JP7263559B2 - 黒参抽出物のジンセノサイドの生体利用率の増大及び吸収促進用組成物 - Google Patents
黒参抽出物のジンセノサイドの生体利用率の増大及び吸収促進用組成物 Download PDFInfo
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- JP7263559B2 JP7263559B2 JP2021569187A JP2021569187A JP7263559B2 JP 7263559 B2 JP7263559 B2 JP 7263559B2 JP 2021569187 A JP2021569187 A JP 2021569187A JP 2021569187 A JP2021569187 A JP 2021569187A JP 7263559 B2 JP7263559 B2 JP 7263559B2
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- black ginseng
- ginsenosides
- ginseng extract
- hours
- black
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Description
ただし、下記製造例、実験例は、本発明を例示するためのものに過ぎず、本出願の内容が下記製造例及び実験例により限定されるものではない。
1-1.紅参(黒参1蒸曝)の製造
4年根水参の根をスクリュー洗浄機(三角fmc、韓国)に入れ、土又は異物を除去するために3分間1回洗浄した。洗浄された水参の根を蒸熟器に入れた後、予熱時間を除いて98℃で水蒸気を用いて2時間1次蒸熟した。蒸熟器の温度を55℃に冷却させた後、蒸熟された水参の根を蒸熟器から取り出した。熱風乾燥器の内部温度を55℃に維持させた後、蒸熟された水参の根を入れて約18時間乾燥させた。乾燥させた紅参は、日乾場(自然光による乾燥施設)に移して一ヶ月以上乾燥させた。以上の過程により、紅参(1蒸曝)930kgを製造した。
前記製造例1-1で製造された紅参930kgを、製造例1-1と同一の蒸熟、乾燥条件で蒸熟した。前述した蒸熟及び乾燥工程を繰り返し行い、繰り返しが行われる回数に応じて2蒸曝~9蒸曝と称した。蒸熟及び乾燥を繰り返すにつれて色が次第に黒色に近くなった。前記工程を介して九蒸九曝黒参853.8kgを製造した。下記製造例2の黒参抽出物又は製造例3の黒参濃縮液の製造時には、九蒸九曝黒参を原料として使用した。
製造例1-1で製造された紅参、製造例1-2で製造された黒参284.6kgをそれぞれ取って粉砕した後、粉砕物重量に対して6倍に70%(v/v)エタノール水溶液を投入し、82℃で6時間1次抽出した。1次抽出の際に発生した抽出残渣物を、1次抽出と同一の条件で2次抽出した。2次抽出の際に発生した抽出残渣物重量に対して6倍に50%(v/v)エタノール水溶液を投入し、85℃で6時間3次抽出した。3次抽出の際に発生した抽出残渣物重量に対して6倍に30%(v/v)エタノール水溶液を投入し、90℃で6時間4次抽出した。4次抽出の際に発生した抽出残渣物重量に対して6倍の精製水を投入し、95℃で6時間5次抽出した。
製造例2で得た1次~4次紅参抽出物、1次~4次黒参抽出物をそれぞれ濾過して、抽出物と抽出残渣を分離した。各原料に対して分離した抽出物のみを濃縮器に入れ、55℃で減圧濃縮器で4時間濃縮した。70Brix%である紅参濃縮液231kgを得て、70Brix%である黒参濃縮液212kgを得た。得た紅参濃縮液、黒参濃縮液を、下記の成分分析及び生体利用率の増大及び吸収速度実験の試料として使用した。
蒸熟時間を1時間、2時間、3時間に変更した以外には、製造例1-1の蒸熟条件と同様にして9蒸曝した。製造された紅参は、製造例2及び製造例3により抽出して濃縮した。ジンセノサイドの含量は、下記実験例3の方法と同様に測定した。
製造例3で得た黒参濃縮液を2mlバイアルに入れ、ウォーターバスで70℃、80℃、90℃でそれぞれ8時間熟成し、時間によるジンセノサイド含量変化を測定した。ジンセノサイドの含量は、実験例3の方法と同様に測定した。
製造例3で得た紅参濃縮液又は黒参濃縮液に存在するジンセノサイド成分の分析は、HPLCを用いて実施した。試料は、前記製造例3から得た黒参濃縮液をメタノールに1/30の割合で希釈した後、0.45umのフィルターで濾過した溶液を使用した。HPLCは、HPLC1260(DAD)(Agilent、USA)を使用した。コラムは、VENUSIL XBPC18(4.6MM×250MM、5.0um)を使用し、移動相は、LC級水をA溶媒に、アセトニトリルをB溶媒に使用した。コラム温度は、35℃に維持した。勾配条件は、0~3.8分間1ml/分の流速でB溶媒を30%に維持し、3.81分から4.5分間0.5ml/分の流速でB溶媒を30%に維持し、4.51分から6分間1.2ml/分の流速でB溶媒を30%から42%に増加させ、7.5分から13分間0.5ml/分の流速でB溶媒を42%に維持し、18分まで0.5ml/分の流速でB溶媒を47.4%に増加させ、18.1分から25分間1ml/分の流速でB溶媒を55%に増加させ、30分から32分まで1ml/分の流速でB溶媒を60%に増加させ、42分から50分まで1ml/分の流速でB溶媒を60%から90%に増加させ、50.1分から53分まで1ml/分の流速でB溶媒を30%に維持した。成分分析の結果を下記に示す(単位mg/g)。
製造例3で得た紅参濃縮液、黒参濃縮液に存在する酸性多糖体成分の分析は、カルバゾール硫酸比色法を介して測定した。試料は、前記製造例3で得た各濃縮液300mgずつ10mlの蒸溜水に希釈して使用した。希釈液を90℃で3時間湯煎した後、冷却して遠心分離(3000rpm、10分)し、上澄液から1mlを取り、ここに4mlのエタノールを添加して白色沈殿物を形成した。白色沈殿物を得るために遠心分離(3000rpm、10分)した後、上澄液を除去し、4mlの蒸溜水を添加して白色沈殿物を溶解させた後、n-ブタノールとCHCl3を1:4で混合した溶液1mlを入れて撹拌し、再び遠心分離(3000rpm、10分)した。これから上澄液である水抽出液4mlを取り、ソニケーションを進行した。
製造例3で得た紅参濃縮液、黒参濃縮液に存在するポリフェノールの含量を、Microplate Reader(Powerwave XS、BioTek、USA)を用いて測定した。炭酸ナトリウム(Sigma223484、CAS No.497-19-8)2gを100mlの定容フラスコに取った後、蒸溜水で100ml定容して2%炭酸ナトリウム試薬を製造する。フォリン-チオカルトー(Folin-Ciocalteu’s)フェノール試薬(SigmaF9252-1L)と蒸溜水を1:1の割合で混合し、50%フォリン-チオカルトーフェノール試薬を製造し、光が透過されないようにアルミニウムホイルで覆った。
[式1]
総ポリフェノールの含量(mg/ml)=(A×B×C)/D
*A:試験溶液の全量(ml)、B:希釈倍数、C:試験溶液中の総ポリフェノールの濃度(mg/ml)、D:試料採取量(ml)
製造例3で製造された黒参濃縮液は、ジンセノサイドRk1、Rg5、酸性多糖体、ポリフェノールの含量が、製造例3で製造された紅参濃縮液に比べて高く、これにより製造例3の黒参濃縮液を標準化黒参濃縮液と命名した。前記標準化黒参濃縮液をジンセノサイドの生体利用率の測定に使用した。
実験例6では、健康な男性志願者を対象にして、黒参濃縮液と紅参濃縮液とを単回経口投与した後、ジンセノサイド(ginsenoside)の薬動学的(生体利用率)特性を比較評価した。
実験対象者2人で紅参濃縮液と黒参濃縮液とを2週間隔で摂取した後、ジンセノサイド(Ginsenoside)14種(Rb1、Rb2、Rc、Rd、Rg3(s)、Rk1、Rg5、CK、Rh2(s)、PPD(s)、Re、Rg1、Rh1(s)、PPT(s))に対して吸収率分析を行った。吸収率分析の結果、以下の表4に示したように、紅参濃縮液に対して黒参濃縮液の吸収率(AUC基準)は、ジンセノサイド(Ginsenoside)14種の合計は1.8倍、Rg3は15.7倍、Rh2は11.7倍にそれぞれ増大された。特に、紅参濃縮液摂取群で、Rk1は吸収が0であるのに対し、黒参濃縮液摂取群では、Rk1(AUC基準)は32ug・hr/mlの摂取率が測定された。全てのジンセノサイドを合わせた総吸収率で、紅参は456ug・hr/mlであり、黒参は770ug・hr/mlで、1.7倍増加した。また、吸収速度の側面で、紅参濃縮液は、総ジンセノサイド最高含量の到達時間が投与後の11時間(Tmax 11hr)であるのに対し、黒参濃縮液は、総ジンセノサイド最高含量の到達時間は投与後の8.5時間(Tmax 8.5hr)であるので、吸収速度も黒参濃縮液に優れた。
4-1.小麦粉食品の製造
本発明の黒参抽出物0.5~5.0重量部を小麦粉に添加し、この混合物を用いてパン、ケーキ、クッキー、クラッカー及び麺類を製造した。
本発明の黒参抽出物0.2~5.0重量部をスープ及び肉汁に添加して健康増進用肉加工製品、麺類のスープ及び肉汁を製造した。
本発明の黒参抽出物10重量部をグラウンドビーフに添加して健康増進用グラウンドビーフを製造した。
本発明の黒参抽出物5~10重量部を牛乳に添加し、前記牛乳を用いてバター及びアイスクリームのような多様な乳製品を製造した。
玄米、麦、もち米、鳩麦を公知の方法でアルファ化させて乾燥させたものを焙煎した後、粉砕機で粒度60メッシュの粉末に製造した。
液状果糖(0.5%)、オリゴ糖(2%)、砂糖(2%)、食塩(0.5%)、水(75%)のような副材料と、本発明の黒参抽出物5gとを均質に配合して瞬間殺菌した後、これをガラス瓶、ペット瓶などの小包装容器に包装して製造した。
本発明の黒参抽出物5gを、トマト又はニンジンジュース1,000mlに加えて野菜ジュースを製造した。
本発明の黒参抽出物1gを、リンゴ又はブドウジュース1,000mlに加えて果物ジュースを製造した。
5-1.散剤の製造
本発明の黒参抽出物2g
乳糖1g
前記成分を混合し、気密包に充填して散剤を製造した。
本発明の黒参抽出物100mg
トウモロコシ澱粉100mg
乳糖100mg
ステアリン酸マグネシウム2mg
前記成分を混合した後、通常の錠剤の製造方法により打錠して錠剤を製造した。
本発明の黒参抽出物100mg
トウモロコシ澱粉100mg
乳糖100mg
ステアリン酸マグネシウム2mg
前記成分を混合した後、通常のカプセル剤の製造方法によりゼラチンカプセルに充填してカプセル剤を製造した。
本発明の黒参抽出物1g
乳糖1.5g
グリセリン1g
キシリトール0.5g
前記成分を混合した後、通常の方法により1丸当り4gになるように製造した。
本発明の黒参抽出物150mg
大豆抽出物50mg
ブドウ糖200mg
澱粉600mg
前記成分を混合した後、30%エタノール100mgを添加し、摂氏60℃で乾燥して顆粒を形成した後、包に充填した。
Claims (8)
- ジンセノサイドRk1及びRg5を含む黒参(black ginseng)抽出物を含み、
前記ジンセノサイドRk1及びRg5の含量の合計は、ジンセノサイドRb1、Rb2、Rc、Rd、Re、Rg1、Rg3(s)、Rk1、Rg5及びRh1(s)の含量の合計100重量部に対して20重量部~90重量部で含み、
前記Rk1及びRg5の含量の合計は、9mg/g以上であり、
前記黒参抽出物は、さらにポリフェノールを含み、前記ポリフェノールの含量は、15mg/g以上のものである、黒参抽出物のジンセノサイドの生体利用率の増大及び吸収促進用組成物。 - 黒参抽出物を9g投与する際に、ジンセノサイドRb1、Rb2、Rc、Rd、Rg3(s)、Rk1、Rg5、CK、Rh2(s)、PPD(s)、Re、Rg1、Rh1(s)、及びPPT(s)のAUC(Area under the plasma level-time curve)値の合計は、500~1200ug・hr/mlである、請求項1に記載の黒参抽出物のジンセノサイドの生体利用率の増大及び吸収促進用組成物。
- 黒参抽出物を9g投与する際に、ジンセノサイドRk1のAUC(Area under the plasma level-time curve)値は、20~40ug・hr/mlである、請求項2に記載の黒参抽出物のジンセノサイドの生体利用率の増大及び吸収促進用組成物。
- 黒参抽出物を9g投与する際に、ジンセノサイドRg3のAUC(Area under the plasma level-time curve)値は、40~80ug・hr/mlである、請求項2に記載の黒参抽出物のジンセノサイドの生体利用率の増大及び吸収促進用組成物。
- 黒参抽出物を9g投与する際に、ジンセノサイドRh2のAUC(Area under the plasma level-time curve)値は、250~400ug・hr/mlである、請求項2に記載の黒参抽出物のジンセノサイドの生体利用率の増大及び吸収促進用組成物。
- 黒参抽出物のジンセノサイドRb1、Rb2、Rc、Rd、Rg3(s)、Rk1、Rg5、CK、Rh2(s)、PPD(s)、Re、Rg1、Rh1(s)、及びPPT(s)のTmax値は、5時間~10時間である、請求項1に記載の黒参抽出物のジンセノサイドの生体利用率の増大及び吸収促進用組成物。
- 前記黒参抽出物は、酸性多糖体をさらに含むものである、請求項1に記載の黒参抽出物のジンセノサイドの生体利用率の増大及び吸収促進用組成物。
- 前記組成物は、食品又は薬学組成物である、請求項1に記載の黒参抽出物のジンセノサイドの生体利用率の増大及び吸収促進用組成物。
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CN116459267A (zh) * | 2022-01-11 | 2023-07-21 | 大连富生天然药物开发有限公司 | 人参皂苷Rd的抗炎应用 |
KR20240037390A (ko) | 2022-09-13 | 2024-03-22 | 주식회사 진생바이팜 | 인삼류 추출물 및 스테로이드계 화합물을 유효 성분을 함유하는 항바이러스 조성물 |
KR20240081512A (ko) | 2022-11-28 | 2024-06-10 | 농업회사법인 주식회사 함초록 | 산양삼 또는 인삼으로부터 저분자 진세노사이드를 유효성분으로 함유하도록 발효한 흑삼 추출물을 이용한 흑삼 음료의 제조방법 |
KR102549962B1 (ko) * | 2023-01-18 | 2023-06-30 | 최광태 | 진세노사이드 Rg3, Rg5 및 Rk1의 함량이 증가된 흑삼 및 그 제조방법 |
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US20220249587A1 (en) | 2022-08-11 |
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