METHOD FOR ESTIMATING PRODUCTION LOCATION
20210025817 ยท 2021-01-28
Inventors
Cpc classification
G01N21/718
PHYSICS
International classification
Abstract
The stable isotope ratios of carbon and hydrogen contained in a measurement target object collected from an animal are measured. An area in which the animal has grown is estimated from the measured stable isotope ratios of carbon and hydrogen (S3).
Claims
1.-11. (canceled)
12. A geographical origin estimation method comprising: measuring a stable isotope ratio of carbon contained in a measurement target object collected from an animal; measuring a stable isotope ratio of a constituent element of water contained in the measurement target object; and estimating an area where the animal has grown based on the stable isotope ratio of carbon and the stable isotope ratio of the constituent element of water.
13. The geographical origin estimation method according to claim 12, wherein measuring the stable isotope ratio of carbon contained in the measurement target object comprises: washing the measurement target object with a neutral detergent; drying the measurement target object after washing the measurement target object; performing a degreasing process by dipping the measurement target object in ethanol after drying the measurement target object; and measuring the stable isotope ratio of carbon contained in the measurement target object after the degreasing process.
14. The geographical origin estimation method according to claim 12, wherein measuring the stable isotope ratio of the constituent element of water contained in the measurement target object comprises: freeze-drying the measurement target object; and measuring the stable isotope ratio of the constituent element of water contained in the measurement target object while the measurement target object is freeze-dried.
15. The geographical origin estimation method according to claim 12, wherein estimating the area where the animal has grown comprises: estimating a rough area where the animal has grown from the stable isotope ratio of the constituent element of water; estimating feed used to grow the animal from the stable isotope ratio of carbon; and narrowing down the rough area to an area using the feed estimated from the stable isotope ratio of carbon; and estimating that the area using the feed is the area where the animal has grown.
16. The geographical origin estimation method according to claim 15, wherein th estimating the rough area comprises: estimating the rough area from the stable isotope ratio of the constituent element of water according to an equation, the equation indicating a relationship between a difference and a stable isotope ratio D, the difference being between a stable isotope ratio of a constituent element of water contained in precipitation at a predetermined reference point and a stable isotope ratio of a constituent element of water contained in precipitation in an estimation target area, and the stable isotope ratio D being of a constituent element of water contained in a measurement target object collected from an animal having grown in the estimation target area.
17. The geographical origin estimation method according to claim 16, wherein the equation is:
D=1.41+198.
18. The geographical origin estimation method according to claim 12, wherein measuring the stable isotope ratio of carbon or measuring the stable isotope ratio of the constituent element of water comprises using a laser spectroscopic stable isotope ratio analyzer configured to: analyze a stable isotope ratio of carbon or the constituent element of water contained in the measurement target object by irradiating a carbon dioxide gas and water vapor generated by burning the measurement target object with a laser beam; and analyzing an absorption amount of the laser beam.
19. The geographical origin estimation method according to claim 12, wherein the measurement target object is animal hair.
20. The geographical origin estimation method according to claim 12, wherein the measurement target object is hair of a cashmere goat.
21. The geographical origin estimation method according to claim 12, wherein the constituent element of water is hydrogen.
22. The geographical origin estimation method according to claim 12, wherein the constituent element of water is oxygen.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
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DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS
Best Mode for Carrying Out Embodiments of the Invention
[0041] An embodiment of the present invention will be explained in detail below with reference to the accompanying drawings.
Principle of Embodiment
[0042] A geographical origin estimation method as one embodiment of the present invention is characterized by estimating the growth area of an animal from the stable isotope ratios of carbon and a constituent element of water contained in a measurement target object collected from the animal.
[0043] More specifically, as shown in
[0044] An experiment to be described later revealed that the stable isotope ratio of a constituent element of water contained in a measurement target object collected from an animal depends on the stable isotope ratio of a constituent element of water drunk by the animal. Water drunk by an animal is water of precipitation in the area. The stable isotope ratio of a constituent element of water contained in precipitation has an area dependence. Accordingly, the stable isotope ratio of a constituent element of water contained in a measurement target object collected from an animal shows the area dependence. It was, therefore, found by research conducted by the inventor of embodiments of the present invention that the growth areas of an animal can be narrowed down from the stable isotope ratio of a constituent element of water contained in a measurement target object collected from the animal.
[0045] Also, as will be apparent from another experiment to be described later, the stable isotope ratio of carbon contained in a measurement target object collected from an animal depends on the kind of a plant eaten by the animal. The present inventor focused on the fact that a plant or feed eaten by an animal changes in accordance with the growth area of the animal. That is, it was found that the growth areas of an animal eating a given plant can be narrowed down by estimating the kind of the plant from the stable isotope ratio of carbon and checking the distribution of habitats of the plant.
[0046] The important point of this geographical origin estimation method is to narrow down, based on the above findings, the growth areas of an animal, from which a measurement target object is collected, by using the stable isotope ratios of a plurality of elements.
[0047] Based on the above-described principle, step S3 of estimating the growth area of an animal can be executed by a procedure shown in
[0048] When a database indicating the relationship between the stable isotope ratios of both carbon and a constituent element of water and the growth area of an animal is prepared beforehand, it is possible to immediately obtain the estimation result of the growth area of the animal from the measurement results of the stable isotope ratios of carbon and the constituent element of water in steps S1 and S2. However, if this database is formed by taking account of areas using feed, the estimation method using the database is practically the same as the estimation method including steps S31 to S33.
[0049] Note that step S1 of measuring the stable isotope ratio of carbon and step S2 of measuring the stable isotope ratio of the constituent element of water described above can be executed in the order of steps S1 and S2 or vice versa, i.e., in the order of steps S2 and S1. If possible, the stable isotope ratios of carbon and hydrogen may also be measured in parallel by simultaneously executing steps S1 and S2.
Definition of Stable Isotope Ratio
[0050] Next, the definition of the stable isotope ratio will be explained. The ratios of nonradioisotopes existing in nature are used as the stable isotope ratios of carbon and hydrogen to be measured in this embodiment. The ratio of .sup.12C to .sup.13C is used as the stable isotope ratio of carbon. The ratio of .sup.1H to D(.sup.2H) is used as the stable isotope ratio of hydrogen. However, each of these stable isotope ratios is normally expressed not as an absolute ratio but as a 5 value as a millesimal deviation from the isotope ratio of a standard sample.
[0051] The stable isotope ratio of carbon can be indicated as .sup.13C by equation (1) below. The stable isotope ratio of hydrogen can be indicated as D by equation (2) below. Note that in these equations, SAMP indicates the isotope ratio in an analytical sample, and STD indicates the isotope ratio in a standard sample.
[0052] A standard sample of the stable isotope ratio of hydrogen is normally represented by using VSMOW (Vienna Standard Mean Ocean Water). A standard sample of the stable isotope ratio of carbon is normally represented by using VPDB (Vienna Pee Dee Belemnite).
[0053] Measurements of Stable Isotope Ratios
[0054] The measurements of the stable isotope ratios will be explained below. Animal hairs of 10 animals of the same kind grown in the same stock farm by being fed with grass classified as a C3 plant are prepared as samples. In this embodiment, the hairs of cashmere goats are used as the animal hairs.
[0055] The stable isotope ratio of carbon of each of these 10 prepared animal hair samples is measured by using laser spectroscopic stable isotope ratio analysis. First, a carbon dioxide gas is generated by burning the 10 animal hair samples. The generated carbon dioxide gas is irradiated with a laser beam. The stable isotope ratio of carbon is measured by measuring and analyzing the laser beam absorption amount (the optical absorption intensity) by laser spectroscopy.
[0056] The measurement value thus obtained is the stable isotope ratio for all carbon elements contained in, e.g., an organic substance (keratin) forming the animal hair and dirt sticking to the animal hair. Rhombic points in
[0057] On the other hand, a pre-process is performed to remove dirt such as sebum and dandruff from the abovementioned 10 animal hair samples. First, the 10 animal hair samples are washed with an arbitrary neutral detergent. The washed 10 animal hair samples are dried. A degreasing process is performed by dipping the dried 10 animal hair samples in ethanol for 10 min. The stable isotope ratio of carbon of each degreased sample is analyzed by laser spectroscopic stable isotope ratio analysis.
[0058] By thus washing and degreasing the animal hair, carbon contained in the dirt sticking to the animal hair is removed. This makes it possible to measure the stable isotope ratio of only carbon contained in an organic substance (e.g., keratin) sticking to the animal hair. Square points in
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[0060] Like the stable isotope ratio of carbon, the stable isotope ratio of hydrogen can also be analyzed by laser spectroscopic stable isotope ratio analysis. While animal hair samples are stored and transported, however, the animal hair samples absorb hydrogen atoms contained in the surrounding environment as water, so the stable isotope ratio of hydrogen of each animal hair sample changes with time. In addition, the animal hair samples absorb washing water used when washing the animal hair samples and containing a neutral detergent, and this sometimes influences the measurement result of the stable isotope ratio of hydrogen of each animal hair sample. This makes it difficult to estimate a geographical origin by comparison of the stable isotope ratios of hydrogen. To prevent this, water in the animal hair samples is removed by vaporization by using freeze-drying. Consequently, it is possible to measure only the stable isotope ratio of hydrogen contained in an organic substance forming each animal hair sample.
[0061]
[0062] As indicated by symbol p, the variation of the stable isotope ratios (.sup.13C) of carbon was 1 when the degreasing process (ethanol treatment) was not performed. On the other hand, as indicated by symbol q, the variation of the stable isotope ratios (.sup.13C) of carbon was 0-4 when the degreasing process (ethanol treatment) was performed. Accordingly, the degreasing process reduced the variation produced by the individual differences between the stable isotope ratios of carbon from 1 to 0.4.
[0063] As indicated by symbol r, the variation of the stable isotope ratios (D) of hydrogen was 10.3 when the freeze-drying process was not performed. On the other hand, as indicated by symbol s, the variation of the stable isotope ratios (D) of hydrogen was 5.1 when the freeze-drying process was performed. Accordingly, the freeze-drying process reduced the variation produced by the individual differences between the stable isotope ratios of hydrogen from 10.3 to 5.1. These pre-processes make it possible to measure the stable isotope ratios of hydrogen and carbon of animal hair samples in the same geographical origin more accurately.
[0064] Estimation of Feed from Stable Isotope Ratio of Carbon
[0065] Animal hair sample A of an animal grown by being fed with grass classified as a C3 plant as feed, animal hair sample B of an animal grown by being fed with both feed containing a plant classed as a C4 plant such as corn and grass classified as a C3 plant as feed, and animal hair sample C of an animal grown by being fed with a plant classified as the C4 plant such as corn as feed are prepared.
[0066] The abovementioned washing, drying, and degreasing processes are performed on each of these three types of samples A to C. The stable isotope ratio of carbon of each of the three types of samples A to C having undergone the washing, drying, and degreasing processes is analyzed by laser spectroscopic stable isotope ratio analysis.
[0067] The stable isotope ratios of carbon increase in the order of samples A, B, and C. The difference between the stable isotope ratios of carbon of samples A and B is 5, and the difference between the stable isotope ratios of carbon of samples B and C is 3.
[0068] It is known that the stable isotope ratio of carbon of the C3 plant is smaller than the stable isotope ratio of carbon of the C4 plant. On the other hand, the stable isotope ratio of carbon of animal hair sample A of the animal eating the C3 plant as staple feed is smaller than the stable isotope ratio of carbon of animal hair sample C of the animal eating the C4 plant as staple feed. Also, the stable isotope ratio of carbon of animal hair sample B of the animal eating both the C3 and C4 plants as staple feed is intermediate between samples A and C. From the foregoing, there is a correlation between the type of feed and the stable isotope ratio of carbon of animal hair. Based on this correlation, it is possible to estimate feed eaten by an animal from which a measurement target object (animal hair) is collected, by measuring the stable isotope ratio of carbon contained in the measurement target object (animal hair).
[0069] When experiments of this kind were repetitively performed, the stable isotope ratio of carbon contained in the hair of the animal grown by the C3 plant was 21 or less (range a), the stable isotope ratio of carbon contained in the hair of the animal grown by the feed mixture containing the C3 and C4 plants was 21 to 16 (range b), and the stable isotope ratio of carbon contained in the hair of the animal grown by the C4 plant such as corn was 16 or less (range c).
[0070] Estimation of Geographical Origin from Stable Isotope Ratio of Hydrogen: Estimation Example 1
[0071] For each of water samples D, E, F, and G of areas D, E, F, and G arranged in descending order of the latitude, the stable isotope ratios of hydrogen and oxygen are measured by laser spectroscopic stable isotope ratio analysis. First, water samples D to G are evaporated. The generated water vapor is irradiated with a laser beam. The stable isotope ratios of hydrogen and oxygen are measured by measuring and analyzing the laser beam absorption amount (the optical absorption intensity) by laser spectroscopy.
[0072] As the latitude of the area increases, the stable isotope ratio (D) of hydrogen and the stable isotope ratio (.sup.18O) of oxygen decrease. A straight line shown in
[0073]
[0074] Then, animal hair sample H of an animal grown in area H, animal hair sample I of an animal grown in area I, animal hair sample J of an animal grown in area J, and animal hair sample K of an animal grown in area K are prepared. The stable isotope ratio of hydrogen of each of these animal hair samples is measured by laser spectroscopic stable isotope ratio analysis.
[0075]
[0076] As shown in
[0077] Estimation of Geographical Origin from Stable Isotope Ratio of Hydrogen: Estimation Example 2
[0078] <Difference Between Stable Isotope Ratios of Hydrogen Contained in Precipitation>
[0079] Assuming that an arbitrary point is a reference point, a difference between the stable isotope ratio of hydrogen contained in precipitation at this reference point and the stable isotope ratio of hydrogen contained in precipitation in an arbitrary area is represented by the sum of a difference A calculated from the latitude difference between the reference point and the arbitrary area and a difference B calculated from the altitude difference between the reference point and the arbitrary area (see, e.g., non-patent references 6 and 7).
[0080] The difference A calculated from the latitude difference is indicated by equation (3):
A()=bY(3)
[0081] where Y is the latitude difference (), and b is a constant. In this equation, the constant b is 2.8 to 4.4 as a value suitable for cashmere geographical origin estimation.
[0082] The difference B calculated from the altitude difference is indicated by equation (4)
B()=0.0065cZ(4)
[0083] where Z is the altitude difference (m), and c is a constant. In this equation, the constant c is 3.1.
[0084] From equations (3) and (4), the difference between the stable isotope ratios of hydrogen contained in precipitation at the reference point and in the arbitrary area is indicated by equation (5) below:
()=A+B=bY0.0065cZ(5)
[0085] Equation (5) represents that at a point at which the latitude differs by Y from the reference point (an arbitrary point) and the altitude differs by Z from the reference point, the stable isotope ratio of hydrogen contained in precipitation differs by from the reference point.
[0086] Equation Indicating Relationship Between and D
[0087] The stable isotope ratios of hydrogen contained in animal hair samples collected from animals grown in a plurality of areas are measured. In the following explanation, the animals are cashmere goats, and the animal hairs (animal hair samples) collected from the animals are cashmere, unless otherwise specified.
[0088] From the latitude and the altitude of the geographical origin (an area where a cashmere goat has grown) of a collected animal hair sample when it is assumed that each of the latitude and the altitude of the reference point is 0, the difference between the stable isotope ratio of hydrogen contained in precipitation at the reference point and the stable isotope ratio of hydrogen contained in precipitation of the geographical origin of the collected animal hair sample is calculated by using abovementioned equation (5). This calculation of the difference is performed for the geographical origins (the abovementioned plurality of areas) of all the animal hair samples for which the stable isotope ratio of hydrogen is measured.
[0089] A graph is formed by plotting the difference between the stable isotope ratios of hydrogen contained in precipitation at the reference point and in the animal hair sample geographical origin on the abscissa, and plotting the stable isotope ratio D of hydrogen contained in the animal hair sample for which the difference is calculated on the ordinate (see
D=a+(6)
[0090] where and are constants.
[0091] Note that equation (6) is derived by using average values if the stable isotope ratios of hydrogen contained in animal hair samples of the same geographical origin vary, or if the stable isotope ratios of hydrogen contained precipitation in the same area vary.
[0092] Equation (7) is obtained by substituting equation (5) into equation (6):
D=(bY0.0065*cZ)+(7)
[0093] D in equation (7) is a function of the latitude (Y) and the altitude (Z).
[0094] When deriving equation (7), data of stable isotope ratios of hydrogen in at least three areas is necessary. The number of areas is desirably as large as possible because the accuracy of the equation increases. In
[0095] When b=4.0 and c=3.1 in equation (7), =1.41 and =198. That is, when b=4.0 and c=3.1, abovementioned equation (6) is represented by equation (6a):
D=1.41+198(6a)
[0096] Note that this calculation was performed by referring to the values of corresponding to the altitude, latitude, and longitude of the abovementioned area in a database (see, e.g., non-patent literature 8) disclosed on the Internet. However, it is also possible to actually measure the stable isotope ratio of hydrogen contained in precipitation collected in the growth area of a cashmere goat, and calculate the values of a and in equation (6) by using the measured value.
[0097] <Geographical Origin Estimation Using Equation Indicating Relationship Between and D>
[0098] A case in which a measurement target object is cashmere and whether the geographical origin of this cashmere is Chifeng in the Inner Mongolia Autonomous Region is discriminated will be explained below. The range of values which the stable isotope ratio of hydrogen contained in cashmere can take is calculated by substituting the latitude (Y) and the altitude (Z) of Chifeng into equation (7). The stable isotope ratio of hydrogen contained in the measurement target object (cashmere) is measured. If the measured stable isotope ratio of hydrogen falls within the value range obtained by the calculation, it is determined that the geographical origin of the measurement target object can be Chifeng. If the stable isotope ratio falls outside the range, it is determined that the geographical origin of the measurement target object cannot be Chifeng.
[0099] In addition, for each of Outer Mongolia, Tibet of China, and the Inner Mongolia Autonomous Region of China known as cashmere geographical origins, the range of values which the stable isotope ratio D of hydrogen contained in cashmere is calculated by substituting the latitude (Y) and the altitude (Z) of the country or the area into equation (7). Consequently, as shown in
[0100] Note that as described above, the values of a and in equation (6) are obtained from the stable isotope ratio of hydrogen contained in each of precipitation (growth water) and cashmere collected in the growth area of a cashmere goat. The range of the stable isotope ratio D of hydrogen contained in cashmere is also obtained for each geographical origin by using equation (6) in which the values of a and are set, instead of equation (7). Accordingly, the geographical origin can be estimated by measuring the stable isotope ratio D of hydrogen contained in a measurement target object (cashmere), and collating the measurement result with the ranges obtained by equation (6).
[0101] Narrowing-Down of Geographical Origins from Stable Isotope Ratios of Hydrogen and Carbon
[0102] A method of estimating, from an animal hair sample as a measurement target object, the growth area (the geographical origin of animal hair) of an animal from which the animal hair sample was collected will be explained with reference to
[0103] First, the stable isotope ratio of hydrogen contained in the animal hair sample is measured (step S101 in
[0104] A rough area where an animal from which the animal hair sample was collected has grown is estimated from the measured stable isotope ratio of hydrogen (step S102 in
[0105] When adopting Estimation Example 2 described previously, a rough area where an animal from which an animal hair sample was collected has grown is estimated from the equation (equation (6), (6a), or (7)) indicating the relationship between the difference between the stable isotope ratios of hydrogen contained in precipitation at a predetermined reference point and in an estimation target area (an area where an animal from which a measurement target object was collected may have grown), and the stable isotope ratio D of hydrogen contained in an animal hair sample collected from the animal having grown in the estimation target area, and from the stable isotope ratio of hydrogen measured in step S101.
[0106] Then, the stable isotope ratio of carbon contained in the animal hair sample is measured (step S103 in
[0107] From the measured stable isotope ratio of carbon, the type of feed of the animal from which the animal hair sample was collected is estimated (step S104 in
[0108] Subsequently, the rough area estimated in step S102 is narrowed down to an area using the feed estimated in step S104, and it is estimated that this narrowed-down area is the growth area of the animal from which the animal hair sample was collected (step S105 in
[0109]
[0110]
[0111] That is, as shown in
[0112] In an actual process, the stable isotope ratios of carbon and hydrogen of a plurality of animal hair samples whose geographical origins are already known are measured, and a table showing the relationship between the stable isotope ratios of carbon and hydrogen and the collection place (geographical origin) as shown in
[0113] Use of Stable Isotope Ratio of Oxygen
[0114]
[0115] Use of Laser Spectroscopy
[0116] The methods of stable isotope ratio analysis include IRMS (Isotope Ratio Mass Spectrometry) and NMR (Nuclear Magnetic Resonance Spectroscopy). Unfortunately, these methods require large expensive measurement apparatuses requiring complicated operations, and hence take a long time and a high cost. This makes it difficult to introduce stable isotope ratio analysis as a testing item in a distribution process in a city.
[0117] In this embodiment, therefore, a laser spectroscopic stable isotope ratio analyzer can be used when measuring the stable isotope ratios of oxygen, hydrogen, and carbon. This laser spectroscopic stable isotope ratio analyzer can easily analyze the stable isotope ratios of oxygen, hydrogen, and carbon contained in a measurement target object by irradiating a carbon dioxide gas and water vapor generated by burning the measurement target object with a laser beam, and analyzing the absorption amount of the laser beam (see, e.g., non-patent literature 5).
[0118] The laser spectroscopic stable isotope ratio analyzer is inexpensive, compact, and easy to operate when compared to the measurement apparatuses of IRMS and NMR. In addition, chemical pre-processes required for stable isotope ratio analysis by IRMS are unnecessary, so geographical origin estimation can be performed within a short time (e.g., within 1 hr). Accordingly, the use of the laser spectroscopic stable isotope ratio analyzer can reduce the time and the cost. Note that the laser spectroscopic stable isotope ratio analyzer can also be used in only one of the measurement of the stable isotope ratios of oxygen/hydrogen, and the measurement of the stable isotope ratio of carbon.
[0119] Determination of Validity of Geographical Origin Label from Stable Isotope Ratio of Hydrogen
[0120]
Y=6(XX)+Y(8)
[0121] When the stable isotope ratio (Y) of hydrogen of animal hair whose geographical origin (the latitude X) is certain is obtained as basic data, whether the geographical origin label of arbitrary animal hair is correct can be determined. First, the stable isotope ratio (Y) of hydrogen of arbitrary animal hair is measured. Then, the latitude (X) is obtained by substituting the value (Y) into equation (8). If the obtained latitude (X) matches the value of the latitude of the geographical origin indicated by the label, it is possible to determine that the geographical origin label is correct.
[0122] Estimation of Staple Feed from Stable Isotope Ratio of Carbon
[0123] When the stable isotope ratio of carbon contained in animal hair is smaller than 21 (range a in
[0124] Application to Computer
[0125] The above described geographical origin estimation method can also be performed by using a computer. As shown in
[0126] A laser spectroscopic stable isotope ratio analyzer 20 is connected to the input/output interface 13. The laser spectroscopic stable isotope ratio analyzer 20 sequentially outputs the measurement results of the stable isotope ratio of hydrogen or oxygen and the stable isotope ratio of carbon contained in a measurement target object (animal hair) to the computer 10. Note that when the laser spectroscopic stable isotope ratio analyzer 20 is not connected to the computer 10, an operator may also manually input the measurement results to the computer 10. A monitor 30 is connected to the input/output interface 14.
[0127] A program necessary to execute step S3 in
[0128] Extension of Embodiment
[0129] The embodiment of the present invention has been explained above, but the present invention is not limited to the above embodiment. Various changes understandable by those skilled in the art within the technical scope of the present invention can be made on the arrangements and details of the invention.
[0130] For example, as described above, when measuring the stable isotope ratios of hydrogen and carbon, it is possible to obtain more accurate measurement results and estimate a geographical origin more reliably by performing the pre-processes (steps S111 and S131 to S133). However, these pre-processes may also be omitted if rough estimation results need only be obtained.
[0131] Also, the case in which the geographical origin of animal hair, particularly, cashmere hair as a measurement target object is estimated has been explained. However, embodiments of the present invention is also usable when estimating, e.g., the geographical origin of wool.
EXPLANATION OF THE REFERENCE NUMERALS AND SIGNS
[0132] 10 . . . computer, 11 . . . CPU, 12 . . . memory, 13, 14 . . . input/output interface, 15 . . . bus, 20 . . . laser spectroscopic stable isotope ratio analyzer, 30 . . . monitor.