KR20230154368A - A health functional food composition for improving muscle loss or motor ability, including extract of Catalpa bignonioides - Google Patents
A health functional food composition for improving muscle loss or motor ability, including extract of Catalpa bignonioides Download PDFInfo
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- KR20230154368A KR20230154368A KR1020220053738A KR20220053738A KR20230154368A KR 20230154368 A KR20230154368 A KR 20230154368A KR 1020220053738 A KR1020220053738 A KR 1020220053738A KR 20220053738 A KR20220053738 A KR 20220053738A KR 20230154368 A KR20230154368 A KR 20230154368A
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- paulownia
- extract
- extraction
- food composition
- muscle
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- Medicines Containing Plant Substances (AREA)
Abstract
본 발명은 꽃개오동 추출물을 포함하는 근감소 개선용 또는 운동능력 개선용 식품 조성물에 관한 것으로, 더욱 상세하게는 초음파 추출법으로 추출함에 따라 유용성분을 고수율로 추출한 꽃개오동 추출물 또는 이의 분획물을 포함하는 근감소 개선용 또는 운동능력 개선용 식품 조성물에 관한 것이다.The present invention relates to a food composition for improving muscle loss or improving exercise capacity containing an extract of Paulownia perennialis. More specifically, it relates to an extract of Paulownia perennialis or a fraction thereof in which useful components are extracted in high yield by extraction using an ultrasonic extraction method. It relates to a food composition for improving muscle loss or improving exercise ability.
Description
본 발명은 꽃개오동 추출물을 포함하는 근감소 개선용 또는 운동능력 개선용 식품 조성물에 관한 것으로, 더욱 상세하게는 꽃개오동 열매로부터 유용성분을 효율적으로 추출하는 추출방법 및 상기 추출방법으로 추출한 꽃개오동 열매 추출물 또는 이의 분획물을 포함하는 근감소 개선용 또는 운동능력 개선용 식품 조성물에 관한 것이다.The present invention relates to a food composition for improving muscle loss or improving exercise capacity containing a Paulownia flower extract, and more specifically, to an extraction method for efficiently extracting useful components from Paulownia flower fruit and the fruit of Paulownia flower extracted using the above extraction method. It relates to a food composition for improving muscle loss or improving exercise capacity containing an extract or a fraction thereof.
최근 전세계적으로 고령사회에 진입하여 65세 이상 인구의 비중이 높아지고 초고령사회를 목전에 둔 국내 상황에 따라, 고령인구의 근육 약화로 인한 무기력, 더 나아가 근감소증에 대한 경각심이 높아지고 있다. Recently, as the world has entered an aging society, the proportion of the population over 65 years of age has increased, and as Korea is on the verge of becoming a super-aged society, awareness of lethargy due to muscle weakness in the elderly population, and further sarcopenia, is increasing.
근감소 개선 관련 기술은 크게 '골격근의 구조 및 근육성장 과정', '근육 신생과정에서 Myogenic regulatory factors(MRFs)의 발현조절이 미치는 영향', '근육 신생과정에서 단백질 합성 경로의 조절이 미치는 영향', '근육 신생과정에서 단백질 분해 기전의 조절이 미치는 영향'으로 나뉘어 기초연구가 진행되고 있다.Technologies related to improving muscle loss are largely 'Skeleton Muscle Structure and Muscle Growth Process', 'Effect of Regulating the Expression of Myogenic Regulatory Factors (MRFs) in the Muscle Regeneration Process', and 'Effect of Regulation of the Protein Synthesis Pathway in the Muscle Regeneration Process'. , Basic research is being conducted under the category ‘Effect of regulation of protein degradation mechanism in the process of muscle growth.’
한편, 골격근(Skeletal muscle)은 성인 포유류에서 총 체질량의 약 40%를 구성하고, 일반적으로 방추형 형태로 양끝은 뼈에 부착되어 있으며, 근육분화는 Myo D, Myf-5, MRF4 등 각 분화단계에 영향을 미치는 전사인자인 Myogenic regulatory factor(MRFs)에 의해 조절되며, 또한 분화 후기에는 myotube의 주요 구조 단백질인 Myosin light chain(MYLC), Myosin heavy chain(MYHC)의 발현 수준이 증가하는 것으로 알려져 있다.Meanwhile, skeletal muscle makes up about 40% of the total body mass in adult mammals, and is generally spindle-shaped with both ends attached to bone. Muscle differentiation occurs at each differentiation stage, such as Myo D, Myf-5, and MRF4. It is regulated by Myogenic regulatory factors (MRFs), which are transcription factors that influence the expression of Myosin light chain (MYLC) and Myosin heavy chain (MYHC), the main structural proteins of myotube, in the late stages of differentiation.
이와 같이, 고령화 및 건강한 노화가 세계적인 이슈로 대두하고 있는 만큼 국내 기능성 식품 시장에서도 운동수행능력 향상과 관련된 건강기능식품에 대한 관심이 지속적으로 증가하고 있으며 식품 영양 부분에서의 개선 필요성이 강조되고 있는 추세이다.As aging and healthy aging are emerging as global issues, interest in health functional foods related to improving exercise performance is continuously increasing in the domestic functional food market, and the need for improvement in food nutrition is being emphasized. am.
본 특허는 중소벤처기업부(과제번호: S3092691) 발주의 지역특화산업육성사업으로 진행되었으며, 연구과제명은 근육 개선 효과를 가진 천연물 소재 및 식품개발이고, 연구기간은 2021.04.01.~2023.12.31.이다.This patent was carried out as a regional specialized industry development project ordered by the Ministry of SMEs and Startups (Project Number: S3092691). The research project title is Development of natural materials and foods with muscle-improving effects, and the research period is 2021.04.01. ~ 2023.12.31. .am.
본 발명은 꽃개오동으로부터 유용성분을 효과적으로 추출하는 추출 방법을 확립하고, 상기 방법에 따라 추출한 꽃개오동 추출물을 포함함으로써 근육감소 개선 효과 및 운동능력 향상 효과를 나타내는 건강기능식품 조성물을 제공하고자 한다.The present invention seeks to establish an extraction method for effectively extracting useful components from Paulownia perennials, and to provide a health functional food composition that exhibits the effect of improving muscle loss and improving exercise capacity by containing the Paulownia flower extract extracted according to the above method.
본 발명은 꽃개오동 추출물을 포함하는 근감소 개선용 식품 조성물을 제공한다.The present invention provides a food composition for improving muscle loss containing a Paulownia flower extract.
본 발명의 조성물에 있어서, 상기 조성물은, 근감소 개선을 통해 운동능력이 향상된 것일 수 있다.In the composition of the present invention, the composition may improve exercise ability by improving muscle loss.
본 발명의 조성물에 있어서, 상기 꽃개오동 추출물은, 에탄올을 추출용매로 추출한 것일 수 있다. In the composition of the present invention, the Paulownia sinus extract may be extracted with ethanol as an extraction solvent.
본 발명의 조성물에 있어서, 상기 추출은, 초음파 추출한 것일 수 있다. In the composition of the present invention, the extraction may be ultrasonic extraction.
본 발명의 조성물에 있어서, 상기 조성물은, 과립형일 수 있다. In the composition of the present invention, the composition may be in granular form.
본 발명은 꽃개오동으로부터 유용성분을 효율적으로 추출하는 추출방법을 제공하며, 추출된 꽃개오동 추출물을 유효성분으로 포함하여 근감소 개선 및 운동능력 개선에 우수한 효과를 나타내는 건강기능식품 조성물을 제공한다.The present invention provides an extraction method for efficiently extracting useful ingredients from Paulownia perennials, and provides a health functional food composition that contains the extracted Paulownia perilla extract as an active ingredient and exhibits excellent effects in improving muscle loss and exercise ability.
도 1은 꽃개오동 추출물의 항산화 효능 평가 결과를 나타낸 것이다.
도 2는 꽃개오동 추출물의 항염 평가 결과를 나타낸 것이다.
도 3은 꽃개오동 추출물의 추출공정에 따른 수율을 나타낸 그래프이다.
도 4는 꽃개오동 추출물의 추출공정에 따른 DPPH 라디칼 소거능을 확인한 결과를 나타낸 그래프이다.
도 5는 꽃개오동 추출물의 세포독성 평가 결과를 나타낸 것이다.
도 6은 꽃개오동 추출물의 근세포 증식 효능 평가 결과를 나타낸 것이다.
도 7은 근력 개선 관련 인자에 효능을 나타내는 MyoD, Myogenin, p-PI3K 및 p-Akt를 꽃개오동 70% 에탄올 추출물을 처리하여 분석한 결과를 나타낸 것이다.
도 8은 운동 수행 능력 관련 인자에 효능을 나타내는 Glut4와 p-AMPKα를 꽃개오동 70% 에탄올 추출물을 처리하여 분석한 결과를 나타낸 것이다.
도 9는 Pinoresinol과 vanillic acid의 근력개선 및 운동수행능력개선 관련인자 개선 효능을 확인한 결과이다.
도 10은 꽃개오동 70% 에탄올 추출물의 근육세포 분화 관련 인자 개선 효능을 확인한 결과이다.
도 11은 꽃개오동 70% 에탄올 추출물의 운동수행능력개선 관련 인자 개선 효능을 확인한 결과이다.
도 12는 꽃개오동 열매 추출물을 포함하는 과립 제조를 위한 공정도를 나타낸 것이다.
도 13은 제조예 1 내지 3의 안정성 시험 결과(DPPH Assay)를 나타낸 것이다.
도 14는 제조예 1 내지 3의 미생물 시험 결과를 나타낸 것이다.Figure 1 shows the results of evaluating the antioxidant efficacy of the Paulownia columbine extract.
Figure 2 shows the results of anti-inflammatory evaluation of Paulownia columbine extract.
Figure 3 is a graph showing the yield according to the extraction process of the Paulownia flower extract.
Figure 4 is a graph showing the results of confirming the DPPH radical scavenging ability according to the extraction process of the Paulownia flower extract.
Figure 5 shows the results of cytotoxicity evaluation of the Paulownia sinensis extract.
Figure 6 shows the results of evaluating the muscle cell proliferation efficacy of the Paulownia sinensis extract.
Figure 7 shows the results of analysis of MyoD, Myogenin, p-PI3K, and p-Akt, which show efficacy on factors related to muscle strength improvement, by treatment with 70% ethanol extract of Paulownia perilla.
Figure 8 shows the results of analysis of Glut4 and p-AMPKα, which show efficacy on factors related to exercise performance, by treatment with 70% ethanol extract of Paulownia chinensis.
Figure 9 shows the results of confirming the efficacy of pinoresinol and vanillic acid in improving factors related to muscle strength and exercise performance improvement.
Figure 10 shows the results confirming the efficacy of the 70% ethanol extract of Paulownia paulownia in improving factors related to muscle cell differentiation.
Figure 11 shows the results of confirming the efficacy of the 70% ethanol extract of Paulownia flower in improving factors related to improving exercise performance.
Figure 12 shows a process chart for manufacturing granules containing Paulownia paulownia fruit extract.
Figure 13 shows the stability test results (DPPH Assay) of Preparation Examples 1 to 3.
Figure 14 shows the microbiological test results of Preparation Examples 1 to 3.
최근 전세계적으로 고령사회에 진입하여 65세 이상 인구의 비중이 높아짐에 따라, 고령인구의 근육 약화로 인한 무기력, 더 나아가 근감소증에 대한 경각심이 높아지고 있어, 국내 기능성 식품 시장에서도 운동수행능력 향상과 관련된 건강기능식품에 대한 관심이 지속적으로 증가하고 있다.Recently, as the world has entered an aging society and the proportion of the population over 65 years of age has increased, awareness of lethargy due to muscle weakness in the elderly population, and further sarcopenia, has increased, and the domestic functional food market has also seen improvements in exercise performance and Interest in related health functional foods continues to increase.
이에, 본 발명에서는 꽃개오동 추출물을 포함하여 근감소 개선 및 운동능력 개선에 우수한 효과를 나타내는 식품 조성물을 제공하고자 한다.Accordingly, the present invention seeks to provide a food composition that shows excellent effects in improving muscle loss and exercise ability, including an extract of Paulownia perilla.
꽃개오동(Catalpa bignonioides)은 꿀풀목 능소화과의 나무로 북아메리카 원산의 10~30m 정도까지 자라는 낙엽수이다. 꽃개오동은 대체로 개오동과 비슷하나, 개오동보다 키가 크고 잎과 잎자루가 길며, 개오동의 꽃은 연노란색에 가까운 반면, 꽃개오동의 꽃은 하얀색으로 차이가 있다. 꽃개오동은 6~7월에 가지 끝의 원추꽃차례에 흰색 꽃이 피고 안쪽에 노란색 선이 두 줄 나 있으며, 자갈색 반점이 있고, 열매는 삭과로 10월에 열며 길이 20~45 cm 정도로 가늘고 아래로 늘어진다. 꽃개오동은 소화불량 개선, 통증 완화, 결막염 개선에 효과가 있다고 알려져 있다. Catalpa bignonioides is a deciduous tree native to North America that grows to about 10 to 30 m, belonging to the Lamiaceae family. Gaeo-dong is generally similar to Gae-o-dong, but it is taller than Gae-o-dong and has longer leaves and petioles. Its flowers are closer to light yellow, whereas its flowers are white. White flowers bloom in panicles at the ends of branches in June to July, with two yellow lines and purplish spots on the inside, and the fruit is a capsule that opens in October and is about 20 to 45 cm long, thin and downward. It stretches. It is known to be effective in improving indigestion, relieving pain, and improving conjunctivitis.
본 발명은 꽃개오동 추출물을 포함하는 근감소 개선용 식품 조성물을 제공한다. 한편, 상기 조성물은, 근감소 개선을 통해 운동능력이 향상된 것일 수 있다.The present invention provides a food composition for improving muscle loss containing a Paulownia flower extract. Meanwhile, the composition may improve exercise ability by improving muscle loss.
본 발명에 있어서, 상기 꽃개오동 추출물은, 바람직하게 정제수, 탄소수 1~4개의 무수 또는 함수 저급 알코올, 아세톤, 에틸아세테이트, 부틸아세테이트, 글리세린, 부틸렌글리콜, 프로필렌글리콜, 디클로로메탄, 헥산 및 그의 혼합물로부터 선택되는 어느 하나의 추출용매로 추출한 것일 수 있다. 더욱 바람직하게는 에탄올을 추출용매로 추출하는 것이 좋으며, 가장 바람직하게는 70% 에탄올을 추출용매로 추출하는 것이 좋다.In the present invention, the extract of Paulownia flower, Preferably, it is extracted with any one extraction solvent selected from purified water, anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms, acetone, ethyl acetate, butyl acetate, glycerin, butylene glycol, propylene glycol, dichloromethane, hexane, and mixtures thereof. You can. More preferably It is best to extract with ethanol as the extraction solvent, and most preferably with 70% ethanol as the extraction solvent.
본 발명에 있어서, 상기 추출은, 천연식물로부터 유용성분을 추출하기 위해 일반적으로 사용되는 열수추출법, 상온교반추출법, 환류냉각추출법, 초음파추출법, 고온가압추출법 및 저온고압추출법 등을 이용할 수 있는데, 바람직하게는 초음파 추출하는 것이 좋다. In the present invention, the extraction may be performed by using a hot water extraction method, a room temperature stirring extraction method, a reflux cooling extraction method, an ultrasonic extraction method, a high temperature pressure extraction method, and a low temperature high pressure extraction method, which are generally used to extract useful components from natural plants. Ultrasonic extraction is best.
또한, 본 발명에 있어서, 상기 조성물은, 꽃개오동 추출물 외에 상수리나무 추출물, 산돌배나무 추출물, 참당귀 추출물과 같은 천연물 추출물을 더 포함할 수 있으며, 바람직하게는 꽃개오동 추출물 140~160 중량부, 상수리나무 추출물 40~60 중량부, 산돌배나무 추출물 40~60 중량부, 참당귀 추출물 70~90 중량부를 포함하는 것이 좋다. 이때, 상기 조성물은 바람직하게 과립형인 것이 좋은데, 식품 제형을 과립 형태로 제조할 경우 과립 입자들은 다공질 구조가 되어 습윤성이 좋아지기 때문에 체내에서 분산 및 침강 속도가 증가하게 되어 제형에 함유되어 있는 기능성 유효성분들의 체내 흡수가 용이해지기 때문에 타깃으로 하는 생리활성 작용이 활발해지는 등의 장점이 있다. In addition, in the present invention, the composition may further include natural product extracts such as Quercus chinensis extract, Pear pear extract, and Angelica root extract in addition to Paulownia chinensis extract, preferably 140 to 160 parts by weight of Paulownia chinensis extract and Quercus aureus extract. It is recommended to include 40 to 60 parts by weight of tree extract, 40 to 60 parts by weight of Sandol pear extract, and 70 to 90 parts by weight of Angelica root extract. At this time, the composition is preferably in granular form. When the food formulation is manufactured in granular form, the granular particles have a porous structure and improve wettability, so the dispersion and sedimentation rate in the body increases, thereby reducing the functional effectiveness contained in the formulation. Since it is easily absorbed into the body, it has the advantage of increasing the targeted physiological activity.
한편, 본 발명에 있어서, 상기 조성물은, 꽃개오동, 참당귀 등의 원재료 및 부재료와 함께 과립 제조에 필요한 결합제, 부형제 등을 더 혼합하여 제조할 수 있다. 이때, 과립 제조시 사용되는 통상의 결합제, 부형제를 사용하여도 무방하나, 바람직하게는 sodium phosphate mono, sodium phosphate Di, Fungamyl 800L, Pectinex XXL, 탄산나트륨, 덱스트린을 더 혼합하여 제조하며, 더욱 바람직하게는 sodium phosphate mono 15~16 중량부, sodium phosphate Di 28~32 중량부, Fungamyl 800L 1~2 중량부, Pectinex XXL 1~2 중량부, 탄산나트륨 150~200 중량부, 덱스트린 420~168 중량부를 이용하는 것이 좋다. Meanwhile, in the present invention, the composition can be prepared by further mixing raw materials and auxiliary materials such as Paulownia paulownia and Angelica gigas, as well as binders and excipients necessary for producing granules. At this time, common binders and excipients used in granule production may be used, but it is preferably prepared by further mixing sodium phosphate mono, sodium phosphate Di, Fungamyl 800L, Pectinex XXL, sodium carbonate, and dextrin, and more preferably. It is recommended to use 15-16 parts by weight of sodium phosphate mono, 28-32 parts by weight of sodium phosphate Di, 1-2 parts by weight of Fungamyl 800L, 1-2 parts by weight of Pectinex XXL, 150-200 parts by weight of sodium carbonate, and 420-168 parts by weight of dextrin. .
한편, 본 발명에 있어서, 상기 조성물은, 꽃개오동 추출물을 분획한 분획물을 포함할 수도 있다.Meanwhile, in the present invention, the composition may include a fraction obtained by fractionating the Paulownia sinensis extract.
이하, 본 발명의 내용을 하기 실시예 및 실험예를 통해 더욱 상세히 설명하고자 한다. 다만, 본 발명의 권리범위가 하기 실시예 및 실험예에만 한정되는 것은 아니고, 그와 등가의 기술적 사상의 변형까지를 포함한다.Hereinafter, the contents of the present invention will be described in more detail through the following examples and experimental examples. However, the scope of the present invention is not limited to the following examples and experimental examples, and includes modifications of the technical idea equivalent thereto.
[실시예 1 : 꽃개오동 추출물 제조][Example 1: Preparation of Paulownia paulownia extract]
꽃개오동 열매 100 g에 10배의 70% 에탄올을 가한 후, 25±2℃의 실온에서 교반기(Hanbak Scientific Co., Bucheon, Korea)를 이용하여 150 rpm으로 24시간, 3회 반복 추출하여 실시예 1의 꽃개오동 추출물 시료를 제조하였다.After adding 10 times 70% ethanol to 100 g of Paulownia paulownia fruit, extraction was repeated three times for 24 hours at 150 rpm using a stirrer (Hanbak Scientific Co., Bucheon, Korea) at room temperature of 25 ± 2°C. A sample of the Paulownia flower extract of 1 was prepared.
[실험예 1 : 꽃개오동 추출물의 DPPH 라디칼 소거능 확인][Experimental Example 1: Confirmation of DPPH radical scavenging ability of Paulownia paulownia extract]
본 실험에서는 상기 실시예 1에서 제조한 꽃개오동 추출물의 DPPH 라디칼 소거능을 측정하여 항산화 효능을 평가하였다. In this experiment, the antioxidant efficacy of the Paulownia perilla extract prepared in Example 1 was measured by measuring the DPPH radical scavenging ability.
DPPH radical 소거능의 측정은 Blois의 방법을 변형하여 수행하였으며, 시료 0.4 mL에 0.4 mM DPPH(2,2-Diphenyl-1-picrylhydrazyl radical, Sigma-Aldrich Co.) 에탄올 용액 0.8 mL을 진탕 혼합하고, 10분간 방치 후 분광광도계를 사용하여 525 nm에서 흡광도를 측정하여 계산식, DPPH radical scavenging ability(%)=100-[(OD of sample/OD of control) ×100]에 의하여 활성을 산출하였다.Measurement of DPPH radical scavenging ability was performed by modifying Blois' method, mixing 0.8 mL of 0.4 mM DPPH (2,2-Diphenyl-1-picrylhydrazyl radical, Sigma-Aldrich Co.) ethanol solution with 0.4 mL of sample by shaking, and mixing for 10 minutes. After standing for a minute, the absorbance was measured at 525 nm using a spectrophotometer, and the activity was calculated using the formula: DPPH radical scavenging ability (%) = 100 - [(OD of sample/OD of control) × 100].
상기 표 1은 꽃개오동 추출물의 DPPH radical 소거능 측정 결과를 나타낸 것으로, 꽃개오동 추출물은 43.1%의 DPPH radical 소거능을 나타내는 점을 확인하였다.Table 1 above shows the results of measuring the DPPH radical scavenging ability of the Paulownia flower extract. It was confirmed that the Paulownia flower extract had a DPPH radical scavenging ability of 43.1%.
[실험예 2 : 꽃개오동 추출물의 총 폴리페놀 함량 분석][Experimental Example 2: Analysis of total polyphenol content of Paulownia paulownia extract]
본 실험에서는 상기 실시예 1에서 제조한 꽃개오동 추출물의 총 폴리페놀 함량을 분석하고자 하였다. 실험은 상기 실시예 1에서 제조한 꽃개오동 추출물을 0.1, 1, 10 mg/㎖ 농도로 제조하여 진행하였다.In this experiment, the total polyphenol content of the Paulownia sinensis extract prepared in Example 1 was analyzed. The experiment was conducted by preparing the Paulownia sinensis extract prepared in Example 1 at concentrations of 0.1, 1, and 10 mg/ml.
총 폴리페놀 함량은 Folin-Denis(20) 방법에 따라 실험하였다. 시료 1 mL에 0.2 N Folin-Ciocalteu’s phenol reagent(Sigma-Aldrich Co., St. Louis, MO, USA) 1 mL를 가하여 실온에서 3분간 반응시킨 후, 7.5% Na2CO3 1 mL를 가하여 암소에서 1시간 동안 방치한 다음 765 nm에서 흡광도를 측정하고, gallic acid (SigmaAldrich Co.)를 0~200 ppm 농도로 조제하여 작성한 검량선에 의하여 총 폴리페놀 함량을 산출하였다.Total polyphenol content was tested according to the Folin-Denis (20) method. To 1 mL of sample, 1 mL of 0.2 N Folin-Ciocalteu's phenol reagent (Sigma-Aldrich Co., St. Louis, MO, USA) was added and reacted at room temperature for 3 minutes, then 1 mL of 7.5% Na 2 CO 3 was added and incubated in the dark. After leaving for 1 hour, the absorbance was measured at 765 nm, and the total polyphenol content was calculated using a calibration curve prepared with gallic acid (SigmaAldrich Co.) at a concentration of 0 to 200 ppm.
상기 표 2는 꽃개오동 추출물의 총 폴리페놀 함량 분석 결과를 나타낸 것으로, 꽃개오동 추출물은 411.2 mg/g로 높은 폴리페놀 함량을 나타내는 점을 확인할 수 있었다.Table 2 above shows the results of analysis of the total polyphenol content of the Paulownia perilla extract, and it was confirmed that the extract of Paulownia perilla flower had a high polyphenol content of 411.2 mg/g.
[실험예 3 : 꽃개오동 추출물의 플라보노이드 함량 분석][Experimental Example 3: Analysis of flavonoid content of Paulownia paulownia extract]
본 실험에서는 상기 실시예 1에서 제조한 꽃개오동 추출물의 플라보노이드 함량을 분석하고자 하였다. 실험은 상기 실시예 1에서 제조한 꽃개오동 추출물을 0.1, 1, 10 mg/㎖ 농도로 제조하여 진행하였다.In this experiment, we attempted to analyze the flavonoid content of the Paulownia chinensis extract prepared in Example 1. The experiment was conducted by preparing the Paulownia sinensis extract prepared in Example 1 at concentrations of 0.1, 1, and 10 mg/ml.
총 플라보노이드 함량은 Moreno 등의 방법에 따라 추출물 0.5 mL에 10% aluminum nitrate 0.1 mL, 1 M potassium acetate 0.1 mL, ethanol 4.3 mL를 차례로 가하여 실온에서 40분간 반응한 후 415 nm에서 spectrophotometer (UV 1600 PC, Shimadzu, Tokyo, Japan)로 흡광도를 측정하였으며, 검량선은 Quercetin(Sigma Co., USA)를 표준물질로 하여 0 ~ 100 ug/mL의 농도 범위에서 얻어진 값으로 작성하여 추출물의 총 플라보노이드 함량을 계산하였다.The total flavonoid content was determined by sequentially adding 0.1 mL of 10% aluminum nitrate, 0.1 mL of 1 M potassium acetate, and 4.3 mL of ethanol to 0.5 mL of the extract according to the method of Moreno et al., reacting at room temperature for 40 minutes, and measuring it with a spectrophotometer at 415 nm (UV 1600 PC, Absorbance was measured using (Shimadzu, Tokyo, Japan), and a calibration curve was created using Quercetin (Sigma Co., USA) as a standard and the values obtained in the concentration range of 0 to 100 ug/mL to calculate the total flavonoid content of the extract. .
상기 표 3은 플라보노이드 함량 분석 결과를 나타낸 것으로, 꽃개오동 추출물은 43.7 mg/g으로 높은 플라보노이드 함량을 나타내는 점을 확인할 수 있었다.Table 3 shows the results of flavonoid content analysis, and it was confirmed that the Paulownia sinus extract had a high flavonoid content of 43.7 mg/g.
[실험예 4 : 꽃개오동 추출물의 항산화 효능 평가][Experimental Example 4: Evaluation of Antioxidant Efficacy of Paulownia Flower Extract]
본 실험에서는 상기 실시예 1에서 제조한 꽃개오동 추출물에 대하여 C2C12 cell을 이용한 항산화 효능 평가 실험을 진행하였다.In this experiment, an experiment was conducted to evaluate the antioxidant efficacy of the Paulownia sinensis extract prepared in Example 1 above using C 2 C 12 cells.
1) 세포 독성 평가1) Cytotoxicity evaluation
마우스 유래 C2C12 세포를 1 × 105 cells/㎖의 농도로 96well plate에 180 ㎕씩 분주 후 24 시간 배양하고, 시료 처리 후 24시간 배양하였다. 그런 다음, MTT 시약 처리 후 3시간 배양하고 상층액 제거 후 DMSO로 생성된 formazan을 용해하여 450 nm에서 흡광도를 측정하였다.Mouse-derived C 2 C 12 cells were distributed at a concentration of 1 × 10 5 cells/ml in 180 ㎕ of each 96-well plate and cultured for 24 hours. After sample treatment, the cells were cultured for 24 hours. Then, after treatment with MTT reagent, the cells were cultured for 3 hours, the supernatant was removed, the produced formazan was dissolved with DMSO, and the absorbance was measured at 450 nm.
2) 항산화 평가2) Antioxidant evaluation
마우스 유래 C2C12 세포를 1 × 105 cells/㎖의 농도로 96well plate에 180 ㎕씩 분주 후 24 시간 배양하고, 시료 처리 2시간 후 H2O2 10 ㎕를 처리(최종 0.5 mM)하고 24시간 배양 후 MTT assay를 진행하였다. MTT 시약 처리 후 3시간 배양하고 상층액 제거 후 DMSO로 생성된 formazan을 용해하여 450 nm에서 흡광도를 측정하였다.Mouse-derived C 2 C 12 cells were distributed at a concentration of 1 × 10 5 cells/ml in 180 ㎕ of each 96-well plate and cultured for 24 hours. After 2 hours of sample treatment, 10 ㎕ of H 2 O 2 was added (final concentration of 0.5 mM). After 24 hours of incubation, MTT assay was performed. After treatment with MTT reagent, the culture was incubated for 3 hours, the supernatant was removed, the formazan produced was dissolved with DMSO, and the absorbance was measured at 450 nm.
3) 실험 결과3) Experiment results
H2O2를 세포 내에 처리하게 되면 세포 내에 과도한 산화적 스트레스가 유발되고 결과적으로 세포 사멸이 이루어져 생존율이 감소하게 되는데, 실험 결과 H2O2 처리군에서는 H2O2 미처리군(시료만 처리) 대비 세포 생존율이 61.3%로 평가되었다. 세포 생존율이 낮아진다는 것은 항산화능이 낮아짐을 의미하기 때문에, H2O2와 시료를 함께 처리하였을 때의 세포 생존율을 항산화능으로 평가하였다. 도 1은 꽃개오동 추출물의 항산화 효능 평가 결과를 나타낸 것으로, 꽃개오동은 농도별(50, 100, 200 ㎍/ml)로 각각 71%, 82.4%, 94.8%의 세포 생존율을 보여 농도 의존적으로 항산화 효능을 나타내는 점을 확인하였다.When H 2 O 2 is treated within cells, excessive oxidative stress is induced within the cells, resulting in cell death and a decrease in survival rate. As a result of the experiment, the H 2 O 2 treated group was significantly different from the H 2 O 2 untreated group (sample only treated). ), the cell survival rate was evaluated at 61.3%. Since lower cell viability means lower antioxidant capacity, the cell viability when H 2 O 2 and the sample were treated together were evaluated in terms of antioxidant capacity. Figure 1 shows the results of evaluating the antioxidant efficacy of a Paulownia perilla extract, showing cell survival rates of 71%, 82.4%, and 94.8% at different concentrations (50, 100, and 200 μg/ml), respectively, showing a concentration-dependent antioxidant effect. It was confirmed that the point represents .
[실험예 5 : 꽃개오동 추출물의 미백 평가][Experimental Example 5: Whitening evaluation of Paulownia paulownia extract]
본 실험에서는 상기 실시예 1에서 제조한 꽃개오동 추출물의 tyrosinase 활성 억제능(%)을 확인하여 미백 평가 실험을 진행하였다.In this experiment, a whitening evaluation experiment was conducted by confirming the tyrosinase activity inhibition ability (%) of the Paulownia sinus extract prepared in Example 1.
tyrosinase는 tyrosine으로부터 melanin의 생합성에서 가장 중요한 촉매작용을 일으키는 효소로 이를 억제하는 효능을 평가함에 따라 미백 평가를 진행하였다. tyrosinase 억제효과는 dopachrome 방법을 이용하여 측정하였으며, Mushroom tyrosinase(2000 unit/ml)는 사용하기 직전에 용해하여 준비하며, L-tyrosine은 1.5 mM 농도로 제조하였다. 시료 30 ㎕에 L-tyrosine 60 ㎕, 0.1 M phosphate buffer (pH 6.5) 190 ㎕를 넣고, 준비된 tyrosinase 20 ㎕를 최종 첨가하여 37 ℃에서 15분간 반응시킨 다음, 490 nm에서 spectrophotometer (UV 1600 PC, Shimadzu, Tokyo, Japan)로 흡광도를 측정하였다.Tyrosinase is an enzyme that causes the most important catalytic action in the biosynthesis of melanin from tyrosine, and whitening evaluation was conducted to evaluate the efficacy of inhibiting it. The tyrosinase inhibitory effect was measured using the dopachrome method. Mushroom tyrosinase (2000 unit/ml) was prepared by dissolving immediately before use, and L-tyrosine was prepared at a concentration of 1.5 mM. Add 60 ㎕ of L-tyrosine and 190 ㎕ of 0.1 M phosphate buffer (pH 6.5) to 30 ㎕ of sample, add 20 ㎕ of prepared tyrosinase, react at 37°C for 15 minutes, and then measure at 490 nm using a spectrophotometer (UV 1600 PC, Shimadzu). , Tokyo, Japan), and the absorbance was measured.
상기 표 4는 꽃개오동 추출물의 tyrosinase 활성 억제능(%)을 확인한 결과를 나타낸 것으로, 꽃개오동 추출물은 55.0%를 나타내어 유의미한 억제능을 확인할 수 있었다.Table 4 above shows the results of confirming the tyrosinase activity inhibition ability (%) of the Paulownia flower extract. The Paulownia flower extract showed 55.0%, confirming significant inhibitory ability.
[실험예 6 : 꽃개오동 추출물의 항균 평가][Experimental Example 6: Antibacterial evaluation of Paulownia paulownia extract]
본 실험에서는 상기 실시예 1에서 제조한 꽃개오동 추출물의 항균 평가 실험을 진행하였다.In this experiment, an antibacterial evaluation experiment was conducted on the Paulownia chinensis extract prepared in Example 1 above.
실험은 E.coli, S.aureus, C.albican 및 P.aeruginosa 균주의 glycerol stock을 TSB agar에 streaking 후, 37℃에서 24h 배양하여 진행하였다. 5 ml tube에 멸균된 0.85% NaCl용액을 한 개 튜브에는 2 ml, 나머지 3개 튜브에는 1 ml 씩 넣은 후, 각각 자란 균의 colony를 10개 정도 NaCl용액에 풀어 접종한 후, vortexing 하고 630 nm에서 흡광도를 측정하여 균액의 농도를 정하였다. Muller-Hinton agar (90 mm plate에 17 ml씩 분주, 높이는 4 mm)를 90°씩 회전하며 8번 골고루 도말하며, 이때, 면봉에 물기는 최대한 제거 후 사용하였다. 멸균된 DISC 4개를 균일한 간격으로 놓고 DISC에 시료 20 ㎕씩 분주하고, 37℃에서 24h 배양한 후, 각 시료에 따른 균 성장 억제 영역의 지름을 측정하였다.The experiment was conducted by streaking glycerol stocks of E. coli , S. aureus , C. albican , and P. aeruginosa strains onto TSB agar and culturing them at 37°C for 24 h. Into a 5 ml tube, add 2 ml of sterilized 0.85% NaCl solution to one tube and 1 ml to the other three tubes, then inoculate about 10 colonies of each grown bacteria in the NaCl solution, vortex them, and inoculate them at 630 nm. The concentration of the bacterial solution was determined by measuring the absorbance. Muller-Hinton agar (17 ml each on a 90 mm plate, height 4 mm) was spread evenly eight times, rotating 90° each. At this time, the cotton swab was used after removing as much moisture as possible. Four sterilized DISCs were placed at even intervals, 20 ㎕ of sample was dispensed into each DISC, and cultured at 37°C for 24 h, and then the diameter of the bacterial growth inhibition area for each sample was measured.
상기 표 5는 꽃개오동 추출물의 항균 효능을 확인한 결과를 나타낸 것으로, 꽃개오동 추출물은 E.coli에 대한 항균 능력을 나타내는 점을 확인할 수 있었다.Table 5 above shows the results of confirming the antibacterial efficacy of the Paulownia flower extract, and it was confirmed that the Paulownia flower extract exhibits antibacterial ability against E. coli .
[실험예 7 : 꽃개오동 추출물의 항염 평가][Experimental Example 7: Anti-inflammatory evaluation of Paulownia sinensis extract]
본 실험에서는 상기 실시예 1에서 제조한 꽃개오동 추출물의 항염 평가 실험을 진행하였다.In this experiment, an anti-inflammatory evaluation experiment of the Paulownia sinensis extract prepared in Example 1 was conducted.
1) 세포 독성 평가1) Cytotoxicity evaluation
Raw 264.7 cell을 well 당 5 × 104 농도로 24 well plate에 seeding하고 24시간 배양 후 시료 처리 후 24시간 배양하였다. 그런 다음, MTT 시약 처리 후 3시간 배양하고 상층액 제거 후 DMSO로 생성된 formazan을 용해하여 450 nm에서 흡광도를 측정하였다.Raw 264.7 cells were seeded in a 24-well plate at a concentration of 5 × 10 4 per well, cultured for 24 hours, and then cultured for 24 hours after sample treatment. Then, after treatment with MTT reagent, the cells were cultured for 3 hours, the supernatant was removed, the produced formazan was dissolved with DMSO, and the absorbance was measured at 450 nm.
2) NO production assay2) NO production assay
Raw 264.7 cell을 well 당 5 × 104 농도로 24 well plate에 seeding하고 24시간 배양 후 시료와 LPS(1 ㎍/ml)를 처리 24시간 배양한 후 media 상등액을 150 ㎕를 취해 96 well plate로 옮긴 후, Griess A/B(1:1) reagent를 동량 분주 후 차광상태로 15분간 반응시킨 후 540 nm에서 흡광도를 측정하였다.Raw 264.7 cells were seeded in a 24-well plate at a concentration of 5 After that, an equal amount of Griess A/B (1:1) reagent was dispensed, reacted for 15 minutes under light blocking conditions, and the absorbance was measured at 540 nm.
3) 실험 결과3) Experiment results
도 2는 꽃개오동 추출물의 항염 평가 결과를 나타낸 것으로, 꽃개오동 추출물은 200 ㎍/ml에서 72%의 NO 생성 억제 효과를 나타내는 점을 확인할 수 있었다.Figure 2 shows the results of the anti-inflammatory evaluation of the Paulownia perilla extract, and it was confirmed that the extract of Paulownia perilla shows a 72% NO production inhibition effect at 200 ㎍/ml.
이와 같이, 상기 실험예 1 내지 7을 통해 꽃개오동 추출물은 DPPH 라디칼 소거능을 나타내고 총 폴리페놀 함량 및 플라보노이드 함량이 우수하면서, 항산화, tyrosinase 활성 억제, 항균, 항염에 모두 효과를 발휘하는 점을 확인할 수 있었다.In this way, through the above Experimental Examples 1 to 7, it can be confirmed that the Paulownia paulownia extract exhibits DPPH radical scavenging ability, has excellent total polyphenol content and flavonoid content, and is effective in antioxidant, tyrosinase activity inhibition, antibacterial, and anti-inflammatory effects. there was.
이에, 하기 실시예 2에서는 꽃개오동의 유용 성분을 효율적으로 추출하기 위해 추출방법을 달리하여 꽃개오동 추출물을 제조하였으며, 이들의 항산화 활성을 비교 분석하여 최적 추출공정을 확립하고자 하였다.Accordingly, in Example 2 below, extracts of Paulownia perilla were prepared using different extraction methods in order to efficiently extract the useful components of Paulownia perennials, and an attempt was made to establish an optimal extraction process by comparatively analyzing their antioxidant activities.
[실시예 2 : 추출 방법을 달리하여 꽃개오동 추출물 제조][Example 2: Preparation of Paulownia paulownia extract using different extraction methods]
본 실시예에서는 꽃개오동 열매를 각각 상온교반추출, 환류냉각추출, 초음파추출하여 추출방법을 달리한 꽃개오동 추출물을 제조하였다.In this example, paulownia paulownia extracts were prepared using different extraction methods by extracting paulownia flowers at room temperature, extracting them by stirring at room temperature, extracting them under reflux cooling, and extracting them with ultrasonic waves.
1) 상온교반추출(SE, stirrer extraction)1) Room temperature stirrer extraction (SE)
시료 100 g에 10배의 70% 에탄올을 가한 후, 25±2℃의 실온에서 교반기(Hanbak Scientific Co., Bucheon, Korea)를 이용하여 150 rpm으로 24시간, 3회 반복 추출하여 꽃개오동 추출물을 제조하였다.After adding 10 times 70% ethanol to 100 g of the sample, extraction was repeated three times for 24 hours at 150 rpm using a stirrer (Hanbak Scientific Co., Bucheon, Korea) at a room temperature of 25 ± 2°C to obtain a Paulownia flower extract. Manufactured.
2) 환류냉각추출(RE, reflux extraction)2) Reflux extraction (RE)
시료 100 g에 10배의 70% 에탄올을 가한 후, 냉각관을 부착한 60℃의 항온수조에서 3시간, 3회 반복 추출하여 꽃개오동 추출물을 제조하였다.After adding 10 times 70% ethanol to 100 g of the sample, extraction was repeated three times for 3 hours in a constant temperature water bath at 60°C equipped with a cooling tube to prepare a Paulownia paulownia extract.
3) 초음파추출(USE, ultrasonification extraction)3) Ultrasonic extraction (USE, ultrasonification extraction)
시료 100 g에 10배의 70% 에탄올을 가한 후 시료를 넣은 유리병을 초음파 추출기(AUG-R3-900, ASIA ULTRASONIC, Gyeonggi, Korea)를 이용하여 120 kHz의 조건으로 6시간, 3회 반복 추출하여 꽃개오동 추출물을 제조하였다.After adding 10 times 70% ethanol to 100 g of the sample, the glass bottle containing the sample was extracted repeatedly 3 times for 6 hours at 120 kHz using an ultrasonic extractor (AUG-R3-900, ASIA ULTRASONIC, Gyeonggi, Korea). Thus, an extract of Paulownia flower was prepared.
[실험예 8 : 추출방법을 달리하여 제조한 꽃개오동 추출물의 항산화 활성 확인][Experimental Example 8: Confirmation of antioxidant activity of paulownia flower extract prepared using different extraction methods]
본 실험에서는 상기 실시예 2에서 제조한 추출방법에 따른 꽃개오동 추출물의 최적 추출공정을 확립하고자, 수율, 총 폴리페놀 함량, 총 플라보노이드 함량, DPPH 라디칼 소거 활성을 측정하였다. In this experiment, the yield, total polyphenol content, total flavonoid content, and DPPH radical scavenging activity were measured to establish the optimal extraction process of the Paulownia sinensis extract according to the extraction method prepared in Example 2.
1) 추출공정에 따른 수율 확인1) Check yield according to extraction process
상기 실시예 2에서 추출방법을 달리하여 제조한 각각의 꽃개오동 추출물을 감압여과펌프(KNF Laboport Pressure Pump, ColeParmer, Vernon Hills, IL, USA)와 Whatman사(Piscataway, NJ, USA)의 20~25 μm 여과지를 이용하여 여과한 후, 여과한 추출물들을 감압회전농축기(Rotary Vacuum Evaporator N-N series, EYELA, Rikakikai Co., Tokyo, Japan)를 이용해 농축한 다음 동결건조기(PVTFA 10AT, ILSIN)를 이용하여 건조하였다. 이때, 각 추출물의 수율은 동결건조 후 건물 중량을 구한 다음 원료 건물량에 대한 백분율로 나타내었다. Each of the Paulownia paulownia extracts prepared by different extraction methods in Example 2 was filtered using a reduced pressure filtration pump (KNF Laboport Pressure Pump, ColeParmer, Vernon Hills, IL, USA) and a Whatman company (Piscataway, NJ, USA) at 20-25. After filtration using μm filter paper, the filtered extracts were concentrated using a reduced pressure rotary evaporator (Rotary Vacuum Evaporator N-N series, EYELA, Rikakikai Co., Tokyo, Japan) and then dried using a freeze dryer (PVTFA 10AT, ILSIN). did. At this time, the yield of each extract was calculated by calculating the dry matter weight after freeze-drying and then expressed as a percentage of the dry matter amount of the raw material.
도 3은 꽃개오동 추출물의 추출공정에 따른 수율을 나타낸 그래프로, 70% 에탄올을 용매로 사용하여 초음파추출공정(USE)으로 추출한 꽃개오동 추출물에서 18.8%로 가장 높은 수율을 나타내었으며, 상온교반추출공정(SE)으로 추출한 꽃개오동 추출물에서 7.6%로 가장 낮은 수율을 나타내는 점을 확인하였다.Figure 3 is a graph showing the yield according to the extraction process of the Flower of Paulownia extract. The highest yield of 18.8% was shown in the Flower of Paulownia extract extracted by ultrasonic extraction process (USE) using 70% ethanol as a solvent, and the highest yield of 18.8% was obtained by room temperature stirring extraction. It was confirmed that the Paulownia sinus extract extracted through the process (SE) showed the lowest yield at 7.6%.
이와 같이, 상온교반추출물 대비 초음파추출물의 수율이 높게 나타나는 것은 초음파 조사시 형성된 공동현상(cavitation)에 의한 높은 압력으로 세포 내부조직이 파괴되어 추출물의 이동거리가 짧아지고 확산이 용이하게 일어나기 때문으로 판단할 수 있었다.In this way, the reason why the yield of the ultrasonic extract is higher compared to the extract stirred at room temperature is believed to be because the internal tissue of the cells is destroyed by the high pressure caused by cavitation formed during ultrasonic irradiation, which shortens the travel distance of the extract and facilitates diffusion. Could.
또한, 고분자 등 원자 간의 결합이 끊어지게 되어 기존의 상온교반추출 및 환류냉각추출 방법으로 용출이 어려웠던 성분의 용출 가능성 및 초음파 에너지에 의한 조직의 파괴에 기인되는 것으로 판단되며, 초음파추출공정을 통한 추출물에 더욱 많은 유효성분이 함유되어 추출물의 생리활성에도 영향을 끼칠 것으로 판단된다.In addition, it is believed that this is due to the possibility of elution of components that were difficult to elute using the existing room temperature stirring extraction and reflux cooling extraction methods due to the breakage of bonds between atoms such as polymers, and the destruction of tissue by ultrasonic energy, and the extract through the ultrasonic extraction process It is believed that the more active ingredients it contains, the more it will affect the physiological activity of the extract.
2) 총 폴리페놀 및 총 플라보노이드 함량 확인2) Check total polyphenol and total flavonoid content
총 폴리페놀 함량은 Folin-Denis(20) 방법에 따라 실험하였다. 시료 1 mL에 0.2 N Folin-Ciocalteu’s phenol reagent(Sigma-Aldrich Co., St. Louis, MO, USA) 1 mL를 가하여 실온에서 3분간 반응시킨 후, 7.5% Na 2CO 3 1 mL를 가하여 암소에서 1시간 동안 방치한 다음 765 nm에서 흡광도를 측정하였으며, 총 폴리페놀 함량은 gallic acid(SigmaAldrich Co.)를 0~200 ppm 농도로 조제하여 작성한 검량선에 의하여 산출하였다.Total polyphenol content was tested according to the Folin-Denis (20) method. To 1 mL of sample, 1 mL of 0.2 N Folin-Ciocalteu's phenol reagent (Sigma-Aldrich Co., St. Louis, MO, USA) was added and reacted at room temperature for 3 minutes. Then, 1 mL of 7.5% Na 2 CO 3 was added and incubated in the dark for 1 mL. After standing for a while, the absorbance was measured at 765 nm, and the total polyphenol content was calculated using a calibration curve prepared by adjusting gallic acid (SigmaAldrich Co.) at a concentration of 0 to 200 ppm.
총 플라보노이드 함량은 Moreno 등의 방법에 따라 추출물 0.5 mL에 10% aluminum nitrate 0.1 mL, 1 M potassium acetate 0.1 mL, ethanol 4.3 mL를 차례로 가하여 실온에서 40분간 반응한 후 415 nm에서 spectrophotometer (UV 1600 PC, Shimadzu, Tokyo, Japan)로 흡광도를 측정하였다. 검량선은 Quercetin(Sigma Co., USA)를 표준물질로 하여 0 ~ 100 ug/mL의 농도 범위에서 얻어진 값으로 작성하여 추출물의 총 플라보노이드 함량을 계산하였다.The total flavonoid content was determined by sequentially adding 0.1 mL of 10% aluminum nitrate, 0.1 mL of 1 M potassium acetate, and 4.3 mL of ethanol to 0.5 mL of the extract according to the method of Moreno et al., reacting at room temperature for 40 minutes, and measuring it with a spectrophotometer at 415 nm (UV 1600 PC, Absorbance was measured using Shimadzu, Tokyo, Japan). A calibration curve was created using Quercetin (Sigma Co., USA) as a standard and values obtained in the concentration range of 0 to 100 ug/mL to calculate the total flavonoid content of the extract.
하기 표 6은 추출방법을 달리하여 제조한 꽃개오동 추출물의 추출공정별 총 폴리페놀 및 플라보노이드 함량을 분석한 결과를 나타낸 것이다.Table 6 below shows the results of analyzing the total polyphenol and flavonoid content by extraction process of Paulownia sinensis extracts prepared using different extraction methods.
표 6에서 확인할 수 있듯이, 총 폴리페놀 함량은 USE, RE, SE의 순으로 각각 702.58±0.79 GAE mg/g, 522.05±1.94 GAE mg/g, 401.32±9.79 GAE mg/g으로 나타나 USE(초음파추출공정)으로 추출한 추출물에서 가장 높았으며, 플라보노이드 함량도 USE, RE, SE가 각각 98.98±1.83 mg/g, 78.71±0.71 mg/g, 65.17±2.89 mg/g을 나타내 총 폴리페놀 함량과 유사한 경향을 나타냈다.As can be seen in Table 6, the total polyphenol content was 702.58±0.79 GAE mg/g, 522.05±1.94 GAE mg/g, and 401.32±9.79 GAE mg/g in the order of USE, RE, and SE, respectively, indicating USE (ultrasonic extraction process), and the flavonoid content was 98.98±1.83 mg/g, 78.71±0.71 mg/g, and 65.17±2.89 mg/g for USE, RE, and SE, respectively, showing a similar trend to the total polyphenol content. showed.
3) DPPH 라디칼 소거 활성 측정3) Measurement of DPPH radical scavenging activity
DPPH radical 소거능의 측정은 Blois의 방법을 변형하여 수행하였으며, 시료 0.4 mL에 0.4 mM DPPH(2,2-Diphenyl-1-picrylhydrazyl radical, Sigma-Aldrich Co.) 에탄올 용액 0.8 mL을 진탕 혼합하고, 10분간 방치 후 분광광도계를 사용하여 525 nm에서 흡광도를 측정하여 계산식(DPPH radical scavenging ability(%)=100-[(OD of sample/OD of control)×100])에 의하여 활성을 산출하였다.Measurement of DPPH radical scavenging ability was performed by modifying Blois' method, mixing 0.8 mL of 0.4 mM DPPH (2,2-Diphenyl-1-picrylhydrazyl radical, Sigma-Aldrich Co.) ethanol solution with 0.4 mL of sample by shaking, and mixing for 10 minutes. After standing for a minute, the absorbance was measured at 525 nm using a spectrophotometer, and the activity was calculated using the formula (DPPH radical scavenging ability (%) = 100 - [(OD of sample/OD of control) x 100]).
도 4는 꽃개오동 추출물의 추출공정에 따른 DPPH 라디칼 소거능을 확인한 결과를 나타낸 그래프로, 초음파추출공정(USE)을 이용하여 추출시 10, 50, 100, 500, 1,000 ㎍/ml의 농도에서 각각 14.20, 21.24, 29.61, 65.21, 88.91%로 가장 높게 나타나 상온교반추출(SE) 및 환류냉각추출(RE) 공정을 이용한 경우에 비해 우수하였다.Figure 4 is a graph showing the results of confirming the DPPH radical scavenging ability according to the extraction process of the Paulownia flower extract. When extracted using an ultrasonic extraction process (USE), the concentration of 10, 50, 100, 500, and 1,000 ㎍/ml was 14.20, respectively. , 21.24, 29.61, 65.21, and 88.91%, which were the highest, and were superior to those using room temperature stirred extraction (SE) and reflux extraction (RE) processes.
이와 같이, 본 실험에서 초음파추출공정(USE)을 이용한 꽃개오동 추출물이 상온교반추출(SE) 또는 환류냉각추출(RE) 공정으로 추출한 추출물 대비 많은 생리활성 성분이 용출되며 추출 수율이 우수하고, 총 폴리페놀 및 플라보노이드에서도 높은 함량 증가를 나타냈으며, DPPH 라디칼 소거능 역시 가장 우수함을 확인할 수 있었다.As such, in this experiment, the Paulownia chinensis extract using the ultrasonic extraction process (USE) eluted more bioactive ingredients and had an excellent extraction yield compared to extracts extracted using the room temperature stirred extraction (SE) or reflux extraction (RE) process. A high increase in polyphenol and flavonoid content was also observed, and the DPPH radical scavenging ability was also confirmed to be the best.
이에, 하기 실험예 9 내지 11에서는 추출 수율, 총 폴리페놀 및 플라보노이드 함량이 우수하게 나타난 초음파추출공정을 이용하여 추출한 꽃개오동 추출물의 근세포 증식 효능, 운동수행능력 및 근력개선 효능을 확인하고자 하였다.Accordingly, in Experimental Examples 9 to 11 below, we attempted to confirm the muscle cell proliferation efficacy, exercise performance, and muscle strength improvement efficacy of the Paulownia sinensis extract extracted using an ultrasonic extraction process, which showed excellent extraction yield, total polyphenol, and flavonoid content.
[실험예 9 : 꽃개오동 추출물의 근세포 증식 확인][Experimental Example 9: Confirmation of muscle cell proliferation of Paulownia sinensis extract]
본 실험에서는 꽃개오동 추출물의 근세포 증식 효능을 확인하기 위해 에탄올 농도(0, 30, 50, 70, 100 %)에 따른 세포 독성 평가 및 세포 증식 효능 평가를 진행하였다.In this experiment, cytotoxicity and cell proliferation efficacy were evaluated according to ethanol concentration (0, 30, 50, 70, and 100%) to confirm the muscle cell proliferation efficacy of the Paulownia paulownia extract.
1) 세포 독성 평가1) Cytotoxicity evaluation
마우스 유래 C2C12 myoblasts를 1 × 105 cells/ml의 농도로 well-plate에 분주 후 24시간 동안 배양하고 시료 처리 후 24시간 배양한 다음, MTT 시약 처리 후 3시간 배양하고 상층액 제거 후 DMSO로 생성된 formazan을 용해하여 450 nm에서 흡광도를 측정하였다.Mouse-derived C 2 C 12 myoblasts were dispensed into well-plates at a concentration of 1 The formazan produced was dissolved in DMSO and the absorbance was measured at 450 nm.
2) 세포 증식 효능 평가2) Evaluation of cell proliferation efficacy
마우스 유래 myoblasts인 C2C12 세포를 5 × 104 cells/ml의 농도로 24 well-plate에 분주하고, myocytes로 분화시키기 위하여 48시간 간격으로 2% Horse serum과 1% Antibiotic-Antimycotic이 함유된 DMEM 배지로 교환하며 동시에 시료를 같이 처리하였다. 분화 유도 6일 후, 증식한 세포내 DNA strands에 결합한 BrdU를 검출해내는 5-bromo-2’-deoxyuridine (BrdU) assay kit를 활용하여 세포 증식을 측정하였다.C 2 C 12 cells, which are mouse-derived myoblasts, were distributed in 24 well-plates at a concentration of 5 The medium was replaced with DMEM and the samples were processed at the same time. Six days after differentiation induction, cell proliferation was measured using the 5-bromo-2'-deoxyuridine (BrdU) assay kit, which detects BrdU bound to DNA strands within proliferated cells.
3) 실험 결과3) Experiment results
도 5는 꽃개오동 추출물의 세포독성 평가 결과를 나타낸 것으로, 에탄올 농도 0%, 30%, 50%, 70% 및 100% 모두 최고 농도인 100 ㎍/ml에서 세포독성이 나타나지 않는 점을 확인하였다. 도 6은 꽃개오동 추출물의 근세포 증식 효능 평가 결과를 나타낸 것으로, 고농도(25 ㎍/ml)에서는 꽃개오동 100% 에탄올 추출물에 비해 50% 및 70% 에탄올 추출물이 비슷한 근세포 증식활성을 보였고, 저농도(6.25 ㎍/ml)에서는 꽃개오동 100% 에탄올 추출물의 경우 근세포 증식 효능이 나타나지 않았으나, 꽃개오동 50% 에탄올 추출물 및 꽃개오동 70% 에탄올 추출물에서는 근세포 증식이 각각 23%, 30% 증가하였다.Figure 5 shows the cytotoxicity evaluation results of the Paulownia sinensis extract, and it was confirmed that no cytotoxicity was observed at ethanol concentrations of 0%, 30%, 50%, 70%, and 100%, the highest concentration of 100 μg/ml. Figure 6 shows the results of evaluating the muscle cell proliferation efficacy of the Paulownia flower extract. At high concentration (25 ㎍/ml), the 50% and 70% ethanol extracts showed similar myocyte proliferation activity compared to the 100% ethanol extract of Paulownia flower, and at low concentration (6.25 ㎍/ml). ㎍/ml), the 100% ethanol extract of Paulownia flower did not show any effect on muscle cell proliferation, but the proliferation of muscle cells increased by 23% and 30% in the 50% ethanol extract of Paulownia flower and 70% ethanol extract of Paulownia flower, respectively.
이에, 하기 실험예 10 및 11에서는 근세포 증식효능이 가장 우수하게 나타난 꽃개오동 70% 에탄올 추출물을 이용하여 꽃개오동 추출물의 운동수행능력 및 근력개선 효능을 각각 in vitro, in vivo 상에서 평가하였다.Accordingly, in Experimental Examples 10 and 11 below, the 70% ethanol extract of Paulownia paulownia, which showed the best muscle cell proliferation effect, was used to evaluate the exercise performance and muscle strength improvement effects of the Paulownia paulownia extract in vitro and in vivo, respectively.
[실험예 10 : 꽃개오동 추출물의 in vitro 세포 모델에서 운동수행능력 및 근력개선 효능 확인][Experimental Example 10: Confirmation of exercise performance and muscle strength improvement efficacy of Paulownia sinensis extract in in vitro cell model]
본 실험에서는 꽃개오동 추출물의 in vitro 세포 모델에서의 운동수행능력 및 근력개선 효능을 평가하고자 하였다.In this experiment, we aimed to evaluate the efficacy of Paulownia columbine extract in improving exercise performance and muscle strength in an in vitro cell model.
1) 근육 세포 분화 관련 인자 변화 평가1) Evaluation of changes in factors related to muscle cell differentiation
마우스 유래 myoblasts인 C2C12 세포를 5 × 104 cells/ml의 농도로 24 well-plate에 분주하고 myocytes로 분화시키기 위하여 48시간 간격으로 2% Horse serum과 1% Antibiotic-Antimycotic이 함유된 DMEM 배지로 교환하며 동시에 시료를 같이 처리하였다. 분화 유도 6일 후, 세포 내 단백질을 용해시킨 후 Western blot 분석법으로 근육분화를 유도하는 MyoD, Myogenin, p-mTOR 등의 항체와 근육분화를 억제하는 Smad 계열의 항체를 사용하여 각 단백질의 변화를 확인하였다.C 2 C 12 cells, which are mouse-derived myoblasts, were distributed in 24 well-plates at a concentration of 5 The medium was exchanged and the samples were processed at the same time. Six days after differentiation induction, intracellular proteins were dissolved and changes in each protein were measured using Western blot analysis using antibodies such as MyoD, Myogenin, and p-mTOR, which induce muscle differentiation, and Smad series antibodies, which inhibit muscle differentiation. Confirmed.
2) 운동 수행 능력 관련 인자 변화 평가2) Evaluation of changes in factors related to exercise performance
마우스 유래 myoblasts인 C2C12 세포를 5 × 104 cells/ml의 농도로 24 well-plate에 분주하고 myocytes로 분화시키기 위하여 48시간 간격으로 2% Horse serum과 1% Antibiotic-Antimycotic이 함유된 DMEM 배지로 교환하며 동시에 시료를 같이 처리하였다. 분화 유도 6일 후, 세포 내 단백질을 용해시킨 후 Western blot 분석법으로 운동 수행 능력 향상에 도움을 주는 단백질인 Glut4, p-AMPKα, p-SirT1, PGC-1α 등의 항체를 사용하여 각 단백질의 변화를 확인하였다.C 2 C 12 cells, which are mouse-derived myoblasts, were distributed in 24 well-plates at a concentration of 5 The medium was exchanged and the samples were processed at the same time. After 6 days of inducing differentiation, intracellular proteins were dissolved and changes in each protein were analyzed using Western blot analysis using antibodies against Glut4, p-AMPKα, p-SirT1, and PGC-1α, which are proteins that help improve exercise performance. was confirmed.
3) 실험 결과3) Experiment results
도 7은 근력 개선 관련 인자에 효능을 나타내는 MyoD, Myogenin, p-PI3K 및 p-Akt를 꽃개오동 70% 에탄올 추출물을 처리하여 분석한 결과를 나타낸 것으로, 꽃개오동 70% 에탄올 추출물 25 ㎍/ml 농도에서 근육 분화를 유도하는 MyoD, Myogenin, p-PI3K 및 p-AKT의 발현이 증가하는 것을 확인하였다. 도 8은 운동 수행 능력 관련 인자에 효능을 나타내는 Glut4와 p-AMPKα를 꽃개오동 70% 에탄올 추출물을 처리하여 분석한 결과를 나타낸 것으로, 꽃개오동 70% 에탄올 추출물 처리시 운동 수행 능력에 도움을 주는 단백질인 Glut4와 p-AMPKα의 발현이 증가한 점을 확인하였다. 또한, 꽃개오동의 유효성분인 Pinoresinol과 vanilic acid의 운동수행능력 및 근증식 작용기전을 확인한 결과, 근육 분화를 유도하는 단백질인 MyoD, Myogenin, p-Akt 및 p-mTOR를 증가시키고, 운동수행능력을 증가시켜주는 단백질인 Glut4과 p-AMPkα를 증가시키는 것을 확인하였다(도 9).Figure 7 shows the results of analysis of MyoD, Myogenin, p-PI3K, and p-Akt, which show efficacy on factors related to muscle strength improvement, by treatment with 70% ethanol extract of Paulownia chinensis, at a concentration of 25 ㎍/ml. It was confirmed that the expression of MyoD, Myogenin, p-PI3K, and p-AKT, which induce muscle differentiation, increased. Figure 8 shows the results of analysis of Glut4 and p-AMPKα, which show efficacy on factors related to exercise performance, by treatment with 70% ethanol extract of Paulownia chinensis. Proteins that help exercise performance when treated with 70% ethanol extract of Paulownia chinensis. It was confirmed that the expression of Glut4 and p-AMPKα increased. In addition, as a result of confirming the exercise performance and muscle proliferation mechanism of Pinoresinol and vanilic acid, which are active ingredients of Phalaenopsis perilla, they increased MyoD, Myogenin, p-Akt and p-mTOR, proteins that induce muscle differentiation, and exercise performance. It was confirmed that Glut4 and p-AMPkα, proteins that increase , were increased (Figure 9).
[실험예 11 : 꽃개오동 추출물의 in vivo 동물 모델에서 운동수행능력 및 근력개선 효능 확인][Experimental Example 11: Confirmation of the efficacy of Paulownia sinensis extract in improving exercise performance and muscle strength in an in vivo animal model]
본 실험에서는 꽃개오동 추출물의 in vivo 동물 모델에서의 운동수행능력 및 근력개선 효능을 평가하고자 하였다.In this experiment, we aimed to evaluate the efficacy of Paulownia columbine extract in improving exercise performance and muscle strength in an in vivo animal model.
1) 실험 동물 사육 조건 및 투여1) Experimental animal rearing conditions and administration
6주령의 ICR 마우스를 구입하여 1주간 순화 후 사용하였으며, 23±2℃의 온도와 50%의 습도를 유지하고, 12시간 간격으로 조명의 on/off를 조절하여 낮과 밤의 환경을 유지시켰다. 사료나 식수는 자유롭게 섭취할 수 있도록 하며 식이 섭취량 및 체중 증가량은 주 당 1회 일정한 시간에 측정하였고, 시료는 10주간 50 mg/kg과 200 mg/kg을 경구 투여하였다.Six-week-old ICR mice were purchased and used after acclimatization for one week. The temperature and humidity were maintained at 23±2°C and humidity at 50%, and the day and night environment was maintained by turning lights on/off at 12-hour intervals. . Feed and water were freely available, and dietary intake and weight gain were measured once a week at regular times, and samples were orally administered 50 mg/kg and 200 mg/kg for 10 weeks.
2) 꽃개오동 추출물의 운동수행능력 개선 및 근력 개선 평가2) Evaluation of exercise performance and muscle strength improvement of Paulownia sinensis extract
3일간 실험동물을 트레드밀에 적응시킨 후 운동수행능력을 평가하였다. 탈진 시까지 달리기 시험은 일정 시간 간격을 두고 속도를 증가시키고, 더 이상 트레드밀 속도를 따라가지 못할 때까지의 트레드밀 속도, 거리와 시간을 측정하였으며, 악력테스트는 트레드밀 적응 기간 중 악력테스트기(Chatillon digital force gauge, Chatillon)를 사용하여 평가하였다.After adapting the experimental animals to the treadmill for 3 days, their exercise performance was evaluated. In the running test until exhaustion, the speed was increased at regular intervals and the treadmill speed, distance, and time were measured until the treadmill speed could no longer be followed. The grip test was performed using a grip tester (Chatillon digital force) during the treadmill adaptation period. It was evaluated using gauge, Chatillon).
3) 실험 결과3) Experiment results
도 10은 꽃개오동 70% 에탄올 추출물과 양성대조군(creatine)을 10주간 매일 투여 후 몸무게 변화, 간 무게, 혈청 내 ALT/AST 변화를 측정한 결과를 나타낸 것으로, 도 10에서 보듯이 꽃개오동 추출물과 양성대조군 간 유의적인 차이가 나타나지 않아 꽃개오동 추출물은 독성이 없음을 확인하였다. 한편, 도 11은 꽃개오동 70% 에탄올 추출물의 운동수행능력개선 및 근력개선 효능을 확인한 결과를 나타낸 것으로, 꽃개오동 70% 에탄올 추출물과 양성대조군(creatine)을 10주간 매일 투여 후 탈진 시까지 걸리는 시간을 측정한 결과, control군은 2,272초인 반면 양성대조군은 2,691초, 꽃개오동 추출물 저농도군(50 mg/kg)은 2,636초, 꽃개오동 추출물 고농도군(200 mg/kg)은 2,630초로 측정되었다. 즉, 꽃개오동 추출물 저농도 및 고농도 군에서 탈진 시까지 걸리는 시간이 약 16% 증가하여 18% 증가한 양성대조군과 거의 유사한 것을 알 수 있다. 또한, 탈진 시까지의 거리는 control군은 624 m인 반면, 양성대조군은 821 m, 꽃개오동 추출물 저농도군은 793 m, 꽃개오동 추출물 고농도군은 788 m로 측정되어 꽃개오동 추출물 저농도 및 고농도군은 양성대조군과 유사하였으며, 탈진 시까지의 트레드밀 속도는 control군은 26.1 m/min인 반면, 양성대조군은 29.4 m/min, 꽃개오동 추출물 저농도군은 29.1 m/min, 꽃개오동 추출물 고농도군은 29.0 m/min으로 측정되어, 꽃개오동 추출물 저농도 및 고농도군에서 탈진 시까지의 트레드밀 속도가 약 11% 증가하여 약 13% 증가한 양성대조군과 유사한 점을 확인할 수 있다(도 11). 한편, 악력테스트 결과 control군은 0.424 lbf인 반면, 양성대조군은 0.538 lbf, 꽃개오동 추출물 저농도군은 0.537 lbf, 꽃개오동 추출물 고농도군은 0.529 lbf로 측정되어 꽃개오동 추출물 저농도 및 고농도군과 양성대조군이 거의 유사하였다(도 11).Figure 10 shows the results of measuring body weight change, liver weight, and ALT/AST change in serum after daily administration of 70% ethanol extract of Paulownia paulownia and positive control group (creatine) for 10 weeks. As shown in Figure 10, the extract of Paulownia flower and the change in ALT/AST in serum are shown. There was no significant difference between the positive control groups, confirming that the Paulownia sinensis extract was not toxic. Meanwhile, Figure 11 shows the results of confirming the exercise performance improvement and muscle strength improvement effect of the 70% ethanol extract of Flowering Paulownia and the time taken to exhaustion after daily administration of the 70% ethanol extract of Flowering Paulownia and positive control group (creatine) for 10 weeks. As a result of measuring, the control group was measured at 2,272 seconds, while the positive control group was measured at 2,691 seconds, the Paulownia perilla extract low concentration group (50 mg/kg) was measured at 2,636 seconds, and the Paulownia perilla extract high concentration group (200 mg/kg) was measured at 2,630 seconds. In other words, it can be seen that the time until exhaustion increased by about 16% in the low- and high-concentration groups of Paulownia paulownia extract, which was almost similar to the positive control group, which increased by 18%. In addition, the distance to exhaustion was measured at 624 m for the control group, while the positive control group was 821 m, the low-concentration Paulownia perilla extract group was 793 m, and the high-concentration Paulownia perilla extract group was measured at 788 m. The low- and high-concentration groups of Paulownia perilla sinus extract were measured at 788 m, respectively. The treadmill speed until exhaustion was 26.1 m/min for the control group, 29.4 m/min for the positive control group, 29.1 m/min for the low-concentration Paulownia perilla extract group, and 29.0 m/min for the high-concentration Paulownia perilla extract group. As measured, it can be seen that the treadmill speed until exhaustion increased by about 11% in the low and high concentration groups of Paulownia sinensis extract, which is similar to the positive control group, which increased by about 13% (FIG. 11). Meanwhile, as a result of the grip test, the control group was measured at 0.424 lbf, while the positive control group was measured at 0.538 lbf, the low-concentration Paulownia perilla extract group was measured at 0.537 lbf, and the high-concentration Paulownia sinus extract group was measured at 0.529 lbf. It was almost similar (Figure 11).
[제조예 1 내지 3 : 꽃개오동 추출물을 포함하는 과립 제조][Preparation Examples 1 to 3: Preparation of granules containing Paulownia paulownia extract]
본 제조예에서는 근감소 개선에 우수한 효능을 가지는 꽃개오동, 참당귀 등의 원재료 및 부재료와 함께 과립 제조에 필요한 결합제, 부형제 등을 혼합하여 과립 제형의 식품 제조예 1 내지 3을 제조하였다. In this production example, granule-type food production examples 1 to 3 were prepared by mixing raw materials and auxiliary materials such as Paulownia paulownia and Angelica gigas, which have excellent efficacy in improving muscle loss, with binders and excipients necessary for granule production.
아래 표 7은 꽃개오동 추출물(70% 에탄올, 초음파 추출)을 포함하는 제조예 1 내지 3 과립의 배합비를 나타낸 것으로, 도 12의 공정도에 따라 제조예 1 내지 3의 과립을 제조하였다.Table 7 below shows the mixing ratio of the granules of Preparation Examples 1 to 3 containing Paulownia paulownia extract (70% ethanol, ultrasonic extraction), and the granules of Preparation Examples 1 to 3 were manufactured according to the process diagram of FIG. 12.
한편, 표 7에 따라 제조한 제조예 1 내지 3의 성상 확인 결과, 제조예 1의 경우 과립 형성이 균일하게 잘 이루어졌으나, 제조예 2 및 3은 제조예 1 대비 흡습성이 높아 색이 짙어지고 뭉치는 현상이 발생하였다. 또한, 제조예 1 내지 3을 10 mg/ml의 농도로 DPPH 소거능을 통해 1개월 동안 가혹 조건(40℃, 70%)에서 보관한 결과, 제조예 1 내지 3 과립 모두 항산화 효능이 안정적으로 유지되는 점을 확인하였으며(도 13), 미생물 시험 결과 총 세균수가 미검출되어 안전함을 확인하였다(도 14). Meanwhile, as a result of checking the properties of Preparation Examples 1 to 3 prepared according to Table 7, in Preparation Example 1, granules were formed uniformly, but Preparation Examples 2 and 3 had higher hygroscopicity compared to Preparation Example 1, so the color became darker and lumps formed. A phenomenon occurred. In addition, as a result of storing Preparation Examples 1 to 3 at a concentration of 10 mg/ml under harsh conditions (40°C, 70%) for 1 month through DPPH scavenging ability, the antioxidant efficacy of all granules of Preparation Examples 1 to 3 was stably maintained. This point was confirmed (FIG. 13), and as a result of the microbiological test, the total number of bacteria was not detected, confirming that it was safe (FIG. 14).
Claims (5)
A food composition for improving muscle loss containing a paulownia flower extract.
상기 조성물은,
근감소 개선을 통해 운동능력이 향상된 것을 특징으로 하는 식품 조성물.
According to paragraph 1,
The composition is,
A food composition characterized by improved exercise capacity through improved muscle loss.
상기 꽃개오동 추출물은,
에탄올을 추출용매로 추출한 것을 특징으로 하는 식품 조성물.
According to paragraph 1,
The extract of Paulownia flower is,
A food composition characterized in that it is extracted with ethanol as an extraction solvent.
상기 추출은,
초음파 추출한 것을 특징으로 하는 식품 조성물.
According to paragraph 3,
The extraction is,
A food composition characterized by ultrasonic extraction.
상기 조성물은,
과립형인 것을 특징으로 하는 식품 조성물.
According to any one of claims 1 to 3,
The composition is,
A food composition characterized in that it is granular.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101723022B1 (en) | 2015-12-21 | 2017-04-06 | 한국과학기술연구원 | Composition for preventing or alleviating hangover comprising extracts of catalpa ovata g. don |
| KR102119421B1 (en) | 2017-04-25 | 2020-06-05 | 재단법인 경기도경제과학진흥원 | Composition for Anti-Allergy Using an Extract of Catalpa ovata |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101723022B1 (en) | 2015-12-21 | 2017-04-06 | 한국과학기술연구원 | Composition for preventing or alleviating hangover comprising extracts of catalpa ovata g. don |
| KR102119421B1 (en) | 2017-04-25 | 2020-06-05 | 재단법인 경기도경제과학진흥원 | Composition for Anti-Allergy Using an Extract of Catalpa ovata |
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