Barley–3
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GERMINATION
The ability of barley to germinate fully and with vigor
is essential to normal malting practice (see Malt-2F).
The measurement of this has been designated
“Germinative Energy.” A number of methods have been
devised to evaluate this property and, predictably, the
performance of the grain if introduced into malting at
the time the test is made (Refs. 1–6). Some of these tests
involve a short pregermination steep and wash. Some
place the dry kernels directly onto moistened absorbent
paper (filter paper or blotting paper). All involve sub-
jecting the moistened kernels to optimum growth condi-
tions and measuring response over a period of several
days.
With adequate spacing between kernels on the mois-
tened absorbent paper, no advantage was found for pre-
germination steeping (Ref. 3) and a simple, reproducible
procedure was found adequate (Ref. 4).
The loss of germinative energy or germinative
capacity often is the result of the environment in
which the barley was grown or stored. Especially in
the time period immediately following harvesting of
the barley, it may be observed that the grain
germinates poorly or erratically. It is useful then to
determine “Germinative Capacity” by evaluating
viability of the kernels and thus the potential
germinative energy at some future date when
dormancy has ceased to be a factor. Two procedures
have been found useful: hydrogen peroxide treatment
(Refs. 2,5,6) and use of vital stains, such as 2,3,5-
triphenyl tetrazolium chloride (Refs. 5,6).
A. GERMINATIVE ENERGY
The method recommended (Ref. 4) involves placing
dry kernels on moistened absorbent paper and observing
germination after 2 and 3 days of incubation under opti-
mum growth conditions. It is similar to the “Aubry
Method” described in Ref. 5, p. EI9.
Reagent
(a) Tap water. Water temperature may vary within a
limited range without affecting germination results
on normal barley. Room temperature (20–23°C) is a
convenient temperature for performing this test.
Apparatus
(a) Germinator. Mangelsdorf, Minnesota, Master, or
console types of germinators can all be used for bar-
ley germination. Heating or refrigerating capa-
bilities are optional to user. Temperature ranges of
15.5–30.0°C do not affect results.
(b) Blotting paper. 120 lb/ream blotting paper provides
excellent moisture retention. If lighter weight stock
must be used, two lower blotters may be necessary
to maintain even moisture retention (see Note 1).
Method
Count out 2 × 100 kernels from representative sample
of barley.
Immerse two blotters in pan of clean, fresh, room-
temperature water for 3 min. Remove blotters from pan
and allow to drain 2 min. Place one blotter on germina-
tor tray and place the 100 kernels on blotter. Provide
adequate spacing between kernels. Place other blotter on
top of kernels. Press down evenly so that kernels are
firmly sandwiched between moist blotters. Check daily
to be sure that blotters are sufficiently damp.
After 44 hr (48 hr, if preferred) in germinator, count
germinated kernels and discard them. Return ungermi-
nated kernels to germinator. Be sure that kernels are
firmly sandwiched between moist blotters. Make final
germination count at end of 72 hr from beginning of
germination (see Note 2).
Notes
1. A 5-in. (12.7-cm) by 7-in. (17.8-cm) sheet of blot-
ting paper is adequate for 100 well-spaced kernels. Usu-
ally, it can be ordered cut to size.
2. It may be observed that, while acrospires and root-
lets usually grow simultaneously, on occasion there is
acrospire growth without rootlet growth. This would be
considered a viable kernel. If abnormalities in growth
are excessive, or if there is marked lack of uniformity of
growth, report such observations.
Calculation
Report 44-hr (48-hr) and 72-hr germinations as %.
Example
Germination at 44 hr = 68%; at 72 hr = 97%.
In collaborative tests (Ref. 4) , within-laboratory error
(Sr) varied from 0.85 to 3.8% and between-laboratory
error (Sb) from 0.21 to 0.55%.
References
1. American Society of Brewing Chemists. Report of Subcommit-
tee on Methods of Barley Analysis. Proc. 1946, p. 92.
2. Ibid., Proc. 1947, p. 121.
3. Ibid., Journal 34:106, 1976.
4. Ibid., Journal 35:114, 1977.
5. European Brewery Convention. Analytica-EBC, 5th ed.
Schweizer Brauerei-Rundschau, CH 8047, Zurich, Switzerland,
1998.
6. Institute of Brewing. Recommended Methods of Analysis.
Institute of Brewing, 33 Clarges St., London, England W1Y
8EE, 1982.
1958, rev. 1977
doi: 10.1094/ASBCMOA-Barley-3
Barley–3
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B. GERMINATIVE CAPACITY—HYDROGEN
PEROXIDE METHOD
(International Method)
Hydrogen peroxide is used as a medium for overcom-
ing latent germinative response in freshly harvested bar-
leys (Refs. 1–3).
Reagent
(a) Hydrogen peroxide. Keep in refrigerator, 30%,
stabilized, analytical reagent grade.
Apparatus
(a) Beakers, 150 mL.
(b) Watch glasses, approx. 65 mm diameter.
(c) Tea strainer, or pieces of clean cheesecloth.
Method
Prepare 0.75% solution of peroxide (H2O2) by diluting
5 mL 30% H2O2 (reagent a) to 200 mL with distilled
water.
Count 100 kernels of barley from each sample to be
tested, and place each 100 kernels in separate 150-mL
beakers. Add 100 mL 0.75% H2O2 solution to each
beaker, pushing down any floating kernels. Cover beak-
ers with watch glasses.
After 48 hr since beginning of experiment, drain off
H2O2 solution through a tea strainer or piece of cheese-
cloth. Count the number of kernels in each sample that
show evidence of chitting (examination of kernels after
18–24 hr may show extensive chitting and provide use-
ful information before 48 hr has elapsed).
Return only those kernels that show no evidence of
chitting to their respective beakers and add 100 mL
freshly prepared 0.75% H2O2 solution.
After 96 hr since beginning of experiment, drain as
above (g) and count additional kernels that have chitted.
Calculation
Record total number of kernels that have chitted in
time period selected.
Example
98% (48 hr).
In collaborative tests (Ref. 1), within-laboratory error
(Sr) varied from 0.9 to 1.7 and combined-laboratory
error (Sc) from 1.8 to 2.8.
References
1. American Society of Brewing Chemists. Report of Subcommit-
tee on Barley Analysis. Journal 36:114, 1978.
2. European Brewery Convention. Analytica-EBC, 4th ed.
Schweizer Brauerei-Rundschau: Zurich, Switzerland, 1987.
3. Institute of Brewing. Recommended Methods of Analysis. The
Institute: London, 1982.
1978, rev. 1981
C. GERMINATIVE ENERGY, GERMINATIVE
CAPACITY, AND WATER SENSITIVITY—
SIMULTANEOUS DETERMINATION
This method is designed to simultaneously determine
percent germinative energy, germinative capacity, and
water sensitivity. This method was found to be accept-
able when compared with Barley-3A, Barley-3B, and
EBC Method 3.6.2 (Ref. 3.)
Reagents
(a) Tap, distilled, or deionized water, containing <0.2
mg/L of free chlorine.
(b) Hydrogen peroxide, 30% (w/v), stabilized analytical
grade.
(c) Hydrogen peroxide, 0.75% (w/v). Prepare by dilut-
ing 5 mL of hydrogen peroxide (reagent b) in a 200-
mL volumetric flask with distilled water and mix.
Apparatus
(a) Glass petri dishes, 90-mm.
(b) Filter papers, white, Whatman No. 1 or equivalent,
90-mm.
(c) Dark cabinet, controlled at 18–21°C.
(d) Volumetric pipets, 10-mL, graduated.
(e) Polyethylene bags, to cover petri dishes.
(f) Volumetric flask, 200-mL.
Method
For each barley sample to be tested, prepare four petri
dishes by placing two filter papers in the bottom of each
dish. Place 100 kernels of each sample to be tested into
each dish. Into two of the dishes, labeled A and B, add 4
mL of water, and into the remaining two dishes, labeled C
and D, add 8 mL of water. Ensure that the kernels are
uniformly distributed over the surface of the filter paper.
Cover all dishes with lids, place into a polyethylene bag,
and seal. Incubate in the cabinet, and remove chitted
kernels after 24, 48, and 72 hr of steeping to avoid
excessive moisture uptake by the early-germinated kernels.
For each sample, record the percentage chitted after 72 hr.
For dishes A and B, place all ungerminated kernels
back in the petri dish, ventral side touching the paper.
Add 2 mL of 0.75% hydrogen peroxide (reagent c).
Incubate an additional 48 hr (120 hr total since steep-
ing), and record percent chitted for each sample.
Calculations
Germinative energy
Calculate the percent germinative energy (GE) as:
A+ B
GE, % =
2
in which
 Barley–3
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A = number of kernels germinated in 4-mL dish A Example

after 72 hr; After 72 hr, dishes A and B contain an average of 99
B = number of kernels germinated in 4-mL dish B germinated kernels, and dishes C and D contain an aver-
after 72 hr. age of 97 germinated kernels. After 120 hr, 100 kernels
Report results to nearest whole percent. (average) have germinated in dishes A and B.
GE, % = 99
Germinative capacity GC, % = 100
Calculate the percent germinative capacity (GC) as: WS, % = 99–97 = 2
A′ + B ′
GC, % = Precision
2
in which Based on a collaborative study (Ref. 2), repeatability
A′ = no. of kernels germinated in 4-mL dish A, incu- coefficients of variation of 0.6–1.7% and reproducibility
bated 48 hr with H2O2 (peroxide). coefficients of variation of 0.7–2.1% can be expected for
B′ = no. of kernels germinated in 4-mL dish B, incu- GE.
bated 48 hr with H2O2 (peroxide). Repeatability coefficients of variation of 0.6–1.4%
Report results to nearest whole percent. and reproducibility coefficients of variation of 0.6–1.8%
can be expected for GC.
Water sensitivity Repeatability coefficients of variation of 22.7–54.1%
Calculate the percent water sensitivity (WS) as: and reproducibility coefficients of variation of 39.8–
A+ B C + D
58.2% can be expected for WS.
WS, % = −
2 2 References
in which 1. American Society of Brewing Chemists. Report of Subcommit-
A = number of kernels germinated in 4-mL dish A tee on Water Sensitivity in Barley. Journal 54:257, 1996.
2. American Society of Brewing Chemists. Report of Subcommit-
after 72 hr; tee on Simultaneous Determination of Germination Energy,
B = number of kernels germinated in 4-mL dish B Water Sensitivity, and Germination Capacity in Barley. Journal
after 72 hr; 55:179, 1997.
C = number of kernels germinated in 8-mL dish C 3. European Brewery Convention. Analytica-EBC. Method 3.6.2.
BRF, 1987.
after 72 hr;
D = number of kernels germinated in 8-mL dish D 1997
after 72 hr.
Report results to the nearest whole percent.