JP7548542B2 - How to identify alcohol - Google Patents
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- JP7548542B2 JP7548542B2 JP2020094046A JP2020094046A JP7548542B2 JP 7548542 B2 JP7548542 B2 JP 7548542B2 JP 2020094046 A JP2020094046 A JP 2020094046A JP 2020094046 A JP2020094046 A JP 2020094046A JP 7548542 B2 JP7548542 B2 JP 7548542B2
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 title claims description 51
- 238000000034 method Methods 0.000 claims description 22
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 16
- 229910052799 carbon Inorganic materials 0.000 claims description 16
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 13
- 239000008103 glucose Substances 0.000 claims description 13
- 238000004811 liquid chromatography Methods 0.000 claims description 9
- 238000000926 separation method Methods 0.000 claims description 8
- 235000016127 added sugars Nutrition 0.000 claims description 4
- 241000196324 Embryophyta Species 0.000 description 13
- 235000000346 sugar Nutrition 0.000 description 10
- 235000013334 alcoholic beverage Nutrition 0.000 description 9
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 7
- 238000005259 measurement Methods 0.000 description 7
- 150000008163 sugars Chemical class 0.000 description 7
- 241000209094 Oryza Species 0.000 description 6
- 235000007164 Oryza sativa Nutrition 0.000 description 6
- 235000009566 rice Nutrition 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 235000020083 shōchū Nutrition 0.000 description 5
- 239000002994 raw material Substances 0.000 description 4
- 240000000111 Saccharum officinarum Species 0.000 description 3
- 235000007201 Saccharum officinarum Nutrition 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 238000002485 combustion reaction Methods 0.000 description 2
- 238000002307 isotope ratio mass spectrometry Methods 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- CHQMHPLRPQMAMX-UHFFFAOYSA-L sodium persulfate Chemical compound [Na+].[Na+].[O-]S(=O)(=O)OOS([O-])(=O)=O CHQMHPLRPQMAMX-UHFFFAOYSA-L 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 244000099147 Ananas comosus Species 0.000 description 1
- 235000007119 Ananas comosus Nutrition 0.000 description 1
- 241000219357 Cactaceae Species 0.000 description 1
- GXCLVBGFBYZDAG-UHFFFAOYSA-N N-[2-(1H-indol-3-yl)ethyl]-N-methylprop-2-en-1-amine Chemical compound CN(CCC1=CNC2=C1C=CC=C2)CC=C GXCLVBGFBYZDAG-UHFFFAOYSA-N 0.000 description 1
- 101150038791 Pak1 gene Proteins 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 244000098338 Triticum aestivum Species 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 235000012907 honey Nutrition 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
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- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Alcoholic Beverages (AREA)
Description
特許法第３０条第２項適用 掲載日 令和元年１１月２７日 掲載アドレス ｈｔｔｐｓ：／／ｗｗｗ．ｎａｔｕｒｅ．ｃｏｍ／ａｒｔｉｃｌｅｓ／ｓ４１５９８－０１９－５４１６２－６ 集会名 ７ｔｈ Ｃｏｎｆｅｒｅｎｃｅ ｏｆ ｔｈｅ Ｆｏｒｅｎｓｉｃ Ｉｓｏｔｏｐｅ Ｒａｔｉｏ Ｍａｓｓ Ｓｐｅｃｔｒｏｍｅｔｒｙ Ｎｅｔｗｏｒｋ，Ｓａｎ Ｍｉｃｈｅｌｅ ａｌｌ’ Ａｄｉｇｅ（ＴＮ），Ｉｔａｌｙ 開催日 令和元年９月１６～１９日 掲載日 令和２年５月１５日 掲載アドレス ｈｔｔｐ：／／ｃｏｎｆｅｒｅｎｃｅ．ｗｄｃ－ｊｐ．ｃｏｍ／ｊｓａｃ／ｔｏｕｒｏｎ／８０／ｐｒｏｇｒａｍ／ｃｏｎｔｅｎｔｓ／ｃｏｍｍｏｎ／ｐｄｆ／８０＿ｐｒｏｇｒａｍ．ｐｄｆ 掲載日 令和２年５月１日 掲載アドレス ｈｔｔｐ：／／ｗｗｗ．ｍｓｓｊ．ｊｐ／ｃｏｎｆ／６８／ｐｒｏｇｒａｍ／２Ｃ－Ｏ７－０９０５．ｈｔｍｌ 発行者名 読売新聞社 刊行物名 読売新聞秋田版 発行日 令和２年１月２２日Article 30, Paragraph 2 of the Patent Act applies Date of publication: November 27, 2019 Address of publication: https://www. nature. com/articles/s41598-019-54162-6 Conference Name 7th Conference of the Forensic Isotope Ratio Mass Spectrometry Network, San Michele all' Adige (TN), Italy Event date: September 16-19, 2019 Publication date: Reiwa May 15, 2020 Publication address: http://conference. wdc-jp. com/jsac/touron/80/program/contents/common/pdf/80_program.pdf Date of publication May 1, 2020 Address http://www.mssj.jp/conf/68/program/2C-O7 -0905.html Publisher: Yomiuri Shimbun Publication name: Yomiuri Shimbun Akita edition Publication date: January 22, 2020
本発明は、酒（日本酒等）の識別方法に関する。 The present invention relates to a method for identifying alcoholic beverages (such as Japanese sake).
食品の識別を食品に含まれる軽元素（炭素、窒素、酸素、水素等）の安定同位体比率を用いて行う技術は、例えば非特許文献１に記載されている。ここでは、植物は光合成の方式の違いによりＣ３植物（米、麦、芋等）、Ｃ４植物（サトウキビ、トウモロコシ等）、ＣＡＭ植物（サボテン、パイナップル等）に分類でき、それぞれにおける重い炭素（１３Ｃ）の取り込みやすさの違いに起因して、炭素の同位体比（１３Ｃ／１２Ｃ）が異なることが用いられる。焼酎におけるアルコールの原料となる植物はＣ３植物、Ｃ４植物であるため、焼酎の種類によって、含まれるアルコール中の炭素における同位体比は異なり、この同位体比を、焼酎の種類の識別の指標として用いることができる。 A technology for identifying foods using the stable isotope ratio of light elements (carbon, nitrogen, oxygen, hydrogen, etc.) contained in foods is described in, for example, Non-Patent Document 1. Here, plants can be classified into C3 plants (rice, wheat, potato, etc.), C4 plants (sugarcane, corn, etc.), and CAM plants (cactus, pineapple, etc.) based on the difference in the photosynthetic method, and the carbon isotope ratio ( 13 C/ 12 C) is different due to the difference in the ease of uptake of heavy carbon ( 13 C) in each of them. Since the plants that are the raw material for alcohol in shochu are C3 plants and C4 plants, the carbon isotope ratio in the alcohol contained in the shochu varies depending on the type of shochu, and this isotope ratio can be used as an index for identifying the type of shochu.
日本酒は伝統的なアルコール飲料であり、その中には、原料を米、米麹、水のみとした純米酒、上記の原料に加えてサトウキビ等を原料とする醸造アルコールが添加された吟醸酒、更にこれに糖類、有機酸が添加された普通酒がある。このうち、純米酒は最も高価であるため、偽装品の対象となりやすく、この偽装品においては、本来は添加されない醸造アルコール等が添加されている。このため、特に日本酒を高精度で識別することができる技術が求められた。 Sake is a traditional alcoholic beverage, and there are three types: junmai sake, made only from rice, rice malt, and water; ginjo sake, which is made from the above ingredients plus brewer's alcohol made from sugarcane and other raw materials; and futsushu, which has sugars and organic acids added to it. Of these, junmai sake is the most expensive and is therefore the most likely to be counterfeited, and counterfeit products contain brewer's alcohol and other ingredients that are not normally added. For this reason, there was a demand for technology that could identify sake with a high degree of accuracy.
この場合において、米はＣ３植物、醸造アルコールの原料となるサトウキビはＣ４植物であるため、非特許文献１に記載されたような、アルコール中の炭素の同位体比をこの識別の指標として用いることができる。しかしながら、この場合における識別の精度は充分ではなく、更なる高精度の識別方法が求められた。 In this case, rice is a C3 plant, and sugarcane, the raw material for brewer's alcohol, is a C4 plant, so the carbon isotope ratio in the alcohol can be used as an indicator for this identification, as described in Non-Patent Document 1. However, the accuracy of identification in this case is not sufficient, and a more accurate identification method is required.
このため、Ｃ３植物を原料とした酒の識別を高精度で行う技術が求められた。 Therefore, there was a demand for technology that could accurately identify alcoholic beverages made from C3 plants.
本発明は、かかる問題点に鑑みてなされたものであり、上記問題点を解決する発明を提供することを目的とする。 The present invention was made in consideration of these problems, and aims to provide an invention that solves the above problems.
本発明は、上記課題を解決すべく、以下に掲げる構成とした。
本発明の酒の識別方法は、Ｃ３植物を原料として製造される酒の識別方法であって、前記酒から生成された試料から、液体クロマトグラフ（ＬＣ）によってエタノール成分とグルコース成分を分離する分離工程と、前記エタノール成分における炭素同位体比δ１３Ｃであるδ１３Ｃｅｔｈと、前記グルコース成分における炭素同位体比δ１３Ｃであるδ１３Ｃｇｌｕを安定同位体比質量分析計（ＩＲＭＳ）によって測定する同位体比測定工程と、を具備し、δ１３Ｃｅｔｈとδ１３Ｃｇｌｕに基づいて、前記酒の種類を識別することを特徴とする。
本発明の酒の識別方法において、前記酒は日本酒であり、識別される前記酒の種類は、純米酒、吟醸酒、普通酒の３種であることを特徴とする。
本発明の酒の識別方法は、前記酒が吟醸酒又は普通酒であると認識された場合において、醸造アルコールの添加量をδ１３Ｃｅｔｈの値に基づいて算出することを特徴とする。
本発明の酒の識別方法は、前記酒が普通酒であると認識された場合において、糖類の添加量をδ１３Ｃｇｌｕの値に基づいて算出することを特徴とする。
In order to solve the above problems, the present invention has the following configuration.
The method for identifying alcohol of the present invention is a method for identifying alcohol produced using a C3 plant as a raw material, and includes a separation step of separating an ethanol component and a glucose component from a sample produced from the alcohol by liquid chromatography (LC), and an isotope ratio measurement step of measuring δ 13 C eth , which is the carbon isotope ratio δ 13 C of the ethanol component, and δ 13 C glu , which is the carbon isotope ratio δ 13 C of the glucose component, by a stable isotope ratio mass spectrometer (IRMS), and is characterized in that the type of alcohol is identified based on δ 13 C eth and δ 13 C glu .
In the method for identifying alcoholic beverages of the present invention, the alcoholic beverage is Japanese sake, and the types of alcoholic beverages to be identified are three types: Junmai sake, Ginjo sake, and Ordinary sake.
The method for identifying alcohol of the present invention is characterized in that, when the alcohol is recognized to be ginjo-shu or futsu-shu, the amount of brewer's alcohol added is calculated based on the value of δ 13 C eth .
The method for identifying alcoholic beverages of the present invention is characterized in that, when the alcoholic beverage is recognized as ordinary alcoholic beverage, the amount of added sugars is calculated based on the δ 13 C glu value.
本発明は以上のように構成されているので、Ｃ３植物を原料とした酒の識別を高精度で行うことができる。 As the present invention is configured as described above, it is possible to identify alcoholic beverages made from C3 plants with high accuracy.
本発明の実施の形態に係る酒（日本酒）の識別方法について説明する。この識別方法においても、非特許文献１に記載の技術と同様に炭素の同位体比が測定されるが、非特許文献１に記載の技術においては焼酎の中における炭素同位体比が測定されたのに対し、この識別方法においては日本酒中の特定成分（化合物）中における炭素同位体比が測定される。 This section describes a method for identifying alcohol (Japanese sake) according to an embodiment of the present invention. In this identification method, the carbon isotope ratio is measured in the same way as in the technology described in Non-Patent Document 1, but whereas the technology described in Non-Patent Document 1 measures the carbon isotope ratio in shochu, this identification method measures the carbon isotope ratio in specific components (compounds) in sake.
この識別方法においては、川島洋人、「安定同位体比を用いた食品の産地識別と偽和判定の研究動向」、Ｊｏｕｒｎａｌ ｏｆ ｔｈｅ Ｍａｓｓ Ｓｐｅｃｔｒｏｍｅｔｒｙ Ｓｏｃｉｅｔｙ ｏｆ Ｊａｐａｎ、第６７巻、第２号（２０１９年）に記載されたＬＣ（液体クロマトグラフ）／ＩＲＭＳ（同位体分析）が用いられる。このため、この識別方法においても、液体クロマトグラフによって分離された物質（化合物）に対して、同位体分析が行われる。ここで同位体分析によって算出されるのは炭素の同位体比δ１３Ｃである。ここで、試料におけるδ１３Ｃは、「（（試料における１３Ｃ／１２Ｃの同位体比）／（国際標準化物質における１３Ｃ／１２Ｃの同位体比）－１）×１０００（‰：パーミル）」で定義される。 In this identification method, LC (liquid chromatography)/IRMS (isotope analysis) described in Hiroto Kawashima, "Research Trends in Identifying the Origin of Food and Determining Adulteration Using Stable Isotope Ratios," Journal of the Mass Spectrometry Society of Japan, Vol. 67, No. 2 (2019) is used. Therefore, in this identification method, isotope analysis is also performed on the substance (compound) separated by liquid chromatography. Here, the isotope ratio δ 13 C of carbon is calculated by isotope analysis. Here, δ 13 C in the sample is defined as "((isotope ratio of 13 C/ 12 C in the sample)/(isotope ratio of 13 C/ 12 C in the international standardized material)-1)×1000 (‰: per mille)".
ここで、識別対象となる日本酒は、純米酒、吟醸酒、普通酒の３種類に大別される。純米酒は、原料を米、米麹、水のみとした日本酒であり、吟醸酒はこれに醸造アルコール（サトウキビ原料）のみが添加されたものであり、普通酒はこれに更に糖類、有機酸が添加されたものである。Ｔ．Ｅ．Ｃｅｒｌｉｎｇ、Ｊ．Ｍ．Ｈａｒｒｉｓ、Ｂ．Ｊ．ＭａｃＦａｄｄｅｎ、Ｍ．Ｇ．Ｌｅａｋｅｙ、Ｊ．Ｑｕａｄｅ、Ｖ．Ｅｉｓｅｎｍａｎｎ ａｎｄ Ｊ．Ｒ．Ｅｈｌｅｒｉｎｇｅｒ、「Ｇｌｏｂａｌ Ｖｅｇｅｔａｔｉｏｎ Ｃｈａｎｇｅ Ｔｈｒｏｕｇｈ ｔｈｅ Ｍｉｏｃｅｎｅ／Ｐｌｉｏｃｅｎｅ Ｂｏｕｎｄａｒｙ」、Ｎａｔｕｒｅ、Ｖｏｌ．３８９、ｐ１５３（１９９７）に記載されるように、Ｃ３植物である米におけるδ１３Ｃは－３０～－２２‰程度であり、Ｃ４植物であるサトウキビにおけるδ１３Ｃは－１４～－１０‰程度であり、後者の方が１３Ｃ比率が高い。このため、δ１３Ｃをこれらの識別のための指標の一つとすることができる。 Here, sake to be identified is broadly divided into three types: Junmai sake, Ginjo sake, and Futsushu sake. Junmai sake is sake made only from rice, rice malt, and water, Ginjo sake is sake to which only brewer's alcohol (made from sugarcane) is added, and Futsushu sake is sake to which sugars and organic acids are further added. T. E. Cerling, J. M. Harris, B. J. MacFadden, M. G. Lakey, J. Quade, V. Eisenmann and J. R. Ehleringer, "Global Vegetation Change Through the Miocene/Pliocene Boundary", Nature, Vol. 389, p. 153 (1997), the δ 13 C in rice, a C3 plant, is about -30 to -22‰, while the δ 13 C in sugarcane, a C4 plant, is about -14 to -10‰, with the latter having a higher 13 C ratio. Therefore, δ 13 C can be used as one of the indices for distinguishing between these.
この識別方法においては、液体クロマトグラフによって、試料（日本酒）において上記のδ１３Ｃの測定の対象となる２種類の化合物が液体クロマトグラフによって分離される（分離工程）。ここで分離されるのは、エタノール（Ｅｔｈａｎｏｌ）とグルコース（Ｇｌｕｃｏｓｅ）の２つである。その後、このエタノールとグルコースに対してそれぞれ前記のδ１３Ｃが測定される（同位体比測定工程）。 In this identification method, two types of compounds to be measured for δ 13 C in a sample (sake) are separated by liquid chromatography (separation step). The two compounds separated here are ethanol and glucose. Then, the δ 13 C of each of the ethanol and glucose is measured (isotope ratio measurement step).
この場合の実際の測定結果について以下に説明する。ここで用いられたＬＣ／ＩＲＭＳの装置構成は、Ｈ．Ｋａｗａｓｈｉｍａ、Ｍ．Ｓｕｔｏ ａｎｄ Ｎ．Ｓｕｔｏ、「Ｄｅｔｅｒｍｉｎａｔｉｏｎ ｏｆ Ｃａｒｂｏｎ Ｉｓｏｔｏｐｅ Ｒａｔｉｏｓ ｆｏｒ Ｈｏｎｅｙ Ｓａｍｐｌｅｓ ｂｙ Ｍｅａｎｓ ｏｆ ａ Ｌｉｑｕｉｄ Ｃｈｒｏｍａｔｇｒａｐｈｙ／Ｉｓｏｔｏｐｅ Ｒａｔｉｏ Ｍａｓｓ Ｓｐｅｃｔｒｏｓｃｏｐｙ Ｓｙｓｔｅｍ Ｃｏｕｐｌｅｄ ｗｉｔｈ ａ Ｐｏｓｔ－Ｃｏｌｕｍｎ Ｐｕｍｐ］、Ｒａｐｉｄ Ｃｏｍｍｕｎｉｃａｔｉｏｎｓ ｉｎ Ｍａｓｓ Ｓｐｅｃｔｒｏｍｅｔｒｙ、Ｖｏｌ．３２、ｐ１２７１（２０１８）に記載されている。 The actual measurement results in this case are described below. The LC/IRMS device configuration used here is the same as that described in H. Kawashima, M. Suto and N. This is described in "Determination of Carbon Isotope Ratios for Honey Samples by Means of a Liquid Chromatography/Isotope Ratio Mass Spectroscopy System Coupled with a Post-Column Pump" by Suto, Rapid Communications in Mass Spectrometry, Vol. 32, p. 1271 (2018).
まず、試料からエタノールとグルコースを分離するための液体クロマトグラフ（分離工程）について説明する。ここでは、配位子交換カラム（製品名Ｓｕｇａｒ Ｐａｋ１、Ｗａｔｅｒｓ社製）が用いられ、試料が混合される溶出剤は超純水とされ、脱気処理が行われ、カラム流量は０．５ｍｌ／ｍｉｎ、ポストカラム流量は０．３ｍｌ／ｍｉｎとされ、カラム温度は８０℃とされた。 First, the liquid chromatograph (separation process) for separating ethanol and glucose from the sample will be described. Here, a ligand exchange column (product name Sugar Pak1, manufactured by Waters) was used, the eluent with which the sample was mixed was ultrapure water, which was degassed, the column flow rate was 0.5 ml/min, the post-column flow rate was 0.3 ml/min, and the column temperature was 80°C.
図１は、この場合におけるクロマトグラムを示す。ここで、初めにみられる５つのパルスは校正用のＣＯ２ガスに対応し、その後でグルコース、エタノールが明確に分離されて見られる。このため、上記の条件では１０００ｓｅｃ以内の範囲で、測定対象となるグルコースとエタノールの分離ができることが確認できる。 Figure 1 shows the chromatogram in this case. The first five pulses correspond to the CO2 gas used for calibration, and then glucose and ethanol are clearly separated. Therefore, it can be confirmed that the glucose and ethanol to be measured can be separated within 1000 seconds under the above conditions.
その後、これによって分離されグルコースとエタノールは、それぞれ酸化剤となるペルオキソ二硫酸ナトリウム、緩衝液となるリン酸と混合された後に、９９℃に加熱されることによって燃焼してＣＯ２が生成される。このＣＯ２が膜分離によってＨｅと混合されて、安定同位体比質量分析計（ＩＲＭＳ）によって炭素の同位体（１３Ｃ、１２Ｃ）分析が行われた（同位体比測定工程）。なお、この場合には試料から分離されたグルコースとエタノールによって生成されたＣＯ２が炭素同位体分析の対象となるが、その他に、カラムを経由せず分離処理が行われない試料に対しても上記と同様の処理が行われ、同様に燃焼によって生成されたＣＯ２に対しても、同様の測定が行われた。 Then, the glucose and ethanol separated in this way are mixed with sodium peroxodisulfate as an oxidizing agent and phosphoric acid as a buffer solution, respectively, and then heated to 99°C to generate CO 2 through combustion. This CO 2 is mixed with He by membrane separation, and carbon isotope ( 13 C, 12 C) analysis is performed by a stable isotope ratio mass spectrometer (IRMS) (isotope ratio measurement process). In this case, the CO 2 generated by glucose and ethanol separated from the sample is the subject of carbon isotope analysis, but the same process as above is also performed on samples that do not pass through a column and are not subjected to separation processing, and the same measurement is also performed on the CO 2 generated by combustion.
上記のように、純米酒（計１３種）、吟醸酒（計１５種）、普通種（計１２種）の計４０種の各々について上記の測定を行い、分離処理の行われない場合、分離されたグルコース、分離されたエタノールの３種類についてδ１３Ｃをそれぞれ測定し、以下では、これらの測定結果をそれぞれδ１３Ｃｂｕｌｋ、δ１３Ｃｇｌｕ、δ１３Ｃｅｔｈとする。 As described above, the above measurements were carried out for a total of 40 types of sake, including junmai sake (13 types in total), ginjo sake (15 types in total), and regular sake (12 types in total). When no separation treatment was performed, the δ 13 C of the three types of separated glucose and separated ethanol were measured. In the following, these measurement results are referred to as δ 13 C bulk , δ 13 C glu , and δ 13 C eth , respectively.
図２は、各試料におけるこれらのδ１３Ｃの平均値、差分を示す表である。また、図３は、全ての試料における、δ１３Ｃｂｕｌｋとδ１３Ｃｅｔｈの関係（ａ）、δ１３Ｃｂｕｌｋとδ１３Ｃｇｌｕの関係（ｂ）をそれぞれ示す。 Fig. 2 is a table showing the average values and differences of δ 13 C for each sample. Also, Fig. 3 shows the relationship between δ 13 C bulk and δ 13 C eth (a) and the relationship between δ 13 C bulk and δ 13 C glu (b) for all samples.
図３より、δ１３Ｃｂｕｌｋとδ１３Ｃｅｔｈ（図３（ａ））に強い正の相関関係があり、δ１３Ｃｂｕｌｋとδ１３Ｃｇｌｕ（図３（ｂ））においては特に明確な相関関係は認められない。 FIG. 3 shows that there is a strong positive correlation between δ 13 C bulk and δ 13 C eth (FIG. 3(a)), but no particularly clear correlation is observed between δ 13 C bulk and δ 13 C glu (FIG. 3(b)).
一方、図４は、各試料におけるδ１３Ｃｅｔｈとδ１３Ｃｇｌｕの相関関係を示す図である。この図においては、２次元の領域として、δ１３Ｃｅｔｈ≦－２６．３‰かつδ１３Ｃｇｌｕ＜－２１．９‰である領域Ａ、δ１３Ｃｅｔｈ＞－２６．３‰かつδ１３Ｃｇｌｕ＜－２１．９‰である領域Ｂ、δ１３Ｃｅｔｈ＞－２６．３‰かつδ１３Ｃｇｌｕ≧－２１．９‰である領域Ｃが定義できる。ここで、点線で囲まれた普通酒の４種は、糖類が添加された普通酒であり、これら以外の普通酒には、糖類が添加されていない。 On the other hand, Fig. 4 shows the correlation between δ 13 C eth and δ 13 C glu for each sample. In this figure, the following two-dimensional regions can be defined: Region A where δ 13 C eth ≦-26.3‰ and δ 13 C glu <-21.9‰, Region B where δ 13 C eth >-26.3‰ and δ 13 C glu <-21.9‰, and Region C where δ 13 C eth >-26.3‰ and δ 13 C glu ≧-21.9‰. The four types of futsushu surrounded by dotted lines are futsushu to which sugar has been added, and the other futsushu have no added sugar.
このため、図４において、純米酒は領域Ａ内にあり、醸造アルコールが添加された吟醸酒、及び醸造アルコールが添加され、かつ糖類が添加されない普通酒は領域Ｂ内にあり、糖類が添加された普通酒は領域Ｃ内にある。このような識別を、測定されたδ１３Ｃｅｔｈとδ１３Ｃｇｌｕの相関関係によって行うことができる。この際、図４において、点線で囲まれた普通酒の４種と、これら以外の普通酒においては、δ１３Ｃｇｌｕが大きく異なるのに対し、δ１３Ｃｅｔｈは同等である。このため、δ１３Ｃｇｌｕは糖類の添加によって大きく上昇し、δ１３Ｃｅｔｈは糖類の添加によって影響を受けない。 For this reason, in Fig. 4, junmai sake is in region A, ginjo sake to which brewing alcohol has been added and futsu-shu to which brewing alcohol has been added but no added sugars are in region B, and futsu-shu to which sugars have been added is in region C. Such discrimination can be made based on the correlation between the measured δ 13 C eth and δ 13 C glu . In Fig. 4, the 4 types of futsu-shu surrounded by dotted lines and the other futsu-shu have significantly different δ 13 C glu , but the δ 13 C eth are equivalent. For this reason, δ 13 C glu increases significantly with the addition of sugars, while δ 13 C eth is not affected by the addition of sugars.
また、純米酒（δ１３Ｃｅｔｈ＝－２８．４±０．１‰、δ１３Ｃｇｌｕ＝－２７．５±０．４‰）に対する醸造アルコールの添加量を変えて、δ１３Ｃｅｔｈとδ１３Ｃｇｌｕを測定した結果を図５に示す。横軸ｘは、添加された醸造アルコールのアルコールの全体の量に対する比率（％）である。この結果より、δ１３Ｃｇｌｕは醸造アルコール添加量には依存せず略一定であるのに対して、δ１３Ｃｅｔｈは醸造アルコール添加量に対して直線的に変化し、この特性は、ｙ（δ１３Ｃｅｔｈ：単位‰）＝０．１４９×ｘ－２８．４０５と近似できる。このため、この関係式から、δ１３Ｃｅｔｈを用いて醸造アルコールの添加量を算出することができる。 FIG. 5 shows the results of measuring δ 13 C eth and δ 13 C glu when the amount of brewed alcohol added to junmai sake (δ 13 C eth =-28.4±0.1‰, δ 13 C glu =-27.5±0.4‰) was changed. The horizontal axis x represents the ratio (%) of the added brewed alcohol to the total amount of alcohol. From these results, δ 13 C glu is approximately constant and does not depend on the amount of brewed alcohol added, whereas δ 13 C eth changes linearly with the amount of brewed alcohol added, and this characteristic can be approximated as y (δ 13 C eth : unit ‰) = 0.149 x - 28.405. Therefore, from this relational equation, the amount of brewed alcohol added can be calculated using δ 13 C eth .
また、普通酒において、δ１３Ｃｅｔｈにおける糖類の添加量の依存性は小さく、δ１３Ｃｇｌｕにおける糖類の添加量依存性が大きいため、同様に、普通酒においては、δ１３Ｃｇｌｕを用いて糖類の添加量を算出することもできる。 Furthermore, in ordinary sake, since δ 13 C eth has little dependence on the amount of sugars added, and δ 13 C glu has great dependence on the amount of sugars added, it is also possible to calculate the amount of sugars added in ordinary sake using δ 13 C glu .
このような醸造アルコール添加量とδ１３Ｃｅｔｈの関係、糖類添加量とδ１３Ｃｇｌｕの関係は、これらが添加されない場合の酒（純米酒に対応）のδ１３Ｃｅｔｈ、δ１３Ｃｇｌｕの値等に応じ、予め実験によって求めることができる。 The relationship between the amount of brewer's alcohol added and δ 13 C eth , and the relationship between the amount of sugar added and δ 13 C glu can be determined in advance by experiment according to the values of δ 13 C eth and δ 13 C glu of sake (corresponding to pure rice sake) to which these substances are not added.
上記の例では、識別の対象が米を原料とする日本酒であったが、他のＣ３植物を原料とした場合においても、上記の識別方法を同様に適用できることは明らかである。また、上記の例では、分離工程において共通の試料からエタノール成分とグルコース成分を分離するために液体クロマトグラムが用いられたが、同様にこれらが分離できる限りにおいて、他の方法を用いてもよい。 In the above example, the subject of identification was sake made from rice, but it is clear that the above identification method can be similarly applied when other C3 plants are used as the raw material. Also, in the above example, liquid chromatography was used to separate the ethanol component and the glucose component from the common sample in the separation process, but other methods may be used as long as they can be similarly separated.
Claims (4)
前記日本酒から、液体クロマトグラフ（ＬＣ）によってエタノール成分とグルコース成分を分離する分離工程と、
前記エタノール成分における炭素同位体比δ１３Ｃであるδ１３Ｃｅｔｈと、前記グルコース成分における炭素同位体比δ１３Ｃであるδ１３Ｃｇｌｕを安定同位体比質量分析計（ＩＲＭＳ）によって測定する同位体比測定工程と、
を具備し、
δ１３Ｃｅｔｈとδ１３Ｃｇｌｕに基づいて、前記日本酒の種類を識別することを特徴とする日本酒の識別方法。 A method for identifying sake , comprising:
A separation step of separating an ethanol component and a glucose component from the sake by liquid chromatography (LC);
an isotope ratio measuring step of measuring the carbon isotope ratio δ 13 C eth of the ethanol component and the carbon isotope ratio δ 13 C glu of the glucose component by a stable isotope ratio mass spectrometer (IRMS);
Equipped with
A method for identifying sake , comprising identifying the type of sake based on δ 13 C eth and δ 13 C glu .
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| JP2014224717A (en) | 2013-05-15 | 2014-12-04 | 独立行政法人酒類総合研究所 | Method for determining presence/absence of alcohol addition in drink |
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| ANA I CABAERO、他２名，Isotope ratio mass spectrometry coupled to liquid and gas chromatography for wine ethanol characterization，Rapid Commun Mass Spectrom，2008年10月22日，Vol.20，Page.3111-3118， doi: 10.1002/rcm.3711，PMID: 18798196 |
| ANA I CABAERO、他２名，Simultaneous stable carbon isotopic analysis of wine glycerol and ethanol by liquid chromatography coupled to isotope ratio mass spectrometry，J Agric Food Chem，2010年01月27日，Vol.58、No.2，Page.722-728，DOI: 10.1021/jf9029095，PMID: 20025274 |
| FUMIKAZU AKAMATSU、他２名，Separation and Purification of Glucose in Sake for Carbon Stable Isotope Analysis，Food Analytical Methods，Vol.13,No.6，2020年 |
| 堀井幸江、他４名，清酒および焼酎におけるC4植物由来原材料比率の推測，醸造学会誌，2011年，Vol.106,No.1，Page.45-49 |
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