CN113670959A - Novel method for analyzing relative crystallinity of starch - Google Patents
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- 229920002472 Starch Polymers 0.000 title claims abstract description 148
- 235000019698 starch Nutrition 0.000 title claims abstract description 148
- 239000008107 starch Substances 0.000 title claims abstract description 147
- 238000000034 method Methods 0.000 title claims abstract description 48
- 238000001228 spectrum Methods 0.000 claims abstract description 31
- 238000002425 crystallisation Methods 0.000 claims abstract description 9
- 230000008025 crystallization Effects 0.000 claims abstract description 9
- 238000002441 X-ray diffraction Methods 0.000 claims description 12
- 229940100486 rice starch Drugs 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 4
- 238000004040 coloring Methods 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 239000000839 emulsion Substances 0.000 claims description 3
- 238000000227 grinding Methods 0.000 claims description 3
- 239000002244 precipitate Substances 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 3
- 239000012498 ultrapure water Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 238000007873 sieving Methods 0.000 claims description 2
- 235000013305 food Nutrition 0.000 description 4
- 238000013507 mapping Methods 0.000 description 4
- 241000431987 Neocheiropteris fortunei Species 0.000 description 3
- 229920000945 Amylopectin Polymers 0.000 description 2
- 229920000856 Amylose Polymers 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 241000209094 Oryza Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 240000008467 Oryza sativa Japonica Group Species 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000009499 grossing Methods 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
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- G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
- G01N23/20—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by using diffraction of the radiation by the materials, e.g. for investigating crystal structure; by using scattering of the radiation by the materials, e.g. for investigating non-crystalline materials; by using reflection of the radiation by the materials
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- G01N23/20008—Constructional details of analysers, e.g. characterised by X-ray source, detector or optical system; Accessories therefor; Preparing specimens therefor
- G01N23/2005—Preparation of powder samples therefor
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N23/20—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by using diffraction of the radiation by the materials, e.g. for investigating crystal structure; by using scattering of the radiation by the materials, e.g. for investigating non-crystalline materials; by using reflection of the radiation by the materials
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Abstract
The invention discloses a novel method for analyzing relative crystallinity of starch, which comprises the following steps: preparing amorphous starch by using natural starch, and acquiring natural starch and amorphous starch spectrum; drawing by taking natural starch as a background, ensuring that the 2 theta angle positions of the two types of starch are overlapped at 4 degrees, deforming the peak intensity of amorphous starch, and searching a second overlapped 2 theta angle position; drawing a straight line at the position of 4-40 degrees of an amorphous starch spectrogram, counting the relative areas of a crystallization peak and an amorphous peak by using software, and calculating the relative crystallinity of the starch. The method completes the calculation of the relative crystallinity of the starch by the X-ray diffractometer, analyzes the relative crystallinity of the starch after reducing the peak area based on the principle that the amorphous starch crystallization peak disappears, can effectively avoid the deviation caused by manually searching the anchor point by the traditional method, is more scientific, credible and accurate, can be implemented aiming at the starch of different plant sources, and has wide application range.
Description
Technical Field
The invention relates to a method for analyzing the crystallinity of starch, in particular to a novel method for analyzing the relative crystallinity of starch.
Background
Starch is a major carbohydrate source in humans and plays an important role in the daily diet. The plant natural starch has a semi-crystalline structure and is composed of amylose and amylopectin molecules. Amylose forms a loose disordered structure, also called an amorphous region, while amylopectin molecules form a more densely structured crystalline region in the form of a double helix. Natural starch in nature can be divided into three types of A- (cereal), B- (tuber) and C-type starch (beans and yam tubers) according to the difference of the peak positions of X-ray diffraction patterns, and the amorphous areas and the crystallized areas of different types of starch have different characteristics and proportions. The ratio of the two has great influence on the physical and chemical properties of the starch, wherein the amorphous area is easily damaged by acid and enzyme, the digestion speed is higher, and the crystalline area has stronger resistance to the acid and the enzyme and has lower digestion speed. Therefore, accurate calculation of the relative proportion of crystalline regions in starch granules, i.e. the relative crystallinity, and definition of the granule structural characteristics of starch, is of great importance for the use of starch in the food and non-food industries. The existing methods for calculating the relative crystallinity of starch mainly comprise a software peak fitting method, a software curve method, a curve mapping method, a straight line mapping method and the like, and the methods relate to smoothing the wave spectrum and artificially adding anchor points of a crystallization peak base line, so that the calculated relative crystallinity value has a large error with an actual value, particularly, when the relative crystallinity of the starch of the same type is compared, the result of fitting an objective rule is difficult to obtain, and the judgment of the proportion of the starch crystalline region by people is influenced.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to provide a novel method for scientifically, accurately, simply and conveniently analyzing the relative crystallinity of starch.
The technical scheme is as follows: the invention provides a novel method for analyzing relative crystallinity of starch, which comprises the following steps:
(1) preparing amorphous starch by using natural starch, and acquiring natural starch and amorphous starch spectrum;
(2) drawing by taking natural starch as a background, ensuring that the 2 theta angle positions of the two types of starch are overlapped at 4 degrees, deforming the peak intensity of amorphous starch, and searching a second overlapped 2 theta angle position;
(3) drawing a straight line at the position of 4-40 degrees of an amorphous starch spectrogram, counting the relative areas of a crystallization peak and an amorphous peak by using software, and calculating the relative crystallinity of the starch.
Further, the preparation method of the step (1) comprises the following steps: mixing rice starch and ultrapure water to prepare starch emulsion, heating, centrifuging, removing supernatant, drying precipitate, grinding dried sample, and sieving to obtain amorphous starch.
Further, the 2 theta angle position of the two types of starch is ensured to be overlapped in the step (1). The 4 degrees are the starting points of the spectrums, and no crystallization peak is formed in the area, so that the overlapping of the 4 degrees can effectively avoid the difference of the spectrum areas of the two types of starch, and ensure that the spectrum of the amorphous starch is completely separated from the natural starch. Further, the rice starch is A-type starch, B-type starch or C-type starch.
Further, in the step (1), the natural starch and the amorphous starch are placed on an X-ray diffractometer, and a spectrum scanning is carried out at an angle of 2 theta of 4 degrees to 40 degrees, so as to obtain an X-ray diffraction pattern of the starch.
Furthermore, in the step (3), a straight line is drawn at two points of 4-40 degrees of the amorphous starch spectrum by using Photoshop software, two ends of the straight line are ensured to be positioned at the center of the spectrum, a spacing region between the upper part of the amorphous spectrum and the natural starch and a lower region of the amorphous spectrum are colored respectively, and the relative area values of the two points are counted by using Image-Pro Plus software, so that the areas of a crystalline region and an amorphous peak region can be obtained.
Further, the method for calculating the relative crystallinity of the starch in the step (3) is as follows:
relative crystallinity of starch ═ crystalline peak relative area × 100%/(crystalline peak relative area + amorphous peak relative area).
The invention uses natural starch to prepare amorphous starch, and obtains the X-ray diffraction spectrum of the starch; utilizing Origin software to complete the difference reduction of the natural starch spectrum and the amorphous starch spectrum; and drawing a baseline of the amorphous spectrum, counting the relative areas of a crystalline peak and an amorphous peak by using software, and calculating the relative crystallinity of the starch. Compared with the prior method, the method has the advantages of simple and convenient operation, high accuracy and wide application range. The method prepares the amorphous starch corresponding to the natural starch, determines the amorphous component in the natural starch map according to the characteristic peak difference of the X-ray diffraction maps of the two types of starch, completes the subtraction of the amorphous starch map by taking the X-ray diffraction maps of the natural starch as the background, takes the spectrum of the amorphous starch as the boundary, and has the upper part corresponding to the crystalline area and the lower part corresponding to the amorphous area.
Has the advantages that: compared with the prior art, the invention has the following advantages:
1. the operation process is simple: this is done by preparing amorphous starch and analyzing the X-ray diffraction pattern of the starch using software.
2. The result is scientific and accurate: the method is developed based on the principle that the crystallization peak of the amorphous starch X-ray diffraction pattern is lost, and the difference of an amorphous area can be reduced by deforming the peak intensity of the amorphous starch with the natural starch pattern as the background. The analysis result is more scientific, credible and accurate, and has important application prospect in the food and non-food processing field.
3. The application range is wide: the method can be implemented for starch of different species based on the characteristic that different species have similar amorphous starch patterns, so that the method can be implemented for A-, B-or C-type starch and has wide range.
Drawings
FIG. 1X-ray diffraction patterns of native and amorphous rice starches, (A) native rice starch pattern; (B) amorphous starch map;
FIG. 2 is a graph of an amorphous starch against the background of the X-ray diffraction pattern of native starch;
FIG. 3X-ray diffraction pattern after adding straight lines using Photoshop software;
FIG. 4 is an X-ray diffraction pattern obtained by coloring the crystalline region and the amorphous region using Photoshop software;
FIG. 5 relative areas of crystalline and amorphous regions analyzed using Image-Pro Plus, (A) area of crystalline region; (B) the area of the amorphous region;
FIG. 6 the relative areas of crystalline and amorphous regions of drynaria fortunei starch (C-type starch) analyzed using Image-Pro Plus, (A) the area of the crystalline region; (B) the area of the amorphous region;
FIG. 7 is a comparison of the results of the present method and the prior art methods, (A) the relative crystallinity measured by the present method; (B) relative crystallinity as measured by graph plotting.
Detailed Description
The technical solution of the present invention is described in detail below, but the scope of the present invention is not limited to the embodiments.
The specific implementation of the method is illustrated below by taking high-yield and high-quality japonica rice 'Nipponbare' starch (A-type starch) as an example.
(1) Preparing amorphous starch: preparing starch emulsion according to the proportion of 100mg starch and 3mL ultrapure water, heating for 1h at 95 ℃, centrifuging for 10min at 5000g, removing supernatant, spreading precipitate in a glass culture dish, and drying in an oven at 42 ℃. Grinding the dried sample by a mortar, and screening the ground sample by a metal mesh screen of 100 meshes to obtain amorphous starch;
(2) obtaining an X-ray diffraction pattern of starch: and (3) placing the natural starch and the amorphous starch on an X-ray diffractometer, and performing spectrum scanning at the 2 theta angle of 4-40 degrees with the step length of 0.6s to obtain the X-ray diffraction spectrum of the starch. It can be seen from the figure that the rice native starch has stronger diffraction peaks at 2 theta angles of 15 degrees, 17 degrees, 18 degrees, 19 degrees and 23 degrees, while the amorphous starch has only one diffraction peak at 19 degrees (figure 1);
(3) treating the natural starch and the amorphous starch map: since amorphous starch crystalline regions are lost, crystalline regions can be derived from the principle of subtracting amorphous spectra from the natural spectra. The specific method is that natural starch is used as a background to be drawn in Origin software, the curve thickness is set to be 0.2 (the area occupied by the curve can be effectively reduced), the starting point with the 2 theta angle of 4 degrees is aligned, the peak intensity height of the amorphous spectrum is deformed, the second overlapping 2 theta angle position is searched, two overlapping points of two spectra are ensured, and finally the spectrum is derived. The aligned amorphous starch is contained within the native starch spectrum, and the boundary between the two can be used as a reference edge to distinguish crystalline and amorphous peaks (fig. 2):
(4) area analysis of crystalline and amorphous peaks: drawing a straight line at two points of 4-40 degrees of an amorphous starch map by using Photoshop software, ensuring that two ends of the straight line are positioned at the center of the spectrum, and ensuring that the heights of resonance peaks at the upper end and the lower end of the straight line are consistent (figure 3); coloring the blank positions at the upper part and the lower part of the spectrum peak of the amorphous pattern respectively, and corresponding to the crystallization area and the amorphous area respectively, wherein the area occupied by the amorphous area is larger, and the area occupied by the crystallization area is smaller (figure 4); the relative area values of two sites, i.e. the crystalline peak and amorphous peak areas, were calculated using Image-Pro Plus software as 284749 and 582308 (fig. 5), respectively;
(5) calculation of the relative crystallinity of starch: relative crystallinity of starch (relative area of crystalline peak x 100%/(relative area of crystalline peak + relative area of amorphous peak) 284749 x 100%/(284749 +582308) 32.84%;
(6) the method can also be applied to the calculation of relative crystallinity of other types of starch, taking wild starch plant resource drynaria fortunei starch (C-type starch) as an example, the areas of a crystalline region and an amorphous region can be obtained by the method, which are 231737 and 649557 (figure 6), respectively, and the relative crystallinity is 231737 multiplied by 100%/(231737 +649557) ═ 26.30%;
(7) the method is compared with the prior art. The method selects natural starch of drynaria fortunei, starch pretreated at 55 ℃, 75 ℃ and 95 ℃ (amorphous starch), and compares the curve mapping method with the measurement result of the method (figure 7), and can be seen that the conclusion obtained by the two methods is consistent, namely the natural starch has the highest crystallinity and the amorphous starch has the lowest crystallinity, which also indicates that the crystal structure of the starch is lost in the continuous heating process. In addition, the relative crystallinity value of the amorphous starch can still be measured by using a curve mapping method, which is obviously inconsistent with the crystal structure characteristics of the amorphous starch, and the relative crystallinity value of the amorphous starch measured by the method is zero, thereby conforming to the objective rule of the starch structure. Therefore, the comparison of the two methods shows that the measurement result of the method is more reliable, the deviation generated by implementation is small, and the practicability is wider.
Claims (7)
1. A novel method for analyzing the relative crystallinity of starch, characterized by: the method comprises the following steps:
(1) preparing amorphous starch by using natural starch, and acquiring natural starch and amorphous starch spectrum;
(2) drawing by taking natural starch as a background, ensuring the position overlapping of 2 theta angles of the two types of starch, deforming the peak intensity of amorphous starch, and searching a second overlapping 2 theta angle position;
(3) drawing a straight line at the position of 4-40 degrees of an amorphous starch spectrogram, counting the relative areas of a crystallization peak and an amorphous peak by using software, and calculating the relative crystallinity of the starch.
2. The novel method of analyzing the relative crystallinity of starch according to claim 1, characterized in that: the preparation method of the step (1) comprises the following steps: mixing rice starch and ultrapure water to prepare starch emulsion, heating, centrifuging, removing supernatant, drying precipitate, grinding dried sample, and sieving to obtain amorphous starch.
3. The novel method of analyzing the relative crystallinity of starch according to claim 2, characterized in that: the rice starch is A-type starch, B-type starch or C-type starch.
4. The novel method of analyzing the relative crystallinity of starch according to claim 1, characterized in that: in the step (1), the 2 theta angle of the two types of starch is ensured to be overlapped by 4 degrees.
5. The novel method of analyzing the relative crystallinity of starch according to claim 1, characterized in that: in the step (1), the natural starch and the amorphous starch are placed on an X-ray diffractometer, and spectrum scanning is carried out at an angle of 2 theta of 4-40 degrees to obtain an X-ray diffraction pattern of the starch.
6. The novel method of analyzing the relative crystallinity of starch according to claim 1, characterized in that: and (3) drawing a straight line at two points of 4-40 degrees of the amorphous starch map by using Photoshop software, ensuring that two ends of the straight line are positioned at the center of the spectrum, respectively coloring a spacing region between the upper part of the amorphous spectrum and the natural starch and a lower region of the amorphous spectrum, and counting the relative area values of the two positions by using Image-Pro Plus software to obtain the areas of a crystalline region and an amorphous peak region.
7. The novel method of analyzing the relative crystallinity of starch according to claim 1, characterized in that: the method for calculating the relative crystallinity of the starch in the step (3) comprises the following steps:
relative crystallinity of starch ═ crystalline peak relative area × 100%/(crystalline peak relative area + amorphous peak relative area).
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| CN104062309A (en) * | 2014-07-17 | 2014-09-24 | 扬州大学 | Method for measuring relative crystallinity of starch |
| CN106770395A (en) * | 2016-03-29 | 2017-05-31 | 天津商业大学 | The dynamic (dynamical) method of starch retrogradation is studied using peak area is selected |
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Patent Citations (2)
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| CN104062309A (en) * | 2014-07-17 | 2014-09-24 | 扬州大学 | Method for measuring relative crystallinity of starch |
| CN106770395A (en) * | 2016-03-29 | 2017-05-31 | 天津商业大学 | The dynamic (dynamical) method of starch retrogradation is studied using peak area is selected |
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