Rice Science, 2008, 15(1): 6972

Copyright 2008, China National Rice Research Institute. Published by Elsevier BV. All rights reserved

Accurate Measurement of Pasting Temperature by the Rapid Visco-Analyser: a Case Study Using Rice Flour
BAO Jin-song
(Institute of Nuclear-Agricultural Sciences/Key Laboratory of Chinese Ministry of Agriculture for Nuclear-Agricultural Sciences, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310029, China)

Abstract: Pasting properties are among the most important characteristics of starch, determining its applications in food processing and other industries. Pasting temperature derived from the Rapid Visco-analyser (RVA) (Newport Scientific), in most cases, is overestimated by the Thermocline for Windows software program. Here, two methods facilitating accurate measurement of pasting temperature by RVA were described. One is to change parameter setting to screen the true point where the pasting viscosity begins to increase, the other is to manually record the time (T1) when the pasting viscosity begins to increase and calculate the pasting temperature with the formula of (45/3.8)(T11)+50 for rice flour. The latter method gave a manually determined pasting temperature which was significantly correlated with the gelatinization temperature measured by differential scanning calorimetry. Key words: rice; starch; gelatinization temperature; pasting temperature; methodology

In addition to amylose content, pasting and gelatinization properties are among the most important physicochemical properties of starch that determine its applications in food industry and other uses [1]. Among rice grain quality, amylose content is believed to be the most important parameter in determining the eating and cooking quality of cooked rice [1-3], and it is agreed that pasting viscosity properties and gelatinization temperature (GT) also contribute to differences in cooked rice quality [4-5]. Pasting viscosity properties have been used to predict the end-use quality of various products, e.g. cooked rice texture [4,6] and noodles [7]. In many food industries, analysis of pasting viscosity and gelatinization temperature is still necessary for samples at different stages of processing. Pasting properties of starches are measured by using various instruments such as Rapid Visco Analyser (RVA, Newport Scientific, Warriewood, Australia), Micro ViscoAmylograph (Brabender, Duisberg, Germany), etc [8]. GT is the critical temperature at which starch granules irreversibly lose their birefringence and crystalline order during heating. GT can be directly measured under a polarized light microscopy with a heating stage to record the temperature when the birefringence is lost. GT can also be measured by differential scanning calorimetry (DSC) [9], or indirectly by the alkali spreading value test for rice grains [10]. Some studies also measured the pasting temperature (PT) of starch from RVA analysis [3, 6, 8, 11]. If the PT obtained from RVA were equivalent to the GT from DSC, information on pasting viscosity and gelatinization temperature
Received: 2 August 2007; Accepted: 9 October 2007 Corresponding author: BAO Jin-song (jsbao@zju.edu.cn) This is an English version of the paper published in Chinese in Chinese Journal of Rice Science, Vol. 21. No. 5, 2007, Pages 543546.

could be simultaneously obtained from a single test on the RVA, thus saving time and resources. Unfortunately, the PT measured from RVA is always overestimated, being much higher than the GT derived from DSC. In some cases, the RVA derived PT of rice samples was higher than 90C [3, 6]. Hence, the PT from RVA will be misleading if it is not interpreted properly. In the present paper, we report how to accurately estimate the gelatinization temperature using RVA.

MATERIALS AND METHODS

Rice materials A total of 15 rice varieties (or breeding lines) representing a wide range of amylose content (2% to 30%) and gelatinization temperature were used in this study. The rice was grown in Hainan Province, China from November 2003 to April 2004. Rough rice grains were dehulled, and milled to white rice with a Satake mill (Satake Corp. Japan) and then ground to flour with a Cyclone sample mill (UDY Corp., Fort Collins, CO). The flour was further passed through a 100-mesh sieve prior to analysis. RVA measurement Rice flour pasting properties were determined using a Rapid Visco-Analyzer model 3D (RVA) (Newport Scientific, Warriewood, Australia) with the software program Thermocline for Windows (TCW) according to AACC method 61-02 [12]. Each rice sample (3 g, 12% m.b.) was mixed with 25 g of distilled water in an RVA sample canister. The idle temperature was set at 50C, and the following 12.5-minute test profile was run: (1) held at 50C for 1.0 min, (2) linearly ramped up to

70 95C in 3.8 min, (3) held at 95C for 2.5 min, (4) linearly ramped down to 50C in 3.8 min and (5) held at 50C for 1.4 min. The peak viscosity, holding viscosity, final viscosity, and pasting temperature were determined by the analysis window of TCW. In the program, the formula to measure pasting temperature is TempAtViscRate (T1,T2,.Tinc,Vinc), which means that the temperature (C) when the rate of change in viscosity (Vinc/Tinc) is first achieved between T1 (min) and T2 (min) of the curves. The viscosity is either in RVA arbitrary unit (RVU) or centiPoise (cP). For rice flour, the recommended total test time is generally 12.5 min (AACC 2000), thereby the first two parameters (T1 and T2) of the formula are generally set as (2, 7), whereas the latter two parameters (Tinc and Vinc) can be changed. DSC analysis The gelatinization temperatures of the rice flour samples were measured with a DSC 2920 Modulated DSC thermal analyser (TA Instruments, Newcastle, DE) equipped with DSC standard and dual sample cells. Rice flour (2.0 mg) was weighed into an aluminum pan, 6 L distilled water was added, and the pan was sealed hermetically with a lid. After equilibration at room temperature for 1 h, the sample was heated at a rate of 10C/min from 30C to 110 C. A sealed empty pan was used as a reference. Onset (To), peak (Tp), and completion (Tc) temperatures of gelatinization were calculated by a Universal Analysis Program, Version 1.9D (TA Instruments, Newcastle, DE). Statistical analysis Students t-test for comparing the means of the paired data

Rice Science, Vol. 15, No. 1, 2008 and analysis of correlation coefficient were carried out in Microsoft Excel (Microsoft Corp, Seattle, WA).

RESULTS AND DISCUSSION

According to the software formula setting, the pasting temperature (PT) was recorded when the rate of change viscosity above the set point was first achieved. In many cases, the viscosity increased rapidly after initial rise, so the formula given by the software, such as TempAtViscRate (2,7,.2,24), accurately measured the PT (Fig. 1-A). For example, the software gave the samples BP003, BP004 and BP095 the PTs with less than 2C deviation when different parameters were tried (Table 1). In some cases, wide deviations in the PTs were found when different formulas were tried. For example, the formula TempAtViscRate (2,7,.1,36) gave BP530 and BP547 the PTs higher than 88C (Table 1), whereas the TempAtViscRate (2,7,.1,10) formula gave them around 73oC (Table 1). The reason was that the pasting viscosity was increasing at a low rate, even lower than the set point for a long time (Fig. 1-B to F). For example, if a samples pasting viscosity increased at a constant rate of 18 cP/s between the time 2 min to 7 min during test, no PT would be recorded with formula TempAtViscRate (2,7,.1,36) or TempAtViscRate (2,7,.1,24) because the actual rate was always less than 36 or 24 cP/s. Thus, the recorded PT would gradually decline as the set rate declined as shown in Fig. 1-B to F. However, too small a change rate setting would also result in incorrect PT, such as TempAtViscRate(2,7,.1,10) gave BP005 66C, much lower than the true value (Table 1). Because of genotypic differences in pasting behavior, accurate

Temperature

Viscosity (cP)

Viscosity

Temperature (C)
Temperature Viscosity

Viscosity

Time (min)

Temperature (C)

(cP )

Viscosity (cP)

Time (min)

Fig. 1. Typical pasting profile of rice flour tested on a Rapid Visco-analyser. A, PT derived from the program formulae of TempAtViscRate (2,7,.2,24) for rice BP003; B to F, PT derived from various program formula of rice BP025; B, TempAtViscRate (2,7,.1,36); C, TempAtViscRate (2,7,.1,24); D, TempAtViscRate (2,7,.1,18); E, TempAtViscRate (2,7,.1,12); F, TempAtViscRate (2,7,.1,10). Peak, Peak viscosity; Hold, Holding viscosity; Final, Final viscosity; PT, Pasting temperature.

BAO Jin-song. Accurate Measurement of Pasting Temperature by the Rapid Visco-Analyser: a Case Study Using Rice Flour

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Table 1. Pasting temperatures of rice flours derived from different methods. PT a (C) Sample BP003 BP004 BP005 BP010 BP011 BP025 BP026 BP086 BP095 BP133 BP225 BP420 BP530 Bp547 Bp551
a

PTm b (1,18) 77.7 77.6 79.2 73.5 79.1 72.9 71.3 81.6 76.8 66.8 69.7 81.5 83.0 84.6 72.9 (1,12) 77.7 76.8 75.2 70.6 78.3 71.9 70.5 69.8 76.8 66.8 69.0 80.7 78.3 72.9 69.7 (1,10) 76.7 76.8 66.0 67.4 75.8 70.6 70.5 68.2 76.0 66.0 68.2 70.5 73.6 72.9 69.7 Time (min) 3.17 3.28 3.47 2.50 3.43 2.73 2.75 2.42 3.20 2.38 2.55 2.48 2.75 2.77 2.63 PT (C) 75.7 77.0 79.2 67.8 78.8 70.5 70.7 66.8 76.1 66.4 68.4 67.6 70.7 70.9 69.3 To 71.2 72.4 74.4 58.3 74.8 63.0 62.9 58.1 70.6 56.3 62.5 58.7 62.9 62.7 59.2

DSC c (C) Tp 75.2 75.8 78.9 67.0 79.4 69.2 69.6 65.2 75.3 63.4 70.3 65.6 69.2 68.9 66.0 Tc 81.0 81.2 83.7 76.1 84.6 76.1 78.0 72.6 81.2 70.9 77.3 72.9 76.1 75.8 73.1

(2,4) d 77.7 77.6 79.9 85.4 79.1 83.8 73.6 83.0 76.8 79.9 69.7 85.4 87.8 87.2 82.2

(2,24) 76.7 76.8 79.2 72.9 78.3 71.9 70.5 69.1 76.0 66.8 69.0 80.7 77.4 84.0 70.5

(2,36) 77.7 77.6 79.2 72.9 79.1 76.0 71.3 81.6 76.8 78.3 69.0 84.0 83.8 85.4 77.6

(1,36) 78.4 78.4 79.9 86.1 79.9 87.9 74.5 87.2 77.6 82.9 71.4 86.2 88.6 89.5 84.5

(1,24) 77.7 77.6 79.9 73.5 79.9 79.1 74.5 82.3 77.6 79.1 69.7 84.6 87.2 86.1 81.5

PT was pasting temperature derived from program software formula, TempAtViscRate (2,7,.Tinc,Vinc); b PTm was pasting temperature derived from manual location of the time when the viscosity began to rise; c Differential scanning calorimetry: To, Onset temperature; Tp, Peak temperature; Tc, Completion temperature; d This was set in the RVU mode, whereas others were in cP mode.

automatic measurement of PT by a single formula was impossible. To analyze with different formulae and check whether the PT was determined at the proper point would help to obtain a correct result for a specific pasting profile. Two inferences could be drawn. One is that an accurate PT is easier to be obtained for higher viscosity samples because of a generally rapid increase in viscosity for these samples. The other is that measurement of PT will be affected by starch or flour concentrations. Lower concentration of a sample results in higher PT when measured by the software formula though it is actually low in gelatinization temperature, whereas higher concentration leads to more accurate measurement of the PT because of the rapid increase in viscosity. Varavinit et al [3] used 1.5 g rice flours instead of 3.0 g to test pasting properties, so the high PTs (>90C) might result from the low concentration. PT could be measured by inspection of individual curves to determine the point at which the pasting viscosity began to increase (Fig. 1-B). Since the temperature was increased linearly from 50C to 95C in 3.8 min during pasting and was

held at 50C for 1.0 min before increasing the temperature, the PT could be calculated by the formula, PTm = (45/3.8) (T11)+50, where T1 is the time when the pasting viscosity began to rise (Table 1). In the TCW software, the time at the point when the mouse arrow lies in the curve will be displayed when the Show Cursor Coordinates is chosen in the Options. This method was very accurate, because in TCW, one can enlarge the profile so as to locate the point at a right place. We recorded the time for all 15 samples and calculated the PT (Table 1). As a result, the manually calculated PTs (PTm) were significantly correlated with those from DSC measurements, whereas all other software formula derivatives gave poor correlation with DSC parameters (Table 2). TempAtViscRate (2,7,.2,24), TempAtViscRate (2,7,.1,12) and TempAtViscRate (2,7,.1,10) gave relatively more accurate PT because of their higher correlation with DSC parameters (Table 2). Under the assumption that the two paired data had the same mean, the Students t-test indicated that PTm was significantly different from To (P<0.0001), Tp (P<0.01) and Tc

Table 2. Correlation coefficients for pasting temperatures derived from different methods with DSC values a. Parameter To Tp Tc (2,4) b -0.35 -0.38 -0.40 (2,24) 0.49 0.47 0.44 (2,36) -0.02 -0.09 -0.18 (1,36) -0.46 -0.49 -0.52* (1,24) -0.11 -0.18 -0.26 (1,18) 0.28 0.25 0.20 (1,12) 0.59* 0.56* 0.52* (1,10) 0.60* 0.54* 0.52* PTm 0.99** 0.97** 0.94**

* and ** are significant at P<0.05 and P<0.01, respectively; a Differential scanning calorimetry: To, Onset temperature; Tp, Peak temperature; Tc, Completion temperature; b This was set in the RVU mode, whereas others were in cP mode.

72 (P<0.0001). However, when the difference between two paired data was set to 1C, the PTm was not significantly different from Tp (P = 0.71), but it was still significantly different from To and Tc (P<0.0001), indicating that the mean PTm (71.7C) was only 1C higher than the mean Tp of DSC (70.6C). Our analysis only focused on the measurement of PT of rice flour tested according to the AACC method 61-02; whether other samples or other methods will encounter the same problem is not known. The manually obtained PT should always resolve the problem, but the formula to calculate the PT could be reestablished for that specific method.
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Juliano B O. Criteria and tests for rice grain quality. In: Juliano B O. Rice: Chemistry and Technology. 2nd ed. St Paul, MN: AACC, 1985: 443524. Varavinit S, Shobsngob S, Varanyanond W, Chinachoti P, Naivikul O. Effect of amylose content on gelatinization, retrogradation and pasting properties of flours from different cultivars of Thai rice. Starch, 2003, 55: 410415. Shu Q Y, Wu D X, Xia Y W, Gao M W, McClung A. Relationship between RVA profile character and eating quality in Oryza sativa L. Sci Agric Sin, 1998, 31 (3): 2529. (in Chinese with English abstract) Bao J S, Shu Q Y, Wu D X, Xia Y W. Comparative study on the RVA profiles from the milled and brown rice flour. Chinese J Rice Sci, 2001, 15(2): 145146. (in Chinese with English abstract) Limpisut P, Jindal V. Comparison of rice flour pasting properties using brabender viscoamylograph and rapid visco analyzer for evaluating cooked rice texture. Starch, 2002, 54: 350357 Bhattacharya M, Zee S Y, Corke H. Physicochemical properties related to quality of rice noodles. Cereal Chem, 1999, 76: 861867. Suh D S, Jane J L. Comparison of starch pasting properties at various cooking conditions using the micro amylograph and the rapid visco analyzer. Cereal Chem, 2003, 80: 745749. Normand F L, Marshall W E. Differential scanning calorimetry of whole grain milled rice and milled rice flour. Cereal Chem, 1989, 66: 317320. Little R R, Hilder G B, Dawson E H, Elsie H. Differential effect of dilute alkali on 25 varieties of milled white rice. Cereal Chem, 1958, 35: 111126. Liang X M, King J M, Shih F F. Pasting property differences of commercial and isolated rice starch with added lipids and -cyclodextrin. Cereal Chem, 2002, 79: 812818. American Association of Cereal Chemists (AACC). Methods 61-02 for RVA. In: Approved Methods of the AACC. 10th ed. St. Paul, MN: AACC, 2000.

CONCLUSIONS
Pasting temperature measured by RVA does not reflect the true gelatinization temperature, such as measured by DSC. Changing parameters in the analysis program could increase the measurement accuracy but still gave great deviation for some samples. Inspecting the curves to record the time (T1) when the pasting viscosity began to rise and calculating the pasting temperature by a specific formula, such as (45/3.8)(T11)+50 for rice flour can measure the pasting temperature (estimate gelatinization temperature) very accurately. The results also indicated that the pasting temperature from RVA was 1C higher than peak temperature of DSC.

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ACKNOWLEDGEMENTS
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This research was financially supported in part by the National High Technology Development Project of China (Grant No. 2006AA10Z193), the National Natural Science Foundation of China (Grant No. 30300227), and the Science and Technology Department of Zhejiang Province (Grant No. 2007C32014).

REFERENCES
1 Bao J, Bergman C J. The functionality of rice starch. In: Eliasson A C. Starch in Food: Structure, function and applications. Cambridge: Woodhead Publishing Ltd., 2004: 258294.