JP2022114263A - Method for evaluating the anti-glycation properties of wine - Google Patents

Abstract

To obtain a result proved by actual feeling of a consumer in anti-glycation evaluation of wine.SOLUTION: A method of evaluating anti-glycation of wine includes steps in which: (step 1) a subject ingests wine; (step 2) information on mind and body of the subject is acquired from the subject by questionnaire; and (step 3) saccharification stress marker and/or stress of the subject are/is quantitatively measured by an invasive method.SELECTED DRAWING: Figure 1

Description

The present invention relates to a method for evaluating the anti-glycation properties of wine. The method includes performing a non-invasive measurement on a wine taster. The method includes an in vitro antiglycation evaluation step for the test wine.

Glycation is a non-enzymatic chemical reaction between amino acids/proteins and reducing sugars, and is also called Maillard reaction. Saccharification has attracted attention in the field of food chemistry because it is a reaction that occurs during heating of food, changes in color, aroma and flavor, and decreases nutritional value during storage. In addition, measurement of glycation reaction product HbA1c in human blood is clinically applied in the field of diabetes treatment as a blood sugar control index. It is known that glycation occurs not only in hemoglobin but also in various proteins in vivo, leading to the formation of advanced glycation endproducts (AGEs) via various intermediates centered on carbonyl compounds.

Studies over the past 20 years or so have revealed that glycation that progresses in the body is associated with various diseases, and that natural products contain compounds that have the function of suppressing glycation. Non-Patent Document 1 describes the relationship between aging phenomena such as arteriosclerosis, diabetes, Alzheimer's disease, osteoporosis, renal disease, cataracts, and skin wrinkles, and glycation, as well as anti-glycation substances contained in foods. summarized.

Recently, many attempts have been made to prevent aging and diseases caused by glycative stress by using food materials with high anti-glycation activity. Attempts have been made to search for more useful anti-glycation substances using the evaluation method described in Non-Patent Document 2.

Among them, wine is attracting attention as a food that is expected to have anti-aging properties while being a luxury item. Until now, there have been various research results regarding anti-glycation substances derived from wine grape varieties, such as those described in Non-Patent Documents 3 to 8 and Patent Document 1.

However, for consumers who purchase and drink wine in anticipation of anti-aging, the in vitro anti-glycation evaluation results described in the prior art documents are not familiar information. Consumers are interested in whether drinking wine has an anti-aging effect on their bodies. Rather than reading the research results of experts published in specialized journals and books, I would like to think that "drinking wine made *** better." There has been a change." The realization and experiences of the customers will increase the desire to buy wine. Wine is not only a health food, but also a traditional luxury item. Consumers' motives for choosing and purchasing wine are not as simple as when choosing medicines or health foods based on the label of efficacy and effects. “Good for the body and delicious” is the motivation for consumers to purchase.

Up until now, the parties that provide wine to the market and consumers have actively carried out sales promotion activities such as tastings at sales floors, winery tours, and wine seminars in order to let consumers experience the taste of wine. However, there have been few opportunities to detect and analyze the anti-glycation effect that occurred in consumers who actually drank wine, and to utilize the analysis results in wine sales promotion activities. Although there are many expert analysis results such as the above-mentioned prior art, they do not touch the consumer's real feeling. For this reason, there was a lack of information directly useful for proposing and predicting the wines that consumers would like.

"Disease due to glycation and anti-glycation foods/materials popular version" CMC Publishing, 2016, ISBN: 978-4-7813-1127-2 "Evaluation of glycative stress and antiglycation effect" Oleoscience Vol. 18 No. 2 (2018), Online ISSN: 2087-3461 “Development of Food Materials by New Extraction Method of Functional Ingredients of Yamabudo” Shokuhin Kogyo, Vol. 51, No. 12 (2008), ISSN: 0559-8990 "Search for anti-glycation substances contained in wine press residue" Journal of Japan Grape and Wine Society, Vol. 23, No. 2, pp. 96-97 (2012), ISSN: 1342-2324 "Search for anti-glycation substances contained in pinot noir wine press residue" Journal of Japan Grape and Wine Society, Vol. 25, No. 2, pp. 79-80 (2014), ISSN: 1342-2324 "Search for AGEs Production Inhibitors Contained in Merlot Wine" Journal of the Japan Grape and Wine Society, Vol. 27, No. 2, pp. 62-63 (2016), ISSN: 1342-2324 "Search for anti-glycation substances contained in Cabernet Sauvignon wine" Journal of the Japan Society of Grape and Wine, Vol. 29, No. 2, pp. 100-101 (2018), ISSN: 1342-2324 "Characterization of phenolics of recovered red grape skins from winemaking by-products and their in vitro reducing capacity and anti-glycation activity" Bioresource Technology, Vol. 140, pp. 263-268 (2013), ISSN: 0960-8524

JP 2016-138042 A

Thus, no case has yet been found in which the anti-glycation properties of wine have been examined and evaluated from both scientific objectivity and consumer subjectivity. Therefore, the present inventor sought a new means for evaluating the anti-saccharification properties of wine, based on the actual experience of consumers, which is likely to gain the sympathy of consumers.

As a result, the present inventor found a method combining a questionnaire and non-invasive measurement as a method for evaluating the anti-saccharification properties of wine. That is, the present invention is as follows.
(Invention 1) A method for evaluating anti-saccharification properties of wine, comprising the following steps. (Step 1) A step in which a subject ingests wine. (Step 2) A step of obtaining information on the subject's mind and body from the subject through a questionnaire. (Step 3) A step of quantitatively measuring glycative stress markers and/or stress in the subject by a non-invasive method.
(Invention 2) The method of Invention 1, wherein the mental and physical information in step 2 is information on one or more selected from mood, sleep, bowel movements, and eating habits.
(Invention 3) The method of Invention 1, wherein in step 3, one or more selected from skin AGEs, body AGEs, heart rate variability, and skin condition is measured by a noninvasive method.
(Invention 4) The method of Invention 1, wherein in the step 2 and/or the step 3, a terminal and/or a measuring device connected to the Internet are used.
(Invention 5) (Step 4) The method of Invention 1, further comprising the step of evaluating the in vitro anti-glycation effect of the wine using an automated device.

The method of evaluating the anti-glycation properties of wine of the present invention is an evaluation method that ensures scientific objectivity and takes into consideration the subject's feelings and subjectivity. According to the method of the present invention, it is possible to obtain analytical results that are objective and easily obtain empathy and understanding from wine purchasers/consumers.

Fig. 2 schematically shows how one example of the method of the present invention is carried out;

[Method for evaluating the anti-glycation property of wine]
The present invention: the "method for evaluating the anti-glycation properties of wine" includes (step 1) a step of ingesting wine by a subject, (step 2) a step of obtaining mental and physical information of the subject from the subject by a questionnaire method. , (Step 3) quantitatively measuring glycative stress markers and/or stress in the subject by a non-invasive method.

Wines to be evaluated by the method of the present invention are not limited. As the wine, preferably used are wines having a high content of polyphenols, which are considered to have high anti-glycation properties, that is, various red wines.

Subjects for the methods of the present invention are not limited as long as they are healthy adults. The subjects are preferably males and females aged 20 to 65 and free from so-called lifestyle-related diseases such as diabetes.

[Step 1]
Step 1 of the method of the present invention is the step of ingesting wine by the subject. The above intake is generally carried out in a so-called cool system. 1 course is, for example, 3 days / week intake (one day of the week is a non-ingestion day) for 6 weeks, 6 days / week intake (one day of the week is a non-ingestion day) ) is defined as 4 weeks. The daily wine intake is generally set in the range of 50 ml to 250 ml for commercially available wine.

[Step 2]
Step 2 of the present invention is a step of obtaining information on the subject's mind and body from the subject by means of a questionnaire. It is preferable that the above-mentioned questionnaire is conducted periodically, for example, every week, during the period from the start to the end of the method of the present invention.

The above mental and physical information is not limited as long as it is an element related to the subject's fatigue or stress. Such psychosomatic information is generally information about mood, sleep, bowel movements, and eating habits. The first questionnaire may include, for example, the following questions and options.

Q01: How are you feeling now? /A01: Better than usual/Same as usual/Worse than usual Q02: How was the quality of your sleep last night? /A02: Better than usual/Same as usual/Worse than usual Q03: How is your bowel movement today? /A03: Normal / Poor Q40: How is your appetite today? /A04: higher than usual/same as usual/lower than usual In the present invention, the questionnaire can be conducted via the Internet. The test subject logs into the questionnaire site using a terminal device such as a smartphone, tablet, or PC connected to the Internet, and inputs an answer. The entered answers are stored in a server on the Internet, tallied by a tally program, and statisticized.

[Step 3]
Step 3 of the present invention is a step of quantitatively measuring glycative stress markers and/or stress in the subject by a non-invasive method. The glycative stress marker and/or stress indicator is not limited as long as it is commonly used in biochemistry and clinical medicine. As such indicators, for example, skin AGEs, body AGEs, heart rate variability, skin conditions, and the like can be used.

Skin AGEs and body AGEs are measured non-invasively with fluorescence sensors. This method utilizes the property that when the skin is irradiated with ultraviolet rays, the fluorescent AGEs accumulated in the tissue are excited and emit specific autofluorescence. As such a fluorescent sensor, for example, AGE Reader (registered trademark), AGE sensor "RQ-AG01J" manufactured by Sharp Life Science Co., Ltd., and the like are commercially available.

Heart rate variability is detected from changes in electrocardiogram and pulse wave. "Vital Monitor VM500" manufactured by Fatigue Science Laboratory Co., Ltd. can be used as a simple device for measuring heart rate variability. This monitor collects electrocardiograms and pulse waves from the fingertip to generate heart rate variability data. The generated heart rate variability data is transmitted to terminal devices such as smartphones, tablets, and PCs, and evaluation results are displayed on the evaluation matrix.

The skin condition is detected as, for example, transepidermal water loss, stratum corneum water content, dermis water content, stratum corneum morphology, and stratum corneum CML content. Various devices are available for measuring transepidermal water loss, stratum corneum water content, and dermal water content. Stratum corneum morphology can be measured by the tape stripping method. In this method, stratum corneum cells are peeled off with an adhesive tape and collected. By this method, the number of sloughing corneocytes, the corneocyte area, the cell mobility, the survival rate of nuclei, etc. can be measured. In addition, the exfoliated stratum corneum can also be analyzed biochemically. The amount of CML (carboxymethyllysine, which is one type of AGE) contained in the stratum corneum extract is measured using the exfoliated stratum corneum extract and a measurement kit according to the ALISA method. Separately, the amount of protein contained in the stratum corneum extract is measured, and the amount of CML (keratin CML) per stratum corneum protein is calculated.

The above step 3 is performed at an appropriate frequency, for example, once every four weeks or once every two weeks, during the period from the start to the end of the method of the present invention.

[Step 4]
The method of the present invention may further comprise step 4: measuring the in vitro anti-glycation activity of the wine using an automated device. The results obtained in step 4 above can be compared and contrasted with the subject data obtained in steps 2 and 3 above to predict the anti-aging performance of various wines for consumers.

In order to measure the in vitro antiglycation effect, model proteins such as human serum albumin (HSA), reducing sugars such as glucose and fructose, and wine to be tested were added to a phosphate buffer (pH 7.4). ) at 60°C for 40 hours. After that, the amount of AGEs in the reaction solution is compared between the presence and absence of the test substance, and the AGE production inhibition rate (%) is calculated. The level of AGEs produced when HSA and glucose were incubated at 60°C for 40 hours corresponds to approximately 60 days at 37°C. In the present invention, operations up to completion of incubation can be performed by an automated apparatus.

Next, the AGEs-derived fluorescence intensity measurement (excitation wavelength 370 nm, fluorescence wavelength 440 nm) can be used for the detection and quantification of the produced AGEs. Fluorescent AGEs include pentosidine (excitation wavelength (ex) 335 nm/fluorescence wavelength (em) 385 nm), crossrin (ex: 379 nm/em: 463 nm), and pyropyridine (ex: 370 nm/em: 455 nm). Since CML, pentosidine, 3-methyldeoxyglucose (3DC), glyoxal (GO), and methylglyoxal (MGO) are also produced in the reaction solution, the amounts of these produced can also be measured by each method. A device that automatically outputs and displays measured values can also be used for the fluorescence intensity measurement. As such a device, Infinite/Signrise series manufactured by TECAM can be used.

When measuring the in vitro anti-glycation effect, after reacting with the concentration of the wine to be tested set to three or more stages, based on the measured values of AGEs and saccharification reaction intermediates in each reaction solution Calculate the 50% production inhibition concentration of the above wine.

[evaluation]
The method of the invention can be performed on a variety of wines and subjects. By accumulating, referring to, and comparing the answers and measurement results obtained in the above step 2 and the above step 3, and more preferably the above step 4, the anti-aging effect and stress relief effect are provided to wine purchasers and consumers. It is possible to select wine brands with anti-glycation properties more accurately and in detail, and to propose wine brands optimized for wine purchasers and consumers.

The anti-glycation properties of red wine were evaluated under the following conditions.

[Wine, Mineral Water] 2016 red wine from Bordeaux, France (alcohol concentration: 13.5%) imported by MOTOX Co., Ltd. was used. Commercially available mineral water was prepared for control experiments.

[Subjects] 30 males and 30 females aged between 20 and 65 years old who are neither diabetic nor prediabetic were selected.

[Start] A briefing session was held for the subjects. Sixty subjects were randomly divided into two groups (Group A and Group B). We distributed wine, mineral water, and glasses to the subjects.

[Step 1 (Subjects of Group A)]
- For 4 weeks from the 2nd to 5th week, the wine distributed at the briefing session was ingested. The wine was consumed 6 times each week (6 days/week). The daily wine intake was 125 ml. No other alcohol was consumed during this period.
- During the 6th and 7th weeks, the subjects lived a normal life without drinking the wine or mineral water distributed at the briefing session. However, no alcohol was consumed during this period.
- For 4 weeks from 8th to 11th week, the subjects took mineral water distributed at the briefing session instead of wine. The above mineral water was ingested 6 times each week (6 days/week). The daily intake of mineral water was 125 ml. No alcohol was consumed during this period.

[Step 1 (Subjects of Group B)]
・For 4 weeks from the 2nd to the 5th week, I took mineral water distributed at the briefing session. The above mineral water was ingested 6 times each week (6 days/week). The daily intake of mineral water was 125 ml. No alcohol was consumed during this period.
- During the 6th and 7th weeks, the subjects lived a normal life without drinking the wine or mineral water distributed at the briefing session. However, no alcohol was consumed during this period.
- For 4 weeks from 8th to 11th week, the wine distributed at the briefing session was ingested. The wine was consumed 6 times each week (6 days/week). Daily wine intake was 125 ml. No other alcohol was consumed during this period.

[Step 2] The following non-invasive measurements were performed on all subjects at the start (the day of the briefing session), the last day of the 5th week, the first day of the 8th week, and the last day of the 11th week.
・Amount of AGEs in the body: Measured by touching an AGEs sensor (AGE sensor “RQ-AG01J” manufactured by Sharp Life Science) with a fingertip.
- Heart rate variability: The degree of stress was measured using "Vital Monitor VM500" manufactured by Fatigue Science Laboratory. The subject's thumb is inserted into the sensor for measurement, and the evaluation result is output to the terminal at the measurement venue.
- Skin condition: Transepidermal water loss was measured using "VAPO METER SWL-5001" manufactured by Delfin. The sensor at the tip of the measuring instrument was brought into contact with the skin on the inside of the subject's left arm for 10 seconds to measure.

[Step 3] Every week from the first week (one week after the briefing session), all subjects responded to a questionnaire asking about general physical conditions such as mood, quality of sleep, and bowel movements by e-mail.

[Step 4] Separately, the in vitro anti-glycation activity was measured for the wine distributed at the briefing session. The pretreatment for reacting the wine, human serum albumin, and reducing sugar in a phosphate buffer was semi-automated. A fluorescence plate reader “Infinite (registered trademark) 200PRO” manufactured by TECAN was used for the detection and quantification of the produced AGEs.

[Evaluation] When the questionnaire results obtained in step 2 above and the measurement results obtained in step 3 were compared and contrasted, it was found that consumption of the wine distributed at the explanatory meeting reduced the amount of AGEs in the subject's body and relieved stress. It became clear that The results obtained in step 4 confirmed this effect.

According to the present invention, it is possible to confirm the anti-saccharification effect of wine, which is supported by consumers' real feelings. By evaluating and selecting wines according to the method of the present invention, wines that are expected to be more satisfying to consumers can be offered to the market. The present invention can be a means of new market development for wine producers and sellers.

101 Group A Wine Consumption Period (Step 1)
102 Group B Wine Consumption Period (Step 1)
2 Questionnaire responses (process 2)
3 Non-invasive measurement 4 In vitro measurement of anti-glycation effect

Claims (5)

A method for evaluating the anti-glycation properties of wine, comprising the steps of:
(Step 1) A step in which a subject ingests wine.
(Step 2) A step of obtaining information on the subject's mind and body from the subject through a questionnaire.
(Step 3) A step of quantitatively measuring glycative stress markers and/or stress in the subject by a non-invasive method. 2. The method according to claim 1, wherein the psychosomatic information in step 2 is information on one or more selected from mood, sleep, bowel movements, and diet. 2. The method according to claim 1, wherein in step 3, one or more selected from skin AGEs, body AGEs, heart rate variability, and skin condition is measured by a non-invasive method. The method according to claim 1, wherein in said step 2 and/or said step 3, terminals and/or measuring equipment connected to the Internet are used. (Step 4) The method according to claim 1, further comprising the step of evaluating the in vitro anti-glycation activity of the wine using an automated device.

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