SEED TESTING METHODS,
PROCEDURES AND
LABORATORY STANDARDS
LEILA DELA PEÑA AND NOEMI AVENIDA
SEED ANALYSTS
BPI-NSQCS 1
National Seed Quality Control Services
SEED TESTING
first seed quality control scheme developed to determine the
planting value of seeds
developed to avoid hazards in crop production due to planting
of inferior seeds
Seed Testing
Procedures for seed testing
in accordance with the rules
and regulations of the
International Seed Testing
Association (ISTA) or what is
sanctioned by the National
Seed Industry Council thru
Technical Working Group on
Seed Certification and
Standards
Reasons for Testing Seeds
1. To determine quality or suitability of seed for sowing purposes.
2. To obtain information for labeling seeds, especially seed stocks.
3. To obtain information for basis of distribution, storage and
disposition of seeds.
4. To obtain information for basis of buying and selling.
Other Reasons
1. For the evaluation of research in seed technology.
2. To determine the processing and blending procedures and to
evaluate the blending process.
3. To determine the potential and actual seed quality problems and
to offer solutions to these problems.
SUBMISSION AND
RECEIPT OF
FILE BULK SEED SAMPLES
SAMPLES
PURITY, OTHER VARIETIES
FRACTIONS
MOISTURE TEST
CHECK
RESULTS
RECORDING OF PURITY TEST
SAMPLES
REPORT RESULTS
GERMINATION TEST
PREPARATION OF WORKING
ISSUE APPROPRIATE
SAMPLES
CERTIFICATES, TAGS, SEALS
Accelerated Aging Test VARIETAL TEST
Scarification
TO PRODUCER PROCESSOR,
FARMER
SEED HEALTH TEST
OTHER TEST
MOISTURE CONTENT
DETERMINATION
IMPORTANCE OF MOISTURE TESTING
Moisture content
used as an index for the proper time of harvesting.
affects the mechanical efficiency and the degree of
mechanical seed damage incurred during harvesting and
processing of grain crops.
Moisture Content (MC)
Very critical determinant of seed longevity and grain
quality during storage.
1% difference in moisture content can increase or
decrease seed longevity two times.
ISTA Rules are used to determine the MC of the seeds.
Moisture Content (MC)
Milling recovery in rice is affected by grain MC
Accurate determination of grain moisture is economically important
For valid interpretation of results in the evaluation of varietal
performance in seed yield and in seed storage (crop research) =
accurate MC determination is a MUST!
METHODS OF MOISTURE
CONTENT DETERMINATION
1. Primary or Oven Method
2. Secondary or Practical Method
Primary or Oven Method
Moisture content is calculated directly from the weight loss
due to removal of moisture.
Secondary or Practical Method
MC is taken by moisture meters wherein a direct reading
scale or print out is obtained.
Secondary or
Practical Method
1. Infra-red Lamp
Secondary or Practical
Method
2. Kett digital and
Kett riceter
Oven Method
Determination of
Moisture Content
Using Oven Method
Oven Method
Objective
To determine the moisture content of seed by methods suitable
for routine use.
Definition
The moisture content of a sample is the loss in weight when it is
dried.
Expressed as a percentage of the weight of the original sample.
Oven Method
Principle
The methods prescribed are designed to reduce
oxidation, decomposition or the loss of other volatile
substances while ensuring the removal of as much
moisture as possible.
Apparatus
1. Adjustable 2. Constant-
Grinding Mill Temperature
Oven
3. Accessories for oven method
a. Desiccator b. Containers c. Tong
4. Analytical Balance
Procedure
1. Submitted sample is accepted for moisture determination
(only if it is in an intact, moisture – proof container from
which as much air as possible has been excluded).
2. Determination shall be started as soon as possible after
receipt.
Prescribed Methods
1. Low Constant Temperature Oven Method
2. High Constant Temperature Oven Method
Low Constant Temperature Oven
Method
103 + 2 °C dry for 17 + 1 hours
1.Working sample must be evenly distributed over the
surface of the container.
2.Weigh the container and its cover before and after filling.
Low Constant Temperature Oven
Method
3.Place the container rapidly on top of its cover, in an oven
maintained at 103 + 2 °C dry for 17 + 1 hours.
4.The drying period begins at the time the oven returns to
the required temperature.
5.At the end of the prescribed period cover the container
and place in a desiccator to cool for 30 – 45 mins.
Low Constant Temperature Oven
Method
6.After cooling, weigh the container with its cover and
contents.
7.The relative humidity of the ambient air in the
laboratory must be less than 70% when the
determination is carried out.
High Constant Temperature Oven
Method
Procedure same as in LCT oven method
Temperature range is 130 – 133°C
Corn dried for 4 hours
Other cereals (rice, sorghum) – 2 hours
Other species – 1 hour
Computation of MC Using Oven
Method
MC = W1 – W2 x 100
W1
MC = _____%
Where:
W1 = weight of seed before drying
W2 = weight of seed after drying
MC = Moisture Content
Tolerance
The difference between 2 replications should be 0.2%.
However, if the results of the two replications are more
than this value, re – testing should be done.
Species for Which Low Constant Temperature
(103oC) Oven Method is Used
Onion Radish All tree sp.
Peanut Eggplant
Mustard Pepper
Pechay Soybean
Cabbage Cotton
Cauliflower
Species for Which High Constant Temperature
(130oC) Oven Method is Used
Celery Tomato Patola
Asparagus Rice Squash
Beets Beans Kangkong
Watermelon Peas Upo
Cucumber Sorghum Ampalaya
Melon Spinach Tobacco
Carrot Wheat Lettuce
Corn
Illustrated Procedure for
Moisture Content
Determination
Weighing of Tin Cans
Grinding of Seeds
Oven Method
Tin Cans in the Dessicator Weighing of Dried Seeds
Working Sheet for MC
Recording Sheet for MC
PHYSICAL PURITY TEST
Physical Purity Test
Separation of Component Parts as Pure Seeds, Other Seeds and Inert Matter
Objective
To determine percentage composition by weight of the
sample being tested and by inference the composition of
the entire seed lot.
To identify the various species of seeds and inert particles
constituting the sample.
Definition
Pure Seed
Refers to the species stated by the sender and shall include
all botanical varieties or cultivars of the species.
Immature, undersized, shriveled, diseased and germinated
provided can be definitely identified and not transformed
into fungal sclerotia, smut balls, nematode galls.
In graminae: Florets with caryopsis, free caryopsis and seed
unit more than half of the original size.
Definition
Other Seed
Refers to seed units of other plant species other than
that of pure seed
Inert Matter
Includes seed units and all other matters and
structures not defined as pure seed or other seed
such as:
a. Seed units that are sterile
b. Florets with caryopsis ½ original size
c. Broken or damaged seed units ½ less original size
Inert Matter
Unattached sterile florets, empty glumes, lemma, palea, chaff,
stem, leaves, fungal bodies like ergot, sclerotia, smut balls, sand,
soil, stone and other non-seed matter
Leguminosae, Brassicaceae, Pinaceae with seed coat entirely
removed
Separated cotyledons of leguminosae
General Principle
The three components are separated as:
1.Pure Seed
2.Other Seeds
3.Inert Matter
Weighed and percentage of each part is determined by
weight
Identify seed and inert matter present
The three components are separated as:
1. Pure Seed 2. Other Seeds 3. Inert Matter
(Other Crop Seeds and Weed Seeds)
Apparatus
b. Seed Blower
a. Sieve
Apparatus
c. Purity Workboard
d. Magnifying Glass
f. Petri Dish
e. Forceps
Procedure
Working Sample
The Purity Analysis for rice seeds shall be 70 grams.
Weigh to the minimum number of decimal places
necessary to calculate the % of component parts to one
decimal place.
Working Sample
Weight of Working Number of Decimal
Sample (grams) Places
Less than 1.0000 4
1.000 to 9.999 3
10.00 to 99.99 2
100.0 to 999.9 1
1000 or more 0
Separation
Separate sample into component parts
by seed blowers/sieves to separate light
seeds from heavy seeds.
Repeat separation manually to identify
the other seeds and inert materials
present in the sample
Weigh the different components in
grams.
Separating sample into
Component Parts by
Seed Blower
Calculation and Expression of
Results
Results should be calculated to one decimal place.
% must be based on the sum of the weight of
components, not on the original weight of working
sample.
Calculation and Expression of
Results
The sum of the weights of components must be
compare with original weight as a check of material loss
or error.
A discrepancy of more than 5% of the original weight, a
re-test must be made.
Reporting Results
Results shall be given to one decimal place and the sum of
components must be equal to 100%.
Components less than 0.05% shall be reported as “Trace”.
Report scientific name of other seed.
Report the kind of inert matter.
Computation:
Purity - 70 g
g %
Pure seed - 67.00 95.7
Other seeds - 2.10 3.0
Weed seed - 0.02 (0.03) Trace
Inert Matter - 0.91 1.3
70.03 g 100.0 %
VARIETAL PURITY TEST
VARIETAL PURITY TEST
Determination of Other Varieties thru Varietal Purity Test.
Objective
To determine the extent to which submitted sample
conforms to species or cultivar claimed for.
To gain information on the amount of other varieties
and identify the mixtures as much as possible.
Definition
Other varieties
includes other kinds, varieties of rice.
Principle
Determination is made by count and expressed as number in
percentage of seeds found in the sample examined.
Seeds are compared with authentic samples.
Apparatus
Purity Workboard
w/
Diaphanoscope
Magnifying Glasses Stereo Microscope
Forceps, Spatula, & Petri Dish
Procedure
The 500 grams working samples for rice and corn are
examined seed by seed.
The number of other varieties is reported as number
per working sample.
Other varieties are determined.
Other Varieties
A. Seeds
Sample is searched for other cultivars side from subject
sample but of same species.
Separate the mixtures or other varieties
Identify the mixtures or other varieties.
Samples of other varieties based on:
GRAIN LENGTH
Samples of other varieties based on:
GRAIN SHAPE
Other Varieties
B. Seedling Characteristics
Germinate / plant seeds of other varieties together with the
authentic sample.
Observe the seedling characteristics.
Calculation and
Expression of Results
Result is expressed as number of seeds per unit
weight as prescribed in the standard.
SUBMISSION AND
RECEIPT OF
FILE BULK SEED SAMPLES
SAMPLES
PURITY, OTHER VARIETIES
FRACTIONS
MOISTURE TEST
CHECK
RESULTS
RECORDING OF PURITY TEST
SAMPLES
REPORT RESULTS
GERMINATION TEST
PREPARATION OF WORKING
ISSUE APPROPRIATE
SAMPLES
CERTIFICATES, TAGS, SEALS
Accelerated Aging Test VARIETAL TEST
Scarification
TO PRODUCER PROCESSOR,
FARMER
SEED HEALTH TEST
OTHER TEST
SEED STANDARDS
Factors Breeder Foundation Registered Certified
Seed Seed Seed Seed
Pure seed (min.) 99% 99% 98% 98%
Weed & other crop 0% 0% .04% .04%
seed (max).
Inert matter (max.) 1% 1% 2% 2%
Other varieties, 0 2 5 20
grains/500g (max.)
Germination (min.)* 85% 85% 85% 85%
Moisture content 14% 14% 14% 14%
(max.)
*based on pure seed only.
GERMINATION TEST
Germination is the emergence and development from the seed
embryo of those essential structures of the different seeds.
Germination Testing (Setting and Evaluation).
Objective:
To determine the maximum germination potential of a seed lot,
which can then in turn be used to compare the quality of different
lots and also estimate the field planting value.
Requirements on Germination Test:
Adequate moisture
Favorable temperature
Three levels of temperature
1.Minimum Temperature
2.Optimum Temperature
3.Maximum Temperature
Suitable substratum
Light
Germination Methods
Source of seed
No. of seed to plant
The seeds shall receive no pre treatment
Germination media or substrate
Paper substrates are used for the
following:
Top of Paper (TP)
Between Paper (BP)
Pleated Paper (PP)
Top of Paper (TP)
Between Paper (BP)
Pleated Paper (PP)
Duration of the Test
varies in different species
1st count - when majority of seedlings have reached the stage of
development wherein proper evaluation is possible
counting of germination tests are done during the preliminary or first
counts and subsequently until the final count
Scientific Name Common Prescription for First Final Additional
Name Substrate Temperature Count Count directions
(0C) including rec. for
breaking dormancy
Ipomoea aquatica Kangkong BP; S 30 4 10 --
Lactuca sativa Lettuce TP; BP 20 4 7 Prechill
Lolium perenne Perennial TP 20-30; 15-25; 5 14 Prechill; KNO3
Luffa acutangula grass BP; S 20 4 14 KNO3
Lycopersicon Patola TP; BP; S 30 5 14 Preheat (500C)
lycopersicum Tomato TP; BP; S 20-30 5 14 soak in
Oryza sativa Rice BP; S 20-30; 25 4 14 H20 or HNO3 (24
Momordica Ampalaya BP; S 20-30; 30 5 9 hrs.)
charantia Lima beans BP; S 20-30; 25 5 9 KNO3
Phaseolus lunatus beans BP; S 20-30; 25; 20 4 7
--
Phaseolus vulgaris Mongo BP; S 20-30; 25; 20 5 8
--
Phaseolus mungo Pea BP; S 20 4 14
--
Pisum sativum Winged TP; BP 20-30; 30 4 10
--
Psophocarpus bean TP; BP; S 25 4 10
--
tetragonolobus Kudzu TP; BP; S 20-30; 20 7 14
--
Pueraria javanica Radish TP; BP 20-30 4 10
H2SO4
Raphanus sativus Eggplant TP; BP 20-30 7 21
Solanum melongena Sorghum TP; BP; S 15; 10 4 8 Prechill
Sorghum vulgare Spinach BP; S 20 5 8 KNO3
Spinacia oleraceae Wheat BP; S 20-30; 25 4 7 Prechill
Triticum aestivum Pole Sitao 20-30; 25; 20 Prechill
Vigna sesquipedalis Corn Preheat (30-350C)
Zea mays Prechill; GA3
Agricultural and Vegetable Seeds --
--
Hypogeal
Germination
Epigeal
Germination
Growth and Development of the Seedling
Hypogeal Germination of Wheat (Triticum aestivum)
Categories of Seedlings and Seeds
1. Normal Seedlings 2. Abnormal Seedlings
Damaged
coleoptile
Shoot deformed Root not sufficiently developed
or missing or totally missing
Categories of Seedling and Seed
3. Fresh Ungerminated
Seeds 4. Hard Seeds 5. Dead Seeds
A. Preliminary or First Count
Normal Seedlings
Doubtful seedlings that have not yet developed
Mold growth/deteriorated
B. Second Counts to Final Counts
Normal Seedlings
Abnormal Seedlings
Fresh ungerminated
Dead seeds
When to Retest?
Out of tolerance
Presence of firm ungerminated seed.
Evidence that the results may not be reliable:
• Improper test condition
• Errors in seedling evaluation
• Presence of fungi and bacteria
• Inaccuracies in counting and recording results
Seedling abnormality due to chemical treatment
Average Average
percent No. of replicates percent No. of replicates
germination of 100 seeds germination of 100 seeds
4 3 2 4 3 2
A B C D E A B C D E
99 2 5 - - 75 26 17 16 14
98 3 6 5 - 74 27 17 16 14
97 4 7 6 6 73 28 17 16 14
96 5 8 7 6 72 29 18 16 14
95 6 9 8 7 71 30 18 16 14
94 7 10 9 8 70 31 18 17 14
93 8 10 9 8 69 32 18 17 14
92 9 11 10 9 68 33 18 17 15
91 10 11 11 9 67 34 19 17 15
90 11 12 11 9 66 35 19 17 15
89 12 12 11 10 65 36 19 17 15
88 13 13 12 10 64 37 19 17 15
87 14 13 12 11 63 38 19 18 15
86 15 14 13 11 62 39 19 18 15
85 16 14 13 11 61 40 19 18 15
84 17 14 13 11 60 41 19 18 15
83 18 15 14 12 59 42 19 18 15
82 19 15 14 12 58 43 19 18 15
81 20 15 14 12 57 44 19 18 15
80 21 16 15 12 56 45 19 18 15
79 22 16 15 13 55 46 20 18 15
78 23 16 15 13 54 47 20 18 16
77 24 17 15 13 53 48 20 18 16
76 25 17 16 13 52 49 20 18 16
Maximum Tolerated Ranges in Germination Percentage
REFERENCE:
Malabanan , Josephine C. and Torio, Elalin T. SEED TESTING METHODS,
PROCEDURES AND LABORATORY STANDARDS
