KR20030024934A - Ascorbic acid derivatives with antioxidnat activity and high stability and preparation method for the same - Google Patents
Ascorbic acid derivatives with antioxidnat activity and high stability and preparation method for the same Download PDFInfo
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Abstract
Description
본 발명은 항산화 효과 및 높은 안정성을 갖는 아스코르브산 유도체 및 그것의 제조방법에 관한 것이다.The present invention relates to an ascorbic acid derivative having an antioxidant effect and high stability and a method for producing the same.
아스코르브산은 매우 높은 항산화력을 갖는 천연 항산화제로서, 토코페롤의 산화를 방지하는 2차 항산화제의 기능도 있다. 또한, 아스코르브산은 치아, 잇몸, 뼈의 필요성분이며, 감염에 대한 저항성과 면역성을 제공하고, 괴혈병과 감기를 예방하며, 철분 흡수에도 도움을 주어 빈혈을 예방한다고 보고되고 있다.Ascorbic acid is a natural antioxidant with very high antioxidant power, and also has the function of a secondary antioxidant that prevents the oxidation of tocopherol. In addition, ascorbic acid is a necessary component of teeth, gums and bones, has been reported to provide resistance and immunity to infections, to prevent scurvy and cold, and to help iron absorption to prevent anemia.
그러나 아스코르브산은 안정성이 매우 낮아 이를 증가시키기 위한 여러 방법들이 고안되어오고 있다. 그중 당전이를 통해 안정화시키는 방법으로는 아스코르브산과 글루코스의 1,6결합과 1,2결합 물질인 α-1,6-글루코실 아스코르베이트와 α-1,2-글루코실 아스코르베이트가 있으며, 아스코르브산과 갈락토오스의 1,6결합인 α-1,6-갈락토실 아스코르베이트가 있다. 1,6결합 아스코르브산 유도체는 아스코르브산 보다 다소의 안정성을 가지고, 항산화력이 있는 반면, 1,2-결합 아스코르브산 유도체는 매우 높은 안정성이 있으나, 아스코르브산의 항산화력을 가지는 사이클로펜틸링에 직접 결합하여 항산화력이 없어지는 문제점이 있다.However, ascorbic acid is very stable, and various methods have been devised to increase it. Among them, the methods of stabilizing by sugar transfer include α-1,6-glucosyl ascorbate and α-1,2-glucosyl ascorbate, which are 1,6 and 1,2 bonds of ascorbic acid and glucose. , Α-1,6-galactosyl ascorbate, which is the 1,6 bond of ascorbic acid and galactose. 1,6-linked ascorbic acid derivatives are somewhat more stable and more antioxidant than ascorbic acid, whereas 1,2-linked ascorbic acid derivatives have very high stability, but are directly directed to cyclopentyling, which has the antioxidant power of ascorbic acid. There is a problem in that the antioxidant power is lost.
따라서, 아스코르브산의 항산화력을 가지면서, 높은 안정성을 갖는 아스코르브산 유도체 및 그 제조방법을 개발할 필요가 있다.Accordingly, there is a need to develop ascorbic acid derivatives having high stability while having ascorbic acid and a method for producing the same.
본 발명의 목적은 항산화 효과뿐만 아니라 안정성이 높은 아스코르브산 유도체를 제공하는 것이다.It is an object of the present invention to provide ascorbic acid derivatives having high stability as well as antioxidant effects.
본 발명의 또다른 목적은 항산화 효과뿐만 아니라 안정성이 높은 아스코르브산 유도체를 제조하는 방법을 제공하는 것이다.It is another object of the present invention to provide a method for preparing ascorbic acid derivatives having high stability as well as an antioxidant effect.
도 1은 α-1,6-말토실 아스코르베이트 생산액의 박막크로마토그라피 사진이다. 도면에서 AA는 아스코르브산의 약자이고, G1-AA는 글루코실 아스로브베이트, G2-AA는 말토실 아스코르베이트를 말한다.1 is a thin film chromatography photograph of α-1,6-maltosyl ascorbate production solution. In the figure, AA stands for ascorbic acid, G1-AA refers to glucosyl aslovate, and G2-AA refers to maltosyl ascorbate.
도 2는 α-1,6-말토실 아스코르베이트 생산액의 고성능 액체크로마토그램이다.2 is a high performance liquid chromatogram of α-1,6-maltosyl ascorbate production liquid.
도 3은 α-1,6-말토실 아스코르베이트의 FAB-MASS 스펙트럼이다.3 is a FAB-MASS spectrum of α-1,6-maltosyl ascorbate.
도 4는 α-1,6-말토실 아스코르베이트의13C 핵자기공명분석 스펙트럼이다.4 is a 13 C nuclear magnetic resonance spectrum of α-1,6-maltosyl ascorbate.
도 5는 α-1,6-말토실 아스코르베이트의 구리이온에 대한 안정성을 나타내는 그래프이다.FIG. 5 is a graph showing stability of copper ions of α-1,6-maltosyl ascorbate. FIG.
도 6은 α-1,6-말토실 아스코르베이트의 아스코르브산 산화효소에 대한 안정성을 나타내는 그래프이다.6 is a graph showing the stability of ascorbic acid oxidase of α-1,6-maltosyl ascorbate.
본 발명은 하기 화학식 1을 갖는 아스코르브산 유도체에 관한 것이다.The present invention relates to an ascorbic acid derivative having the formula (1).
본 발명에 따른 화합물은 우수한 안정성을 갖는, 항산화제로 사용될 수 있다.The compounds according to the invention can be used as antioxidants with good stability.
본 발명은 또한, 당전이성 아밀라제에 의해 아스코르브산에 말토스를 전이시킨 α-1,6-말토실 아스코르베이트를 제조하는 방법에 관한 것이다.The present invention also relates to a method for producing α-1,6-maltosyl ascorbate in which maltose is transferred to ascorbic acid by sugar transfer amylase.
이하, 본 발명에 대해서 상세히 설명한다.EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.
본 발명에서 사용하는 당전이성 아밀라제는 전분을 가수분해하여 주로 말토스를 생산하고, 풀루란을 분해하여 주로 판노스를 생성하며, 사이클로덱스트린을 분해하여 주로 말토스를 생성하며 전분을 분해하며 아카비오신-글루코스를 단당류 또는 이당류 이상의 올리고당에 전이하는 효소를 의미한다.The glycotransfer amylase used in the present invention hydrolyzes starch to produce maltose, and mainly decomposes pullulan to produce pannose, and to decompose cyclodextrin to produce maltose mainly to decompose starch, It refers to an enzyme that transfers the on-glucose to the oligosaccharides of the monosaccharide or the disaccharide or more.
본 발명자들은 이와 같은 효소에 대하여 한국특허 공개공보 제 2000-25033호에서 밝힌 바 있으며, 이 특허출원의 기재 내용은 본 명세서에 포함된다. 예를 들면, 한 예로, 한국특허공고공보 99-186716에 기재된, 바실러스스테아로써머필러스(KFCC-10896)부터 생산되는 아밀라제(이하 BSMA라 함)를 사용하여 탄수화물분해효소를 저해시키는 화합물을 제조할 수 있다. 또다른 예로, 미국특허 5,583,039에 기재된 바실러스 리케니포미스 (Bacillus licheniformis,ATCC 27811), 한국특허 공개공보 제 2000-47164호에 기재된Thermussp. (IM6501 KCTC 0527BP)에서 분리된 아밀라제(이하 ThMA라 함) 등이 상기 당전이성 아밀라제에 속하며, 본 발명의 화합물을 제조하기 위하여 사용될 수 있다.The present inventors have disclosed in Korean Patent Laid-Open Publication No. 2000-25033 for such an enzyme, and the description of this patent application is included herein. For example, a compound for inhibiting carbohydrate degrading enzyme is prepared by using amylase (hereinafter referred to as BSMA) produced from Bacillus stearer merphilus (KFCC-10896) described in Korean Patent Publication No. 99-186716. can do. As another example, Bacillus licheniformis ( ATCC 27811) described in US Pat. No. 5,583,039, Thermus sp. Amylase (hereinafter referred to as ThMA) isolated from (IM6501 KCTC 0527BP) belongs to the glycotransfer amylase and can be used to prepare the compounds of the present invention.
본 발명의 바람직한 방법에서, 전분, 말토트리오스, 또는 말토덱스트린등의 기질에 당전이성 아밀라제를 반응시키면 기질이 분해되어 말토스가 생성되고 이 말토스가 아스코르브산에 전이되어 α-1,6-말토실 아스코르베이트를 효과적으로 생산할 수 있다.In a preferred method of the present invention, the reaction of glycotransfer amylase with a substrate such as starch, maltotriose, or maltodextrin decomposes the substrate to produce maltose, which is transferred to ascorbic acid, and α-1,6- Maltosyl ascorbate can be produced effectively.
본 발명에서 사용가능한 기질로는 당전이성 아밀라제에 의해 말토스를 생성가능한 기질이며 모두 가능하며, 특히 예를 들면 전분, 말토트리오스 또는 말토덱스트린등이 포함된다. 따라서 본 발명의 방법은 아스코르브산 존재하에서 전분에 당전이성 아밀라제를 반응시켜 이루어질 수 있고, 또한 전분에 아밀라제를 반응시켜 얻어지는 산물인 말토스를 아스코르브산 존재하에서 당전이성 아밀라제로 반응시킬 수도 있다.Substrates usable in the present invention are all substrates capable of producing maltose by glycotransfer amylases, and in particular, include, for example, starch, maltotriose or maltodextrin. Therefore, the method of the present invention may be achieved by reacting sugar-transfer amylase with starch in the presence of ascorbic acid, and maltose, which is a product obtained by reacting amylase with starch, may be reacted with sugar-transfer amylase in the presence of ascorbic acid.
상기 방법에 따라 제조된 말토실-아스코르베이트는 박막크로마토그래피, 핵자기공명분석 스펙트럼, 분자량 측정 및 고성능 액체크로마토그래피로 확인한다.Maltosyl-ascorbate prepared according to the above method is confirmed by thin layer chromatography, nuclear magnetic resonance spectra, molecular weight measurement and high performance liquid chromatography.
본 발명의 화합물을 확인하기 위해서 박막크로마토그래피를 수행한 결과, 도 1에서 첫 번째 줄은 아스코르브산, 두 번째 줄은 당표준시약, 세 번째 줄은 실시예1의 반응생성물, 네 번째 줄은 실시예 1에서 얻은 반응생성물을 이온교환수지와 젤 투과 크로마토그래피로 정제하여 얻은 화합물중 α-1,6-글루코실 아스코르베이트 이며, 다섯 번째 줄은 정제된 α-1,6-말토실 아스코르베이트이다. 세 번째 줄에서 알 수 있듯이, U.V.에서 아스코르브산이 전이된 두 가지 새로운 물질이 생성된 것이 확인되었다. α-1,6 결합은 핵자기공명분석 스펙트럼을 이용하여 확인하였다.As a result of performing thin layer chromatography to identify the compound of the present invention, in FIG. 1, the first line is ascorbic acid, the second line is a sugar standard reagent, and the third line is a reaction product of Example 1, and the fourth line is performed. The reaction product obtained in Example 1 was α-1,6-glucosyl ascorbate among the compounds obtained by purification by ion exchange resin and gel permeation chromatography, and the fifth line was purified α-1,6-maltosyl ascorbic acid. It's a bait. As can be seen from the third line, two new substances from U.V. to which ascorbic acid was transferred were identified. α-1,6 binding was confirmed using nuclear magnetic resonance spectra.
본 발명에 따른 화합물의 안정성을 측정한 결과, α-1,6-글루코실 아스코르베이트는 아스코르브산보다 다소의 안정성을 보이나, α-1,6-말토실 아스코르베이트는 아스코르브산보다 매우 높은 안정성을 보임을 알 수 있다.As a result of measuring the stability of the compound according to the present invention, α-1,6-glucosyl ascorbate showed more stability than ascorbic acid, but α-1,6-maltosyl ascorbate was much higher than ascorbic acid. It can be seen that the stability is shown.
아래에서 실시예를 통하여 본 발명을 설명하고자 하나, 이들 실시예에 의하여 본 발명의 범위가 한정되지는 않는다.Hereinafter, the present invention will be described through examples, but the scope of the present invention is not limited by these examples.
[실시예]EXAMPLE
실시예 1. α-1,6-말토실 아스코르베이트의 제조 및 정제Example 1. Preparation and Purification of α-1,6-maltosyl ascorbate
대한민국 서울특별시 서대문구 신촌동 134번지에 위치하는 사단법인 한국종균협회에 수탁번호 KFCC-10896으로 기탁된,Bacillus stearothermophilus균주를 LB 배지 (0.5 % 효모 추출물, 1 % 박토 트립톤, 그리고 0.5 % NaCl) 에 접종하여 37℃에서 전배양한 후 같은 배지에서 10시간 배양하고 원심분리하여 균체를 얻었다. 회수한 균체를 pH 7.5의 50 mM Tris-HCl 완충용액으로 현탁시킨 다음 초음파로 파괴한 뒤 원심분리하여 상등액을 취하였다. 상등액을 컬럼크로마토그래피로 Ni-NTA 컬럼을 통과시켜 정제된 아밀라제를 얻었다. Bacillus stearothermophilus strain, deposited with KFCC-10896, with the accession No. KFCC-10896, to the Korean Breeders Association located at 134, Sinchon-dong, Seodaemun-gu, Seoul, Korea After incubation at 37 ℃ incubated for 10 hours in the same medium and centrifuged to obtain the cells. The recovered cells were suspended in 50 mM Tris-HCl buffer at pH 7.5 and then destroyed by ultrasonication and centrifuged to obtain supernatant. The supernatant was passed through a Ni-NTA column by column chromatography to obtain purified amylase.
50 mM 시트레이트 완충용액(pH 6.0)에 전분 10%(w/v)와 아스코르브산60%(w/v)를 용해시켜 기질로 사용하였다. 기질 용액을 미리 55 ℃ 수조에 10분간 방치한 후 전분 1 mg 당 1 U의 BSMA를 첨가하여 48시간 동안 반응시켰다. 반응 후 반응액을 pH 2.0으로 적정하여 냉장보관하였다. 아스코르브산의 음전하를 이용하여 반응액을 이온 교환수지를 통과시켜 두가지 생성물과 아스코르브산을 다른 부산물과 분리하고, BioGel P-2 컬럼(1.6×90cm)을 통과시켜 반응생성물을 분리정제하였다.10% (w / v) of starch and 60% (w / v) of ascorbic acid were dissolved in 50 mM citrate buffer (pH 6.0) and used as a substrate. The substrate solution was previously left in a 55 ° C. water bath for 10 minutes and then reacted for 48 hours by adding 1 U of BSMA per 1 mg of starch. After the reaction, the reaction solution was titrated to pH 2.0 and refrigerated. Using the negative charge of ascorbic acid, the reaction solution was passed through an ion exchange resin to separate the two products and ascorbic acid from other byproducts, and the reaction product was separated and purified through a BioGel P-2 column (1.6 × 90 cm).
실시예 2: 박막크로마토그라피를 이용한 반응생성물의 분석Example 2 Analysis of Reaction Products Using Thin Film Chromatography
실시예 1의 반응에서 얻어진 반응생성물을 분석하기 위해서 박막크로마토그라피를 수행하였다. 반응 종결 후, 각 시료를 Whatman K5F TLC 플레이트(20cm×20cm)에 1μL씩 로딩하여 n-부탄올: 아세트산: 증류수 3:1:1 전개액에서 전개시켰다. 전개 후에 플레이트를 잘 건조시켜 먼저 254nm U.V.에서 확인하고, 메탄올에 0.3%(w/v) N-(1-나프틸)-에틸렌디아민과 5%(v/v) 황산을 용해시킨 발색액에 담갔다가 꺼낸 후 110℃ 오븐에서 10 분 동안 발색시켰다. 얻어진 결과를 도 1에 나타냈다. 박막크로마토그라피를 이용하여 반응생성물을 분석하였으며 그 결과를 도 1에 나타낸다. 세 번째 줄에서 알 수 있듯이, U.V.에서 아스코르브산이 전이된 두 가지 새로운 물질이 생성된 것이 확인되었다.Thin layer chromatography was performed to analyze the reaction product obtained in the reaction of Example 1. After completion of the reaction, each sample was loaded into Whatman K5F TLC plate (20 cm × 20 cm) by 1 μL and developed in n-butanol: acetic acid: distilled water 3: 1: 1 developing solution. After development, the plate was well dried, first checked at 254 nm UV, and then immersed in a coloring solution in which 0.3% (w / v) N- (1-naphthyl) -ethylenediamine and 5% (v / v) sulfuric acid were dissolved in methanol. After evaporation, the color was developed for 10 minutes in an oven at 110 ° C. The obtained result is shown in FIG. The reaction product was analyzed using thin layer chromatography and the results are shown in FIG. 1. As can be seen from the third line, two new substances from U.V. to which ascorbic acid was transferred were identified.
실시예 3. 고성능액체크로마토그라피를 이용한 α-1,6-말토실 아스코르베이트의 분석Example 3 Analysis of α-1,6-maltosyl Ascorbate Using High Performance Liquid Chromatography
실시예 1에서 얻은 생성물을 고성능액체크로마토그라피(High Performance Liquid Chromatography, HPLC)를 이용하여 분석하였다. 시료는 실시예 1에서 얻은반응생성물을 이온교환수지와 젤 투과 크로마토그래피로 정제하여 얻은 분리된 화합물을 초순수로 희석하여 0.45μm 박막여과지로 여과하여 20 μL를 주입하였고 유속은 0.7 mL/분으로 하여 분석하였다. HPLC 분석은 삼성사(한국)의 SLC-100 그래디언트 펌프와 자외선 검출기 및 C18 컬럼을 이용하여 시행하였으며, 용매는 100 mM(pH 2.0) 칼륨인산완충용액을 사용하였다. 검출파장은 265nm에서 시행하였으며, 결과를 도 2에 나타낸다. 피크 1은 아스코르브산, 피크 2는 α-1,6-글루코실 아스코르베이트, 피크 3은 α-1,6-말토실 아스코르베이트를 나타낸다.The product obtained in Example 1 was analyzed using High Performance Liquid Chromatography (HPLC). The sample was purified by ion exchange resin and gel permeation chromatography from the reaction product obtained in Example 1, diluted with ultrapure water, filtered through 0.45μm thin film filter, and injected into 20μL, and the flow rate was 0.7mL / min. Analyzed. HPLC analysis was performed using SLC-100 gradient pump, UV detector and C18 column of Samsung (Korea), and 100 mM (pH 2.0) potassium phosphate buffer solution was used as a solvent. The detection wavelength was performed at 265 nm, and the results are shown in FIG. 2. Peak 1 represents ascorbic acid, peak 2 represents α-1,6-glucosyl ascorbate, and peak 3 represents α-1,6-maltosyl ascorbate.
실시예 4. FAB-MASS(Xe-gas)를 이용한 α-1,6-말토실 아스코르베이트 의 분자량 측정Example 4 Determination of Molecular Weight of α-1,6-maltosyl Ascorbate Using FAB-MASS (Xe-gas)
FAB-MASS(Xe-gas)를 이용하여, 상기 실시예 3에서 얻어진 도 1의 래인 4와 5에 나타난 화합물의 분자량을 측정하였다. 시스템은 JMS-LCmate(일본 JEOL사)를 사용하였으며, 글리세롤을 매트릭스로 사용하였다. 그 실험결과를 도 3a 및 도 3b에 나타냈다.The molecular weight of the compounds shown in lanes 4 and 5 of Figure 1 obtained in Example 3 was measured using FAB-MASS (Xe-gas). The system used JMS-LCmate (Jeol, Japan) and glycerol was used as a matrix. The experimental results are shown in FIGS. 3A and 3B.
도 3a에서 α-1,6-글루코실 아스코르베이트는 361 ([M+H]+)와 383 ([M+Na]+)에 나타나는 이온 피크로부터 시료의 분자량이 360으로 측정되었고, 도 3b에서 α-1,6-말토실 아스코르베이트 는 523 ([M+H]+)와 545 ([M+Na]+)에 나타나는 이온 피크로부터 시료의 분자량이 522으로 측정되었다. 이것은 각각 글루코실 아스코르브산와 말토실 아스코르브산 수소와 소디움 글루코실과 말토실 아스코르브산의 분자량과 일치하였으며, 이로부터 실시예 1에 의한 주요 화합물이 글루코실 아스코르브산과 말토실 아스코르브산임을 확인하였다.In FIG. 3A, α-1,6-glucosyl ascorbate has a molecular weight of 360 measured from the ion peaks shown at 361 ([M + H] + ) and 383 ([M + Na] + ), and FIG. 3B. In α-1,6-maltosyl ascorbate, the molecular weight of the sample was determined to be 522 from the ionic peaks appearing at 523 ([M + H] + ) and 545 ([M + Na] + ). This coincided with the molecular weights of glucosyl ascorbic acid, maltosyl hydrogen ascorbate, sodium glucosyl and maltosyl ascorbic acid, respectively. From this, it was confirmed that the main compounds according to Example 1 were glucosyl ascorbic acid and maltosyl ascorbic acid.
실시예 5:Example 5: 1313 C NMR을 이용한 α-1,6-말토실 아스코르베이트 의 구조 분석Structural Analysis of α-1,6-maltosyl Ascorbate Using C NMR
본 발명의 주요 전이물인 α-1,6-글루코실과 말토실 아스코르베이트의 구조를 확인하기 위하여13C 핵자기공명분석 (NMR; JNM LA-400 FT-NMR 스펙트로미터(Jeol, 일본))을 수행하였다. 시료는 실시예 1에서 순수 정제한, α-1,6-글루코실 아스코르베이트 와 α-1,6-말토실 아스코르베이트 20mg을 D2O에 각각 용해시킨 것을 사용하였다. 결과를 표 1 및 도 4에 나타낸다. α-1,6-글루코실 아스코르베이트왁 α-1,6-말토실 아스코르베이트는 DEPT를 통해서 아스코르브산의 6번 탄소가 이동하였음을 알 수 있었다. 13 C nuclear magnetic resonance analysis (NMR; JNM LA-400 FT-NMR spectrometer (Jeol, Japan)) to confirm the structure of α-1,6-glucosyl and maltosyl ascorbate, the main transitions of the present invention Was performed. As a sample, 20 mg of α-1,6-glucosyl ascorbate and α-1,6-maltosyl ascorbate purified in Example 1 were dissolved in D 2 O, respectively. The results are shown in Table 1 and FIG. 4. α-1,6-glucosyl ascorbate Wax α-1,6-maltosyl ascorbate was found to move 6 carbon of ascorbic acid through DEPT.
상기 표에서, AA는 아스코르브산의 약자이고, G1-AA는 글루코실 아스로브베이트, G2-AA는 말토실 아스코르베이트를 말한다. 그리고 고리 A 및 B는 아스코르브산의 고리를 말한다.In the above table, AA stands for ascorbic acid, G1-AA refers to glucosyl aslovate, and G2-AA refers to maltosyl ascorbate. And rings A and B refer to rings of ascorbic acid.
실시예 6: α-1,6-말토실 아스코르베이트의 안정성 측정Example 6 Determination of Stability of α-1,6-maltosyl Ascorbate
안정성 측정은 구리이온에 대한 안정성과 아스코르브산 산화제에 대한 안정성을 측정하였다. 구리이온에 대한 안정성 측정시 구리이온의 농도는 10μM을 사용하였고, 아스코르브산 산화효소에 대한 안정성 측정시는 아스코르브산 산화효소 200mU를 사용하였으며, 각 시료는 pH5.6 아세트산 나트륨에 용해시켜 측정하였다. 실험결과를 도 5에 나타냈다. α-1,6-글루코실 아스코르베이트는 아스코르브산보다 다소의 안정성을 보이나, α-1,6-말토실 아스코르베이트는 아스코르브산보다 매우 높은 안정성을 보임을 알 수 있다.Stability measurements measured stability to copper ions and ascorbic acid oxidizer. 10 μM of copper ion was used for measuring stability against copper ions, and 200 mU of ascorbic oxidase was used for measuring stability against ascorbic acid oxidase. Each sample was measured by dissolving in pH5.6 sodium acetate. The experimental results are shown in FIG. 5. Although α-1,6-glucosyl ascorbate shows more stability than ascorbic acid, α-1,6-maltosyl ascorbate has much higher stability than ascorbic acid.
실시예 7: α-1,6-말토실 아스코르베이트의 항산화력 측정Example 7 Determination of Antioxidant Capacity of α-1,6-maltosyl Ascorbate
항산화력 측정은 DPPH(1,1-Dipheny-1-picrylhydrazyl radical)의 수소와 화합하는 활성을 이용하여 측정하였다. DPPH를 99%에탄올에 1.0x10-4M로 만들고, 시료농도는 20mM이 되도록 한다. 암실에서 30분간 반응시킨후 517nm에서 흡광도 차이를 측정하였고, 각 측정값은 3번 반복치의 평균이며, SD는 표준편차를 나타낸다. 측정값은 표2과 같다.Antioxidant activity was measured using the activity of DPPH (1,1-Dipheny-1-picrylhydrazyl radical) in combination with hydrogen. Make DPPH 1.0x10 -4 M in 99% ethanol and make the sample concentration 20mM. After reacting for 30 minutes in the dark room, the difference in absorbance was measured at 517 nm. The measured values are shown in Table 2.
본 발명에 의해 항산화력을 가지는 동시에 높은 안정성을 가지는 α-1,6-말토실 아스코르베이트를 제공할 수 있다.According to the present invention, α-1,6-maltosyl ascorbate having antioxidant stability and high stability can be provided.
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