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Roslen Anacleto
Current position: Senior Associate Scientist Education and training
2009, Projects in Controlled Environments (PRINCE2), Singapore 1997, MS in Computer Science, University of the Philippines, Los Baos, Laguna 1991, BS in Computer Science, University of the Philippines, Los Baos, Laguna
Work experience
2009 2005 2003 2000 1998 1991 present, Senior Associate Scientist, Grain Quality and Nutrition Center, IRRI 2009, Programmer, Experiment Station, IRRI 2005, Assistant Professor and Head, MIS Unit, University of the Philippines Open University 2003, IT Consultant, various local and international clients 2000, Academic Head, Systems Technology Institute, Cagayan De Oro City 1998, Assistant Professor, Central Mindanao University, Musuan, Bukidnon
Research highlights
Keyless data entry for grain quality evaluation Implemented barcoding for sample labeling and tracking at the quality evaluation laboratory Currently working on a LIMS implementation for GQNC Member of the IRRI Experiment Station ISO 14001:2004 certification working group Conversion of various databases at the Experiment Station from MS Access silos to a true relational database
Tools to measure quality do not give breeders accurate enough data about eating quality.
Current tools to measure amylose, gel temp and gel consistency are not globally standardised. Consumers do not have consistent adjectives to describe the quality of rice they like. New analytical technologies facilitate a surge on new understanding of sensory quality. Improved information and communication technologies make global collaboration routine rather than a challenge. We are in an era where genotyping is becoming routine. Serious investment into new, accurate phenotyping tools would greatly help genotyping work.
3
The INQR
80 members from almost every rice quality evaluation program. NARES
ARI
The amylose project then began with 30 labs, and by the time it was concluded in 2010, there were 45 labs and the International Standards Organisation involved.
Compound granules
7
Goami 2
35 30 25 20 15 10 5 0 -5 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
IR 24
IR 64
Sample Number
11
30 25 20 15 10 5 0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
Sample Number
12
Amylose content using 2 brands of potato amylose, done in one laboratory, by one person
40 35
30 25 20 15 10 5 0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 13 17
Brand 1 Brand 2
Sample Number
14
35 30 25 20 15 10 5 0 -5 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
Sample number
16
Testing the relationship between amylose content by SEC and by iodine using both calibration methods
Calibration values by iodine
35
30
30
25
25
20
20
15
15
10
10
5
5
0
0
-5.0 0.0 -5 5.0 10.0 15.0 20.0 25.0 30.0
0 -5
10
15
20
25
30
35
30
20
10
IR 24
0
0 1 2 3 4 5 6 7 8 9 10
IR 64
11 12 13 14 15 16 17
AM
0.4
Absorbance
0.3
90% AP
0.2
70% AP
0.1 0 480 520 560 600 640 680 720 760 800
Wavelength (nm)
19
Technically, the best method should be the one that more accurately reports amylose. This means calibration by SEC and making the AP/iodine complex invisible, so a wavelength of 720 nm. AM-I and AP-I signatures fade through time, thus a standing time of 0 min was chosen.
0.7
Normalized DRI
0.3
0.4
0.5
0.6
0.1
0.2
AM
AP
10000 10000
1000 1000
100 100
10 10
11
20
Chain-Length
Performance
SEC calibrated standards measured at different wavelengths at 0 min
Goami 2 Goami 2
IR 24
IR 64
IR 24
IR 64
21
20 15 10 5 0 0 1 2 3 4 5
22
Standard
Using the same test method Measuring on identical material At the same laboratory By the same operator Using the same equipment At different laboratories By different operators Using different equipment
23
Procedure
1 5 SEC calibrated standards, 720 nm wavelength, 0 min standing time 18 samples from the same source distributed to 44 participating labs Agreed time frame for conducting the experiment Each lab assigned the same technician and equipment
24
Repeatability conditions
Reproducibility conditions
Using the same test method Measuring on identical material At the same laboratory By the same operator Using the same equipment At different laboratories By different operators Using different equipment
Boxplot of results
IR 74 IR 64 Seraup 27
RC 18
Sample
25
26
ISO 5725-2:1994(E)
Excel worksheets
Form B, Form C 5
Discard or correct outliers Check the results for consistency and outliers**
Outliers found?
No **ISO 5725 suggests two ways of checking for consistency and outliers: 1. Graphical Mandels k and h statistics 2. Numerical Cochrans and Grubbs tests 7
Compute the general mean, repeatability std. dev., and reproducibility std. dev., etc.
27
28
4.50
4.00
1% significance 5% significance
Mandels k-statistic
3.50
k=2.10
3.00
k=1.72
2.50
2.00
1.50
1.00
0.50
0.00 lab1 lab2 lab4 lab6 lab7 lab10 lab11 lab12 lab13 lab15 lab17 lab18 lab21 lab22 lab23 lab24 lab25
Laboratory
29
4.50
4.00
1% significance 5% significance
Mandels k-statistic
3.50
Sample 4
k=2.10 2.10
3.00
1.72 k=1.72
2.50
2.00
1.50
1.00
0.50
0.00 lab26 lab27 lab28 lab29 lab30 lab32 lab33 lab36 lab37 lab38 lab41 lab42 lab46 lab47 lab54 lab64 lab65
Laboratory
30
4.50
4.00
1% significance 5% significance
Mandels k-statistic
3.50
Sample 4
k=2.10
3.00
k=1.72
2.50
2.00
1.50
1.00
0.50
0.00 lab4 lab6 lab10 lab13 lab26 lab28 lab30 lab47 lab54 lab65
Laboratory
31
5.00
4.00
3.00
Mandels h-statistic
2.00
Sample 4 Sample 5
1.00
0.00
Sample 9 Sample 10
-1.00
Sample 11
lab1
lab2
lab4
lab6
lab7
lab10
lab11
lab12
lab13
lab15
lab17
lab18
lab21
lab22
lab23
lab24
lab25
Sample 12 Sample 13
-2.00
Sample 14 Sample 15
-3.00
k=-1.91 h=-2.45
-4.00
-5.00
-6.00
Laboratory
32
5.00
4.00
3.00
Mandels h-statistic
2.00
Sample 4 Sample 5
1.00
0.00
Sample 9 Sample 10
-1.00
Sample 11
lab26
lab27
lab28
lab29
lab30
lab32
lab33
lab36
lab37
lab38
lab41
lab42
lab46
lab47
lab54
lab64
lab65
Sample 12 Sample 13
-2.00
-3.00
k=-1.91 h=-2.45
Sample 17 Sample 18
-4.00
-5.00
-6.00
Laboratory
33
h=2.45 k=1.91
1% significance 5% significance
Sample 1 Sample 2 Sample 3 Sample 4
Mandels h-statistic
2.00 1.00 0.00 -1.00 lab12 -2.00 -3.00 -4.00 lab26 lab30 lab54 lab65
Sample 5 Sample 6 Sample 7 Sample 8 Sample 9 Sample 10 Sample 11 Sample 12 Sample 13 Sample 14 Sample 15
k=-1.91 h=-2.45
-5.00 -6.00
Laboratory
34
17 14 Total within-lab outliers - 33 (10 unique labs) Total between-lab outliers - 25 (5 unique labs) Total outliers - 58
16
11
36
23.635 29.792
Note on the suffixes: - c denotes rejection due to Cochrans statistic - g denotes rejection due to Grubbs statistic 37
-82.071 11.670
3 3 3 2 3 28c, 12g, 54c, 47c, 54g, 26c, 30g 26c, 30c, 47c 65g 65g 30g, 65c
Note on the suffixes: - c denotes rejection due to Cochrans statistic - g denotes rejection due to Grubbs statistic
38
Summarized results
Laboratories with repeatability problems -> lab 26, 28, 30, 47, 54, 65 Laboratories with reproducibility problems -> lab 12, 26, 30, 54, 65 Laboratories with both problems! -> lab 26, 30, 54, 65
4 out of 34
11.76%
39
With repeatability issues : 26, 28, 30, 47, 54,65 With reproducibility issues : 12, 26, 30, 54, 65
IRRI GQNC
40
VL
VH
So then
Previously the categories were waxy, very low, low, intermediate, high. That will not change. Current work is done in establishing the relationship between the values obtained using both methods This relationship would be determined through rigorous computational methods on precise data sets Any immutable relationship will be used as correction factor should breeders/rice scientists refer to old values
43
44
Conclusions
Rigorous analytical and computational procedures were done to move from apparent to actual amylose content A platform for collaborative work is in place --- INQR Current classifications for amylose content: waxy, very low, low, high, and very high will not change Work is in progress to determine the relationship between old and new amylose values A series of other collaborative work will be undertaken to improve existing tools for measuring quality Current research activities at the Grain Quality and Nutrition Laboratory progressively improve our understanding of rice quality
45
Acknowledgement
The authors thank The INQR participating laboratories and other contributing members PBGB breeders and GRC for providing quality samples The GQNC technicians: Teodie, Johnny, Boy, Dennis, Leah, Ferdie, and Lucy for their usual outstanding support
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