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 [email protected]; 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: [email protected]
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 [email protected])
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: [email protected]
A4
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
r-.....-n!:Sll-=:;;::;::--TI;"?""""""~ '° __ ""'"'"""....,.. __ _ 0 ' i.» 0 0
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: [email protected], or me at 301/975-3120; e-mail: [email protected]; 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.
C1
Rou
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L L
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y R
esul
ts
All R
esul
ts in
µg/
mL
Tota
l Ret
inol
tra
ns-R
etin
ol
Ret
inyl
Pal
mita
te
α-Tocopherol
γ/β-Tocopherol
δ-Tocopherol
C2
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
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6.53
6.8
9 2.
61 2
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2.40
1.9
1 2.
55
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0.65
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1.04
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6.78
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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