KR102129595B1 - Method for producing beverage for relieving menopausal symptoms using complex fermented Pleuropterus multflorus extract and medicinal herbs extract - Google Patents
Method for producing beverage for relieving menopausal symptoms using complex fermented Pleuropterus multflorus extract and medicinal herbs extract Download PDFInfo
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- KR102129595B1 KR102129595B1 KR1020190016652A KR20190016652A KR102129595B1 KR 102129595 B1 KR102129595 B1 KR 102129595B1 KR 1020190016652 A KR1020190016652 A KR 1020190016652A KR 20190016652 A KR20190016652 A KR 20190016652A KR 102129595 B1 KR102129595 B1 KR 102129595B1
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- A—HUMAN NECESSITIES
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- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
본 발명은 (1) 하수오에 글루코스(glucose) 및 수크로스(sucrose)를 첨가한 후 멸균한 하수오 혼합물에 표고균사체(Lentinula edodes)를 접종한 후 배양한 하수오 발효물을 건조 및 분쇄하여 하수오 발효 분말을 제조하는 단계; (2) 상기 (1)단계의 제조한 하수오 발효 분말에 물을 첨가한 후 추출 및 여과하고 농축하여 하수오 발효 농축액을 제조하는 단계; (3) 상기 (2)단계의 제조한 하수오 발효 농축액에 물, 에탄올, 아세토박터 아세티(Acetobacter aceti) 및 아세토박터 파스테리아누스(Acetobacter pasteurianus)를 첨가한 후 초산 발효하여 하수오 초산 발효물을 제조하는 단계; (4) 홍화자, 속단, 당귀 및 오미자에 각각 물을 첨가한 후 추출 및 여과하여 홍화자 추출물, 속단 추출물, 당귀 추출물 및 오미자 추출물을 제조하는 단계: 및 (5) 상기 (3)단계의 제조한 하수오 초산 발효물과 상기 (4)단계의 제조한 속단 추출물, 홍화자 추출물, 당귀 추출물 및 오미자 추출물을 혼합한 혼합물을 가열하는 단계를 포함하여 제조하는 것을 특징으로 하는 하수오 혼합 음료의 제조방법 및 상기 방법으로 제조된 하수오 혼합 음료에 관한 것이다.The present invention (1) added to glucose (glucose) and sucrose (sucrose) in sewage, and sterilized mycelia ( Lentinula ) in the sterilized sewage mixture edode s) after inoculation, drying and grinding the cultured sewage fermentation product to prepare a sewage fermentation powder; (2) adding water to the sewage fermentation powder prepared in step (1), followed by extraction, filtration, and concentration to prepare a sewage fermentation concentrate; (3) Water, ethanol, acetobacter aceti ( acetobacter ) in the sewage fermentation concentrate prepared in step (2) aceti ) and acetobacter pasteurianus , followed by acetic acid fermentation to produce sewage acetic acid fermentation product; (4) Safflower, Sokdan, Angelica and Omija, respectively, after adding water and extracting and filtering to prepare Safflower extract, Sokdan extract, Angelica extract and Omija extract: and (5) sewage prepared in the step (3) Method for producing a sewage mixed beverage, characterized in that it comprises a step of heating a mixture of acetic acid fermentation product and the soybean extract, safflower extract, Angelica extract and Omija extract prepared in step (4) and the method It relates to a sewage mixed beverage prepared.
Description
본 발명은 복합 발효에 의한 하수오 초산 발효물과 속단 추출물, 홍화자 추출물, 당귀 추출물 및 오미자 추출물을 혼합한 혼합물을 가열하는 단계를 포함하여 제조하는 것을 특징으로 하는 하수오 혼합 음료의 제조방법 및 상기 방법으로 제조된 하수오 혼합 음료에 관한 것이다.The present invention is a method for producing a sewage mixed beverage, characterized in that it comprises a step of heating a mixture of a fermented sewage acetic acid by complex fermentation and a rapid extract, safflower extract, Angelica extract and Omija extract, and the above method. It relates to a sewage mixed beverage prepared.
내분비계 호르몬의 하나인 에스트로겐은 체내에서 콜레스테롤로부터 합성되며, 분비원은 주로 난소의 여포 및 황체이다. 뇌하수체전엽에서 분비되는 성선자극호르몬(gonadotropin)이 생식기관에 신호를 전달하면 월경주기를 조절하고 임신을 가능케 하는 호르몬인 여포자극호르몬(follicle stimulating hormone; FSH)과 황체형성호르몬(Lutenizing hormone; LH)이 분비되고, 이는 다시 난소에 작용하여 에스트로겐과 프로게스테론을 분비하게 한다. 상기 에스트로겐은 에스트라디올, 에스트론, 에스트리올 등을 총칭한다.Estrogen, one of the hormones of the endocrine system, is synthesized from cholesterol in the body, and the secretion source is mainly the follicle and luteum of the ovaries. When gonadotropin secreted from the anterior pituitary gland transmits signals to the reproductive organs, follicle stimulating hormone (FSH) and luteinizing hormone (LH) are hormones that regulate the menstrual cycle and enable pregnancy. It is secreted, which in turn acts on the ovaries to secrete estrogen and progesterone. The estrogen generically refers to estradiol, estrone, estriol, and the like.
폐경 전의 여성에 있어 난소는 에스트로겐의 주요 공급원이다. 여성이 폐경기에 접어들면, 난소가 노화됨에 따라 하수체성 성선자극 호르몬(여포자극 호르몬 및 황체형성 호르몬)에 대한 반응이 감소하고, 이로 인해 초기 여포기가 더욱 단축되어 월경 주기가 더욱 단축되며, 배란기가 더욱 단축되고, 프로게스테론 생성이 감소되며, 사이클이 더욱 불규칙하게 된다. 결국, 여포가 반응하지 못하게 되어 에스트로겐을 생성하지 못하게 된다. 에스트로겐의 분비 감소는 여성의 비뇨생식계통 뿐만 아니라 신체 전반에 걸쳐 큰 영향을 미치게 되며, 배란이 중단되어 월경이 사라지는 폐경 또는 그 이전에 호르몬의 감소로 인한 폐경기 증상이 나타나게 된다.In premenopausal women, the ovaries are the main source of estrogen. When a woman enters menopause, the response to the pituitary gonadotropin (follicle stimulating hormone and luteinizing hormone) decreases as the ovaries age, which further shortens the initial follicle cycle, shortens the menstrual cycle and ovulates. Is shorter, progesterone production is reduced, and the cycle becomes more irregular. Eventually, the follicle becomes unable to react and estrogen is not produced. Estrogen secretion decreases not only in the genitourinary system of women, but also throughout the body, and menopause in which ovulation ceases and menstruation disappears, or menopausal symptoms appear due to a decrease in hormones before.
폐경기 이후 여성에서는 에스트로겐의 급속한 감소로 인하여, 심리학적 및 감성적 징후, 예컨대 피로, 흥분, 불면, 집중력 저하, 우울증, 기억 상실, 두통, 근심 및 신경과민이나 소심증 등이 발생할 우려가 현저히 높아지게 되며, 재발성 안면 홍조에 의한 수면 방해에 따른 피로와 흥분, 간헐성 현기증, 감각이상, 심계항진 및 빈맥, 메스꺼움, 변비, 설사, 관절통, 근육통, 수족 냉증 및 체중 증가 현상의 발생 우려 또한 현저히 높아지게 된다. 또한, 골다공증이 쉽게 유발될 뿐 아니라, 심장병, 고혈압, 뇌졸중 등의 심혈관계 질환으로 인한 사망률도 증가한다. 또한, 피부 및 비뇨생식계통의 변화를 초래하며, 자가면역성 질환, 백내장 및 대장암 등의 발병 가능성이 높아지게 된다.In postmenopausal women, the rapid decrease in estrogen causes a significant increase in the risk of psychological and emotional signs, such as fatigue, excitement, insomnia, decreased concentration, depression, memory loss, headache, anxiety and neurosensitivity or timidity. Fatigue and excitement, intermittent dizziness, sensory abnormalities, cardiac hyperactivity and tachycardia, nausea, constipation, diarrhea, joint pain, muscle soreness, coldness of the feet and weight gain are also significantly increased. In addition, not only is osteoporosis easily induced, but mortality from cardiovascular diseases such as heart disease, hypertension, and stroke also increases. In addition, it causes changes in the skin and genitourinary system, and the likelihood of developing autoimmune diseases, cataracts and colon cancer is increased.
현재, 상기 갱년기 증상을 개선하는데 있어서 가장 효과적인 공지의 방법은 에스트로겐을 투여하여 결핍된 에스트로겐을 보충해 주는 것으로 알려져 있다.Currently, the most effective and known method for improving the menopausal symptoms is known to supplement estrogen by administering estrogen.
그러나, 합성 에스트로겐의 장기간에 걸친 투여는 유방암과 자궁암을 유발할 수 있다는 심각한 문제점이 있다. 최근 들어서는 장기간에 걸친 합성 에스트로겐 대체 요법이 유방암을 유발할 가능성이 높다는 믿을 만한 주장이 다수 제기되면서 학자들 간에는 물론, 사회 전반에 걸쳐 논란이 가열되고 있다. 아직 확실히 증명된 것은 아니지만, 합성 에스트로겐 대체 요법을 받은 여성에 있어서의 유방암 발생 위험은 시간 경과에 따른 에스트로겐 노출 용량과 관련이 있는 것으로 믿어지고 있다.However, there is a serious problem that long-term administration of synthetic estrogen can cause breast cancer and uterine cancer. In recent years, controversy has heated up among scholars as well as across society, with a number of credible claims that long-term synthetic estrogen replacement therapy is more likely to cause breast cancer. Although not yet clearly demonstrated, it is believed that the risk of developing breast cancer in women receiving synthetic estrogen replacement therapy is related to the estrogen exposure dose over time.
따라서, 상기 호르몬 대체 요법에 사용되는 합성 에스트로겐의 보다 안전한 대체물로서 식물성 에스트로겐인 피토에스트로겐을 찾고자 하는 연구가 최근 활발히 진행되고 있는 추세에 있다. 상기 피토에스트로겐은 동물의 체내에서 에스트로겐성 효과를 발휘하는 식물 중에 존재하는 비스테로이드성 화합물을 지칭하며, 항암 작용과 심장병에 효과가 있는 것으로 알려져 있는 곡물, 과일, 채소들의 대부분은 피토에스트로겐을 미량 포함하고 있다.Therefore, research to find phytoestrogens, which are phytoestrogens, as safer alternatives to synthetic estrogens used in the hormone replacement therapy has recently been actively conducted. The phytoestrogen refers to a non-steroidal compound present in a plant that exerts an estrogen effect in the body of an animal, and most of the grains, fruits, and vegetables known to have anti-cancer activity and heart disease include phytoestrogens in trace amounts Doing.
즉, 여성의 갱년기 증상을 개선하는데 효과적이면서도 유방암, 자궁암 등을 유발하지 않는 등 안전한 약품 또는 건강기능식품에 대한 필요성이 꾸준히 제기되어 오고 있다.In other words, the need for safe drugs or health functional foods, such as effective in improving menopausal symptoms in women, but not causing breast cancer and uterine cancer, has been steadily raised.
한국공개특허 제2017-0080946호에는 여성 갱년기 증상 개선 효능이 있는 한방 복합 추출물 홍삼 음료의 제조방법이 개시되어 있으며, 한국공개특허 제2013-0083692호에는 대두배아 발효물을 포함하는 갱년기 증상 개선용 음료 조성물의 제조방법이 개시되어 있으나, 본 발명의 복합 발효 하수오 추출물과 한약재 추출물을 이용한 갱년기 증상 개선용 음료의 제조방법과는 상이하다.Korean Patent Publication No. 2017-0080946 discloses a method of manufacturing a herbal complex extract red ginseng beverage having the effect of improving menopausal symptoms, and Korean Patent Publication No. 2013-0083692 discloses a drink for improving menopausal symptoms, including fermented soybean germ. Although a method for preparing the composition is disclosed, it is different from a method for preparing a beverage for improving menopausal symptoms using the complex fermentation sewage extract and the herbal medicine extract of the present invention.
본 발명은 상기와 같은 요구에 의해 안출된 것으로서, 본 발명의 목적은 여성 갱년기 증상을 개선하기 위한 약재 선정, 약재 전처리 및 배합비 등의 제조조건을 최적화하여, 음용이 용이하면서도 기호도를 높이고 더불어 갱년기 증상 개선의 건강 증진 효과를 기대할 수 있는 새로운 형태의 건강 기능성 음료의 제조방법을 제공하는 데 있다.The present invention was devised by the above-mentioned demands, and the object of the present invention is to optimize the manufacturing conditions such as selection of medicines to improve the symptoms of women's menopause, pretreatment of the medicines, and mixing ratio, so as to facilitate drinking, increase preference, and improve menopausal symptoms. It is to provide a method of manufacturing a health functional beverage of a new type that can be expected to improve health.
상기 과제를 해결하기 위해, 본 발명은 (1) 하수오에 글루코스(glucose) 및 수크로스(sucrose)를 첨가한 후 멸균한 하수오 혼합물에 표고균사체(Lentinula edodes)를 접종한 후 배양한 하수오 발효물을 건조 및 분쇄하여 하수오 발효 분말을 제조하는 단계; (2) 상기 (1)단계의 제조한 하수오 발효 분말에 물을 첨가한 후 추출 및 여과하고 농축하여 하수오 발효 농축액을 제조하는 단계; (3) 상기 (2)단계의 제조한 하수오 발효 농축액에 물, 에탄올, 아세토박터 아세티(Acetobacter aceti) 및 아세토박터 파스테리아누스(Acetobacter pasteurianus)를 첨가한 후 초산 발효하여 하수오 초산 발효물을 제조하는 단계; (4) 홍화자, 속단, 당귀 및 오미자에 각각 물을 첨가한 후 추출 및 여과하여 홍화자 추출물, 속단 추출물, 당귀 추출물 및 오미자 추출물을 제조하는 단계: 및 (5) 상기 (3)단계의 제조한 하수오 초산 발효물과 상기 (4)단계의 제조한 속단 추출물, 홍화자 추출물, 당귀 추출물 및 오미자 추출물을 혼합한 혼합물을 가열하는 단계를 포함하여 제조하는 것을 특징으로 하는 하수오 혼합 음료의 제조방법을 제공한다.In order to solve the above problems, the present invention (1) after adding glucose (glucose) and sucrose (sucrose) to sewage, sterilized sewage mixture inoculated with shiitake mycelium ( Lentinula edode s) and cultured sewage fermentation Drying and grinding to prepare a sewage fermentation powder; (2) adding water to the sewage fermentation powder prepared in step (1), followed by extraction, filtration, and concentration to prepare a sewage fermentation concentrate; (3) the above (2) water in the manufacture of concentrated liquid sewage fermentation step, ethanol, acetonitrile bakteo Oh Shetty (Acetobacter aceti) and acetonitrile bakteo Pas terrier Taunus (Acetobacter pasteurianus ), followed by acetic acid fermentation to produce sewage acetic acid fermentation product; (4) Safflower, Sokdan, Angelica and Omija, respectively, after adding water and extracting and filtering to prepare Safflower extract, Sokdan extract, Angelica extract and Omija extract: and (5) sewage prepared in the step (3) It provides a method for producing a sewage mixed beverage, characterized in that it comprises a step of heating a mixture of acetic acid fermentation product and the soybean extract, safflower extract, Angelica extract and Omija extract prepared in step (4).
또한, 본 발명은 상기 방법으로 제조된 하수오 혼합 음료를 제공한다.In addition, the present invention provides a sewage mixed beverage prepared by the above method.
본 발명의 여성 갱년기 증상을 개선시킬 수 있는 천연물 소재 중 최적의 건강 지향적 기능성 소재를 발굴하여, 재료 특유의 풋내는 제거하면서 유효성분이 증진될 수 있도록 재료 전처리, 추출 및 배합 등을 최적화함으로써, 여성 갱년기 증상을 효과적으로 개선시킬 수 있으며, 재료 특유의 이취가 없어 음용이 용이한 이점이 있다.By discovering the optimal health-oriented functional material among natural materials that can improve the symptoms of women's menopause of the present invention, by removing the peculiar to the ingredients, the material pre-treatment, extraction, and formulation are optimized to improve the active ingredients, so that women climacteric Symptoms can be effectively improved, and there is an advantage in that it is easy to drink because there is no characteristic odor.
도 1은 본 발명의 하수오 발효 분말의 제조공정을 도식화한 것이다,
도 2는 본 발명의 하수오 발효 농축액의 제조공정을 도식화한 것이다.
도 3은 본 발명의 하수오 초산 발효물의 제조공정을 도식화한 것이다.
도 4는 홍화자, 속단, 당귀 및 오미자 추출물의 제조공정을 도식화한 것이다.
도 5는 각 처리구별 실험 랫트(rat)의 주당 체중 변화를 비교한 그래프이다.
도 6은 실험 종료 후 각 처리구별 실험 랫트(rat)의 체중 증가율(%)을 비교한 그래프이다.
도 7은 각 처리구별 실험 랫트(rat)의 주당 식이량을 비교한 그래프이다.
도 8은 실험 종료 후 각 처리구별 실험 랫트(rat)의 총 식이량을 비교한 그래프이다.
도 9는 실험 종료 후 각 처리구별 실험 랫트(rat)의 식이 효율을 비교한 그래프이다.
도 10은 실험 종료 후 각 처리구별 실험 랫트(rat)의 간 조직 무게를 비교한 그래프이다.
도 11은 실험 종료 후 각 처리구별 실험 랫트(rat)의 비장 조직 무게를 비교한 그래프이다.
도 12는 실험 종료 후 각 처리구별 실험 랫트(rat)의 신장 조직 무게를 비교한 그래프이다.
도 13은 실험 종료 후 각 처리구별 실험 랫트(rat)의 자궁 및 난소 조직 무게를 비교한 그래프이다.
도 14는 실험 종료 후 각 처리구별 실험 랫트(rat)의 자궁 형태를 비교한 사진이다.
도 15는 실험 종료 후 각 처리구별 실험 랫트(rat)의 자궁관 상부 단면을 현미경으로 관찰한 사진이다.
도 16은 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 총 단백질 농도를 비교한 그래프이다.
도 17은 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 알부민 농도를 비교한 그래프이다.
도 18은 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 칼슘 농도를 비교한 그래프이다.
도 19는 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 APL(alkaline phosphatase) 농도를 비교한 그래프이다.
도 20은 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 에스트로겐 농도를 비교한 그래프이다.
도 21은 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 에스트라디올 농도를 비교한 그래프이다.
도 22는 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 테스토스테론 농도를 비교한 그래프이다.
도 23은 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 글루코스 농도를 비교한 그래프이다.
도 24는 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 AST 농도를 비교한 그래프이다.
도 25는 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 ALT 농도를 비교한 그래프이다.
도 26은 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 총 콜레스테롤 농도를 비교한 그래프이다.
도 27은 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 중성지방 농도를 비교한 그래프이다.
도 28은 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 고밀도지단백 콜레스테롤 농도를 비교한 그래프이다.
도 29는 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 저밀도지단백 콜레스테롤 농도를 비교한 그래프이다.
도 30은 실험 종료 후 각 처리구별 실험 랫트(rat)의 혈청 내 초저밀도지단백 콜레스테롤 농도를 비교한 그래프이다.
도 5 내지 30의 일반대조군: 난소제거 여성갱년기 모델 무처리, OVX모델+estradiol: 난소제거 여성갱년기 모델+에스트라디올 E2 섭취(양성대조군), OVX모델+saline: 난소제거 여성갱년기 모델+생리식염수 섭취(음성대조군), OVX모델+하수오생약초복합물: 난소제거 여성갱년기 모델+시료 음료 섭취(실험군) 처리구를 의미한다.1 is a schematic view showing the manufacturing process of the fermentation powder of sewage of the present invention,
Figure 2 is a schematic view showing the manufacturing process of the sewage fermentation concentrate of the present invention.
Figure 3 is a schematic view showing the manufacturing process of the fermented sewage of the present invention.
Figure 4 is a schematic drawing of the production process of safflower, Sokdan, Angelica and Omija extract.
5 is a graph comparing the weight change per week of the experimental rats for each treatment group.
Figure 6 is a graph comparing the weight gain (%) of the experimental rats (rat) for each treatment group after the end of the experiment.
7 is a graph comparing the amount of diet per week of experimental rats for each treatment group.
Figure 8 is a graph comparing the total diet of the experimental rats (rat) for each treatment after the end of the experiment.
9 is a graph comparing the dietary efficiency of each experimental rat after each experiment.
10 is a graph comparing the liver tissue weight of each experimental rat after each experiment.
11 is a graph comparing the spleen tissue weight of each experimental rat after each experiment.
12 is a graph comparing the renal tissue weight of each experimental rat after each experiment.
13 is a graph comparing the weight of uterine and ovarian tissues in each experimental rat after treatment.
Figure 14 is a photograph comparing the uterus shape of the experimental rat (rat) for each treatment group after the end of the experiment.
Figure 15 is a photograph of the upper section of the uterine tube of the experimental rat (rat) for each treatment group after the end of the experiment observed with a microscope.
16 is a graph comparing the total protein concentration in serum of the experimental rats for each treatment group after the end of the experiment.
17 is a graph comparing albumin concentrations in serum of experimental rats for each treatment group after the end of the experiment.
18 is a graph comparing the concentration of calcium in the serum of the experimental rats for each treatment group after the end of the experiment.
FIG. 19 is a graph comparing the concentration of alkaline phosphatase (APL) in serum of experimental rats for each treatment group after the end of the experiment.
20 is a graph comparing the concentration of estrogen in the serum of the experimental rats for each treatment group after the end of the experiment.
21 is a graph comparing the concentration of estradiol in the serum of the experimental rats for each treatment group after the end of the experiment.
22 is a graph comparing testosterone concentrations in serum of experimental rats for each treatment group after the end of the experiment.
23 is a graph comparing the concentration of glucose in the serum of each experimental rat after each experiment.
24 is a graph comparing the concentration of AST in the serum of experimental rats for each treatment group after the end of the experiment.
25 is a graph comparing the concentration of ALT in serum of the experimental rats for each treatment group after the end of the experiment.
26 is a graph comparing total cholesterol concentrations in serum of experimental rats for each treatment group after the end of the experiment.
27 is a graph comparing triglyceride concentrations in serum of experimental rats for each treatment group after the end of the experiment.
28 is a graph comparing the concentration of high-density lipoprotein cholesterol in the serum of experimental rats for each treatment group after the end of the experiment.
29 is a graph comparing the low-density lipoprotein cholesterol concentration in the serum of the experimental rat for each treatment group after the end of the experiment.
30 is a graph comparing the concentration of ultra-low density lipoprotein cholesterol in the serum of experimental rats for each treatment group after the end of the experiment.
General control of Figs. 5 to 30: ovarian removal climacteric model no treatment, OVX model +estradiol: ovarian removal female climacteric model + estradiol E2 intake (positive control), OVX model + saline: ovarian removal female climacteric model + physiological saline intake (Eumseong Control), OVX model + Sewage Herbal Herb Complex: Ovariectomy female climacteric model + sample drink intake (experimental group).
본 발명의 목적을 달성하기 위하여, 본 발명은In order to achieve the object of the present invention, the present invention
(1) 하수오에 글루코스(glucose) 및 수크로스(sucrose)를 첨가한 후 멸균한 하수오 혼합물에 표고균사체(Lentinula edodes)를 접종한 후 배양한 하수오 발효물을 건조 및 분쇄하여 하수오 발효 분말을 제조하는 단계;(1) Glucose (glucose) and sucrose (sucrose) were added to sewage, followed by inoculation of shiitake mycelium ( Lentinula edode s) into a sterilized sewage mixture, followed by drying and grinding of the cultured sewage fermentation to prepare sewage fermentation powder. To do;
(2) 상기 (1)단계의 제조한 하수오 발효 분말에 물을 첨가한 후 추출 및 여과하고 농축하여 하수오 발효 농축액을 제조하는 단계;(2) adding water to the sewage fermentation powder prepared in step (1), followed by extraction, filtration, and concentration to prepare a sewage fermentation concentrate;
(3) 상기 (2)단계의 제조한 하수오 발효 농축액에 물, 에탄올, 아세토박터 아세티(Acetobacter aceti) 및 아세토박터 파스테리아누스(Acetobacter pasteurianus)를 첨가한 후 초산 발효하여 하수오 초산 발효물을 제조하는 단계;(3) Water, ethanol, acetobacter aceti ( acetobacter ) in the sewage fermentation concentrate prepared in step (2) aceti ) and acetobacter pasteurianus , followed by acetic acid fermentation to produce sewage acetic acid fermentation product;
(4) 홍화자, 속단, 당귀 및 오미자에 각각 물을 첨가한 후 추출 및 여과하여 홍화자 추출물, 속단 추출물, 당귀 추출물 및 오미자 추출물을 제조하는 단계: 및(4) Safflower, Sokdan, Angelica and Omija, respectively, after adding water and extracting and filtering to prepare Safflower extract, Sokdan extract, Angelica extract and Omija extract: and
(5) 상기 (3)단계의 제조한 하수오 초산 발효물과 상기 (4)단계의 제조한 속단 추출물, 홍화자 추출물, 당귀 추출물 및 오미자 추출물을 혼합한 혼합물을 가열하는 단계를 포함하여 제조하는 것을 특징으로 하는 하수오 혼합 음료의 제조방법을 제공한다.(5) characterized in that it comprises a step of heating the mixture of the fermented sewage acetic acid prepared in step (3) and the Sokdan extract, safflower extract, Angelica extract and Omija extract prepared in step (4). It provides a method for producing a mixed drink of sewage.
본 발명의 하수오 혼합 음료의 제조방법에서, 상기 (1)단계의 하수오 발효 분말은 바람직하게는 하수오 450~550 g에 하수오 중량 대비 글루코스(glucose) 0.8~1.2% 및 수크로스(sucrose) 0.8~1.2%를 첨가한 후 110~130℃에서 10~20분 동안 멸균한 하수오 혼합물에 표고균사체(Lentinula edodes) 8~12 mL를 접종한 후 22~28℃에서 25~35일 동안 배양한 하수오 발효물을 75~85℃에서 10~14시간 동안 건조 및 분쇄하여 제조할 수 있으며, 더욱 바람직하게는 하수오 500 g에 하수오 중량 대비 글루코스(glucose) 1% 및 수크로스(sucrose) 1%를 첨가한 후 121℃에서 15분 동안 멸균한 하수오 혼합물에 표고균사체(Lentinula edodes) 10 mL를 접종한 후 25℃에서 30일 동안 배양한 하수오 발효물을 80℃에서 12시간 동안 건조 및 분쇄하여 제조할 수 있다.In the method for producing a mixed drink of sewage of the present invention, the fermentation powder of sewage in step (1) is preferably from 450 to 550 g of sewage, 0.8 to 1.2% of glucose to gross weight, and 0.8 to 1.2 of sucrose. Shiitake mycelium ( Lentinula) in a sewage mixture sterilized for 10-20 minutes at 110~130℃ after adding% edode s) can be prepared by inoculating 8-12 mL and drying and grinding the fermented sewage fermented at 22-28 °C for 25-35 days for 10-14 hours at 75-85°C, more preferably After adding 1% of glucose and 1% of sucrose to 500 g of sewage, shiitake mycelium ( Lentinula) in a sewage mixture sterilized at 121°C for 15 minutes. edode s) After inoculating 10 mL, the fermented sewage cultured at 25° C. for 30 days can be prepared by drying and grinding at 80° C. for 12 hours.
또한, 본 발명의 하수오 혼합 음료의 제조방법에서, 상기 (2)단계의 하수오 발효 농축액은 바람직하게는 하수오 발효 분말에 물을 18~22배(v/w) 첨가한 후 60~100℃에서 1~15시간 동안 추출 및 여과하고 28~32 brix가 될 때까지 농축하여 제조할 수 있으며, 더욱 바람직하게는 하수오 발효 분말에 물을 20배(v/w) 첨가한 후 60~100℃에서 1~15시간 동안 추출 및 여과하고 30 brix가 될 때까지 농축하여 제조할 수 있다.In addition, in the method for producing a mixed drink of sewage of the present invention, the sewage fermentation concentrate of step (2) is preferably added at 18 to 22 times (v/w) of water to the sewage fermentation powder at 60 to 100°
또한, 본 발명의 하수오 혼합 음료의 제조방법에서, 상기 (3)단계의 하수오 초산 발효물은 바람직하게는 하수오 발효 농축액에 물, 에탄올, 아세토박터 아세티(Acetobacter aceti) 및 아세토박터 파스테리아누스(Acetobacter pasteurianus)를 첨가한 후 28~32℃에서 10~14일 동안 초산 발효하여 제조할 수 있으며, 더욱 바람직하게는 하수오 발효 농축액에 물, 에탄올, 아세토박터 아세티(Acetobacter aceti) 및 아세토박터 파스테리아누스(Acetobacter pasteurianus)를 첨가한 후 30℃에서 12일 동안 초산 발효하여 제조할 수 있다.Further, in the manufacturing method of the sewage mixed beverage of the present invention, the (3) step of the acetic acid fermentation sewage water is preferably in the sewage water fermentation concentrate, ethanol, acetonitrile bakteo Oh Shetty (Acetobacter aceti ) and acetobacter After adding pasteurianus ), it can be prepared by acetic acid fermentation at 28~32℃ for 10~14 days, more preferably, water, ethanol, acetobacter aceti and acetobacter pasteria in sewage fermentation concentrate ( Acetobacter pasteurianus ) can be prepared by fermenting acetic acid at 30°C for 12 days.
상기 (1) 내지 (3)단계에 거쳐 하수오 초산 발효물을 제조하는 것이 하수오 특유의 이취는 제거되고 순하고 부드러우면서 갱년기 개선 효과가 더욱 증진된 하수오 초산 발효물로 제조할 수 있었다.Preparing the fermented product of sewage through the steps (1) to (3) was able to be produced as a fermented product of sewage produced by removing the odor peculiar to sewage and being mild and smooth, and further improving menopausal effect.
또한, 본 발명의 하수오 혼합 음료의 제조방법에서, 상기 (4)단계는 바람직하게는 홍화자, 속단, 당귀 및 오미자에 각각 물을 18~22배(v/w) 첨가한 후 70~90℃에서 2~4시간 동안 추출 및 여과하여 홍화자 추출물, 속단 추출물, 당귀 추출물 및 오미자 추출물을 제조할 수 있으며, 더욱 바람직하게는 홍화자, 속단, 당귀 및 오미자에 각각 물을 20배(v/w) 첨가한 후 80℃에서 3시간 동안 추출 및 여과하여 홍화자 추출물, 속단 추출물, 당귀 추출물 및 오미자 추출물을 제조할 수 있다. 상기와 같은 조건으로 제조된 추출물은 유효성분 함량이 높고 품질이 우수하고, 순하면서 풍미가 증진되어 음용이 용이하면서 향미가 우수하여 음료 제조에 적합한 약용식물 추출액으로 제조할 수 있었다.In addition, in the method for producing a mixed drink of sewage of the present invention, step (4) is preferably added at 18 to 22 times (v/w) of water to safflower, Sokdan, Angelica and Omija at 70-90° C. Extraction and filtration for 2-4 hours can produce safflower extract, quick-release extract, Angelica extract and Schisandra chinensis extract, more preferably, 20 times (v/w) of water is added to safflower, quick-set, Angelica and Schizandra, respectively After extraction and filtering for 3 hours at 80 ℃ it can be prepared safflower extract, Sokdan extract, Angelica extract and Omija extract. The extract prepared under the above conditions was able to be prepared as a medicinal plant extract suitable for beverage production because the active ingredient content is high, the quality is excellent, the flavor is enhanced while being easy to drink, and the flavor is excellent.
또한, 본 발명의 하수오 혼합 음료의 제조방법에서, 상기 (5)단계의 혼합물은 바람직하게는 혼합물 총 중량 기준으로, 하수오 초산 발효물 24~28 중량%, 속단 추출물 16~20 중량%, 홍화자 추출물 16~20 중량%, 당귀 추출물 18~22 중량% 및 오미자 추출물 16~20 중량%를 혼합한 혼합물일 수 있으며, 더욱 바람직하게는 혼합물 총 중량 기준으로, 하수오 초산 발효물 26 중량%, 속단 추출물 18 중량%, 홍화자 추출물 18 중량%, 당귀 추출물 20 중량% 및 오미자 추출물 18 중량%를 혼합한 혼합물일 수 있다. 상기와 같은 재료 및 배합비로 혼합한 혼합물은 갱년기 개선에 더욱 효과를 나타내면서 기호도가 우수한 음료 제조에 적합한 혼합물로 준비할 수 있었다.In addition, in the method for producing a mixed drink of sewage of the present invention, the mixture of step (5) is preferably based on the total weight of the mixture, 24 to 28% by weight of fermented sewage acetate, 16 to 20% by weight of fast extract, safflower extract It may be a mixture of 16 to 20% by weight, 18 to 22% by weight of Angelica extract and 16 to 20% by weight of Omija extract, and more preferably, based on the total weight of the mixture, 26% by weight of fermented sewage acetate, 18% by rapid extract It may be a mixture of weight%, safflower extract 18% by weight, Angelica extract 20% by weight, and Omija extract 18% by weight. The mixture mixed with the above-described materials and compounding ratios could be prepared as a mixture suitable for beverage preparation with excellent preference while exhibiting more effect in improving menopause.
본 발명의 하수오 혼합 음료의 제조방법은, 보다 구체적으로는The method for producing a mixed drink of sewage of the present invention, more specifically
(1) 하수오 450~550 g에 하수오 중량 대비 글루코스(glucose) 0.8~1.2% 및 수크로스(sucrose) 0.8~1.2%를 첨가한 후 110~130℃에서 10~20분 동안 멸균한 하수오 혼합물에 표고균사체(Lentinula edodes) 8~12 mL를 접종한 후 22~28℃에서 25~35일 동안 배양한 하수오 발효물을 75~85℃에서 10~14시간 동안 건조 및 분쇄하여 하수오 발효 분말을 제조하는 단계;(1) After adding 0.8 to 1.2% of glucose and 0.8 to 1.2% of sucrose to 450 to 550 g of sewage weight, the mixture is sterilized at 110 to 130°C for 10 to 20 minutes, and then elevation is applied to the sewage mixture. Mycelium ( Lentinula edode s) preparing a sewage fermentation powder by inoculating 8-12 mL and drying and pulverizing the sewage fermentation product cultured at 22-28 °C for 25-35 days for 10-14 hours at 75-85°C;
(2) 상기 (1)단계의 제조한 하수오 발효 분말에 물을 18~22배(v/w) 첨가한 후 60~100℃에서 1~15시간 동안 추출 및 여과하고 28~32 brix가 될 때까지 농축하여 하수오 발효 농축액을 제조하는 단계;(2) When 18-22 times (v/w) water is added to the sewage fermentation powder prepared in the step (1), extraction and filtration at 60-100°C for 1-15 hours, and when it becomes 28-32 brix Concentrating to prepare a sewage fermentation concentrate;
(3) 상기 (2)단계의 제조한 하수오 발효 농축액에 물, 에탄올, 아세토박터 아세티(Acetobacter aceti) 및 아세토박터 파스테리아누스(Acetobacter pasteurianus)를 첨가한 후 28~32℃에서 10~14일 동안 초산 발효하여 하수오 초산 발효물을 제조하는 단계;(3) Water, ethanol, acetobacter aceti ( acetobacter ) in the sewage fermentation concentrate prepared in step (2) aceti ) and acetobacter pasteurianus , followed by acetic acid fermentation at 28-32° C. for 10-14 days to produce a fermented sewage acetic acid;
(4) 홍화자, 속단, 당귀 및 오미자에 각각 물을 18~22배(v/w) 첨가한 후 70~90℃에서 2~4시간 동안 추출 및 여과하여 홍화자 추출물, 속단 추출물, 당귀 추출물 및 오미자 추출물을 제조하는 단계: 및(4) After adding 18-22 times (v/w) of water to safflower, Sokdan, Angelica and Omija, respectively, extracting and filtering at 70~90℃ for 2-4 hours, and then filtering Safflower extract, Sokdan extract, Angelica extract and Omija Preparing an extract: and
(5) 혼합물 총 중량 기준으로, 상기 (3)단계의 제조한 하수오 초산 발효물 24~28 중량%, 상기 (4)단계의 제조한 속단 추출물 16~20 중량%, 홍화자 추출물 16~20 중량%, 당귀 추출물 18~22 중량% 및 오미자 추출물 16~20 중량%를 혼합한 혼합물을 가열하는 단계를 포함할 수 있으며,(5) Based on the total weight of the mixture, 24 to 28% by weight of fermented sewage acetate produced in step (3), 16 to 20% by weight of extract from Sokdan prepared in step (4), 16 to 20% by weight of safflower extract , It may include the step of heating the mixture of Angelica extract 18-22% by weight and Omija extract 16-20% by weight,
더욱 구체적으로는More specifically
(1) 하수오 500 g에 하수오 중량 대비 글루코스(glucose) 1% 및 수크로스(sucrose) 1%를 첨가한 후 121℃에서 15분 동안 멸균한 하수오 혼합물에 표고균사체(Lentinula edodes) 10 mL를 접종한 후 25℃에서 30일 동안 배양한 하수오 발효물을 80℃에서 12시간 동안 건조 및 분쇄하여 하수오 발효 분말을 제조하는 단계;(1) After adding 1% of glucose and 1% of sucrose to 500 g of sewage weight, shiitake mycelium ( Lentinula) in a sewage mixture sterilized at 121° C. for 15 minutes. edode s) preparing a sewage fermentation powder by inoculating 10 mL and drying and grinding the sewage fermentation product cultured at 25°C for 30 days for 12 hours;
(2) 상기 (1)단계의 제조한 하수오 발효 분말에 물을 18~22배(v/w) 첨가한 후 60~100℃에서 1~15시간 동안 추출 및 여과하고 30 brix가 될 때까지 농축하여 하수오 발효 농축액을 제조하는 단계;(2) After adding 18-22 times (v/w) of water to the sewage fermentation powder prepared in the step (1), extracting and filtering for 1-15 hours at 60-100° C. and concentrating until 30 brix. Preparing a fermentation concentrate by sewage;
(3) 상기 (2)단계의 제조한 하수오 발효 농축액에 물, 에탄올, 아세토박터 아세티(Acetobacter aceti) 및 아세토박터 파스테리아누스(Acetobacter pasteurianus)를 첨가한 후 30℃에서 12일 동안 초산 발효하여 하수오 초산 발효물을 제조하는 단계;(3) Water, ethanol, acetobacter aceti ( acetobacter ) in the sewage fermentation concentrate prepared in step (2) aceti ) and acetobacter pasteurianus , followed by acetic acid fermentation at 30° C. for 12 days to prepare sewage acetic acid fermentation product;
(4) 홍화자, 속단, 당귀 및 오미자에 각각 물을 20배(v/w) 첨가한 후 80℃에서 3시간 동안 추출 및 여과하여 홍화자 추출물, 속단 추출물, 당귀 추출물 및 오미자 추출물을 제조하는 단계: 및(4) After adding
(5) 혼합물 총 중량 기준으로, 상기 (3)단계의 제조한 하수오 초산 발효물 26 중량%, 상기 (4)단계의 제조한 속단 추출물 18 중량%, 홍화자 추출물 18 중량%, 당귀 추출물 20 중량% 및 오미자 추출물 18 중량%를 혼합한 혼합물을 가열하는 단계를 포함할 수 있다.(5) Based on the total weight of the mixture, 26% by weight of fermented sewage acetate produced in step (3), 18% by weight of extract from Sokdan prepared in step (4), 18% by weight of safflower extract, 20% by weight of Angelica extract And heating the mixture of 18% by weight of Omija extract.
본 발명은 또한, 상기 방법으로 제조된 하수오 혼합 음료를 제공한다.The present invention also provides a sewage mixed beverage prepared by the above method.
이하, 본 발명의 제조예 및 실시예를 들어 상세히 설명한다. 단, 하기 제조예 및 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 제조예 및 실시예에 한정되는 것은 아니다.Hereinafter, the production examples and examples of the present invention will be described in detail. However, the following production examples and examples are only to illustrate the present invention, the content of the present invention is not limited to the following production examples and examples.
제조예Manufacturing example 1. 복합 발효 1. Complex fermentation 하수오Ha Shou 추출물과 한약재 추출물을 이용한 갱년기 증상 개선용 For improvement of menopausal symptoms using extracts and herbal extracts 하수오Ha Shou 생약초Herbal medicine 복합물 Complex
(1) 하수오를 분쇄한 분쇄물 500 g에 하수오 분쇄물 대비 글루코스 1%(w/w) 및 수크로스 1%(w/w)를 첨가한 후 121℃에서 15분 동안 멸균하였다. 상기 멸균한 하수오 혼합물을 냉각한 후, 표고균사체(Lentinula edodes KCTC18583P) 10 mL를 접종한 후 25℃에서 30일 동안 배양하였다. 상기 배양한 하수오 발효물을 탈병 및 80℃에서 12시간 동안 건조하고, 분쇄하여 하수오 발효 분말을 제조하였다(도 1).(1) To 500 g of ground crushed sewage, 1% glucose (w/w) and 1% sucrose (w/w) compared to sewage ground were added and sterilized at 121°C for 15 minutes. After cooling the sterilized sewage mixture, shiitake mycelium ( Lentinula edode s KCTC18583P) After inoculating 10 mL, it was incubated at 25°C for 30 days. The cultured sewage fermentation product was dehydrated and dried at 80° C. for 12 hours, followed by grinding to prepare a sewage fermentation powder (FIG. 1 ).
(2) 상기 (1)단계의 제조한 하수오 발효 분말에 물을 20배(v/w) 첨가한 후 60~100℃에서 1~15시간 동안 추출한 후 여과하고 30 brix가 될 때까지 농축하여 하수오 발효 농축액을 제조하였다(도 2).(2) After adding 20 times (v/w) of water to the sewage fermentation powder prepared in the step (1), extract for 1-15 hours at 60~100℃, filter and concentrate until 30 brix, and then sewage. A fermentation concentrate was prepared (FIG. 2).
(3) 상기 (2)단계의 제조한 하수오 발효 농축액에 정제수를 3:7(v:v) 비율로 첨가하고, 여기에 발효 에탄올(알코올을 발효 후 증류)과 초산균(Acetobacter aceti KCCM40229, Acetobacter pasteurianus KCCM40011)을 5 mL 주입하여 30℃에서 12일 동안 초산 발효하여 하수오 초산 발효물을 제조하였다(도 3).(3) the above (2) purified water in a sewage fermentation concentrate prepared in step 3: 7 (v: v) was added at a rate, and (after the fermentation of alcohol distillation) fermentation and ethanol here chosangyun (Acetobacter aceti KCCM40229, Acetobacter pasteurianus KCCM40011) was injected with 5 mL, and acetic acid fermentation was performed at 30° C. for 12 days to prepare a sewage acetic acid fermentation product (FIG. 3 ).
(4) 홍화자, 속단, 당귀 및 오미자에 각각 물을 20배(v/w) 첨가한 후 80℃에서 3시간 동안 추출한 후 4,000 rpm에서 15분 동안 원심분리하여 홍화자 추출물, 속단 추출물, 당귀 추출물 및 오미자 추출물을 제조하였다(도 4).(4) After adding 20 times (v/w) of water to safflower, Sokdan, Angelica and Omija respectively, extracting them at 80°C for 3 hours, and centrifuging at 4,000 rpm for 15 minutes to extract Safflower extract, Sokdan extract, Angelica extract, and Omija extract was prepared (FIG. 4 ).
(5) 상기 (3)단계의 제조한 하수오 초산 발효물 26 중량%와, 상기 (4)단계의 제조한 속단 추출물 18 중량%, 홍화자 추출물 18 중량%, 당귀 추출물 20 중량% 및 오미자 추출물 18 중량%를 혼합한 혼합물을 100℃에서 30분 동안 가열하였다.(5) 26% by weight of fermented sewage acetic acid prepared in the step (3), 18% by weight of the Sokdan extract prepared in the step (4), 18% by weight of the safflower extract, 20% by weight of the Angelica extract, and 18% of Omija extract The mixture of% was heated at 100° C. for 30 minutes.
비교예Comparative example 1. One. 하수오Ha Shou 발효 농축액 Fermentation concentrate
상기 제조예 1의 (1) 및 (2)단계에 거쳐 하수오 발효 농축액을 제조하였다.Through the steps (1) and (2) of Preparation Example 1, a sewage fermentation concentrate was prepared.
비교예Comparative example 2. 2. 하수오Ha Shou 초산 Acetic acid 발효물Fermentation
상기 제조예 1의 (1) 내지 (3)단계에 거쳐 하수오 초산 발효물을 제조하였다.Through the steps (1) to (3) of Preparation Example 1, fermented sewage acetic acid was prepared.
비교예Comparative example 3 내지 5: 3 to 5: 하수오Ha Shou 생약초Herbal medicine 복합물 Complex
상기 제조예 1의 방법으로 하수오 생약초 복합물을 제조하되, (5)단계의 혼합물 혼합 시 하기 표 1의 배합비로 혼합한 혼합물을 이용하여 비교예 3 내지 5의 하수오 생약초 복합물을 제조하였다.In the method of Preparation Example 1 to prepare a composite of herbal medicinal herbs, but when mixing the mixture of step (5), using the mixture mixed in the mixing ratio shown in Table 1 to prepare the medicinal herbs of the composite of Examples 3 to 5.
실험방법Experiment method
1. 폐경모델 제작 및 여성 갱년기 개선 분석1. Menopause model production and improvement of female menopause
(1) 폐경모델 제작(1) Menopause model production
Sprague-Dawley계 암컷 랫트(200~210 g)을 모델을 처치하지 않은 일반대조군과 모델을 처치한 실험군으로 나누며, 실험군은 근육주사로 마취시킨 후 복부의 털을 제거한 다음 무균 상태로 수술을 시행하였다. 먼저 등쪽 하부 양쪽을 최소한으로 절개하여 노출시킨 후 양측의 난소를 제거하고 다시 복부를 봉합하였다. 3일 후 다시 시료를 투여하였다.The Sprague-Dawley female rats (200-210 g) were divided into a general control group that did not kill the model and an experimental group that treated the model, and the experimental group was anesthetized by intramuscular injection, and then the abdominal hair was removed and then operated aseptically. . First, both sides of the dorsal lower side were exposed with minimal incision, and then the ovaries on both sides were removed and the abdomen was closed again. After 3 days, the sample was administered again.
(2) 그룹 설정 및 시료 투여방법(2) Group setting and sample administration method
시료 투여는 200 mg/kg/day 농도로 경구투여하고 음성대조군은 같은 용량의 생리식염수를 투여하였다. 양성대조군은 에스트라디올 E2(estradiol E2)를 0.5 mg/kg/day 농도로 경구투여하였다. 실험군의 모든 시료 투여기간은 폐경모델 처치 3일 후부터 50일간 투여하였다.The sample was administered orally at a concentration of 200 mg/kg/day, and the negative control group received the same amount of physiological saline. The positive control group orally administered estradiol E2 (estradiol E2) at a concentration of 0.5 mg/kg/day. All sample administration periods in the experimental group were administered for 50 days from 3 days after the menopause model treatment.
(3) 체중 및 식이량 측정(3) Weight and dietary measure
폐경모델 처치 3일 후부터 50일간 일주일에 한번 각 그룹의 체중과 식이량을 측정하여 체중증가율 및 식이량 변화를 분석하였다.The weight gain and the amount of diet were analyzed by measuring the weight and diet of each group once a week for 50 days from 3 days after the menopause model treatment.
(4) 혈액 내 생화학적 분석(4) Biochemical analysis in blood
50일간 시료 투여 종료 후 혈청 분리를 위해 실험동물을 해부 전 24시간 절식시킨 후 에테르 마취 하에서 개복한 후 정맥에서 채혈하고 실온에서 30분간 방치 한 후, 3,000 rpm 및 25℃에서 15분간 원심분리하여 혈청을 분리하고, 실험 시까지 -70℃에 보관하였다.After the administration of the sample for 50 days, the experimental animals were fasted for 24 hours prior to dissection, opened under ether anesthesia, collected from a vein, left at room temperature for 30 minutes, and then centrifuged at 3,000 rpm and 25°C for 15 minutes. Was isolated and stored at -70°C until experimentation.
혈청 내 생화학적 지표로 혈청 총 단백질, 알부민, 골대사 지표 칼슘, 알칼리 포스파타아제(alkaline phosphatase), 혈중 여성 호르몬 지표 에스트로겐(estrogen), 에스트라디올(estradiol), 성욕 감쇠 지표 호르몬 테스토스테론(testosterone), 혈당 지표 글루코스(glucose), 간기능 지표 AST 및 ALT, 지질대사 지표 총 콜레스테롤(total cholesterol), HDL, LDL, VLDL 및 트리글리세리드(triacylglycerol)를 각 지표 측정용 키트 및 자동혈액분석기기(Vitros 250 chemistry system (Ortho Clinical Diagnostics))를 사용하여 분석하였다.Serum biochemical indicators include serum total protein, albumin, bone metabolism indicator calcium, alkaline phosphatase, blood female hormone indicator estrogen, estradiol, libido indicator hormone testosterone, blood sugar Indicator glucose and liver function indicators AST and ALT, lipid metabolism indicator total cholesterol, HDL, LDL, VLDL and triglyceride (triacylglycerol) for each indicator measurement kit and automatic blood analyzer (Vitros 250 chemistry system ( Ortho Clinical Diagnostics)).
(5) 조직 분석(5) Organizational analysis
장기 적출을 위해 실험동물을 해부 전 24시간 절식시킨 후 에테르 마취 하에서 개복한 후 혈청 분리 후 0.9% 생리 식염수 용액으로 관류시킨 후 물기를 제거하고, 각 조직(간, 신장, 비장, 자궁-난소)은 분리하며, 조직에 있는 지방을 제거하고 무게를 잰 후 분석할 때 까지 -70℃에 보관하였다.For long-term extraction, the experimental animals were fasted for 24 hours before dissection, opened under ether anesthesia, separated by serum, perfused with 0.9% physiological saline solution, and then drained, and each tissue (liver, kidney, spleen, uterus-ovary) The silver was separated, the fat in the tissue was removed, weighed and stored at -70°C until analysis.
조직염색을 위해 동물은 희생시켜 자궁-난소 조직을 세척 후 디지털 카메라(Canon Power Shot SX40 HS)로 이미지 결과 도출하고 자궁관 조직 상부 약 0.5 cm를 절제하여 동결절편(cryosection)법과 저온유지장치(model KD-2950, Kedee, China)을 이용하여 10 ㎛ 두께로 절개한 후 Hematoxylin 염색을 실시하고 현미경(Leica microscope, DM500, Leica, Germany)으로 관찰하였다.For tissue staining, animals are sacrificed to wash the uterine-ovarian tissue, then derive the image results with a digital camera (Canon Power Shot SX40 HS) and excise about 0.5 cm above the uterine duct tissue to cryosection and cryosection. KD-2950, Kedee, China) was incised to a thickness of 10 μm, and then Hematoxylin staining was performed and observed under a microscope (Leica microscope, DM500, Leica, Germany).
실시예Example 1. 여성갱년기 백서 모델을 이용한 여성갱년기 개선 효능 분석 1. Analyze the efficacy of improving menopause using the menopausal white paper model
(1) 주당 체중 변화 분석(1) Weight change analysis per week
실험동물의 체중 측정은 동물 사육실에서 적응기간(7일)과 난소제거 여성갱년기 모델 처치 후 회복기간(3일)을 제외하고 50일 시료투여 기간 동안 8번 수행되었다. 난소제거 여성갱년기 모델을 처치하지 않은 일반대조군은 처음 평균체중 257.5±16.3 g에서 50일 후 평균체중 322.0±17.0 g으로 평균적으로 64.5 g의 체중변화를 보였다. 난소제거 여성갱년기 모델을 처치한 그룹 중 일반사료 및 생리식염수만 급여한 음성대조군(OVX모델+saline)은 처음 평균체중 233.0±1.4 g에서 50일 후 평균체중 405.0±10.6 g으로 평균적으로 172.0 g의 체중변화를 보였다. 난소제거 여성갱년기 모델을 처치한 그룹 중 에스트라디올 E2를 50일 동안 섭취한 양성대조군은 처음 평균체중 237.0±2.8 g에서 50일 후 평균체중 331.5±9.2 g으로 평균적으로 94.5 g의 체중변화를 보였다. 난소제거 여성갱년기 모델을 처치한 그룹 중 시료 음료를 50일 동안 섭취한 실험군은 처음 평균체중 248.5±0.7 g에서 50일 후 평균체중 353.0±9.9 g으로 평균적으로 104.5 g의 체중변화를 보였다(도 5).Measurement of the weight of the experimental animals was performed 8 times during the 50-day sample administration period, excluding the adaptation period (7 days) and the recovery period (3 days) after treatment with the ovarian defecation model in the animal breeding room. The general control group without treatment of the ovarian defecation model showed an average weight change of 64.5 g from 257.5±16.3 g in the first average to 322.0±17.0 g in 50 days. Among the groups treated with the ovarian removal menopausal model, the negative control group (OVX model + saline), who fed only normal feed and physiological saline, had an average weight of 405.0±10.6 g after 50 days from 233.0±1.4 g of the first average weight of 172.0 g. Showed weight change. Among the groups treated with the ovarian defecation model, the positive control group that consumed estradiol E2 for 50 days showed an average weight change of 94.5 g from 237.0±2.8 g in the first average weight to 331.5±9.2 g in 50 days. Among the groups treated with the ovarian removal menopausal model, the experimental group that consumed sample drinks for 50 days showed an average weight change of 104.5 g from 248.5±0.7 g of the first average weight to 353.0±9.9 g of the average weight after 50 days (FIG. 5 ). ).
이러한 각 그룹의 체중값으로 증가율을 분석한 결과, 시료투여 전 체중을 100%로 할 때 일반대조군은 125.5±14.5 g, 에스트라디올 E2를 섭취한 양성대조군은 139.9±5.5 g, 일반사료 및 생리식염수만 섭취한 음성대조군은 170.6±5.6 g, 음료 시료를 섭취한 실험군은 142.0±3.6 g으로 분석되었다(도 6). 모델처치 후 일반사료 및 생리식염수만 섭취한 그룹에서 체중이 약 71%로 크게 증가되었지만 양성대조군(약 40%)과 실험군(약 42%)에서 유사한 수준으로 체중 증가폭이 감소되었다.As a result of analyzing the increase rate by the weight value of each group, when the body weight was set to 100% before sample administration, 125.5±14.5 g in the general control group and 139.9±5.5 g in the positive control group ingesting estradiol E2, general feed and physiological saline The negative control group ingested only was 170.6±5.6 g, and the experimental group ingesting beverage samples was analyzed to be 142.0±3.6 g (FIG. 6 ). After model treatment, the body weight increased significantly to about 71% in the group that consumed only normal feed and physiological saline, but the weight gain was decreased to a similar level in the positive control group (about 40%) and the experimental group (about 42%).
(2) 주당 식이량 변화 분석(2) Analysis of dietary change per week
일반대조군은 1주부터 7주까지 주당 101 g, 77 g, 69 g, 91 g, 60 g, 81 g, 56 g을 섭취하였고 양성대조군은 1주부터 7주까지 주당 62.5 g, 77.5 g, 63.5 g, 101.5 g, 69 g, 105 g, 60 g을 섭취하였다. 음성대조군(OVX모델+saline)은 1주부터 7주까지 주당 56 g, 51 g, 43 g, 59 g, 45 g, 58 g, 44 g을 섭취하였고, 시료 음료를 섭취한 실험군은 1주부터 7주까지 주당 61.5 g, 73.5 g, 76.5 g, 104 g, 79 g, 115 g, 84.5 g을 섭취하였다(도 7). 총 섭취량은 일반대조군이 1070 g, 양성대조군이 1078 g, 음성대조군이 712 g, 실험군이 1188 g을 섭취하여, 실험군(1188 g) > 양성대조군(1078 g) > 일반대조군(1070 g) > 음성대조군(712 g) 순서로 확인되었다 (도 8).The general control group consumed 101 g, 77 g, 69 g, 91 g, 60 g, 81 g, and 56 g per week from 1 week to 7 weeks, and the positive control group was 62.5 g, 77.5 g, 63.5 per week from 1 week to 7 weeks. g, 101.5 g, 69 g, 105 g, and 60 g were ingested. The negative control group (OVX model + saline) consumed 56 g, 51 g, 43 g, 59 g, 45 g, 58 g, 44 g per week from 1 week to 7 weeks. Up to 7 weeks, 61.5 g, 73.5 g, 76.5 g, 104 g, 79 g, 115 g, and 84.5 g were consumed per week (Fig. 7). The total intake was 1070 g in the general control group, 1078 g in the positive control group, 712 g in the negative control group, and 1188 g in the experimental group, so that the experimental group (1188 g)> positive control group (1078 g)> general control group (1070 g)> negative It was confirmed in the order of control (712 g) (Fig. 8).
(3) 식이효율 변화 분석(3) Analysis of changes in dietary efficiency
주당 식이량과 체중 변화 결과를 활용하여 식이효율 변화와 50일 동안의 최종 식이효율을 분석하였다. 일반대조군의 식이효율은 1~7주까지 차례로 -0.06, 0.48, 0.53, 0.34, 0.68, 0.74, 1.15로 분석되었고 양성대조군의 식이효율은 1~7주까지 차례로 0.26, 0.85, 1.43, 1.11, 1.47, 0.86, 1.58로 분석되었다. 음성대조군(OVX모델+saline)의 식이효율은 1~7주까지 차례로 1.21, 2.25, 3.23, 2.86, 4.04, 3.03, 3.91로 분석되었고 실험군의 식이효율은 1~7주까지 차례로 -0.03, 0.69, 1.16, 0.79, 1.22, 0.90, 1.24로 분석되었다(도 9). 모델처치 그룹 중 식염수만 섭취한 음성대조군에서 식이량 대비 체중 증가량이 높아 비교적 높은 식이효율 수치를 보였고 양성대조군 > 실험군 > 일반대조군 순서로 식이효율이 높게 나타났다. 50일 동안의 최종 식이효율은 일반대조군에서 0.060, 양성대조군에서 0.088, 음성대조군에서 0.242, 실험군에서 0.088로 확인되어 음성대조군 > 실험군 ≥ 양성대조군 > 일반대조군 순서로 나타났다. 따라서, 모델처치 후 생리식염수만 섭취한 그룹의 급격한 체중증가로 식이효율이 높게 나타났고, 모델처치 후 에스트라디올 E2와 시료 음료를 섭취한 두 그룹에서 체중증가가 감소되어 일반대조군과 비교적 유사한 수준으로 식이효율이 나타났다.Changes in diet efficiency and final diet efficiency over 50 days were analyzed using the results of dietary changes per week and weight. The dietary efficiency of the general control group was analyzed as -0.06, 0.48, 0.53, 0.34, 0.68, 0.74, and 1.15 in order from 1 to 7 weeks, and the dietary efficiency of the positive control group was 0.26, 0.85, 1.43, 1.11, 1.47 in sequence from 1 to 7 weeks. , 0.86, 1.58. The negative control group (OVX model + saline) was analyzed as 1.21, 2.25, 3.23, 2.86, 4.04, 3.03, 3.91 in order from 1 to 7 weeks, and the experimental group's dietary efficiency was -0.03, 0.69, in order from 1 to 7 weeks. It was analyzed as 1.16, 0.79, 1.22, 0.90, and 1.24 (Fig. 9). In the model control group, in the negative control group that consumed only saline, the weight gain compared to the diet was high, showing a relatively high dietary efficiency value, and the dietary efficiency was high in the order of positive control group> experimental group> general control group. The final dietary efficiency over 50 days was confirmed as 0.060 in the general control group, 0.088 in the positive control group, 0.242 in the negative control group, and 0.088 in the experimental group, which was in the order of negative control group> experimental group ≥ positive control group> general control group. Therefore, after model treatment, the dietary efficiency was high due to the rapid weight gain of the group that consumed only physiological saline, and after the model treatment, the weight gain was decreased in the two groups that consumed estradiol E2 and sample drink, and the level was relatively similar to that of the general control group. Dietary efficiency appeared.
(4) 조직 중량 변화 분석(4) Analysis of tissue weight change
50일 동안 식이 종료 후 간, 비장, 신장, 자궁-난소 조직을 절개하여 적출하고 조직 중량을 측정하였다. 간의 조직 중량은 일반대조군에서 6.69±0.63 g, 양성대조군에서 6.55±0.45 g, 음성대조군(OVX모델+saline)에서 8.52±0.55 g, 실험군에서 6.65±0.11 g으로 각각 측정되었다. 음성대조군에서 비교적 가장 높았고 다른 3개 그룹에서 유사한 수준으로 나타났다(도 10).After the end of the diet for 50 days, liver, spleen, kidney, and uterine-ovarian tissues were excised and excised, and tissue weight was measured. The liver tissue weight was measured to be 6.69±0.63 g in the general control group, 6.55±0.45 g in the positive control group, 8.52±0.55 g in the negative control group (OVX model + saline), and 6.65±0.11 g in the experimental group. It was relatively highest in the negative control group and showed similar levels in the other 3 groups (FIG. 10).
비장의 조직 중량은 일반대조군에서 0.47±0.05 g, 양성대조군에서 0.64±0.06 g, 음성대조군에서 0.65±0.03 g, 실험군에서 0.54±0.06 g으로 측정되었다 (도 11). 양성대조군과 음성대조군이 유사한 수준으로 비교적 가장 높게 나왔고 실험군의 비장 중량은 일반대조군보다 다소 높은 것으로 나타났다.The tissue weight of the spleen was measured to be 0.47±0.05 g in the general control group, 0.64±0.06 g in the positive control group, 0.65±0.03 g in the negative control group, and 0.54±0.06 g in the experimental group (FIG. 11 ). The positive control group and the negative control group were relatively highest at similar levels, and the spleen weight of the experimental group was slightly higher than that of the general control group.
신장의 조직 중량은 일반대조군에서 0.99±0.10 g, 양성대조군에서 1.01±0.001 g, 음성대조군에서 1.19±0.05 g, 실험군에서 1.08±0.01 g으로 측정되었다(도 12). 음성대조군에서 비교적 가장 높았고 실험군의 신장 중량이 일반대조군과 양성대조군보다 비교적 높은 것으로 나타났다.The tissue weight of the kidneys was measured as 0.99±0.10 g in the general control group, 1.01±0.001 g in the positive control group, 1.19±0.05 g in the negative control group, and 1.08±0.01 g in the experimental group (FIG. 12). It was the highest in the negative control group, and the height of the experimental group was relatively higher than that of the normal control group and the positive control group.
자궁-난소의 조직 중량은 일반대조군에서 0.82±0.22 g, 양성대조군에서 0.22±0.002 g, 음성대조군에서 0.18±0.005 g, 실험군에서 0.21±0.14 g으로 측정되었다(도 13). 모델처치한 3개 그룹에서 일반대조군에 비해 급격한 자궁-난소 조직 중량의 감소를 보였으며, 양성대조군과 실험군에서 자궁-난소 조직 중량이 음성대조군에 비해 다소 증가된 것을 확인하였다.Uterine-ovarian tissue weight was measured to be 0.82±0.22 g in the general control group, 0.22±0.002 g in the positive control group, 0.18±0.005 g in the negative control group, and 0.21±0.14 g in the experimental group (FIG. 13). The three groups treated with the model showed a sharp decrease in uterine-ovarian tissue weight compared to the general control group, and it was confirmed that the uterine-ovarian tissue weight was slightly increased in the positive control group and the experimental group compared to the negative control group.
(5) 자궁-난소 조직 관찰(5) Uterine-ovarian tissue observation
50일 동안 식이 종료 후 절개한 자궁-난소 조직의 중량을 측정 후 이미지 결과를 만들어 관찰하였다. 또한, 자궁관 상부를 단면 두께 10 ㎛로 동결절편을 만들어 관찰하였다. 먼저 자궁-난소 조직의 이미지를 관찰해보면, 난소제거 모델을 처치하지 않은 일반대조군의 자궁-난소 조직에 비해 난소가 제거된 3개 그룹의 자궁 조직 크기가 상당히 감소된 것을 확인하였으며, 에스트라디올 E2와 음료를 섭취한 2개 그룹에서 조직 크기의 감소 정도가 줄었고 음성대조군에 비해 비교적 회복된 조직 형태를 보였다(도 14). 또한, 자궁관 상부 단면을 현미경으로 관찰한 결과, 자궁-난소 조직 크기와 유사한 패턴의 자궁관 크기가 확인되었고 일반대조군의 자궁관에 비해 난소가 제거된 자궁관 내 구성세포의 밀도가 상당히 감소되었고 구성세포가 약화되어 부분적으로 손실된 부분이 관찰되었지만, 에스트라디올 E2와 시료 음료를 섭취한 2개 그룹에서 구성세포의 부분적 손실이 감소된 것을 확인하였다(도 15).After the diet was completed for 50 days, the weight of the incised uterine-ovary tissue was measured, and the result of the image was observed. In addition, the upper section of the uterine tube was observed by making a frozen section with a cross-section thickness of 10 μm. First, observing the image of the uterine-ovarian tissue, it was confirmed that the size of the uterine tissue of the three groups in which the ovaries were removed was significantly reduced compared to the uterine-ovarian tissue of the general control group without the ovarian removal model. The reduction in tissue size was reduced in the two groups that consumed the drink and showed a relatively recovered tissue form compared to the negative control (FIG. 14). In addition, as a result of observing the upper section of the uterine tube under a microscope, the size of the uterine tube with a pattern similar to the size of the uterine-ovarian tissue was confirmed, and the density of constituent cells in the uterine tube with the ovaries removed was significantly reduced compared to that of the general control Partial loss was observed due to weakening of the constituent cells, but it was confirmed that partial loss of constituent cells was reduced in the two groups ingesting estradiol E2 and the sample beverage (FIG. 15).
(6) 혈청 내 총 단백질 및 알부민 변화 분석(6) Analysis of total protein and albumin changes in serum
50일 동안 식이 종료 후 혈액으로부터 얻은 혈청을 이용하여 총 단백질과 혈액단백질인 알부민을 측정하였다. 혈청 내 총 단백질 농도는 일반대조군에서 5.80±0.99 g/dL, 양성대조군에서 5.25±0.35 g/dL, 음성대조군(OVX모델+saline)에서 5.50±0.28 g/dL, 실험군에서 5.25±0.35 g/dL으로 확인되었다(도 16). 혈청 내 알부민 농도는 일반대조군에서 3.05±0.92 g/dL, 양성대조군에서 2.55±0.07 g/dL, 음성대조군에서 2.70±0.14 g/dL, 실험군에서 2.55±0.21 g/dL으로 확인되었다(도 17). 난소제거 모델을 처치하지 않은 일반대조군의 총 단백질 및 알부민 농도에 비해 난소가 제거된 3개 그룹에서 다소 감소된 것을 확인하였고 에스트라디올 E2와 시료 음료를 섭취한 2개 그룹에서 음성대조군보다 다소 낮은 농도를 보였다.After the end of the diet for 50 days, the serum from the blood was used to measure the total protein and blood protein albumin. Total protein concentration in serum was 5.80±0.99 g/dL in the general control group, 5.25±0.35 g/dL in the positive control group, 5.50±0.28 g/dL in the negative control group (OVX model+saline), and 5.25±0.35 g/dL in the experimental group. It was confirmed as (Fig. 16). The albumin concentration in serum was confirmed to be 3.05±0.92 g/dL in the general control group, 2.55±0.07 g/dL in the positive control group, 2.70±0.14 g/dL in the negative control group, and 2.55±0.21 g/dL in the experimental control group (FIG. 17 ). . It was confirmed that the ovarian removal model was slightly reduced in the 3 groups with ovaries removed compared to the total protein and albumin concentrations in the general control group without treatment of the ovary removal model, and was slightly lower than the negative control in the 2 groups with estradiol E2 and sample drinks. Showed.
(7) 혈청 내 골대사 지표 칼슘, 알칼리 포스파타아제(alkaline phosphatase) 농도 변화 분석(7) Analysis of changes in calcium, alkaline phosphatase levels in bone metabolism in serum
혈청 내 칼슘 농도는 일반대조군에서 18.22±0.63 mg/dL, 양성대조군에서 16.11±0.01 mg/dL, 음성대조군(OVX모델+saline)에서 17.29±0.33 mg/dL, 실험군에서 17.96±0.24 mg/dL으로 측정되었다(도 18). 음성대조군에서 칼슘 농도가 감소되었지만 시료 음료를 섭취한 실험군에서 일반대조군과 유사한 수준으로 증가되었고 에스트라디올 E2를 섭취한 양성대조군에서는 칼슘 농도를 증가시키지 못했다.Calcium concentration in serum was 18.22±0.63 mg/dL in the general control group, 16.11±0.01 mg/dL in the positive control group, 17.29±0.33 mg/dL in the negative control group (OVX model + saline), and 17.96±0.24 mg/dL in the experimental group. It was measured (Fig. 18). Calcium concentrations were reduced in the negative control group, but increased to a level similar to the normal control group in the experimental group ingesting the sample beverage, and did not increase the calcium concentration in the positive control group ingested estradiol E2.
혈청 내 ALP 농도는 일반대조군에서 0.58±0.10 mU, 양성대조군에서 0.66±0.02 mU, 음성대조군에서 0.51±0.09 mU, 실험군에서 0.56±0.05 mU로 측정되었다(도 19). 음성대조군에서 ALP 농도가 감소되었지만 양성대조군에서 그 농도가 상당히 증가되었고 음료를 섭취한 실험군에서는 일반대조군과 유사한 수준으로 증가되었다.The serum ALP concentration was measured to be 0.58±0.10 mU in the general control group, 0.66±0.02 mU in the positive control group, 0.51±0.09 mU in the negative control group, and 0.56±0.05 mU in the experimental group (FIG. 19). Although the ALP concentration was decreased in the negative control group, the concentration was significantly increased in the positive control group and increased to a level similar to that of the general control group in the experimental group ingesting the beverage.
(8) 혈청 내 에스트로겐 농도 변화 분석(8) Analysis of changes in estrogen concentration in serum
혈청 내 여성호르몬인 에스트로겐 농도를 측정하기 위해 에스트로겐 표준물질을 0, 12.5, 62.5, 187.5 pg/mL 농도로 준비하고 450 nm에서 표준곡선을 도출하여 각 혈청의 흡광도 값을 산출하였다. 에스트로겐 농도는 일반대조군에서 77.95±1.59 pg/mL, 양성대조군에서 32.55±21.38 pg/mL, 음성대조군(OVX모델+saline)에서 10.48±2.43 pg/mL, 실험군에서 27.63±17.55 pg/mL으로 측정되었다(도 20). 난소제거 처치 후 음성대조군에서 에스트로겐 농도가 급격히 감소되었지만, 에스트라디올 E2와 시료 음료를 섭취한 2개 그룹에서 에스트로겐 농도가 증가된 것을 확인하였다.To measure the concentration of estrogen, a female hormone in serum, estrogen standards were prepared at concentrations of 0, 12.5, 62.5, and 187.5 pg/mL, and a standard curve was derived at 450 nm to calculate the absorbance value of each serum. Estrogen concentrations were measured at 77.95±1.59 pg/mL in the general control group, 32.55±21.38 pg/mL in the positive control group, 10.48±2.43 pg/mL in the negative control group (OVX model + saline), and 27.63±17.55 pg/mL in the experimental group. (Figure 20). After the ovarian removal treatment, the estrogen concentration was rapidly decreased in the negative control group, but it was confirmed that the estrogen concentration was increased in the two groups ingesting estradiol E2 and the sample beverage.
(9) 혈청 내 에스트라디올 농도 변화 분석(9) Analysis of changes in serum estradiol concentration
혈청 내 여성호르몬인 에스트라디올 E2 농도를 측정하기 위해 에스트라디올 E2 표준물질을 0, 3, 10, 30, 100, 300 pg/mL 농도로 준비하고 450 nm에서 표준곡선을 도출하여 각 혈청의 흡광도 값을 산출하였다. 에스트라디올 E2 농도는 일반대조군에서 50.01±15.12 pg/mL, 양성대조군에서 53.09±26.63 pg/mL, 음성대조군(OVX모델+saline)에서 39.42±8.78 pg/mL, 실험군에서 75.35±6.18 pg/mL으로 측정되었다(도 21). 에스트로게 농도와 유사하게 음성대조군에서 농도가 감소되었지만, 에스트라디올 E2를 섭취한 양성대조군에서 다소 증가되었고 시료 음료를 섭취한 실험군에서 상당히 증가된 것을 확인하였다.To measure the concentration of estradiol E2, a female hormone in serum, prepare estradiol E2 standard at concentrations of 0, 3, 10, 30, 100, 300 pg/mL, and derive a standard curve at 450 nm to measure the absorbance value of each serum. Was calculated. Estradiol E2 concentration was 50.01±15.12 pg/mL in the general control group, 53.09±26.63 pg/mL in the positive control group, 39.42±8.78 pg/mL in the negative control group (OVX model + saline), and 75.35±6.18 pg/mL in the experimental group. It was measured (Fig. 21). It was confirmed that the concentration was decreased in the negative control group similar to the estroge concentration, but slightly increased in the positive control group consuming estradiol E2 and significantly increased in the experimental group ingesting the sample beverage.
(10) 혈청 내 테스토스테론(testosterone) 농도 변화 분석(10) Analysis of testosterone concentration change in serum
혈청 내 여성 테스토스테론 호르몬은 성욕에 관련되어 성욕감퇴 진단에 이용하는 지표이며, 테스토스테론 표준물질을 450 nm에서 표준곡선을 도출하여 각 혈청의 흡광도 값을 산출하였다. 테스토스테론 농도는 일반대조군에서 4.51±2.01 ng/mL, 양성대조군에서 1.81±1.62 ng/mL, 음성대조군(OVX모델+saline)에서 1.65±0.30 ng/mL, 실험군에서 4.35±0.56 ng/mL으로 측정되었다(도 22). 음성대조군에서 테스토스테론 농도가 급격하게 감소되었지만 시료를 섭취한 실험군에서 일반대조군과 유사한 수준으로 증가되었다. 에스트라디올 E2를 섭취한 양성대조군에서는 테스토스테론 농도가 증가되지 않았다.The female testosterone hormone in serum is an indicator used in the diagnosis of libido in relation to sexual desire, and a standard curve for testosterone standard substance was derived at 450 nm to calculate the absorbance value of each serum. Testosterone concentrations were measured as 4.51±2.01 ng/mL in the general control, 1.81±1.62 ng/mL in the positive control, 1.65±0.30 ng/mL in the negative control (OVX model + saline), and 4.35±0.56 ng/mL in the experimental group. (Figure 22). The testosterone concentration in the negative control group decreased rapidly, but increased to a level similar to that of the general control group in the experimental group taking the sample. Testosterone concentrations were not increased in the positive control group that consumed estradiol E2.
(11) 혈청 내 글루코스 농도 변화 분석(11) Analysis of changes in glucose concentration in serum
혈청 내 혈당 농도는 일반대조군에서 170.5±24.7 mg/dL, 양성대조군에서 156.5±34.6 mg/dL, 음성대조군(OVX모델+saline)에서 152.0±9.9 mg/dL, 실험군에서 157.5±2.1 mg/dL으로 측정되었다(도 23). 난소제거 처치 후 3개 그룹에서 모두 일반대조군의 혈당 농도보다 낮았으며, 양성대조군, 음성대조군, 실험군에서 유사한 농도를 보였다.Serum blood glucose levels were 170.5±24.7 mg/dL in the general control group, 156.5±34.6 mg/dL in the positive control group, 152.0±9.9 mg/dL in the negative control group (OVX model + saline), and 157.5±2.1 mg/dL in the experimental group. It was measured (Fig. 23). After ovarian removal, all three groups had lower blood sugar levels than the normal control group, and showed similar concentrations in the positive control group, the negative control group, and the experimental group.
(12) 혈청 내 간기능 지표 AST, ALT 활성 변화 분석(12) Analysis of AST and ALT activity changes in serum liver function indicators
혈청 내 간기능 지표 중 AST 활성은 일반대조군에서 170.5±24.7 U/L, 양성대조군에서 156.5±34.6 U/L, 음성대조군(OVX모델+saline)에서 152.0±9.9 U/L, 실험군에서 157.5±2.1 U/L으로 측정되었다(도 24). 양성대조군과 실험군에서 비교적 높은 AST 활성을 보였다.Among the indicators of liver function in serum, AST activity was 170.5±24.7 U/L in the general control group, 156.5±34.6 U/L in the positive control group, 152.0±9.9 U/L in the negative control group (OVX model + saline), and 157.5±2.1 in the experimental group. It was measured in U/L (FIG. 24). The positive control group and the experimental group showed relatively high AST activity.
혈청 내 간기능 지표 중 ALT 활성은 일반대조군에서 170.5±24.7 U/L, 양성대조군에서 156.5±34.6 U/L, 음성대조군에서 152.0±9.9 U/L, 실험군에서 157.5±2.1 U/L으로 측정되었다(도 25). 난소제거 처치한 양성대조군, 음성대조군, 실험군에서 비교적 높은 ALT 활성을 보였다.Among the indicators of liver function in serum, ALT activity was measured to be 170.5±24.7 U/L in the general control group, 156.5±34.6 U/L in the positive control group, 152.0±9.9 U/L in the negative control group, and 157.5±2.1 U/L in the experimental group. (Figure 25). The positive control group, the negative control group, and the experimental group treated with ovarian removal showed relatively high ALT activity.
(13) 혈청 내 지질대사 지표 변화 분석(13) Analysis of changes in lipid metabolism index in serum
혈청 내 지질대사 지표는 총 콜레스테롤(Total cholesterol), 중성지방(Triacylglycerol), 고밀도지단백 콜레스테롤(HDL-cholesterol), 저밀도지단백 콜레스테롤(LDL-cholesterol), 초저밀도지단백 콜레스테롤(VLDL-cholesterol)을 분석하였다. 혈청 내 총 콜레스테롤 농도는 일반대조군에서 74.5±13.4 mg/dL, 양성대조군에서 66.0±14.1 mg/dL, 음성대조군(OVX모델+saline)에서 83.0±2.8 mg/dL, 실험군에서 75.5±0.7 mg/dL으로 측정되었다(도 26). 음성대조군의 총 콜레스테롤 농도가 일반대조군에 비해 다소 증가되었지만, 에스트라디올 E2와 시료 음료를 섭취한 2개 그룹에서 총 콜레스테롤 농도가 감소되었다.Lipid metabolism indicators in serum were analyzed for total cholesterol, triacylglycerol, high density lipoprotein cholesterol (HDL-cholesterol), low density lipoprotein cholesterol (LDL-cholesterol), and ultra low density lipoprotein cholesterol (VLDL-cholesterol). The total cholesterol concentration in the serum was 74.5±13.4 mg/dL in the general control group, 66.0±14.1 mg/dL in the positive control group, 83.0±2.8 mg/dL in the negative control group (OVX model + saline), and 75.5±0.7 mg/dL in the experimental group. It was measured as (Fig. 26). The total cholesterol concentration of the negative control group was slightly increased compared to that of the general control group, but the total cholesterol concentration was decreased in the two groups ingesting estradiol E2 and the sample beverage.
혈청 내 중성지방 농도는 일반대조군에서 54.0±9.9 mg/dL, 양성대조군에서 43.0±0.0 mg/dL, 음성대조군에서 49.5±3.5 mg/dL, 실험군에서 54.0±4.2 mg/dL으로 측정되었다(도 27). 일반대조군과 실험군은 유사한 농도를 보였고 양성대조군과 음성대조군의 중성지방 농도는 일반대조군에 비해 낮은 것으로 나타났다.The triglyceride concentration in serum was measured to be 54.0±9.9 mg/dL in the general control group, 43.0±0.0 mg/dL in the positive control group, 49.5±3.5 mg/dL in the negative control group, and 54.0±4.2 mg/dL in the experimental group (FIG. 27 ). ). The normal control group and the experimental group showed similar concentrations, and the triglyceride concentrations of the positive control group and the negative control group were lower than those of the normal control group.
혈청 내 고밀도지단백 콜레스테롤 농도는 일반대조군에서 32.4±1.3 mg/dL, 양성대조군에서 42.0±4.2 mg/dL, 음성대조군에서 42.5±3.5 mg/dL, 실험군에서 45.0±0.0 mg/dL으로 측정되었다(도 28). 고밀도지단백 콜레스테롤 농도가 난소제거 처치한 3개 그룹에서 비교적 증가되었고 시료 음료를 섭취한 실험군에서 가장 높은 농도를 보였다.The high-density lipoprotein cholesterol concentration in serum was measured to be 32.4±1.3 mg/dL in the general control group, 42.0±4.2 mg/dL in the positive control group, 42.5±3.5 mg/dL in the negative control group, and 45.0±0.0 mg/dL in the experimental control group (FIG. 28). The concentration of high-density lipoprotein cholesterol was relatively increased in the three groups treated with ovary removal and showed the highest concentration in the experimental group consuming the sample beverage.
혈청 내 저밀도지단백 콜레스테롤 농도는 일반대조군에서 52.9±16.7 mg/dL, 양성대조군에서 32.6±18.4 mg/dL, 음성대조군에서 50.4±1.4 mg/dL, 실험군에서 41.3±0.1 mg/dL으로 측정되었다(도 29). 저밀도지단백 콜레스테롤 농도는 음성대조군에서 큰 변화를 보이지 않았지만 에스트라디올 E2와 시료 음료를 섭취한 2개 그룹에서 감소된 결과를 보였다.The low-density lipoprotein cholesterol concentration in serum was measured to be 52.9±16.7 mg/dL in the general control group, 32.6±18.4 mg/dL in the positive control group, 50.4±1.4 mg/dL in the negative control group, and 41.3±0.1 mg/dL in the experimental group (Fig. 29). The low-density lipoprotein cholesterol concentration did not show a significant change in the negative control group, but decreased in the two groups ingesting estradiol E2 and sample beverage.
혈청 내 초저밀도지단백 콜레스테롤 농도는 일반대조군에서 10.8±2.0 mg/dL, 양성대조군에서 8.6±0.0 mg/dL, 음성대조군에서 9.9±0.7 mg/dL, 실험군에서 10.8±0.8 mg/dL으로 측정되었다(도 30). 실험군에서는 큰 변화가 없었지만 음성대조군과 양성대조군에서 감소된 결과를 보였다.The serum low-density lipoprotein cholesterol concentration was measured to be 10.8±2.0 mg/dL in the general control group, 8.6±0.0 mg/dL in the positive control group, 9.9±0.7 mg/dL in the negative control group, and 10.8±0.8 mg/dL in the experimental group ( Fig. 30). Although there was no significant change in the experimental group, the result was decreased in the negative control group and the positive control group.
실시예Example 2: 갱년기 증상 개선효과 확인 2: Confirm the effect of improving menopausal symptoms
갱년기 증상을 나타내는 40~65세의 여성 60명을 각 6개의 군으로 나누어, 상기 제조예 1과 비교예들의 하수오 생약초 복합물을 각각 1일 2회 80 mL를 복용하게 하였고, 30일 복용한 후 주요 갱년기 증상에 대한 개선 정도를 10점 척도법으로 평가하게 하였다.Sixty women aged 40-65 who had menopausal symptoms were divided into six groups, and 80 ml of the herbal medicine complex of Preparation Example 1 and Comparative Examples were taken twice a day, respectively. The improvement of menopausal symptoms was evaluated by a 10-point scale.
주요 갱년기 증상인 안면홍조, 다한증, 오한증, 피로감, 우울감, 수면장애 및 불안감의 7가지 항목에 대한 개선 정도를 평가한 결과, 모든 항목에서 제조예 1의 하수오 생약초 복합물를 섭취한 갱년기 증상을 가진 여성이 8~9점대의 갱년기 증상 개선 효과를 보인 반면, 비교예들은 7점 이하의 점수를 보여, 제조예 1의 하수오 생약초 복합물이 여성 갱년기 증상 개선 효과가 우수함을 확인할 수 있었다.As a result of evaluating the degree of improvement for the seven items of major menopausal symptoms such as hot flashes, hyperhidrosis, chills, fatigue, depression, sleep disorders and anxiety, a woman with menopausal symptoms who consumed the sewage herbal remedies of Preparation Example 1 in all items was evaluated. While the menopausal symptom improvement effect was shown in the 8~9 range, the comparative examples showed a score of 7 or less, and it was confirmed that the sewage herbal remedies of Preparation Example 1 were excellent in the effect of improving menopausal symptoms.
실시예Example 3: 3: 하수오Ha Shou 생약초Herbal medicine 복합물의 관능검사 Sensory test of complex
40~65세의 여성 60명을 대상으로 제조예 1 및 비교예들의 하수오 생약초 복합물을 시음하게 하고 향, 맛, 쓴맛 및 종합 기호도 평가를 실시하여, 5점 척도법(1점: 매우 나쁨, 2점: 나쁨, 3점: 보통, 4점: 좋음, 5점: 매우 좋음)으로 점수를 매겨 그 결과를 표 3에 나타내었다. 쓴맛에 대한 기호도는 쓴맛의 정도에 따라 1점: 쓴맛이 거의 없음, 3점: 쓴맛이 보통, 5점: 쓴맛이 강함으로 평가하였다.A sample of 60 females aged 40 to 65 years old was sampled with the herbal extracts of Sewuo Herbal Medicine of Preparation Example 1 and Comparative Examples, and evaluated for flavor, taste, bitter taste, and overall preference. : Bad, 3 points: Normal, 4 points: Good, 5 points: Very good), and the results are shown in Table 3. The preference for bitterness was evaluated as 1 point: little bitterness, 3 points: moderate bitterness, 5 points: bitterness was strong, depending on the degree of bitterness.
표 3에서 알 수 있는 바와 같이, 제조예 1의 하수오 생약초 복합물이 쓴맛이 약하면서, 다른 비교예들의 하수오 생약초 복합물에 비해 향, 맛 및 종합 기호도에서 평가자들에 의해 더 선호되는 것을 확인할 수 있었다.As can be seen in Table 3, while the bitter taste of the herbal extracts of Preparation Example 1 was weak, it could be confirmed that the preferences were improved by the evaluators in flavor, taste, and overall preference compared to those of other Comparative Examples.
Claims (5)
(2) 상기 (1)단계의 제조한 하수오 발효 분말에 물을 첨가한 후 추출 및 여과하고 농축하여 하수오 발효 농축액을 제조하는 단계;
(3) 상기 (2)단계의 제조한 하수오 발효 농축액에 물, 에탄올, 아세토박터 아세티(Acetobacter aceti) 및 아세토박터 파스테리아누스(Acetobacter pasteurianus)를 첨가한 후 초산 발효하여 하수오 초산 발효물을 제조하는 단계;
(4) 홍화자, 속단, 당귀 및 오미자에 각각 물을 첨가한 후 추출 및 여과하여 홍화자 추출물, 속단 추출물, 당귀 추출물 및 오미자 추출물을 제조하는 단계: 및
(5) 상기 (3)단계의 제조한 하수오 초산 발효물과 상기 (4)단계의 제조한 속단 추출물, 홍화자 추출물, 당귀 추출물 및 오미자 추출물을 혼합한 혼합물을 가열하는 단계를 포함하여 제조하는 것을 특징으로 하는 하수오 혼합 음료의 제조방법.(1) Glucose (glucose) and sucrose (sucrose) were added to the sewage, and then added to the sterilized sewage mixture to the shiitake mycelium ( Lentinula edode s) after inoculation, drying and grinding the cultured sewage fermentation product to prepare a sewage fermentation powder;
(2) adding water to the sewage fermentation powder prepared in step (1), followed by extraction, filtration, and concentration to prepare a sewage fermentation concentrate;
(3) Water, ethanol, acetobacter aceti ( acetobacter ) in the sewage fermentation concentrate prepared in step (2) aceti ) and acetobacter pasteurianus , followed by acetic acid fermentation to produce sewage acetic acid fermentation product;
(4) Safflower, Sokdan, Angelica and Omija, respectively, after adding water and extracting and filtering to prepare Safflower extract, Sokdan extract, Angelica extract and Omija extract: and
(5) characterized in that it comprises a step of heating the mixture of the fermented sewage acetic acid prepared in step (3) and the Sokdan extract, safflower extract, Angelica extract and Omija extract prepared in step (4). Method for producing a mixed drink of sewage.
(1) 하수오에 글루코스(glucose) 및 수크로스(sucrose)를 첨가한 후 멸균한 하수오 혼합물에 표고균사체(Lentinula edodes)를 접종한 후 22~28℃에서 25~35일 동안 배양한 하수오 발효물을 75~85℃에서 10~14시간 동안 건조 및 분쇄하여 하수오 발효 분말을 제조하는 단계;
(2) 상기 (1)단계의 제조한 하수오 발효 분말에 물을 첨가한 후 60~100℃에서 1~15시간 동안 추출 및 여과하고 28~32 brix가 될 때까지 농축하여 하수오 발효 농축액을 제조하는 단계;
(3) 상기 (2)단계의 제조한 하수오 발효 농축액에 물, 에탄올, 아세토박터 아세티(Acetobacter aceti) 및 아세토박터 파스테리아누스(Acetobacter pasteurianus)를 첨가한 후 28~32℃에서 10~14일 동안 초산 발효하여 하수오 초산 발효물을 제조하는 단계;
(4) 홍화자, 속단, 당귀 및 오미자에 각각 물을 18~22배(v/w) 첨가한 후 70~90℃에서 2~4시간 동안 추출 및 여과하여 홍화자 추출물, 속단 추출물, 당귀 추출물 및 오미자 추출물을 제조하는 단계: 및
(5) 혼합물 총 중량 기준으로, 상기 (3)단계의 제조한 하수오 초산 발효물 24~28 중량%, 상기 (4)단계의 제조한 속단 추출물 16~20 중량%, 홍화자 추출물 16~20 중량%, 당귀 추출물 18~22 중량% 및 오미자 추출물 16~20 중량%를 혼합한 혼합물을 가열하는 단계를 포함하여 제조하는 것을 특징으로 하는 하수오 혼합 음료의 제조방법.According to claim 2,
(1) Glucose (glucose) and sucrose (sucrose) were added to the sewage, and then added to the sterilized sewage mixture to the shiitake mycelium ( Lentinula edode s) to inoculate the sewage fermentation product cultured for 22 to 28° C. for 25 to 35 days and incubated for 10 to 14 hours at 75 to 85° C. to prepare a sewage fermentation powder;
(2) After adding water to the sewage fermentation powder prepared in the above step (1), extracting and filtering at 60-100°C for 1-15 hours, and concentrating until 28 to 32 brix to prepare a sewage fermentation concentrate. step;
(3) After adding water, ethanol, acetobacter aceti and acetobacter pasteurianus to the sewage fermentation concentrate prepared in step (2), 10-14 days at 28~32℃ During the production of acetic acid fermentation product by acetic acid fermentation;
(4) After adding 18-22 times (v/w) of water to safflower, Sokdan, Angelica and Omija, respectively, extracting and filtering at 70~90℃ for 2-4 hours, and then filtering Safflower extract, Sokdan extract, Angelica extract and Omija Preparing an extract: and
(5) Based on the total weight of the mixture, 24 to 28% by weight of fermented sewage acetate produced in step (3), 16 to 20% by weight of extract from Sokdan prepared in step (4), 16 to 20% by weight of safflower extract , 18-22% by weight of Angelica extract and 16-20% by weight of Omija extract.
(1) 하수오 450~550 g에 하수오 중량 대비 글루코스(glucose) 0.8~1.2% 및 수크로스(sucrose) 0.8~1.2%를 첨가한 후 110~130℃에서 10~20분 동안 멸균한 하수오 혼합물에 표고균사체(Lentinula edodes)를 접종한 후 22~28℃에서 25~35일 동안 배양한 하수오 발효물을 75~85℃에서 10~14시간 동안 건조 및 분쇄하여 하수오 발효 분말을 제조하는 단계;
(2) 상기 (1)단계의 제조한 하수오 발효 분말에 물을 18~22배(v/w) 첨가한 후 60~100℃에서 1~15시간 동안 추출 및 여과하고 28~32 brix가 될 때까지 농축하여 하수오 발효 농축액을 제조하는 단계;
(3) 상기 (2)단계의 제조한 하수오 발효 농축액에 물, 에탄올, 아세토박터 아세티(Acetobacter aceti) 및 아세토박터 파스테리아누스(Acetobacter pasteurianus)를 첨가한 후 28~32℃에서 10~14일 동안 초산 발효하여 하수오 초산 발효물을 제조하는 단계;
(4) 홍화자, 속단, 당귀 및 오미자에 각각 물을 18~22배(v/w) 첨가한 후 70~90℃에서 2~4시간 동안 추출 및 여과하여 홍화자 추출물, 속단 추출물, 당귀 추출물 및 오미자 추출물을 제조하는 단계: 및
(5) 혼합물 총 중량 기준으로, 상기 (3)단계의 제조한 하수오 초산 발효물 24~28 중량%, 상기 (4)단계의 제조한 속단 추출물 16~20 중량%, 홍화자 추출물 16~20 중량%, 당귀 추출물 18~22 중량% 및 오미자 추출물 16~20 중량%를 혼합한 혼합물을 가열하는 단계를 포함하여 제조하는 것을 특징으로 하는 하수오 혼합 음료의 제조방법.According to claim 3,
(1) After adding 0.8 to 1.2% of glucose and 0.8 to 1.2% of sucrose to 450 to 550 g of sewage weight, the mixture is sterilized at 110 to 130°C for 10 to 20 minutes, and then elevation is applied to the sewage mixture. Mycelium ( Lentinula edode s) to inoculate the sewage fermentation product cultured for 22 to 28° C. for 25 to 35 days and incubated for 10 to 14 hours at 75 to 85° C. to prepare a sewage fermentation powder;
(2) When 18-22 times (v/w) water is added to the sewage fermentation powder prepared in the step (1), extraction and filtration at 60-100°C for 1-15 hours, and when it becomes 28-32 brix Concentrating to prepare a sewage fermentation concentrate;
(3) Water, ethanol, acetobacter aceti ( acetobacter ) in the sewage fermentation concentrate prepared in step (2) aceti ) and acetobacter pasteurianus , followed by acetic acid fermentation at 28-32° C. for 10-14 days to produce a fermented sewage acetic acid;
(4) After adding 18-22 times (v/w) of water to safflower, Sokdan, Angelica and Omija, respectively, extracting and filtering at 70~90℃ for 2-4 hours, and then filtering Safflower extract, Sokdan extract, Angelica extract and Omija Preparing an extract: and
(5) Based on the total weight of the mixture, 24 to 28% by weight of fermented sewage acetate produced in step (3), 16 to 20% by weight of extract from Sokdan prepared in step (4), 16 to 20% by weight of safflower extract , 18-22% by weight of Angelica extract and 16-20% by weight of Omija extract.
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