031

0.01

9 0.

027

0.01

3 0.

018

FSV-

CE

0.48

7 0.

311

0.43

8 0.

308

0.49

1 FS

V-C

F FS

V-C

G

0.58

0 0.

393

0.52

9 0.

360

0.61

8 0.

365

0.49

8 0.

335

0.57

1 0.

548

0.03

1 0.

028

0.03

0 0.

026

0.04

1 0.

035

0.02

1 0.

032

0.02

1 0.

024

FSV-

CH

0.

466

0.28

3 0.

425

0.29

2 0.

427

0.02

4 0.

016

0.02

3 0.

012

0.01

4 FS

V-C

I FS

V-C

K 0.

447

0.43

4 0.

631

0.51

6 0.

859

0.05

2 0.

034

0.06

2 0.

030

0.

027

FSV-

CL

0.

430

0.28

6 0.

399

0.27

9 0.

434

0.02

3 0.

016

0.02

2 0.

011

0.01

5 FS

V-C

R

FSV-

CV

0.56

8 0.

386

0.59

3 0.

388

0.74

1 0.

063

0.02

4 0.

051

0.16

9 0.

032

FSV-

CW

0.

512

0.34

6 0.

485

0.33

8 0.

526

0.32

4 0.

460

0.31

3 0.

492

0.48

8 0.

024

0.02

2 0.

025

0.02

5 0.

034

0.03

1 0.

022

0.03

1 0.

017

0.02

3 FS

V-C

X >0

.36

>0.2

4 >0

.34

>0.2

1 >0

.37

0.24

0 0.

340

0.21

0 0.

370

0.36

0 >0

.02

>0.0

2 >0

.02

>0.0

2 >0

.02

0.02

0 0.

020

0.02

0 0.

020

0.02

0 FS

V-C

Z 0.

600

0.44

0 0.

560

0.43

0 0.

680

FSV-

DB

0.36

0 0.

210

0.36

0 0.

260

0.42

0 FS

V-D

D

FSV-

DF

FSV-

DI

>0.5

08 >

0.31

9 >0

.509

>0.

342

>0.5

12

0.31

9 0.

509

0.34

2 0.

512

0.50

8 FS

V-D

Q

0.55

0 0.

347

0.50

8 0.

307

0.54

7 0.

036

0.02

5 0.

036

0.01

9 0.

025

FSV-

DR

0.

410

0.30

0 0.

400

0.28

0 0.

590

FSV-

DU

>0

.25

>0.3

7 >0

.24

>0.5

5 !0

.40

0.25

0.

37

0.24

0.

55

FSV-

EH

0.42

4 0.

299

0.40

6 0.

297

0.44

7 0.

274

0.37

7 0.

265

0.40

8 0.

395

0.02

9 0.

025

0.02

9 0.

032

0.03

9 0.

036

0.01

5 0.

036

0.01

4 0.

021

FSV-

EQ

0.49

3 0.

316

0.45

3 0.

304

0.50

6 0.

309

0.44

7 0.

295

0.49

5 0.

487

0.00

6 0.

007

0.00

6 0.

009

0.01

1 0.

020

0.01

2 0.

019

0.01

0

0.01

3 FS

V-FB

0.

606

0.41

4 0.

560

0.39

3 0.

659

0.31

4 0.

431

0.29

6 0.

475

0.46

4 0.

142

0.09

9 0.

129

0.09

7 0.

185

0.03

0 0.

020

0.02

7 0.

016

0.

018

FSV-

FJ

1.27

0 0.

910

1.31

0 0.

780

1.38

0 0.

060

0.03

0 0.

050

0.01

0 0.

030

N

Min

M

edia

n M

ax

SD

CV

Npa

st

Med

ianp

ast

SDpa

st

NIS

Ta

NIS

Tb

NN

IST

Mea

n Sr

ep

Shet

Sa

nl

SNIS

T

31

0.36

0 0.

522

1.27

0 0.

060 11

33

0.50

8 0.

069 3

0.50

3 0.

009

0.01

9 0.

100

0.10

2

31

0.21

0 0.

351

0.91

0 0.

045 13

32

0.35

2 0.

035 3

0.34

5 0.

007

0.00

1 0.

042

0.04

3

31

0.36

0 0.

493

1.31

0 0.

063 13

32

0.49

9 0.

046 3

0.47

0 0.

011

0.01

1 0.

074

0.07

6

31

0.26

0 0.

330

0.78

0 0.

049 15

33

0.30

7 0.

063 3

0.32

5 0.

007

0.00

5 0.

032

0.03

3

31

0.42

0 0.

560

1.38

0 0.

080 14

33

0.58

2 0.

078 3

0.53

7 0.

010

0.00

8 0.

036

0.03

8

12

13

13

13

13

0.24

0 0.

340

0.21

0 0.

370

0.36

0 0.

314

0.43

5 0.

301

0.50

9 0.

488

0.41

0 0.

547

0.37

4 0.

624

0.58

1 0.

031

0.04

3 0.

020

0.05

1 0.

030 6

10

10

7 10

17

14

14

17

11

0.31

6 0.

442

0.28

7 0.

535

0.47

0 0.

025

0.03

4 0.

041

0.06

5 0.

047 2

2 2

2 2

0.34

5 0.

494

0.29

2 0.

517

0.52

8 0.

015

0.02

0 0.

016

0.00

6 0.

008

0.00

4 0.

003

0.02

2 0.

003

0.01

3

0.01

5 0.

020

0.02

7 0.

007

0.01

5

9 0.

006

0.02

9 0.

142

0.00

7 26

11

0.02

4 0.

008

9 0.

007

0.02

5 0.

099

0.00

7 27

10

0.02

3 0.

005

9 0.

006

0.02

6 0.

129

0.00

4 17

10

0.02

5 0.

004

9 0.

009

0.02

5 0.

097

0.01

0 42

11

0.02

0 0.

006

9 0.

011

0.03

9 0.

185

0.00

8 21 8

0.04

1 0.

027

27

27

27

27

27

0.02

0 0.

012

0.01

9 0.

009

0.01

1 0.

032

0.02

0 0.

030

0.01

6 0.

022

0.06

3 0.

034

0.06

2 0.

169

0.03

2 0.

010

0.00

6 0.

009

0.00

7 0.

005 25

31

30

30

42

28

24

23

29

22

0.02

9 0.

018

0.03

0 0.

017

0.02

1 0.

007

0.00

4 0.

007

0.00

8 0.

006

3 1

3 1

0.02

1 0.

031

0.02

2 0.

034

0.00

2 0.

001

0.00

3 0.

003

0.00

1 0.

001

0.00

3 0.

002

0.00

4 0.

003

1 1

1 1

1

0.02

0 0.

020

0.02

0 0.

020

0.02

0 0 0

0 0

0

2 2

2 2

2 0.

026

0.01

7 0.

023

0.01

2 0.

016

0.00

0 0.

001

0.00

5 0.

000

0.00

3 0.

001

0.00

1 0.

002

0.00

1 0.

001

0.00

1 0.

002

0.00

5 0.

001

0.00

3

0.43

2 0.

316

0.42

2 0.

302

0.51

1 0.

574

0.37

5 0.

523

0.34

8 0.

562

0.53

1 0.

345

0.49

4 0.

292

0.51

8 0.

043

0.03

1 0.

029

0.05

6 0.

044

nq

0.03

6 nq

0.

029

nq

>0.0

16 0

.032

0.0

21 0

.032

0.0

22

0.01

6 0.

026

0.01

7 0.

023

0.01

2

NAV

N

AU

0.51

3 0.

348

0.48

2 0.

327

0.54

8 0.

508

0.32

9 0.

464

0.29

6 0.

513

0.02

7 0.

024

0.02

6 0.

023

0.04

0 0.

022

0.03

3 0.

021

0.03

0 0.

019

0.10

3 0.

050

0.07

7 0.

049

0.08

1 0.

059

0.04

0 0.

062

0.03

3 0.

054

0.00

6 0.

004

0.00

3 0.

006

0.01

4 0.

008

0.01

1 0.

007

0.01

0 0.

008

All L

ab R

epor

t Pa

ge 2

/ 6

Rou

nd R

obin

L L

abor

ator

y R

esul

ts

All R

esul

ts in

µg/

mL

Tota

l Lyc

open

e tra

ns-L

ycop

ene

Total β-Cryptoxanthin

Total α-Cryptoxanthin

To

tal L

utei

n tra

ns-L

utei

n

C4

Lab

274

275

276

277

278

274

275

276

277

278

274

275

276

277

278

274

275

276

277

278

274

275

276

277

278

274

275

276

277

278

FSV-

BA

0.18

5 0.

129

0.18

1 0.

333

0.14

4 0.

076

0.06

9 0.

081

0.06

2 0.

078

FSV-

BB

0.33

0 0.

207

0.31

1 0.

572

0.22

2 0.

152

0.10

7 0.

141

0.27

3 0.

117

0.05

5 0.

049

0.05

2 0.

049

0.05

2 0.

025

0.02

3 0.

023

0.01

9 0.

024

0.10

2 0.

092

0.09

2 0.

088

0.07

7 FS

V-BD

FS

V-BE

FS

V-BF

0.

374

0.20

8 0.

386

0.76

8 0.

199

0.04

2 0.

036

0.04

3 0.

040

0.03

5 FS

V-BG

0.

362

0.19

2 0.

252

0.54

9 0.

227

0.20

8 0.

126

0.14

3 0.

335

0.13

0 0.

045

0.04

2 0.

043

0.04

8 0.

046

0.08

1 0.

089

0.08

3 0.

086

0.08

9 FS

V-BH

0.

337

0.20

7 0.

307

0.54

2 0.

223

0.09

0 0.

073

0.08

6 0.

090

0.

078

FSV-

BI

0.28

1 0.

172

0.25

6 0.

509

0.18

3 0.

065

0.05

6 0.

062

0.04

8 0.

062

0.09

2 0.

104

0.08

5 0.

097

0.08

1

FSV-

BJ

0.29

0 0.

179

0.26

3 0.

493

0.21

2 0.

053

0.05

1 0.

055

0.04

8 0.

051

0.08

0 0.

074

0.07

6 0.

074

0.06

5 FS

V-BK

FS

V-BL

FS

V-BM

FS

V-BN

0.

456

0.26

7 0.

425

0.82

3 0.

313

0.23

9 0.

146

0.22

6 0.

470

0.17

5 0.

060

0.04

9 0.

057

0.05

2 0.

061

0.02

4 0.

021

0.02

3 0.

018

0.02

7 0.

112

0.10

4 0.

092

0.09

8 0.

087

FSV-

BO

0.36

8 0.

237

0.22

7 0.

722

0.27

0 0.

045

0.04

2 0.

045

0.04

7 0.

044

0.14

7 0.

144

0.13

2 0.

144

0.13

9 FS

V-BP

0.

407

0.27

6 0.

363

0.63

2 0.

297

0.05

5 0.

049

0.05

0 0.

047

0.05

7 FS

V-BQ

FS

V-BR

FS

V-BU

0.

291

0.17

8 0.

281

0.51

2 0.

201

0.04

8 0.

044

0.04

5 0.

041

0.04

6 FS

V-BV

0.

343

0.21

7 0.

335

0.56

7 0.

238

0.03

3 0.

030

0.03

1 0.

027

0.03

2 FS

V-BW

0.

370

0.23

0 0.

340

0.65

0 0.

230

FSV-

BX

0.15

5 0.

113

0.15

1 0.

290

0.12

0 0.

045

0.04

4 0.

042

0.04

1 0.

045

0.09

5 0.

084

0.09

7 0.

081

0.06

9 FS

V-C

B 0.

308

0.18

4 0.

302

0.52

4 0.

198

0.04

9 0.

044

0.04

8 0.

045

0.04

7 0.

094

0.07

9 0.

072

0.07

2 FS

V-C

C

FSV-

CD

0.

299

0.22

0 0.

280

0.49

1 0.

229

0.04

7 0.

043

0.04

4 0.

056

0.04

3 FS

V-C

E FS

V-C

F FS

V-C

G

0.35

3 0.

213

0.32

3 0.

569

0.23

4 0.

177

0.11

7 0.

160

0.30

6 0.

132

0.04

7 0.

038

0.04

3 0.

044

0.04

2 0.

130

0.11

5 0.

119

0.10

7 0.

106

FSV-

CH

0.

266

0.18

0 0.

253

0.40

4 0.

181

FSV-

CI

FSV-

CK

0.32

8 0.

264

0.37

6 0.

697

0.17

3 0.

081

0.05

9 0.

063

0.06

4 0.

048

0.04

2 0.

038

0.04

4 0.

043

0.04

6 0.

099

0.09

9 0.

099

0.09

9 0.

083

FS

V-C

L

0.31

8 0.

200

0.30

5 0.

501

0.22

0 0.

048

0.04

5 0.

049

0.03

7 0.

047

0.02

6 0.

024

0.02

6 0.

019

0.02

6 FS

V-C

R

FSV-

CV

0.47

3 0.

312

0.49

0 0.

927

0.30

4 FS

V-C

W

0.34

6 0.

215

0.33

2 0.

553

0.22

2 0.

164

0.11

3 0.

155

0.28

7 0.

123

0.05

3 0.

046

0.05

1 0.

049

0.04

9 0.

025

0.02

2 0.

024

0.02

1 0.

024

>0.0

84

>0.0

8 >0

.079

>0

.076

>0

.061

0.

080

0.07

9 0.

076

0.06

1 0.

084

FSV-

CX

0.27

0 0.

170

0.25

0 0.

460

0.18

0 0.

140

0.10

0 0.

130

0.25

0 0.

110

0.06

0 0.

050

0.05

0 0.

040

0.05

0 0.

070

0.07

0 0.

070

0.07

0 0.

060

FSV-

CZ

FSV-

DB

0.32

1 0.

204

0.33

4 0.

541

0.20

7 0.

056

0.04

6 0.

052

0.04

5 0.

056

FSV-

DD

FS

V-D

F FS

V-D

I 0.

241

0.16

5 0.

247

0.40

3 0.

172

0.09

5 0.

089

0.09

2 0.

085

0.07

8 FS

V-D

Q

0.40

2 0.

236

0.36

6 0.

599

0.28

0 0.

063

0.06

1 0.

059

0.05

4 0.

065

0.09

5 0.

108

0.10

2 0.

102

0.09

5 FS

V-D

R

FSV-

DU

FS

V-EH

0.

359

0.22

6 0.

349

0.62

2 0.

230

0.19

3 0.

120

0.20

6 0.

373

0.12

4 0.

059

0.05

1 0.

054

0.05

2 0.

048

0.08

8 0.

081

0.08

4 0.

079

0.07

1 FS

V-EQ

0.

337

0.21

3 0.

330

0.57

3 0.

231

0.05

6 0.

047

0.05

5 0.

056

0.05

0 FS

V-FB

0.

340

0.22

0 0.

320

0.58

0 0.

210

0.28

7 0.

187

0.27

0 0.

491

0.17

6 0.

059

0.05

3 0.

054

0.05

1 0.

059

0.06

4 0.

066

0.06

0 0.

060

0.06

0

FSV-

FJ

N

26

26

26

26

26

11

11

11

11

11

25

25

25

25

25

5 5

5 5

5 15

15

15

14

15

1

1 1

1 1

Min

0.1

73 0

.266

0.1

70 0

.227

0.4

04

0.11

0 0.

140

0.10

0 0.

130

0.25

0 0.

032

0.03

3 0.

030

0.03

1 0.

027

0.02

4 0.

024

0.02

1 0.

023

0.01

8 0.

064

0.06

6 0.

060

0.06

0 0.

060

Med

ian

0.22

3 0.

339

0.21

3 0.

322

0.56

8 0.

130

0.18

5 0.

120

0.16

0 0.

333

0.04

9 0.

055

0.04

7 0.

051

0.04

8 0.

026

0.02

5 0.

023

0.02

4 0.

019

0.09

5 0.

089

0.09

2 0.

087

0.07

8 0.

084

0.08

0 0.

079

0.07

6 0.

061

Max

0.3

13 0

.473

0.3

12 0

.490

0.9

27

0.17

6 0.

287

0.18

7 0.

270

0.49

1 0.

078

0.09

0 0.

073

0.08

6 0.

090

0.04

6 0.

042

0.03

8 0.

044

0.04

3 0.

147

0.14

4 0.

132

0.14

4 0.

139

SD 0

.023

0.0

42 0

.026

0.0

49 0

.085

0.

027

0.04

8 0.

018

0.05

1 0.

074

0.00

8 0.

010

0.00

5 0.

008

0.00

6 0.

002

0.00

1 0.

001

0.00

2 0.

001

0.01

2 0.

018

0.01

4 0.

014

0.01

3 C

V 10

12

12

15

15

21

26

15

32

22

17

18

12

15

12

9

5 6

9 8

13

20

15

16

17

Npa

st

29

27

27

29

24

12

12

12

12

11

29

25

25

29

20

7 5

5 7

9 16

15

15

16

14

0

0 0

0 0

Med

ianp

ast

0.22

8 0.

363

0.23

0 0.

330

0.57

5 0.

135

0.18

9 0.

124

0.16

5 0.

380

0.05

0 0.

057

0.05

1 0.

054

0.06

1 0.

029

0.02

7 0.

024

0.02

6 0.

027

0.09

2 0.

089

0.08

8 0.

083

0.08

3 SD

past

0.0

42 0

.046

0.0

37 0

.057

0.1

35

0.01

8 0.

042

0.03

2 0.

015

0.08

3 0.

010

0.00

7 0.

013

0.01

4 0.

016

0.00

6 0.

007

0.00

9 0.

007

0.00

9 0.

017

0.02

5 0.

029

0.01

2 0.

024

NIS

Ta

NIS

Tb

0.37

0 0.

227

0.33

4 0.

621

0.28

8 N

NIS

T 2

2 2

2 2

4 3

3 3

3 2

1 1

1 1

4 4

4 4

4 2

2 2

2 2

Mea

n 0.

282

0.37

0 0.

227

0.33

0 0.

620

0.04

0 0.

052

0.04

8 0.

047

0.04

6 0.

016

0.03

0 0.

030

0.02

9 0.

024

0.09

1 0.

087

0.08

0 0.

091

0.07

7 0.

073

0.08

0 0.

073

0.07

5 0.

067

0.04

8 0.

046

0.04

8 0.

042

0.04

4 0.

108

0.09

4 0.

087

0.10

7 0.

086

0.06

5 0.

058

0.04

9 0.

052

0.04

7 0.

032

0.03

0 0.

030

0.02

9 0.

024

>0.0

76

>0.0

80

>0.0

73

>0.0

75

>0.0

69

0.07

6 0.

080

0.07

3 0.

075

0.06

9

Srep

0.0

10 0

.006

0.0

31 0

.001

0.0

15

0.00

4 0.

003

0.00

4 0.

004

0.00

5 0.

003

0.00

2 0.

004

0.00

0 0.

007

0.00

4 0.

004

0.00

4 0.

003

0.00

6 0.

000

0.00

6 0.

003

0.00

4 0.

003

Shet

0.0

24 0

.004

0.0

29 0

.015

0.0

01

0.03

2 0.

001

0.00

2 0.

000

0.00

3 0.

023

0.00

7 0.

001

0.00

3 0.

004

0.00

9 0.

010

0.00

0 0.

004

0.00

4 0.

006

Sanl

0.

011

0.00

9 0.

001

0.00

7 0.

002

0.02

4 0.

010

0.01

0 0.

022

0.01

3 SN

IST

0.02

6 0.

007

0.04

3 0.

015

0.01

5 0.

034

0.00

9 0.

005

0.00

8 0.

006

0.02

3 0.

026

0.01

1 0.

011

0.02

2 0.

017

0.01

0 0.

006

0.00

5 0.

006

0.00

7

0.02

7 0.

026

0.02

6 0.

023

0.02

4 0.

093

0.08

8 0.

086

0.08

9 0.

077

0.00

3 0.

004

0.00

3 0.

004

0.00

4 0.

026

0.01

8 0.

020

0.02

3 0.

017

NAV

N

AU 0

.252

0.3

54 0

.220

0.3

26 0

.594

0.

130

0.18

5 0.

120

0.16

0 0.

333

0.04

5 0.

053

0.04

8 0.

049

0.04

7 0.

068

0.07

8 0.

053

0.07

2 0.

121

0.02

7 0.

048

0.02

1 0.

051

0.07

4 0.

035

0.01

3 0.

012

0.01

3 0.

012

All L

ab R

epor

t Pa

ge 3

/ 6

Rou

nd R

obin

L L

abor

ator

y R

esul

ts

All R

esul

ts in

µg/

mL

Tota

l Zea

xant

hin

Tota

l Lut

ein&

Zeax

anth

in

Coe

nzym

e Q

10

Phyl

loqu

inon

e (K

1) x

1000

C5

Lab

274

275

276

277

278

274

275

276

277

278

274

275

276

277

278

274

275

276

277

278

FSV-

BA

0.18

7 0.

186

0.16

6 0.

181

0.13

6 FS

V-BB

0.

046

0.03

7 0.

040

0.03

7 0.

048

0.14

8 0.

129

0.13

2 0.

125

0.12

5 FS

V-BD

FS

V-BE

0.

44 1

.04

0.40

0.3

0 1.

24

FSV-

BF

0.13

8 0.

131

0.12

8 0.

122

0.11

9 FS

V-BG

0.

030

0.02

4 0.

026

0.02

1 0.

024

0.10

1 0.

102

0.09

8 0.

098

0.10

4 FS

V-BH

FS

V-BI

0.

039

0.03

1 0.

036

0.02

9 0.

035

0.13

1 0.

135

0.12

1 0.

126

0.11

6 FS

V-BJ

0.

545

0.55

1 0.

480

0.73

3 0.

546

FSV-

BK

FSV-

BL

FSV-

BM

FSV-

BN

0.05

0 0.

030

0.04

2 0.

030

0.03

4 0.

160

0.13

4 0.

133

0.12

7 0.

120

FSV-

BO

0.04

4 0.

036

0.04

2 0.

021

0.05

3 0.

191

0.18

0 0.

174

0.16

5 0.

192

FSV-

BP

0.16

0 0.

140

0.14

0 0.

140

0.12

0 FS

V-BQ

FS

V-BR

FS

V-BU

0.

082

0.12

0 0.

132

0.11

7 0.

110

FSV-

BV

0.12

0 0.

100

0.11

2 0.

095

0.08

9 FS

V-BW

FS

V-BX

0.

031

0.02

4 0.

030

0.02

1 0.

024

0.12

7 0.

108

0.12

8 0.

102

0.09

3 FS

V-C

B 0.

039

0.02

5 0.

034

0.03

4 0.

134

0.10

4 0.

106

0.12

4 0.

106

FSV-

CC

FS

V-C

D

0.11

7 0.

110

0.11

7 0.

105

0.09

8 FS

V-C

E FS

V-C

F FS

V-C

G

0.02

0 0.

020

0.01

8 0.

017

0.01

8 0.

150

0.13

5 0.

137

0.12

4 0.

124

FSV-

CH

0.

562

0.41

5 0.

572

0.44

9

0.54

5 FS

V-C

I 0.

19 0

.59

0.20

0.1

0 0.

59

FSV-

CK

0.04

1 0.

031

0.04

1 0.

032

0.03

8 0.

129

0.12

8 0.

141

0.13

4 0.

135

FS

V-C

L

0.11

3 0.

099

0.10

2 0.

089

0.07

9 FS

V-C

R

FSV-

CV

0.11

6 0.

109

0.10

7 0.

108

0.09

6 0.

873

0.83

0 0.

947

0.93

5 0.

683

FSV-

CW

0.

039

0.02

8 0.

037

0.02

7 0.

031

>0.1

23

>0.1

08

>0.1

16

>0.1

03

>0.0

92

0.94

0 0.

830

0.91

0 0.

960

0.86

0 FS

V-C

X 0.

040

0.03

0 0.

040

0.03

0 0.

040

0.11

0 0.

100

0.11

0 0.

100

0.10

0 FS

V-C

Z 0.

990

0.93

0 0.

920

1.03

0 1.

090

FSV-

DB

0.12

8 0.

117

0.12

3 0.

108

0.10

0 FS

V-D

D

FSV-

DF

FSV-

DI

0.48

0 0.

517

0.60

9 0.

549

0.42

0 0.

28 1

.10

0.32

0.1

1 1.

07

FSV-

DQ

0.

036

0.02

3 0.

029

0.02

0 0.

023

0.13

1 0.

131

0.13

1 0.

122

0.11

8 FS

V-D

R

FSV-

DU

FS

V-EH

0.

026

0.02

2 0.

026

0.02

0 0.

021

0.11

4 0.

103

0.11

0 0.

099

0.09

2 0.

960

0.84

5 0.

858

1.04

3 0.

882

FSV-

EQ

0.20

8 0.

205

0.19

7 0.

198

0.21

8

FSV-

FB

0.03

3 0.

025

0.03

1 0.

022

0.03

0 0.

097

0.09

1 0.

091

0.08

2 0.

090

FS

V-FJ

NIS

Ta

NIS

Tb

NN

IST

0.03

1 0.

015

0.02

6 nq

0.

028

0.13

9 0.

109

0.11

3 >0

.107

0.

114

>0.0

43

>0.0

36

>0.0

37

>0.0

35

>0.0

44

>0.1

19

>0.1

15

>0.1

10

>0.1

10

>0.1

13

4 4

4 2

4 4

4 4

2 4

N

14

14

14

13

14

23

23

23

23

23

7 7

7 7

7 3

3 3

3 3

Min

0.

020

0.02

0 0.

018

0.01

7 0.

018

0.08

2 0.

091

0.09

1 0.

082

0.07

9 0.

420

0.48

0 0.

415

0.48

0 0.

449

0.59

0.1

9 0.

59 0

.20

0.10

M

edia

n 0.

039

0.02

7 0.

035

0.02

2 0.

032

0.12

9 0.

120

0.12

8 0.

122

0.11

0 0.

683

0.87

3 0.

830

0.85

8 0.

935

1.07

0.2

8 1.

04 0

.32

0.11

M

ax

0.05

0 0.

037

0.04

2 0.

037

0.05

3 0.

208

0.20

5 0.

197

0.19

8 0.

218

1.09

0 0.

990

0.93

0 0.

947

1.04

3 1.

24 0

.44

1.10

0.4

0 0.

30

SD

0.00

7 0.

005

0.00

8 0.

007

0.01

0 0.

025

0.02

3 0.

019

0.01

9 0.

019

0.24

2 0.

294

0.22

5 0.

241

0.26

3 C

V 17

20

23

31

30

20

19

15

15

17

35

34

27

28

28

Npa

st

15

13

13

15

10

29

22

22

29

16

0 5

5 0

0 0

0 0

0 0

Med

ianp

ast

0.03

5 0.

024

0.03

4 0.

027

0.03

4 0.

133

0.12

1 0.

132

0.11

4 0.

117

0.95

0 0.

830

SDpa

st

0.00

9 0.

006

0.00

7 0.

005

0.01

1 0.

025

0.02

7 0.

029

0.01

7 0.

030

0.15

1 0.

162

Mea

n 0.

036

0.02

5 0.

031

0.03

5 0.

037

0.12

7 0.

112

0.11

1 0.

110

0.11

3

Srep

0.

003

0.00

3 0.

003

0.00

7 0.

002

0.00

6 0.

003

0.00

5 0.

010

0.00

9

Shet

0.

007

0.00

2 0.

002

0.00

1 0.

004

0.01

5 0.

002

0.00

6 0.

003

0.00

8

Sanl

0.

007

0.01

5 0.

008

0.01

2 0.

017

0.00

5 0.

002

0.00

1

SNIS

T 0.

011

0.01

5 0.

009

0.00

7 0.

013

0.02

4 0.

006

0.00

8 0.

010

0.01

2

NAV

N

AU

0.03

8 0.

026

0.03

3 0.

028

0.03

4 0.

128

0.11

6 0.

120

0.11

6 0.

112

0.68

3 0.

912

0.83

0 0.

858

0.93

5 0.

011

0.01

5 0.

010

0.01

1 0.

014

0.02

7 0.

025

0.02

9 0.

027

0.02

3 0.

242

0.16

6 0.

139

0.24

1 0.

263

All L

ab R

epor

t Pa

ge 4

/ 6

Round Robin L Laboratory Results All Results in µg/mL

Analytes Reported By One Laboratory

Term N

Min Medianpart

Max SD CV

Npast Medianpast

SDpast

NISTa, NISTb NNIST

MeanNIST Srep Shet Sanl

SNIST

NAV

NAU

nd nq >x

!

italics

Analyte Code 274 275 276 277 278 trans-β-Cryptoxanthin NISTb 0.047 0.053 0.048 0.042 0.0160

trans-Zeaxanthin NISTb 0.043 0.036 0.037 0.035 0.076 trans-Lutein&Zeaxanthin NISTb 0.119 0.115 0.110 0.110 0.037

9-cis-β-Carotene FSV-FB 0.02 0.02 0.02 0.02 0.14 13-cis-β-Carotene FSV-FB 0.12 0.08 0.11 0.08 0.04

Phytofluene FSV-CL 0.011 0.032 0.010 0.030 0.113 Phytoene FSV-CL 0.059 0.079 0.058 0.077 0.044

25-hydroxyvitamin D FSV-BN 0.0073 0.0079 0.0258 nq 0.041

Legend

Definition Number of (non-NIST) quantitative values reported for this analyte Minimum (non-NIST) quantitative value reported Median (non-NIST) quantitative value reported Maximum (non-NIST) quantitative value reported Standard deviation for (non-NIST) results: 0.741*(3rd Quartile - 1st Quartile) Coefficient of Variation for (non-NIST) results: 100*SD/Median

Mean of N(s) from past RR(s)

Mean of Median(s) from past RR(s)

Pooled SD from past RR(s)

Mean of all analyses (vials x duplicates) reported by given NIST analyst Number of total vials analyzed in duplicate by NIST analysts Mean of the NIST-analyzed vial means W ithin-vial pooled standard deviation Among-vial pooled standard deviation Between NIST analyst standard deviation

2)0.5Total standard deviation for NIST analyses: (Srep 2 + Shet 2 + Sanl

NIST Assigned Value

= (Medianpart + MeanNIST)/2 for analytes reported by NIST analyst(s)

= Medianpart for analytes reported by ≥ 10 labs but not NIST

2)0.5 NIST Assigned Uncertainty: (S2 + Sbtw S is the maximum of (0.05*NAV, SD, SNIST, eSD) and Sbtw is the standard deviation between Medianpart and MeanNIST. The expected long-term SD, eSD, is defined in: Duewer, et al. Anal Chem 1997;69(7):1406-1413.

Not detected (i.e., no detectable peak for analyte)

Detected but not quantitatively determined

Concentration greater than x

Non-quantitative value: heterogeneous serum, damaged sample, malfunction, etc.

Not explicitly reported but calculated by NIST from reported values

All Lab Report Page 5 / 6

C6

Lab FSV-BA FSV-BB FSV-BD FSV-BE FSV-BF FSV-BG FSV-BH FSV-BI FSV-BJ FSV-BK FSV-BL FSV-BM FSV-BN FSV-BO FSV-BP FSV-BQ FSV-BR FSV-BU FSV-BV

FSV-BW FSV-BX FSV-CB FSV-CC FSV-CD FSV-CE FSV-CF FSV-CG FSV-CH FSV-CI

FSV-CK FSV-CL FSV-CR

Round Robin L Laboratory Results

Comparability Summary

TR tR RP aT g/bT bC tbC aC TLy tLy TbX TLu TZ L&Z 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 2 1 1 1 1 2 1 1 1 1 1 1 1 2 2 1 1 2 3 1 1 3 1 1 1 1 1 1 1 1 1 1

1 1 1 1 1 1 1 1 1 1 1 1 3 1 1 1 1 2 1 1 1 1 1 2 2 1 1 1 2 2 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 2

1 1 1 1 1 2 3 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 2 1 1 1 1

1 1 1 1 1 1 1 3 1 3 1 1 1 1 1 1 1 2 1

1 2 1 1 1 1 1 1 2 2 1 2 2 1 1 1 1 2 1 1 2 1 1 1 1 1 2 1 2 2 1 1 1 2 1 2 1 1 1 1 2 3 3 1 2 1 4 4 2 2 1 1 1 1 1 1 2 1

Label Lab TR tR

RPaT

g/bTbC

tbC aC

TLy tLy

TbX TLu TZ

L&Z

n % 1 % 2 % 3 % 4

Definition Participant code Total Retinol trans-Retinol Retinyl palmitate α-Tocopherol γ/β-Tocopherol Total β-Carotene trans-β-Carotene Total α-Carotene Total Lycopene trans-Lycopene Total β-Cryptoxanthin Total Lutein Total Zeaxanthin Total Lutein&Zeaxanthin

number of participants providing quantitative data Percent of CS = 1 (within 1 SD of medians) Percent of CS = 2 (within 2 SD of medians) Percent of CS = 3 (within 3 SD of medians) Percent of CS = 4 (3 or more SD from medians)

“Comparability Score”

The Comparability Score (CS) summarizes your measurement performance for a given analyte relative to the consensus medians in this study. CS is the average distance (in units of standard deviation) of your measurement performance characteristics from the consensus performance. CS is calculated when the number of quantitative values you reported, Nyou, is at least two and at least six participants reported quantitative values for the analyte.

We define CS as follows:

MINIMUM 4,INTEGER 1

i

2 2CS = C AP+ +

i N

i

NC = Concordance =

you

2N −Median NAU i

1

FSV-CV 4 2 2 1 2 4 3

N −

FSV-CW 2 1 2 1 1 1 1 1 1 1

you

∑ =

−∑ =

you You Median NAU i 1 i

Youi i 1

FSV-DQ 1 1 1 1 1 1 4 NAU = NIST Assigned Uncertainty FSV-DR 1 1 2

For further details, please see FSV-DU 1 1 2 1

FSV-CX 2 1 3 1 1 1 FSV-CZ 1 4 2 FSV-DB 1 2 2 1 1 1 1 1 FSV-DD 2 FSV-DF 1

AP = Apparent Precision = FSV-DI 1 1 2 1 1 2 you

Duewer DL, Kline MC, Sharpless KE, Brown Thomas J, Gary FSV-EH 2 1 2 2 2 2 1 1 1 1 1 KT. Micronutrients Measurement Quality Assurance FSV-EQ 1 1 1 1 2 1 1 FSV-FB 1 2 2 1 1 1 3 1 2 1 2 FSV-FJ 1 2 1 4 2 NISTa 1 1 1 1 1 1 1 1 1 1 NISTb 1 1 1 1 1 1 1 1 1

n 41 10 12 47 27 33 14 29 27 11 27 16 15 24

TR tR RP aT g/bT bC tbC aC TLy tLy TbX TLu TZ L&Z % 1 78 60 75 64 81 64 64 83 89 73 85 81 93 79 % 2 17 40 25 28 19 30 21 10 7 18 11 13 7 13 % 3 0 0 0 4 0 3 14 3 4 9 4 6 0 4 % 4 5 0 0 4 0 3 0 3 0 0 0 0 0 4

Program: Helping participants use interlaboratory comparison exercise results to improve their long-term measurement performance. Anal Chem 1999;71(9):1870-8.

All Lab Report Page 6 / 6

C7

Appendix D. Representative “Individualized Report” for RR15

Each participant in RR15 received an “Individualized Report” reflecting their reported results. Each report included a detailed analysis for analytes that were assayed by at least five participants. The following analytes met this criterion in RR15:

• Total Retinol • trans-Retinol • Retinyl Palmitate • α-Tocopherol • γ/β-Tocopherol • δ-Tocopherol • Total β-Carotene • trans-β-Carotene • Total cis-β-Carotene • Total α-Carotene • Total Lycopene • trans-Lycopene • Total β-Cryptoxanthin • Total α-Cryptoxanthin • Total Lutein • To