JP2006023185A - Discriminating method of processed citrus fruit - Google Patents
Discriminating method of processed citrus fruit Download PDFInfo
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- JP2006023185A JP2006023185A JP2004201465A JP2004201465A JP2006023185A JP 2006023185 A JP2006023185 A JP 2006023185A JP 2004201465 A JP2004201465 A JP 2004201465A JP 2004201465 A JP2004201465 A JP 2004201465A JP 2006023185 A JP2006023185 A JP 2006023185A
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- APSNPMVGBGZYAJ-GLOOOPAXSA-N clematine Natural products COc1cc(ccc1O)[C@@H]2CC(=O)c3c(O)cc(O[C@@H]4O[C@H](CO[C@H]5O[C@@H](C)[C@H](O)[C@@H](O)[C@H]5O)[C@@H](O)[C@H](O)[C@H]4O)cc3O2 APSNPMVGBGZYAJ-GLOOOPAXSA-N 0.000 description 2
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- MRIAQLRQZPPODS-UHFFFAOYSA-N nobiletin Chemical compound C1=C(OC)C(OC)=CC=C1C1=CC(=O)C2=C(OC)C(OC)=C(OC)C(OC)=C2O1 MRIAQLRQZPPODS-UHFFFAOYSA-N 0.000 description 2
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- ZBJWWKFMHOAPNS-UHFFFAOYSA-N loretin Chemical group C1=CN=C2C(O)=C(I)C=C(S(O)(=O)=O)C2=C1 ZBJWWKFMHOAPNS-UHFFFAOYSA-N 0.000 description 1
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Landscapes
- Preparation Of Fruits And Vegetables (AREA)
Abstract
Description
本発明は、カンキツ加工品の識別方法に関し、詳しくはミカン区に属するシイクワシャーや温州ミカンの加工品の原料中にトウキンカン区のカラマンシーを含むものであるかを判別することによりカンキツ加工品を識別する方法に関する。 The present invention relates to a method for identifying a citrus processed product, and more particularly, to a method for identifying a citrus processed product by determining whether the raw material of a processed product of Shikuwasha or Wenzhou mandarin that belongs to the citrus region contains a calamancy of Tokkinkan region. .
沖縄で産出されるシイクワシャー（Citrus depressa HAYATA）は、他のカンキツ類と比べてポリメトキシフラボノイド成分であるノビレチン、タンゲレチン等を多量に含有する特徴的なカンキツである。このポリメトキシフラボノイド成分に発ガン抑制作用、ガン転移抑制作用が認められ（非特許文献１）、さらに、その加工品（果汁・ペースト）も血糖上昇抑制作用を有していることが報告されるに及び、シイクワシャーの生果、加工品に対する需要が高まっている。 Shikushasha (Citrus depressa HAYATA) produced in Okinawa is a characteristic citrus that contains a large amount of nobiletin, tangeretin and the like, which are polymethoxyflavonoid components, compared to other citrus. This polymethoxyflavonoid component has a carcinogenic inhibitory effect and a cancer metastatic inhibitory effect (Non-Patent Document 1), and it is further reported that processed products (fruit juice / paste) also have an anti-glycemic effect. In addition, there is an increasing demand for fresh fruits and processed products of Shikuwasha.
このような状況下、シイクワシャー以外の原料を用いたカンキツ加工品が市販されるようになり、需要者のみならず食品行政上でも問題となっている。 Under such circumstances, processed citrus products using raw materials other than Shikuwasha have become commercially available, and this is a problem not only for consumers but also for food administration.
本発明は、シイクワシャーおよび／または温州ミカンを原料とする加工品について、他のカンキツ類を原料として含有するカンキツ加工品と識別する方法を提供することを目的とする。 An object of the present invention is to provide a method for discriminating a processed product made from shiikuwasha and / or mandarin oranges from a citrus processed product containing other citrus as a raw material.
そこで、本発明者らは、カンキツ属、特に後生カンキツ亜属のミカン区のシイクワシャー、温州ミカンおよびトウキンカン区のカラマンシーに含まれる成分について比較、検討した。その結果、両者はポリメトキシフラボノイド成分の含有量に差が認められること並びに後者にのみカルコン配糖体が含まれていることを見出した。そこで、かかる知見に基づいてカンキツ加工品、特にシイクワシャーや温州ミカンの加工品中のカルコン配糖体の有無を指標としてカンキツ加工品を識別する方法を確立し、本発明を完成するに至った。 Therefore, the present inventors compared and examined the components contained in the citrus genus of the citrus genus, in particular the subgenus genus Citrus shasha, the citrus mandarin orange, and the calamancy of citrus citrus. As a result, they found that there was a difference in the content of the polymethoxyflavonoid component and that only the latter contained chalcone glycosides. Therefore, based on such knowledge, a method for identifying citrus processed products using the presence or absence of chalcone glycosides in processed citrus products, particularly processed products of Shikuwasha and Unshu mandarin oranges, was established, and the present invention was completed.
すなわち、請求項１記載の本発明は、カンキツ加工品中のカルコン配糖体の有無を調べ、その結果から当該加工品の原料を判別することを特徴とするカンキツ加工品の識別方法である。
請求項２記載の本発明は、カンキツ加工品中のカルコン配糖体の有無を調べ、その結果から当該加工品の原料としてカラマンシーが用いられているか否かを判別することを特徴とする請求項１記載のカンキツ加工品の識別方法である。
請求項３記載の本発明は、カンキツ加工品が、シイクワシャー果汁である請求項１または２記載のカンキツ加工品の識別方法である。
請求項４記載の本発明は、カルコン配糖体が3',5'−ジ−C−β−グルコピラノシルフロレチンである請求項１〜３のいずれかに記載のカンキツ加工品の識別方法である。
That is, the present invention according to claim 1 is a method for identifying a citrus processed product, wherein the presence or absence of a chalcone glycoside in the citrus processed product is examined, and the raw material of the processed product is discriminated from the result.
The present invention according to claim 2 is characterized by examining the presence or absence of chalcone glycosides in citrus processed products, and determining whether or not calamancy is used as a raw material of the processed products from the results. 2. A method for identifying a citrus processed product according to 1.
The present invention according to claim 3 is the citrus processed product identification method according to claim 1 or 2, wherein the processed citrus product is shiikuwasha juice.
In the present invention according to claim 4, the chalcone glycoside is 3 ′, 5′-di-C-β-glucopyranosylphloretin. Is the method.
本発明によれば、カンキツ加工品の原料として用いられているカンキツについてシイクワシャーおよび／または温州ミカン以外のものを含んでいるかどうかを識別して、シイクワシャーおよび／または温州ミカンのみを原料とするカンキツ加工品であるか否かを判定することができる。 According to the present invention, the citrus used as a raw material for processed citrus products is identified whether it contains anything other than citrus and / or citrus, and citrus processing using only citrus and / or citrus as a raw material. It can be determined whether it is a product.
請求項１記載の本発明によれば、カンキツ加工品中のカルコン配糖体の有無を調べ、その結果から当該加工品の原料を判別して当該加工品がシイクワシャーおよび／または温州ミカンのみを原料とするものであるか否かを識別することができる。 According to the first aspect of the present invention, the presence or absence of chalcone glycosides in processed citrus products is examined, the raw materials of the processed products are determined from the results, and the processed products are raw materials of shikuwasha and / or mandarin oranges only. Or not can be identified.
請求項２記載の本発明によれば、カンキツ加工品の原料としてカラマンシーが用いられているか否かを判別することにより、当該加工品がシイクワシャーおよび／または温州ミカンのみを原料とするものであるか否かを識別することができる。 According to the second aspect of the present invention, by determining whether or not calamancy is used as a raw material of a citrus processed product, whether the processed product uses only squid squash and / or mandarin orange as a raw material. Whether or not can be identified.
上記したように、本発明は、カンキツ加工品がシイクワシャーおよび／または温州ミカンのみを原料とするものであるか否かを識別することを基本とするものである。特に、原料中にカラマンシーが含まれているか否かを迅速に、かつ簡便な方法で正確に判別することを目的とする。
したがって、本発明が適用されるカンキツ加工品は、分類学的にシイクワシャーや温州ミカンが属するカンキツ属後生カンキツ亜属のミカン区の他に、トウキンカン区に属するカラマンシーを原料として用いる可能性のあるものである。加工品としては、例えば果汁、ドレッシングなどが挙げられる。
As described above, the present invention is based on identifying whether or not a processed citrus product is made from only squid washer and / or mandarin orange. In particular, it is an object to quickly and accurately discriminate whether or not calamancy is contained in a raw material.
Therefore, the processed citrus product to which the present invention is applied may be taxonomically used as a raw material of calamancy belonging to Tokinkankan, in addition to the Citrus and genus Citrus subgenus belonging to the subfamily of Citrus and Wenzhou mandarin. It is. Examples of processed products include fruit juice and dressing.
ミカン区に属するカンキツ類としては、シイクワシャーの他に温州ミカン等が挙げられる。また、トウキンカン区のカンキツ類としては、カラマンシー、シキキツがある。これらは、いずれもカンキツ加工品の原料として用いられるものである。 Citrus citrus belonging to the mandarin orange area includes Wenzhou mandarin oranges in addition to Shikuwasha. In addition, citrus in Tokinkan District includes Karamancy and Shikikitsu. These are all used as raw materials for processed citrus products.
これらのカンキツ類には、ポリメトキシフラボノイド成分が含まれているが、その種類によって、当該ポリメトキシフラボノイド成分の種類や含有量が相違していることが知られている。例えば、シイクワシャーにはノビレチン、タンゲレチンなどのポリメトキシフラボノイド成分が多量に含まれている。
一方、トウキンカン区のカンキツ類であるカラマンシーには、カルコン配糖体が特異的に含まれており、該化合物の具体例が以下に示す3',5'−ジ−C−β−グルコピラノシルフロレチン（以下、フロレチン配糖体と略記することがある）であることが明らかとなった。
These citrus species contain a polymethoxyflavonoid component, and it is known that the type and content of the polymethoxyflavonoid component differ depending on the type. For example, shiikuwasha contains a large amount of polymethoxyflavonoid components such as nobiletin and tangeretin.
On the other hand, calamancy, which is a citrus fruit of Tokinkan, contains chalcone glycosides specifically, and specific examples of the compounds are shown below as 3 ′, 5′-di-C-β-glucopyranosylphenol. It was revealed that it is loretin (hereinafter sometimes abbreviated as phloretin glycoside).
したがって、カンキツ加工品中の上記特定の成分について分析することにより、該カンキツ加工品の原料として用いられているカンキツ類の種類を識別することができる。すなわち、一つの方法として、予めシイクワシャーのみを原料とするカンキツ加工品中に含まれる特定成分の含有量を求めておき、対象とするカンキツ加工品における当該特定成分の含有量を測定し、この値を標準品であるシイクワシャーのみを原料とするカンキツ加工品の測定値と比べることによって、対象のカンキツ加工品がシイクワシャーのみを原料とする製品であるか否かを識別することができる。
これに対して、本発明では、カンキツ加工品中のカルコン配糖体の有無を調べ、その結果から当該加工品の原料を判別して当該加工品がシイクワシャーおよび／または温州ミカンのみを原料とするものであるか否かを識別する方法である。
Therefore, by analyzing the specific component in the processed citrus product, the type of citrus used as the raw material of the processed citrus product can be identified. That is, as one method, the content of a specific component contained in a citrus processed product using only shiikuwasha as a raw material is obtained in advance, and the content of the specific component in the target citrus processed product is measured, and this value Is compared with the measured value of the citrus processed product using only the standard sikuwasher as a raw material, it is possible to identify whether the target citrus processed product is a product using only the sikuwasher as a raw material.
On the other hand, in the present invention, the presence or absence of chalcone glycosides in citrus processed products is examined, the raw materials of the processed products are determined from the results, and the processed products use only squid squash and / or mandarin oranges as raw materials. This is a method for identifying whether or not a thing is present.
以下に、カンキツ加工品が、果汁である場合について述べる。なお、果汁の製造には、完熟果実を用いるのが一般的である。
当該カンキツ加工品中にカルコン配糖体が含まれているか否かを調べるため、分析用の試料を用意する。通常、分析は薄層クロマトグラフィ（TLC）、高速液体クロマトグラフィ（HPLC)で行う。TLC分析を行う場合、まず果汁に0.5〜2.0倍量の溶媒（例えばメタノール、エタノール等）を加え、ホモジナイズしてホモジネート抽出液を得る。予めメタノール、水で洗浄したセップパック（例えば、C18Sep‐Pak) に該抽出液を通過させて固相処理する。同様に、水、10％メタノールも通過させることによって、糖などの展開の阻害要因となる非吸着物を除去する。最後に、メタノールを通過させてセップパック中の吸着物を集める。
得られた通過溶離液を窒素乾固して約1/4程度に濃縮する。これを、TLC分析用試料として用いる。
Below, the case where a processed citrus product is fruit juice is described. In addition, it is common to use a ripe fruit for manufacture of fruit juice.
In order to examine whether or not the citrus processed product contains a chalcone glycoside, a sample for analysis is prepared. Usually, analysis is performed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). When performing TLC analysis, first, 0.5 to 2.0 times the amount of solvent (for example, methanol, ethanol, etc.) is added to the fruit juice, and homogenized to obtain a homogenate extract. The extract is passed through a Sepp pack (for example, C 18 Sep-Pak) previously washed with methanol and water, and subjected to solid phase treatment. Similarly, non-adsorbed substances that inhibit development such as sugar are removed by passing water and 10% methanol. Finally, methanol is passed through to collect the adsorbate in the sep pack.
The obtained passing eluent is dried to nitrogen and concentrated to about 1/4. This is used as a sample for TLC analysis.
TLCによるカルコン配糖体であるフロレチン配糖体の分析は、例えばメルク社製シリカゲルTLCプレートなどの薄層板を展開槽に入れ、クロロホルム、メタノールおよび水で調製した展開溶媒を用いて行うことができる。なお、10％硫酸溶液を用い、加熱処理することにより、目的とする成分の検出を行う。 Analysis of phloretin glycoside, which is a chalcone glycoside by TLC, can be performed using a developing solvent prepared with chloroform, methanol and water, for example, by placing a thin layer plate such as a silica gel TLC plate manufactured by Merck in a developing tank. it can. In addition, the target component is detected by heat treatment using a 10% sulfuric acid solution.
カンキツ加工品（果汁）中にカラマンシーの属するキンカン属特有のフロレチン配糖体が含まれる場合は、TLCプレートのRf0.2の位置に該配糖体によるスポットが出現する。このことから、試料中にカラマンシー果汁が含まれていること、すなわちシイクワシャーおよび／または温州ミカン以外の原料を使用していることが判別できる。一方、Rf0.2の位置にフロレチン配糖体によるスポットが認められない場合は、試料中にカラマンシー果汁が含まれていないこと、すなわちシイクワシャーおよび／または温州ミカンのみを原料としていることが分かる。 When the processed citrus product (fruit juice) contains phloretin glycoside peculiar to the genus Kumquat to which the calamancy belongs, a spot due to the glycoside appears at the position of Rf0.2 on the TLC plate. From this, it can be determined that the sample contains calamancy juice, that is, that a raw material other than Shikuwasha and / or Satsuma mandarin is used. On the other hand, when a spot due to phloretin glycoside is not observed at the position of Rf0.2, it can be seen that the sample does not contain caramancy juice, that is, only Sikusshasha and / or Satsuma mandarin is used as a raw material.
次に、HPLCによるフロレチン配糖体の分析は、果汁に２〜３倍量の抽出溶媒（例えば、メタノール、エタノールなど）を加えた後、超音波処理により抽出する。得られた抽出液を定容・ろ過したものを、HPLC分析用試料として用いる。 Next, the analysis of phloretin glycosides by HPLC is performed by adding ultrasonic solvent (for example, methanol, ethanol, etc.) in an amount of 2 to 3 times to the fruit juice, followed by sonication. A solution obtained by constant volume and filtration of the obtained extract is used as a sample for HPLC analysis.
HPCLによるフロレチン配糖体の分析は、例えばLiChrospher100RP-18などのカラムを使用し、移動相にメタノールおよび20mM リン酸水溶液を用いグラジエント方式、カラム温度40℃で行うことができる。なお、波長286nmにおける吸光度を計測することにより、目的成分の検出を行う。 Analysis of phloretin glycosides by HPCL can be performed using a column such as LiChrospher100RP-18, using a gradient of methanol and 20 mM aqueous phosphoric acid as the mobile phase, and a column temperature of 40 ° C. The target component is detected by measuring the absorbance at a wavelength of 286 nm.
カンキツ加工品(果汁)中にフロレチン配糖体が含まれる場合は、該配糖体によるピーク（Rt25.3分）が認められる。このことから、試料中にカラマンシー果汁が含まれていること、すなわちシイクワシャーおよび／または温州ミカン以外の原料も含有するカンキツ加工品であることが判別できる。一方、Rt25.3分のフロレチン配糖体によるピークが認められない場合は、試料中にカラマンシー果汁が含まれていないこと、すなわちシイクワシャーおよび／または温州ミカンのみを原料とした加工品であることが分かる。 When phloretin glycoside is contained in the citrus processed product (fruit juice), a peak (Rt 25.3 min) due to the glycoside is observed. From this, it can be determined that the sample contains calamancy juice, that is, it is a processed citrus product containing raw materials other than shikuwasha and / or mandarin orange. On the other hand, if the peak due to the phloretin glycoside at Rt 25.3 is not observed, the sample should not contain calamancy juice, that is, it should be a processed product that uses only squid washer and / or mandarin orange as a raw material. I understand.
果汁以外のカンキツ加工品について識別を行う場合も、上記と同様の方法によって識別することができる。 When identifying citrus processed products other than fruit juice, they can be identified by the same method as described above.
以下、実施例により本発明を詳しく説明するが、本発明はこれらによって限定されるものではない。 EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, this invention is not limited by these.
実施例１
材料の調製
基準の100％果汁（ストレート果汁）として、沖縄県経済連農産加工工場から入手したシイクワシャー果汁並びにフィリピン産のカラマンシー果汁を用いた。
また、試料用のシイクワシャー加工品（果汁）として、沖縄県で入手した市販品11種類および福岡市内で入手した市販品１種類を用いた。
Example 1
As a 100% fruit juice (straight fruit juice) as a preparation standard for materials, Shikuwasha fruit juice obtained from the Okinawa Prefectural Economic Federation Agricultural Processing Plant and Karamancy fruit juice from the Philippines were used.
Moreover, 11 types of commercial products obtained in Okinawa Prefecture and 1 type of commercial products obtained in Fukuoka City were used as processed squish washer products (fruit juice).
試薬
カルコン配糖体として、独立行政法人農業・生物系特定産業技術研究機構果樹研究所から分与された3',5'-ジ-C-β-グルコピラノシルフロレチンの標準品（フロレチン配糖体）を用いた。
TLC分析用の展開溶媒として、クロロホルム（和光純薬製）、メタノール（和光純薬製）を用い、固相処理に使用するメタノールは和光純薬製を用いた。HPLC分析において移動相として用いたメタノールは和光純薬製を用い、抽出に使用したエタノールは、和光純薬製を用いた。また、HPLC用フィルターは、酢酸セルロース フィルタ 0.45μｍ（アドバンテック社製）を用いた。
Reagents 3 ', 5'-di-C-β-glucopyranosylphloretin standard product (phloretin) distributed from the National Institute of Agricultural Sciences and Fruits, as a chalcone glycoside Glycoside).
As developing solvents for TLC analysis, chloroform (manufactured by Wako Pure Chemical Industries) and methanol (manufactured by Wako Pure Chemical Industries) were used, and methanol used for solid phase treatment was manufactured by Wako Pure Chemical Industries. Methanol used as a mobile phase in HPLC analysis was manufactured by Wako Pure Chemical Industries, and ethanol used for extraction was manufactured by Wako Pure Chemical Industries. The HPLC filter used was a cellulose acetate filter 0.45 μm (manufactured by Advantech).
実験方法
TLC分析試料の調製
TLC分析のための基準品、シイクワシャーストレート果汁、カラマンシーストレート果汁および試料である市販シイクワシャー果汁加工品から以下の方法で抽出して分析用試料を得た。
まず、果汁10mlに等量のメタノールを添加し、ホモジナイザー（商品名：ポリトロンホモジナイザー P-1型、セントラル科学貿易製）で1分間ホモジナイズした後、試料2mlをメタノールおよび水で洗浄したセップパック（商品名：C18Sep-Pak、ウォーター社製）に通過させた。
次いで、同様に水2ml、10%メタノール2.5mlをセップパックに通過させ、展開阻害要因である糖などの非吸着物を洗浄・廃棄した。最後に、メタノール2mlを通過させ、C18Sep-Pak中の吸着物を回収し、これを窒素乾固により1/4量程度に濃縮後、TLC分析に供した。
experimental method
Preparation of TLC analysis sample
Samples for analysis were obtained by extraction from the standard products for TLC analysis, Shikuwasha straight fruit juice, Karaman Sea straight fruit juice, and commercially available Shikuwasha fruit juice processed products as follows.
First, add an equal amount of methanol to 10 ml of fruit juice, homogenize for 1 minute with a homogenizer (trade name: Polytron homogenizer type P-1, manufactured by Central Science Trade), then wash the sample 2 ml with methanol and water. Name: C 18 Sep-Pak, manufactured by Water).
Subsequently, 2 ml of water and 2.5 ml of 10% methanol were similarly passed through the Sepp pack to wash and discard non-adsorbed substances such as sugars, which are development inhibiting factors. Finally, 2 ml of methanol was passed through to recover the adsorbate in C 18 Sep-Pak, which was concentrated to about 1/4 by nitrogen drying and subjected to TLC analysis.
TLC分析
上記のように調製した試料をTLCに供した。なお、展開溶媒としてクロロホルム/メタノール/水（65:35:5）を用いた。展開槽として蓋付のガラス製容器を使用し、これに加熱乾燥した薄層板（商品名：シリカゲル60F254 20x20cm、メルク社製）を収納した。各試料を展開させた後、各スポットの検出は、10%H2SO4溶液を噴霧した後、加熱により発色させて行った。結果を図１および図２に示す。
TLC analysis Samples prepared as described above were subjected to TLC. In addition, chloroform / methanol / water (65: 35: 5) was used as a developing solvent. A glass container with a lid was used as a developing tank, and a heat-dried thin layer plate (trade name: silica gel 60F 254 20x20 cm, manufactured by Merck & Co., Inc.) was accommodated in the container. After each sample was developed, each spot was detected by spraying a 10% H 2 SO 4 solution and then coloring by heating. The results are shown in FIG. 1 and FIG.
実験結果
図１に示したように、カラマンシーストレート果汁のスポットは、図中左端の標準試薬（フロレチン配糖体）のスポット（Rf0.2）と同位置にある。また、試料Aについても、同位置にスポットが出現し、フロレチン配糖体の存在が確認された。このことから、これらの試料はカラマンシーを原料として用いていることが分かった。一方、シイクワシャーストレート果汁にはフロレチン配糖体の存在を示す位置にスポットが出現しなかった。
また、図２に示したように、試料A〜Gには、カラマンシーストレート果汁の場合と同様に、フロレチン配糖体の存在を示すRf0.2の位置にスポットが認められた。なお、試料H、Iには、フロレチン配糖体の存在を示す位置に肉眼的に判別できるスポットは認められなかった。
しかし、試料J、KおよびLについては、フロレチン配糖体の存在を示すRf0.2の位置にスポットが出現しなかった。このことから、試料J、KおよびLはシイクワシャーおよび／または温州ミカンのみを原料とする果汁であることが分かった。
Experimental Results As shown in FIG. 1, the spot of Karaman Sea straight juice is at the same position as the spot (Rf0.2) of the standard reagent (phloretin glycoside) at the left end in the figure. In addition, for sample A, a spot appeared at the same position, and the presence of phloretin glycoside was confirmed. From this, it was found that these samples used calamancy as a raw material. On the other hand, no spot appeared in the position where the presence of phloretin glycoside was present in Shikuwasha straight fruit juice.
In addition, as shown in FIG. 2, in Samples A to G, a spot was observed at the position of Rf0.2 indicating the presence of phloretin glycoside, as in the case of caraman sea straight juice. In Samples H and I, no spot that could be visually discriminated was found at a position indicating the presence of phloretin glycoside.
However, for Samples J, K, and L, no spot appeared at the position of Rf0.2 indicating the presence of phloretin glycoside. From this, it was found that Samples J, K, and L were fruit juices made only from Shikuwasha and / or Satsuma mandarin.
実施例２
材料の調製と試薬は、実施例１と同じである。
Example 2
The material preparation and reagents are the same as in Example 1.
実験方法
HPLC分析試料の調製
HPLC分析のための、シイクワシャーストレート果汁、カラマンシーストレート果汁および市販シイクワシャー果汁加工品から以下の方法で抽出して分析用試料を得た。
試料果汁3mlにエタノール7mlを添加し、超音波処理（30分間）によって果汁成分の抽出を行った後、定容・ろ過（フィルター）し、HPLC分析に供した。
experimental method
Preparation of HPLC analysis sample
Samples for analysis were obtained by extraction from Shikuwasha straight fruit juice, Karaman Sea straight fruit juice and commercial processed Shikuwasha fruit juice for HPLC analysis by the following method.
After adding 7 ml of ethanol to 3 ml of sample juice and extracting the juice components by sonication (30 minutes), it was subjected to constant volume filtration (filtering) and subjected to HPLC analysis.
HPLC分析
上記のように調製した試料について、以下の条件でHPLC分析を行い、フロレチン配糖体の含有量を測定した。結果を表１に示す。
装置： ポンプ（商品名：島津LC-10、島津製作所製）
UV-VIS検出器（島津製作所製）
カラム： LiChrospher100：RP-18
（粒径5μm、125x4 mm）
カラム温度：40℃
移動相： メタノール（A）：20mM リン酸水溶液（B）
0-5-45分、20-20-45%（A in B）
流速： 0.9ml/分
検出波長： 286nm
分析時間： 45分
溶出方式： グラジエント方式
HPLC analysis The sample prepared as described above was subjected to HPLC analysis under the following conditions to measure the content of phloretin glycoside. The results are shown in Table 1.
Equipment: Pump (Product name: Shimadzu LC-10, manufactured by Shimadzu Corporation)
UV-VIS detector (manufactured by Shimadzu Corporation)
Column: LiChrospher100: RP-18
(Particle size 5μm, 125x4 mm)
Column temperature: 40 ° C
Mobile phase: Methanol (A): 20 mM phosphoric acid aqueous solution (B)
0-5-45 minutes, 20-20-45% (A in B)
Flow rate: 0.9ml / min Detection wavelength: 286nm
Analysis time: 45 minutes Elution method: Gradient method
n.d.：未検出、偽：偽和製品の疑いあり、真：真正製品と認められる
試料のうち、H,I,Lは10%果汁飲料、Kは40%果汁飲料であり、他は全て100%果汁である。
nd: not detected, false: suspected of false product, true: recognized as genuine product H, I, L is 10% juice drink, K is 40% juice drink, all others are 100% Fruit juice.
表１から明らかなように、試料A〜Gには、基準試料であるカラマンシー果汁と同様に、フロレチン配糖体の存在が確認され、原料にカラマンシーを含む果汁であることが分かった。また、試料HとIについても、少量ではあるが、フロレチン配糖体が検出され、これらの果汁も原料にカラマンシーを含むことが分かった。
しかし、試料J、KおよびLからは、カラマンシーに特異的に含まれるフロレチン配糖体が検出されなかった。このことから、試料J、KおよびLはシイクワシャーおよび／または温州ミカンのみを原料とする果汁であることが分かった。
As is clear from Table 1, the presence of phloretin glycosides was confirmed in samples A to G, as in the case of caramancy fruit juice as a reference sample, and it was found that the fruit juice contained caramancy as a raw material. In addition, phloretin glycosides were also detected in samples H and I, although the amount was small, and it was found that these fruit juices also contain calamancy as a raw material.
However, phloretin glycosides specifically contained in calamancy were not detected from samples J, K, and L. From this, it was found that Samples J, K, and L were fruit juices made only from Shikuwasha and / or Satsuma mandarin.
また、シイクワシャーストレート果汁、カラマンシーストレート果汁および試料の市販シイクワシャー果汁加工品FとJについてのHPLC分析のクロマトグラムを図３に示した。この図から、カラマンシーストレート果汁には、フロレチン配糖体によるピーク（Rt25.3分）が認められた。また、試料Fには、二つの高いピークが認められ、これらはフロレチン配糖体によるピークおよびシイクワシャー果汁に見られるヘスペリジンのピークと溶出時間が同じであると認められたことから、試料Fはカラマンシーを原料として含む加工品であることが明らかとなった。
一方、シイクワシャーストレート果汁および試料Jには、フロレチン配糖体によるピークは認められず、ヘスペリジンよるピーク（Rt28.4分）のみが認められた。このことから、これらの果汁は原料としてカラマンシーを使用していないことが分かった。
In addition, FIG. 3 shows chromatograms of HPLC analysis of shiikuwasha straight fruit juice, calaman sea straight fruit juice, and commercially available processed shikuwasha fruit juice products F and J. From this figure, a peak due to phloretin glycoside (Rt 25.3 min) was observed in the Karaman Sea straight juice. In Sample F, two high peaks were observed, and it was confirmed that the elution time was the same as that of the peak of phloretin glycoside and the peak of hesperidin found in Shikuwasha juice. It became clear that it was a processed product containing as a raw material.
On the other hand, the peak due to hesperidin (Rt 28.4 min) was not observed in the squish shear straight juice and the sample J, but a peak due to phloretin glycoside was not observed. From this, it was found that these juices do not use calamancy as a raw material.
本発明は、果汁等のカンキツ加工品の識別に利用され、当該カンキツ加工品が、その原料として、シイクワシャー以外のもの、特にカラマンシー果実を含有するものであるか否かを簡便に、かつ正確に判定して、カンキツ加工品の真偽を識別することができる。 The present invention is used for identifying citrus processed products such as fruit juice, and whether or not the citrus processed product contains anything other than shiikuwasha, particularly calamancy fruit, as its raw material. By judging, the authenticity of the citrus processed product can be identified.
Claims (4)
The method for identifying a processed citrus product according to any one of claims 1 to 3, wherein the chalcone glycoside is 3 ', 5'-di-C-β-glucopyranosylphloretin.
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