AACCI Approved Methods Technical Committee Report:

Collaborative Study on a Method for Determining the

Gelatinization Temperature of Milled Rice Flour
Using the Rapid Visco Analyser
J. M. C. Dang1 and M. L. Bason1,2

GT. Microscopy methods used to determine GT are laborious,

ABSTRACT time-consuming, and subjective (4).
Gelatinization temperature (GT) is commonly measured to assess In the ASV assay the extent of spreading or disintegration of
the cooking and processing quality of rice. Traditional methods of milled rice kernels soaked in a 1.7% solution of potassium
estimating the GT of rice using the amylograph and differential scan- hydroxide is recorded. Grains with a low GT (score of 6–7)
ning calorimetry (DSC) are time-consuming and, in the case of the dissolve completely; those with an intermediate GT spread par-
amylograph, require large quantities of samples, which is not well tially (score of 3–5); and those with a high GT are essentially
suited to rice breeder requirements. The Rapid Visco Analyser (RVA)
unaffected by the alkali (score of 2) (4,12). The ASV assay is
is commonly used in laboratories to measure the pasting properties of
rice and provides the advantages of speed, ease of use, and small sample
suitable for screening but is subjective.
size requirements. A collaborative study was conducted by the AACC In DSC methods two calorimeters are heated at a constant
International Rice Milling and Quality Technical Committee to assess a rate and maintained at the same temperature: one calorimeter
new method for determining the GT of milled rice flour using the RVA. contains the rice flour–water slurry and the other an empty ref-
Twelve laboratories analyzed three rice flour samples to evaluate the erence. The difference in power needed by the calorimeters to
performance of the method. Within-laboratory repeatability (Sr) values cause an identical temperature change is measured; this is the
were ≤0.15 degrees Celsius for the method and between-laboratory heat flow into the sample (4). DSC methods are time-consum-
reproducibility (SR) values were ≤0.25 degrees Celsius. RVA GT results ing and can give inconsistent results due to the complexity of
were similar to but more precise than those obtained from parallel test- the thermal properties of starch and variable measurement con-
ing using the amylograph and DSC. The RVA method showed good
ditions, i.e., water/rice ratio and sample heating rate (4).
precision for use in determining the GT of milled rice flour.
In the amylograph method a 20% milled rice flour–water
slurry is heated at 1.5 degrees Celsius/min, and changes in the
viscosity of the paste over time is monitored. GT is estimated as
Rice cultivars grown around the world vary greatly in their
the temperature at which the viscosity curve crosses the 20 BU
cooking, sensory, and processing qualities, and preferences
line (1). The method is time-consuming (typically 25–35 min)
regarding these qualities vary by ethnic group and geographic
and requires a sample size of 100 g, which is not suitable for
region (4,5,7). Gelatinization temperature (GT) and amylose
early generation selection in rice breeding programs.
content of the component starch are the two most important
Since its development to detect sprout damage in wheat, the
factors determining the eating and processing qualities of rice
Rapid Visco Analyser (RVA; Perten Instruments) has been used
(5). GT is the critical temperature range within which starch
extensively to measure the starch pasting properties of cereals
granules undergo irreversible loss of crystalline order (3,4). Rice
and other products (20). The combination of speed, precision,
cultivars are typically classified as low (<70°C), intermediate
ease of use, and small sample size requirements make the RVA
(70–74°C), and high (>74°C) GT types (4,7,9,12). The GT of
a unique tool suitable for breeding selection, product develop-
rice is affected by variety, growing environment, starch granular
ment, screening, process control, and quality assurance.
morphology and molecular structure, quantity of minor com-
The existing RVA method for testing the pasting properties of
ponents such as phospholipids, and pretreatments such as par-
rice (AACCI Approved Method 61-02.01) (1) includes determi-
boiling (5,6,12,15–17).
nation of pasting temperature (PT), which is the point, closely
Classification methods have been developed that either pre-
following gelatinization, at which the starch granules swell suf-
dict GT or measure other physical phenomena, such as viscosity
ficiently to cause a measurable increase in viscosity (18). The
changes, that occur as starch takes up water and swells (4). These
RVA PT method overestimates GT compared with the amylo-
methods include hot-stage microscopy, alkali spread value (ASV)
graph and DSC methods (3,10,15). The reasons for these differ-
assay (11), amylography (AACCI Approved Method 61-01.01)
ences include the following:
(1), and differential scanning calorimetry (DSC) (14).
In hot-stage microscopy, the starch suspension is viewed
1) DSC methods measure the endotherm of crystalline melt-
under polarized light as it is heated on the microscope stage.
ing of the starch granule, which precedes the swelling of
Ungelatinized starch granules are birefringent when viewed
the granule (pasting) and consequent increase in viscosity
under polarized light, showing a “Maltese cross” pattern (19).
and, therefore, the PT measured (18).
The disappearance of 90–95% of these crosses (loss of birefrin-
2) The amylograph method uses a slower sample heating
gence) as the starch is heated is used to determine the endpoint
rate (1.5 degrees Celsius/min) than that used in the RVA
1 Perten Instruments of Australia Pty. Ltd., Unit 13, 2 Eden Park Drive, Macquarie Park, NSW 2113,
method (12 degrees Celsius/min), resulting in less lag
Australia. between the measured temperature and the pasting of the
2 Corresponding author. E-mail: [email protected]; Tel: +61 2 9870 3400; Fax: +61 2 9870 3401.
sample (8).
http://dx.doi.org/10.1094/CFW-59-1-0031 3) The higher sample concentration (20%) used in the amylo-
©2014 AACC International, Inc. graph method compared with the RVA method (12%) means

CEREAL FOODS WORLD / 31

 that the critical concentration (c*, the limiting concentra- Method. Collaborators were supplied with three ground
tion above which structural flow is apparent) is reached white (milled) rice samples (Pecos, Dixiebell, and Jacinto), rep-
sooner (3,13), bringing the gelatinization and pasting resenting low-, intermediate-, and high-GT varieties, respec-
points closer together. tively. Collaborators were instructed to perform the following
tests, in duplicate, on each of the samples:
A faster and simpler method to accurately determine the GT
of rice using a small quantity of sample has been sought by rice  Moisture content determination (AACCI Approved Meth-
breeders and others in the industry. In 2002 the AACC Inter- od 44-15.02 [1])
national Rice Milling and Quality Technical Committee con-  RVA GT of rice flour (described below)
ducted a collaborative study on the precision of a proposed  Amylograph method for milled rice (AACCI Approved
method optimized for determining the GT of milled rice flour Method 61-01.01 [1])
using the RVA.  DSC evaluation of rice (14)

Collaborative Study
Collaborators. Of the 12 participating laboratories, 11 re-
turned valid RVA data; data from lab 12 was excluded due to
a failure to follow the method (incorrect sample weight used).
In addition, seven of the laboratories returned valid amylograph
data, and six laboratories returned valid DSC data.

Fig. 1. Example of a rice flour tested using the Rapid Visco Analyser
method.

32 / JANUARY–FEBRUARY 2014, VOL. 59, NO. 1

 The RVA GT method involved holding a concentrated slurry c) Repeatability relative standard deviation (RSDr), with-
of rice flour (6.00 g sample corrected to 12% moisture basis) in in-laboratory repeatability
24.0 g of distilled water at 50°C for 5 min, heating it from 50 to d) Reproducibility standard deviation (SR)
95°C at 3 degrees Celsius/min (Table I), and measuring the e) Reproducibility relative standard deviation (RSDR),
viscosity of the sample throughout. In this method the GT, between-laboratory reproducibility
observed at the point of rapid increase in viscosity, is defined as
the temperature at which the viscosity first increases by at least Precision statistics were generated for each instrumental meth-
24 cP (=24 mPa·sec = 2 Rapid Visco units [RVU]) within 6 sec od and sample measurement separately.
(Fig. 1). Because high viscosities were obtained, the tests were Analysis of Precision Data. Sample moisture content data
manually stopped when viscosity reached ≈1,200 cP (100 RVU), are provided in Table II, GT data are provided in Tables III–V,
which occurred between 10 and 15 min into the test. and precision statistics are provided in Table VI. The number of
Statistical Analysis. Data were analyzed according to the outliers removed prior to statistical analysis of RVA data was
AOAC guidelines for collaborative study procedures (2) using within the limit of 2/9 labs specified by the AOAC procedure
templates provided by AACC International (a Microsoft Excel (2). For the RVA GT method, within-laboratory Sr values were
worksheet). The following analyses were performed: ≤0.15 degrees Celsius and between-laboratory SR values were
≤0.25 degrees Celsius (Table VI).
1) Cochran Test—To remove laboratories with poor replica- Comparison of Methods. Amylograph and DSC data and
tion within the laboratory precision statistics are included for comparison with the RVA
2) Single and Double Grubbs Test—To remove laboratories results. The RVA method gave mean GT values similar to the
with a mean value substantially different from the major- amylograph and DSC methods (Table VI). Precision was supe-
ity of the laboratories rior for the RVA method, with between-laboratory SR values
3) Precision Statistics—Analysis of variance (ANOVA) was ≤0.25 degrees Celsius compared with SR values ≤1.92 degrees
performed on the remaining data to give precision values, Celsius for the amylograph method and SR values ≤2.88 degrees
including Celsius for the DSC method (Table VI).
a) Mean
Conclusions
b) Repeatability standard deviation (Sr)
A new method for the determination of GT of rice flour
using the RVA was assessed by a collaborative study conducted
by the AACC International Rice Milling and Quality Technical
Committee. The proposed method showed acceptable preci-
sion for the determination of GT of rice flour. The RVA meth-
od gave similar but more precise measures of GT of rice flour
compared with amylograph and DSC methods. The method is
simple to perform, rapid, and requires only a small amount of
sample (6 g), making it well suited to the requirements of rice
breeders.

Acknowledgments
We thank Cheryl Earp (AACCI Rice Milling and Quality Techni-
cal Committee chair and study coordinator [2002]), Terry Nelsen
(AACCI), and the participating laboratories, Perten Instruments Aus-
tralia; Texas A&M; Cereal Solutions; the USDA-ARS Dale Bumpers
National Rice Research Center; the USDA-ARS NO; RiceTec; the
American Institute of Baking; Riviana Foods; the University of
Nebraska; the California Cooperative Rice Research Foundation; Rice-
land Foods; and the University of Arkansas, for their assistance in this
study.

CEREAL FOODS WORLD / 33

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An ad appeared here in the print version of the journal
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