JP2020193852A - Non-invasive method for estimating serum zinc value - Google Patents

Abstract

To provide a method for reducing a burden on a subject and easily estimating a serum zinc value in the subject.SOLUTION: Means for solving the problem of the present invention are as follows. 1. A method for estimating a serum zinc value from a zinc value in saliva and/or a feeling profile test value. 2. The method for estimating a serum zinc value according to 1., wherein the feeling profile test is POMS or POMS2. 3. A system for estimating a serum zinc value in the subject by using the method for estimating a serum zinc value according to 1. or 2., and providing the subject with a zinc sufficient state and/or an oral composition for compensating for the shortage of zinc. A method for estimating a serum zinc value from a zinc value in saliva and/or a feeling profile test value.SELECTED DRAWING: Figure 1

Description

The present invention relates to a method of estimating serum zinc levels from saliva zinc levels and / or mood profile test values.

Zinc is an essential trace element, and when deficient, it causes zinc deficiency such as growth disorders, hair loss, dysgeusia, skin disorders, sexual dysfunction, immunodeficiency, loss of appetite, and delayed wound healing. About 1.5 to 3 g of zinc is present in a normal-weight human body, and most of it is distributed in skeletal muscle, bone, skin, liver, brain, kidney, hair, etc. in a state of being bound to protein.

In Japan, the recommended intake of zinc and the upper limit of tolerable amount have been set, but it has been pointed out that it is difficult to judge because there is no specific index of zinc nutritional status, especially zinc deficiency status. As an index of the nutritional status of zinc, a method of collecting the blood of a subject and measuring the serum zinc level is common. However, since blood sampling is invasive, the burden on the subject is heavy, and a non-invasive method for measuring serum zinc levels is required.

One of the non-invasive methods for evaluating the nutritional status of zinc is to use the amount of zinc excreted in urine as an index. However, the amount of zinc excreted in urine is very low at about 0.5 mg / day in healthy subjects, and the binding mode of zinc in serum changes in subjects with diseases that present with hypoalbuminemia. Therefore, there is a risk that the amount of zinc excreted in the urine will increase and the nutritional status of a true zinc deficient person will be misdiagnosed as normal. Another method is to measure the zinc concentration in the hair, but the hair is easily affected by the outside, and especially when a hair styling product or shampoo containing zinc is used, the zinc concentration in the hair increases significantly. , Its interpretation becomes difficult.

Although there is a method of estimating the serum zinc level from the results of the dietary frequency survey, the present inventors have found that there is no correlation between the zinc intake estimated from the results of the dietary frequency survey and the serum zinc level. This is also consistent with the report of Non-Patent Document 1.

Methods for estimating serum zinc levels from saliva components have been investigated, but there are few reports.
Non-Patent Document 2 reports that there is no correlation between the activity of TIMP enzyme (zinc-related enzyme) in saliva and serum zinc level in 10 dysgeusia and 10 healthy subjects. ..

In Non-Patent Document 3, a significant positive correlation (R = 0.398, P <0.05) was found between the immunochromatographic measurement value of Gustin (zinc-binding protein) and the serum zinc value using saliva as a sample. I am reporting that there is. However, the correlation is weak and it is insufficient as a method for estimating the serum zinc level.

Patent Document 1 is characterized by collecting oral-derived tissue of a taste-impaired person by a scraping method and associating the expression level of a taste receptor gene in the collected tissue with a serum zinc level. Zinc deficiency taste abnormality. We are proposing an inspection method for. However, in this test method, there is no disclosure about the correlation between the expression level of the taste receptor gene and the serum zinc level, and it is not used for estimating the serum zinc level.

Patent Document 2 estimates the zinc deficiency state by combining blood test values of hemoglobin concentration, hematocrit, albumin concentration, total cholesterol concentration, triglyceride concentration, sodium ion concentration and chlorine ion concentration, which are items of general blood test. We are proposing a device to do this. It is an excellent technique to reduce the physical burden on the subject in that it does not require additional blood sampling due to the serum zinc level that is not included in the general blood test items. However, this method requires general blood test values, and it is difficult for the subject to easily check the zinc deficiency state on a daily basis, and a simpler method is required.

Non-Patent Document 4 reports that zinc intake may improve mood profile test (POMS) values. Specifically, 41 women aged 18 to 21 years were divided into two groups, a group that took capsules containing only vitamins and a group that took capsules containing vitamins and zinc, and took them for 10 weeks. As a result, in the group that took only vitamins, there was no change in the POMS results before and after the intake, and in the group that took vitamins and zinc, the POMS "anger-hostility (AH)" and "depression" were compared before and after the intake. It is reported that the negative mood scores of "-depression (D)" improved significantly (P = 0.009 and P = 0.012, respectively). However, no significant difference was observed between the groups in "anger-hostility (AH)" and "depression-depression (D)", and "confusion (C)", "fatigue (F)", and "tension-" There was no significant difference between the negative mood scale of "anxiety (TA)" and the positive mood scale of "vigor (V)" before and after ingestion, and the relationship between serum zinc level and mood profile test value was shown. Not.

International Publication WO2007 / 074937 Japanese Patent No. 6421398

Janet King, (2018) J Nutr 148; 1399-1401 Journal of the Japanese Association for the Study of Taste and Smell (200.12) Vol. 7 No. 3; 639-640 Journal of the Japanese Association for the Study of Taste and Smell (2013.12) Vol. 20 No. 3; 345-346 Sawada T (2010) Eur J Clin Nutr. 2010 Mar; 64; 331-3

An object of the present invention is to provide a method for easily estimating the serum zinc level of a subject while suppressing the burden on the subject.

The means for solving the problem of the present invention is as follows.
1. 1. A method of estimating serum zinc levels from saliva zinc levels and / or mood profile test values.
2. 2. 1. The mood profile test is POMS or POMS2. The method for estimating the serum zinc level described in 1.
3.1. Or 2. A system for estimating the serum zinc level of a subject using the method for estimating the serum zinc level described in the above, and providing the subject with an oral composition for compensating for zinc sufficiency and / or zinc deficiency.

The present invention is a method of estimating serum zinc levels from saliva zinc levels and / or mood profile test values. The target person (consumer) collects a saliva sample at a store or at home by a simple method using a saliva collection kit, etc., and mails it together with the mood profile test result to obtain his / her serum zinc level and zinc sufficiency. It becomes possible to know. In addition, subjects who are deficient in zinc can be provided with oral compositions such as supplements and drinks that can efficiently take zinc orally in addition to their daily diet. Furthermore, since the present invention does not require blood sampling and the burden on the subject is extremely small, the subject can easily re-examine the course after ingestion of the oral composition even after a certain period of time.

Scatter plot of serum zinc and saliva zinc Scatter plot of serum zinc levels and mood profile test values (AHT scores) Scatter plot of serum zinc level and mood profile test value (CB T score) Scatter plot of serum zinc level and mood profile test value (DDT score) Scatter plot of serum zinc levels and mood profile test values (FIT scores) Scatter plot of serum zinc level and mood profile test value (TAT score) Scatter plot of serum zinc levels and mood profile test values (VAT scores) Scatter plot of serum zinc level and mood profile test value (FT score) Scatter plot of serum zinc level and mood profile test value (TMD T score) Scatter plot of serum zinc level and serum zinc estimate by multiple regression model (X ₁ = saliva zinc level, X ₂ = AHT score) Scatter plot of serum zinc level and serum zinc estimate by multiple regression model (X ₁ = saliva zinc level, X ₂ = CBT score) Scatter plot of serum zinc level and serum zinc estimate by multiple regression model (X ₁ = saliva zinc level, X ₂ = DDT score) Scatter plot of serum zinc level and serum zinc estimate by multiple regression model (X ₁ = saliva zinc level, X ₂ = FIT score) Scatter plot of serum zinc level and serum zinc estimate by multiple regression model (X ₁ = saliva zinc level, X ₂ = TAT score) Scatter plot of serum zinc levels and serum zinc estimates by multiple regression model (X ₁ = salivary zinc levels, X ₂ = VAT scores) Scatter plot of serum zinc level and serum zinc estimate by multiple regression model (X ₁ = saliva zinc level, X ₂ = FT score) Scatter plot of serum zinc level and serum zinc estimate by multiple regression model (X ₁ = saliva zinc level, X ₂ = TMD T score) Estimated serum zinc level and multiple regression model (X ₁ = AH, X ₂ = CB, X ₃ = DD, X ₄ = FI, X ₅ = TA, X ₆ = VA, X ₇ = FT score) Scatter plot Scatter plot of serum zinc levels and multiple regression models (X ₁ = AH, X ₂ = CB, X ₃ = DD, X ₄ = FI, X ₅ = TAT scores) Scatter plot of serum zinc levels and multiple regression models (X ₁ = salivary zinc levels, X ₂ = AH, X ₃ = CB, X ₄ = DD, X ₅ = FI, X ₆ = TAT scores)

Hereinafter, the present invention will be described in detail.
The present invention is a method of estimating serum zinc levels from saliva zinc levels and / or mood profile test values.

(Mood profile test)
The mood profile test is a test method that objectively quantifies the mood state at the time of the survey using a questionnaire, such as a condition check of athletes, a mental health check at work, and a mood check in the fields of care and welfare. It is a test used internationally in various fields such as the course of treatment for mental disorders (depression, anxiety disorder, etc.), the evaluation scale of the therapeutic effect of drugs on mental stress, and the measurement of exercise effect and relaxation effect. .. Among them, POMS (Profile of Mood States) and POMS2 (Profile of Mood States 2nd Edition) are globally trusted mood profile tests. POMS2 is a revised version of POMS developed by McNair et al. (1971).

POMS includes "Anger-Hostility (AH)", "Confusion (C)", "Depression-Depression (D)", "Fatigue (F)", and "Tension". -Five negative mood scales for "Tension-Anxiety (TA)", positive mood scales for "Vigor;V", and seven "Total Mood Distance (TMD)" The scale is quantified.
POMS2 includes "Anger-Hostility (AH)", "Confusion-Behiderment (CB)", "Depression-Depression (DD)", and "Fatigue-Imp". : FI) ”and“ Tension-Anxiety (TA) ”five negative mood scales, and“ Vigor-Action (VA) ”and“ Friendship (F) ” Two positive mood scales and eight scales of "Total Mood Disturbance (TMD)" are quantified. The definition of each mood scale is as described in the manual.

The mood profile test differs depending on the target population, such as for adults and adolescents, and the number of questions asked at the time of the test, such as the full-item version and the shortened version, is different. The inspection method and the number of question items may be appropriately selected according to the situation.

In the present invention, the mood profile test is preferably POMS or POMS2. One or more POMS or POMS2 test items can be used as explanatory variables, and one or more items selected from the negative mood scale (AH, CB, DD, FI, TA) and the comprehensive mood scale (TMD) can be used. It is preferable to use it as an explanatory variable. When the zinc level in saliva and the mood profile test value are combined, the test items of POMS or POMS2 can be used without particular limitation, and the zinc level in saliva and one or more items selected from the test items of POMS or POMS2 can be used. Can be combined with.

Regarding the zinc sufficiency status, according to the "Clinical Guidelines for Zinc Deficiency 2018" (Japan Society for Clinical Nutrition), it is desirable to measure the serum zinc level on an empty stomach in the early morning, even if the serum zinc level is less than 60 μg / dL. For example, there are guidelines for diagnosing zinc deficiency, and latent zinc deficiency if it is less than 60-80 μg / dL. That is, if the serum zinc level is 80 μg / dL or more, it can be said that there is a high possibility that the serum zinc level is satisfied. An oral composition for estimating the serum zinc level of a subject and compensating for zinc sufficiency and / or zinc deficiency can be provided to the subject.

The oral composition provided in the present invention includes pharmaceuticals (including non-pharmaceutical products), nutritional supplements, nutritionally functional foods, foods for specified health uses, foods with functional claims, functional foods such as foods for the sick, and general foods. It can be used as a food and food additive. It is preferable to have, for example, tablets, capsules, granules, powders, pills, chewable tablets, orally disintegrating tablets, drinks, and jelly-like forms so that continuous ingestion can be easily performed. Foods processed into rods, plates, gummy candies, or added to general food forms may be used. Of these, the forms of tablets and capsules are particularly preferable from the viewpoint of ease of ingestion. The preparation having such a dosage form can be adjusted by a conventional method, and may be a rapid-release or sustained-release preparation in which the release of zinc is controlled.

The zinc to be blended in the oral composition of the present invention is not particularly limited as long as it is distributed as a food raw material for supplementing zinc. Preferably, zinc gluconate, zinc yeast, zinc sulfate, zinc chloride, zinc oxide, zinc stearate can be mentioned, and at least one or a combination of two or more selected from these groups can be used. it can.

In addition to zinc, the oral composition provided in the present invention may contain auxiliary components for increasing the bioavailability of zinc, and other ingredients such as vitamins, minerals, amino acids, sugars, polypeptides, and fish oils. It is possible to mix nutrients such as.

<Clinical trial method>
A clinical trial was conducted on healthy male subjects aged 20 to 60 years. Furthermore, 11 people who did not meet the following exclusion criteria were included in the final analysis.
1) Those who cannot collect saliva 2) Those who have oral diseases such as periodontal disease or those who have bleeding in the oral cavity 3) Those who have metabolic diseases such as diabetes 4) Those who have autoimmune diseases 5) Liver diseases 6) Have a history of serious illness. Or those who are undergoing treatment 7) Those who have been taking chelating drugs for a long time 8) Those who have difficulty in collecting blood (People who have been told that it is difficult to collect blood in medical examinations)
9) Persons judged by the investigator or the examiner to be non-conforming as the subject of the examination As an ethical consideration, this examination should be submitted to the Fankel Clinical Research Ethics Review Board from the viewpoint of ethical, scientific and medical validity. After being examined for suitability, comply with the test plan in accordance with the latest "Ethical Principles Based on the Declaration of Helsinki" and "Ethical Guidelines for Medical Examinations for Humans (Ministry of Education, Science and Technology / Ministry of Health, Labor and Welfare)" And carried out. Before conducting this clinical trial, all the subjects were fully informed of the purpose of the trial, and the consent of the participants in writing was obtained at their own discretion.
This test was conducted as an open test. Subjects gathered at a designated study facility to collect early morning fasting blood and saliva.

<Saliva collection>
Human saliva has three glands, parotid gland, submandibular gland, and submandibular gland, and the saliva secreted from each is distinguished from parotid gland saliva, parotid gland saliva, and submandibular gland saliva. , The mixture of the above saliva is called total saliva. For saliva, all saliva was collected using a saliva sampling device (Saliva Collection Aid; SCA manufactured by Funakoshi Co., Ltd.) by the method described in the manual attached to the SCA. The collected saliva was centrifuged at 3000 G for 2 minutes to remove impurities, and stored at −80 ° C. until measurement.

<Measurement method of zinc level in saliva>
The zinc level in saliva was measured using a metalloassay zinc measurement LS kit (manufactured by Metallogenix). Add 6M hydrochloric acid to the collected saliva to a final concentration of 1%, mix using a vortex mixer, and then centrifuge at 1000 rpm for 10 minutes to remove impurities, and use the supernatant as the measurement sample. did. A mixture of 40 μL of the sample and 200 μL of the color-developing solution was added to a 96-well microplate (manufactured by AGC Technograss), incubated at room temperature for 10 minutes, and then the absorbance was measured using a plate reader (manufactured by JASCO Corporation). The results are shown in Table 1.

<Measurement method of serum zinc level>
Blood was collected from the cubital fossa vein using a blood collection tube containing a serum separator. The collected blood was mixed well and then allowed to stand for 30 minutes to 1 hour to confirm a blood clot, and then the serum components were separated by centrifugation under the conditions of 3000 rpm and 10 minutes. The presence or absence of hemolysis was visually confirmed, and the presence or absence of hemolysis was described in the case report. The separated serum components were stored at −80 ° C. until measurement. We requested a clinical laboratory contract institution (LSI Medience Corporation) and measured it by atomic absorption spectroscopy to obtain a serum zinc level. The results are shown in Table 1.

<Method of mood profile test>
The inspection was carried out according to the Japanese manual using the inspection form of "POMS2 Adult Short Edition" (Kaneko Shobo). These total scores were converted into T scores according to the Japanese manual and used as mood profile test values. TMD similarly converted the value obtained by subtracting the VA score from the total score of the negative mood scales of AH, CB, DD, FI, and TA into the T score. The results are shown in Table 1.

(analysis method)
Statistical analysis was performed using statistical processing software JMP14 (SAS Institute Japan). The correlation coefficient R2 and the statistical value P were obtained by regression analysis of the serum zinc level and the zinc level in saliva. The results are shown in FIG. By regression analysis of serum zinc level and mood profile test value, correlation coefficient R2 and statistical value P were similarly obtained. The results are shown in Figures 2-9. The multiple regression model was created by the method of least squares. The results of the multiple regression model using the zinc level in saliva and the mood profile test value as explanatory variables are shown in Table 2, FIGS. 10 to 17. In addition, the explanatory variables of the multiple regression model were 7 items other than TMD, 5 items of negative mood scale (AH, CB, DD, FI, TA), 5 items of negative mood scale, and zinc level in saliva. The results are shown in Table 3 and FIGS. 18 to 20.

<Result>
(Fig. 1: Correlation between serum zinc level and saliva zinc level)
There is a positive correlation between the serum zinc level and the zinc level in saliva, R2 = 0.5734, P = 0.007, approximate formula "serum zinc level (μg / dL) = 3.5811 × (saliva) Zinc value in (μg / dL)) +66.387 ”was obtained.

(Table 1, Figures 2-9; Correlation between serum zinc level and mood profile test value)
There is a negative correlation between serum zinc level and mood profile test value (AH), R2 = 0.4603, P = 0.0217, approximate formula "serum zinc level (μg / dL) = -0.8359". X (AH) +121.54 ”was obtained.
There is a negative correlation between serum zinc level and mood profile test value (CB), R2 = 0.4316, P = 0.0281, approximate formula "serum zinc level (μg / dL) = -1.4434". X (CB) +145.75 ”was obtained.
There is a negative correlation tendency between the serum zinc level and the mood profile test value (DD), R2 = 0.2999, P = 0.0811, and the approximate formula “serum zinc level (μg / dL) = -1. 1989 × (DD) +136.91 ”was obtained.
There is a negative correlation between serum zinc level and mood profile test value (FI), R2 = 0.5488, P = 0.0091, approximate formula "serum zinc level (μg / dL) = -1.0481". X (FI) +129.32 ”was obtained.
There is a negative correlation tendency between the serum zinc level and the mood profile test value (TA), R2 = 0.2438, P = 0.1226, and the approximate formula “serum zinc level (μg / dL) = −0. 674 × (TA) +112.9 ”was obtained.
There is a negative correlation between the serum zinc level and the mood profile test value (TMD), R2 = 0.5875, P = 0.0059, approximate formula "serum zinc level (μg / dL) = -1.317". X (TMD) +141.06 ”was obtained.
No correlation was found between serum zinc levels and mood profile test values (VA, F).

(Table 2, Figures 10-17; Multiple regression model)
An estimation formula for the zinc level in saliva and the serum zinc level using the mood profile test value as an explanatory variable was obtained (Table 2). As a result of performing regression analysis between the estimated serum zinc value calculated from the estimation formula of the serum zinc level and the serum zinc level, 8 of the mood profile test values (AH, CB, DD, FI, TA, VA, F, TMD) It was shown that there was a high correlation in all items (Figs. 10 to 17). Although there was a correlation with serum zinc levels for the mood scales of AH, CB, DD, FI, TA, and TMD (Figs. 2 to 6 and 9), a multiple regression model was used with the zinc level in saliva as an explanatory variable. It was shown that a higher correlation coefficient was obtained and the serum zinc level could be estimated with high accuracy (Figs. 10-14, 17). In addition, the mood scales of VA and F did not correlate with the serum zinc level (Figs. 7 and 8), but the serum zinc level was estimated by using a multiple regression model using the zinc level in saliva as an explanatory variable. It became possible to do so (Figs. 15 and 16).

(Table 3, Figures 18-20; Multiple regression model)
It was shown that the serum zinc level can be estimated with high accuracy (R2 = 0.9768, P = 0.0186) when 7 items other than TMD are used as explanatory variables. When 5 negative mood scale items are used as explanatory variables, R2 = 0.8524 and P = 0.0385, which is less accurate than when 7 items other than TMD are used as explanatory variables, but 5 negative mood scale items are used. When the zinc value in saliva was added as an explanatory variable, R2 = 0.9400 and P = 0.0199, and the accuracy of estimating the serum zinc level was improved.

Claims (3)

A method of estimating serum zinc levels from saliva zinc levels and / or mood profile test values. The method for estimating serum zinc levels according to claim 1, wherein the mood profile test is POMS or POMS2. A subject is provided with an oral composition for estimating the serum zinc level of the subject using the method for estimating the serum zinc level according to claim 1 or 2, and compensating for the zinc sufficiency state and / or the zinc deficiency amount. System to provide to.

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