JP2012247270A - Skin state determination device, skin state determination method, skin state determination program, and recording medium - Google Patents

Abstract

PROBLEM TO BE SOLVED: To accurately determine the state of skin.SOLUTION: A skin state determination system 20 includes a measuring device 1 for measuring fluorescence intensity of tryptophan AGEs (advanced glycation endproducts) in the skin of an individual to be measured, and a determination device 10 for determining the state of skin using the fluorescence intensity measured by the measuring device 1.

Description

  The present invention relates to a determination device and a determination method for determining a skin condition.

  In recent years, with the westernization of eating habits, the number of lifestyle-related diseases has increased, and medical and social problems have become serious. Currently, it is said that there are 8 million diabetics in Japan, and 20 million including the reserve army. The three major complications of diabetes are “retinopathy, nephropathy, neuropathy”. Furthermore, diabetes is a cause of arteriosclerosis, and there is concern about heart disease and brain disease.

  Diabetes mellitus is caused by disturbance of dietary habits, lifestyle habits, fat cell secretions due to obesity, or oxidative stress, resulting in poor pancreatic function, lack of insulin to control blood sugar levels, and reduced insulin efficacy. It develops by doing. Symptoms such as throat urination and frequent urination appear when people have diabetes, but if this is the case, there is no subjective symptom that it is a disease, and it is almost always detected by examinations at hospitals. This is the reason why there are many “silent” diabetics.

  After an abnormal symptom due to complications appears in a hospital or the like, the medical condition has often progressed and it is difficult to cure completely. In particular, complications are often difficult to treat, and prevention is as important as other lifestyle-related diseases. Early detection and treatment effect determination are indispensable for prevention, and there are many tests for diabetes for that purpose.

  When oxidative stress is applied in an environment where an abnormal amount of sugar or lipid is present in the blood, the protein reacts with the sugar or lipid to produce AGEs (Advanced Glycation Endproducts; late glycation reaction products). Is done. AGEs are final products formed by non-enzymatic sugar addition reaction (Maillard reaction) of proteins. They are yellowish-brown, fluorescing substances that bind to nearby proteins to form crosslinks. It has properties.

  In the case of diabetes, AGEs increase with an increase in blood glucose, so early detection of diabetes or progress can be grasped by monitoring AGEs. Moreover, AGEs are also formed in the skin, and it is said that it causes skin aging such as skin tension and reduced elasticity.

  For example, a method described in Patent Literature 1 has been reported as a method for evaluating the health condition and skin condition of an individual by monitoring such AGEs. In this method, a certain region of the skin of a living body is irradiated with excitation light, and a fluorescence spectrum from AGE-converted skin collagen and elastin and a fluorescence spectrum from tryptophan are detected. Then, the obtained fluorescence intensity ratio is compared with a predetermined control intensity ratio. This assesses the individual's health and skin aging without invasion.

  Fluorescence derived from tryptophan decreases with skin aging, and AGEs collagen and elastin increase with skin aging. Therefore, in the method described in Patent Document 1, the ratio between the fluorescence intensity of tryptophan and the fluorescence intensity of AGE-modified collagen and elastin is used as an index of skin aging.

JP 2005-169124 A (released on June 30, 2005)

  However, since tryptophan is not synthesized in vivo, the amount of tryptophan accumulated in the skin tissue is susceptible to external factors (meal). Therefore, in the method described in Patent Document 1, the measurement result is likely to change depending on the measurement timing (fasting, after eating, etc.). In addition, the amount of tryptophan in a living body is lower in a depressed person whose mental state is unstable than in a healthy person. For these reasons, it is difficult for the method described in Patent Document 1 to accurately evaluate the skin condition.

  In addition, since the fluorescence intensity of AGE-modified collagen and elastin increases in proportion to age, it is easily affected by aging, and it is difficult to determine whether the skin condition has deteriorated due to unhealthy eating habits. . In the first place, Patent Document 1 does not describe the problem of evaluating the deterioration of the skin condition due to an unattended diet.

  In addition, in the invention of Patent Document 1, since two types of excitation wavelengths are required for measurement of fluorescence intensity, there are problems that the configuration of the apparatus becomes complicated and the measurement time becomes long.

  The present invention has been made in order to solve the above-described problems, and an object of the present invention is to determine a skin state (particularly, deterioration of the skin state due to an unattended diet) with high accuracy, and It is to provide a determination method.

In order to solve the above problems, the skin condition determination apparatus according to the present invention
A measurement unit for measuring the fluorescence intensity of tryptophan AGEs (Advanced Glycation Endproducts) in the skin of an individual to be measured;
And a determination unit that determines the state of the skin using the fluorescence intensity measured by the measurement unit.

  According to said structure, the fluorescence intensity of the tryptophan AGEs of skin is measured in a measurement part. Then, the skin state is determined by the determination unit using the measured fluorescence intensity of tryptophan AGEs.

  The inventors of the present invention have found that tryptophan AGEs are an indicator of deterioration of skin condition. Since the fluorescence intensity of tryptophan AGEs does not increase significantly with aging and is significantly affected by dietary habits, it is easy to determine the deterioration of the skin condition due to non-consumption. By determining the skin state using the tryptophan AGEs as an index, the skin state can be determined with high accuracy. In particular, it is possible to determine with high accuracy the deterioration of the skin condition due to unhealthy eating habits.

  Moreover, it is preferable that the said determination part determines the said skin state based on the result of having compared the fluorescence intensity measured by the said measurement part, and a predetermined reference value.

  For example, when the fluorescence intensity measured by the measurement unit is higher than the predetermined reference value, the determination unit may determine that the skin condition is out of a normal state.

  With the above configuration, by comparing an actual measurement value with a reference value prepared in advance, how much the actual measurement value is different from the reference value can be obtained, and the skin state can be determined from the result.

  The predetermined reference value is preferably a value based on a statistical value of the fluorescence intensity of tryptophan AGEs in the age group to which the individual belongs.

  With the above configuration, the skin condition can be determined using a reference value corresponding to the age of the individual to be measured, and the determination reliability can be improved.

  Moreover, it is preferable that the said measurement part irradiates the said skin with excitation light, and measures the intensity | strength of the fluorescence generate | occur | produced by the irradiation of the said excitation light.

  With the above configuration, the fluorescence intensity of tryptophan AGEs in the skin can be measured.

  The excitation light preferably has a wavelength suitable for detecting fluorescence of tryptophan AGEs.

  For example, the peak wavelength of the excitation light is preferably included in a wavelength range of 280 nm to 400 nm.

  With the above configuration, tryptophan AGEs can efficiently emit fluorescence, and the fluorescence intensity of tryptophan AGEs can be measured.

  Moreover, it is preferable that the said measurement part detects the fluorescence which has a peak wavelength in the wavelength range of 370 nm or more and 410 nm or less as fluorescence of tryptophan AGEs.

  With the above configuration, the fluorescence intensity of tryptophan AGEs can be detected efficiently.

  Moreover, it is preferable that the said measurement part and the said determination part can be physically separated and can be connected so that communication is possible.

  According to said structure, a measurement part and a determination part are realizable as a separate apparatus, for example, invention can be implement | achieved as a combination of a measurement apparatus and the personal computer as a determination apparatus.

  The technical scope of the present invention also includes a skin condition determination program for causing a computer to function as a determination unit included in the skin condition determination apparatus, and a computer-readable recording medium recording the skin condition determination program.

In order to solve the above problems, the skin condition determination method according to the present invention
A measurement step of measuring fluorescence intensity of tryptophan AGEs (Advanced Glycation Endproducts) in the skin;
And a determination step of determining the skin condition using the fluorescence intensity measured in the measurement step.

  According to said structure, the fluorescence intensity of the tryptophan AGEs of skin is measured in a measurement process. Then, the skin state is determined using the measured fluorescence intensity of tryptophan AGEs.

  The inventors of the present invention have found that tryptophan AGEs are an indicator of deterioration of skin condition. Since the fluorescence intensity of tryptophan AGEs does not increase significantly with aging and is significantly affected by dietary habits, it is easy to determine the deterioration of the skin condition due to non-consumption. By determining the skin state using the tryptophan AGEs as an index, the skin state can be determined with high accuracy. In particular, it is possible to determine with high accuracy the deterioration of the skin condition due to unhealthy eating habits.

  Moreover, it is preferable to further include a correction step of correcting the fluorescence intensity of tryptophan AGEs using the fluorescence intensity that is a standard for measuring the fluorescence intensity.

  In the above configuration, since the fluorescence intensity of tryptophan AGEs is corrected based on the standard fluorescence (reference), the reliability of the measurement value can be improved.

  As described above, the skin condition determination apparatus according to the present invention uses a measurement unit that measures the fluorescence intensity of tryptophan AGEs (Advanced Glycation Endproducts) in the skin of an individual to be measured, and the fluorescence intensity measured by the measurement unit. It is a structure provided with the determination part which determines the state of the said skin.

  The skin condition determination method according to the present invention includes a measurement process for measuring fluorescence intensity of tryptophan AGEs (Advanced Glycation Endproducts) in the skin, a determination process for determining the skin condition using the fluorescence intensity measured in the measurement process, It is the structure containing.

  Therefore, there is an effect that the skin state can be determined with high accuracy.

It is a figure which shows the structure of the skin condition determination system which concerns on one Embodiment of this invention. It is a flowchart which shows an example of the flow of the process in the said skin state determination system. It is a figure which shows the excitation spectrum of fluorescence wavelength 400nm obtained from a different measuring subject. It is a figure which shows the fluorescence peak wavelength of AGEs derived from various amino acids. It is a graph which shows the relationship between the fluorescence intensity derived from tryptophan AGEs, and saccharification time. It is a graph which shows the fluorescence spectrum of tryptophan, and the fluorescence spectrum of tryptophan AGEs synthesize | combined artificially. It is a graph which shows the excitation spectrum of the tryptophan AGEs synthesize | combined artificially. It is a figure which shows the result of having incubated tryptophan in the presence of glucose of normal blood sugar concentration of a healthy person, and having measured the fluorescence spectrum.

  The following describes one embodiment of the present invention with reference to FIGS.

[Technical idea of the present invention]
First, the technical idea of the present invention will be described.

  The inventor of the present invention has found that among AGEs derived from amino acids present in the living body, tryptophan AGEs serve as an index indicating deterioration of skin condition (aging), and have come up with the present invention.

  AGEs have been used as an index indicating skin aging, but examples of selectively detecting tryptophan AGEs and using them as an index of skin condition have so far been known to the inventors of the present invention. Not.

  In particular, in the present invention, attention is focused on deterioration of skin condition due to unhealthy eating habits (for example, chronic hyperglycemia state, excessive sugar intake, etc.) rather than aging skin with aging, and tryptophan AGEs are used as an index thereof. Used. As far as the inventor of the present invention knows, there has never been an attempt to evaluate the deterioration of the skin condition due to an unhealthy diet.

  Thus, the technical idea of using tryptophan AGEs as an index of the skin condition is novel.

  By using tryptophan AGEs as an index of the skin condition, it is possible to accurately determine the deterioration of the skin condition due to unhealthy eating habits. The fluorescence intensity of tryptophan AGEs does not increase significantly with aging and is significantly affected by dietary habits. Tryptophan AGEs are considered to have a low possibility of accumulation when having a healthy diet.

  Therefore, by measuring tryptophan AGEs, it is possible to determine the deterioration of the skin condition due to an unhealthy diet, independently of skin aging associated with aging.

  In addition, tryptophan AGEs, unlike free tryptophan, do not significantly change the amount accumulated in the skin before and after a meal. This is because it takes time to generate tryptophan AGEs. Therefore, there is little possibility that the measurement result varies depending on the measurement timing.

  Moreover, since the fluorescence of tryptophan AGEs reflects glycation of blood vessels with high accuracy, the health condition can also be evaluated.

[Configuration of Skin Condition Determination System 20]
Next, the structure of the skin condition determination system (skin condition determination apparatus) 20 of this embodiment will be described. FIG. 1 is a diagram illustrating a configuration of the skin condition determination system 20. As shown in FIG. 1, the skin condition determination system 20 includes a measurement device (measurement unit) 1 and a determination device (determination unit) 10.

(Configuration of measuring device 1)
The measuring device 1 is a fluorometer that irradiates the skin (skin) of an individual to be measured with excitation light and measures the intensity of fluorescence generated by the irradiation of the excitation light. More precisely, when the fluorescent material contained in the skin is irradiated with excitation light, fluorescence is emitted from the fluorescent material. By measuring the intensity of this fluorescence, it is possible to measure the amount of accumulated fluorescent substance contained in the skin.

  In particular, the measuring apparatus 1 measures the fluorescence intensity of tryptophan AGEs (Advanced Glycation Endproducts) in the skin of an individual to be measured.

  The portions to be irradiated with the excitation light are arms, wrists, fingers, palms, cheeks, ears, and the like, and the excitation light is irradiated to specific portions or specific positions included in these measurement target portions.

  As shown in FIG. 1, the measuring apparatus 1 includes an excitation light source 2, a detector 3, an incident optical fiber 4, and a reflective optical fiber 5.

(Excitation light source 2)
The excitation light source 2 is a light source that generates excitation light that irradiates the measurement target unit 30. This excitation light is for detecting fluorescence derived from tryptophan AGEs, and has a wavelength range suitable for measuring tryptophan AGEs. Specifically, the excitation light emitted from the excitation light source 2 has a peak wavelength in the range of about 280 nm to about 400 nm. If it is the excitation light of this range, tryptophan AGEs can be excited and fluorescence can be emitted.

  As the type of the light source used as the excitation light source 2, a tube type such as a halogen or xenon light source, an LED, an LD, or the like can be used. Excitation light emitted from the excitation light source 2 is guided to the measurement target unit 30 through the incident optical fiber 4.

(Detector 3)
The detector 3 receives the fluorescence generated by irradiating the surface (skin) of the measurement target portion 30 with the excitation light through the reflection optical fiber 5 and measures the intensity for each wavelength of the fluorescence. That is, the detector 3 measures the intensity of which wavelength of fluorescence is detected.

  As the detector 3, a semiconductor detector such as a charge-coupled device (CCD) array or a metal-oxide semiconductor (CMOS) image sensor, a photomultiplier tube (PMT), a channeltron detector, or the like can be used. However, in order to increase the portability of the measuring apparatus 1, it is advantageous to use a semiconductor detector.

  Since the fluorescence has a longer wavelength than the excitation light, the detector 3 only needs to be able to detect light in the range of 320 to 500 nm, but the fluorescence also has a range of wavelengths that are detected depending on the type of AGEs. Therefore, any device capable of detecting the range of 320 to 900 nm can be used.

  In particular, the detector 3 preferably detects fluorescence having a peak wavelength in the wavelength range of 370 nm to 410 nm as the fluorescence of tryptophan AGEs. The fluorescence intensity of tryptophan AGEs can be measured by detecting fluorescence having a fluorescence peak intensity within this wavelength range.

(Incoming optical fiber 4 / reflective optical fiber 5)
The incident optical fiber 4 is a light guide unit (probe) that guides the excitation light emitted from the excitation light source 2 to the measurement target unit 30. The reflection optical fiber 5 is a light guide unit that receives fluorescence generated in the measurement target unit 30 and guides it to the detector 3. The leading end of the incident optical fiber 4 and the leading end of the reflecting optical fiber 5 coincide with each other. That is, the incident optical fiber 4 and the reflective optical fiber 5 are two-branch optical fibers.

(Configuration of determination apparatus 10)
The determination device 10 determines the skin state of the individual to be measured using the fluorescence intensity measured by the measurement device 1. The determination device 10 and the measurement device 1 are realized as physically separated individual devices, and are connected to be communicable with each other by wire or wirelessly. The determination device 10 may be a personal computer. Note that the measurement device 1 and the determination device 10 may be integrated.

  The determination device 10 includes a data acquisition unit 11, a determination unit 12, a storage unit 13, a display unit 14, and an operation unit 15.

  The data acquisition unit 11 acquires measurement data indicating the measurement result of the fluorescence intensity from the measurement device 1 and analyzes the measurement data. Analysis of the measurement data includes peak separation and extraction, peak position calculation, and the like. For analysis of the measurement data, commercially available software such as Peak Fit can be used.

  The determination unit 12 determines the state of the skin based on the result of comparing the fluorescence intensity of tryptophan AGEs (referred to as measured fluorescence intensity) indicated by the acquired measurement data with a predetermined reference value (referred to as reference fluorescence intensity). . The reference fluorescence intensity is a value serving as a standard for evaluating the measured fluorescence intensity. For example, the reference fluorescence intensity is a value of at least one stage calculated based on a statistical value (for example, an average value) of the fluorescence intensity of tryptophan AGEs of healthy persons calculated in advance for each age group. Further, the reference fluorescence intensity may be in the range of the fluorescence intensity of tryptophan AGEs of healthy persons.

  The determination unit 12 determines that the skin state is out of the normal state when the measured fluorescence intensity is higher than the reference fluorescence intensity. A plurality of reference fluorescence intensities may be provided, and determination results such as normal, slightly deteriorated, and significantly deteriorated may be output in multiple stages. Such a determination result is displayed on the display unit 14.

(Storage unit 13)
The storage unit 13 is a non-volatile storage device such as a hard disk or a flash memory, and stores various types of information such as measurement data, reference fluorescence intensity, and user setting information.

(Display unit 14)
The display unit 14 is a display device that displays the determination result of the determination unit 12, and is, for example, a liquid crystal display.

(Operation unit 15)
The operation unit 15 is an input device that receives an input operation from a user, and includes, for example, a keyboard, a mouse, an input button, and the like.

[Flow of processing in skin condition determination system 20]
Next, an example of a processing flow in the skin condition determination system 20 will be described. FIG. 2 is a flowchart illustrating an example of a process flow in the skin condition determination system 20 (particularly, the determination apparatus 10).

  First, the user (measurement target person) places the tips of the incident optical fiber 4 and the reflecting optical fiber 5 on the skin part where measurement is desired. When excitation light (for example, 290 nm) having a wavelength suitable for the measurement of tryptophan AGEs is emitted from the excitation light source 2, the excitation light is emitted from the tip of the incident optical fiber 4 to the skin (excitation light emission step).

  When tryptophan AGEs contained in the skin are irradiated with excitation light, fluorescence is emitted. This fluorescence enters from the tip of the reflecting optical fiber 5 and is guided to the detector 3.

  When receiving the fluorescence (fluorescence light receiving step), the detector 3 measures the fluorescence intensity for each wavelength of the fluorescence, and outputs measurement data indicating the measurement result to the data acquisition unit 11 of the determination device 10.

  When the data acquisition unit 11 receives the measurement data (S1), the data acquisition unit 11 performs analysis such as peak separation / extraction on the measurement data, stores the analysis result in the storage unit 13, and receives the measurement data. Is output to the determination unit 12.

  The determination unit 12 extracts the fluorescence intensity at the peak wavelength (eg, 390 nm) of tryptophan AGEs from the analysis result stored in the storage unit 13 (S2).

  Then, the determination unit 12 determines the degree of deterioration of the skin condition by comparing the extracted fluorescence intensity (measured fluorescence intensity) with the reference fluorescence intensity stored in the storage unit 13 (determination step) (S3). ).

  As the reference fluorescence intensity, for example, an average value of the fluorescence intensity of tryptophan AGEs in the skin is obtained for each age group such as teens and 20s, and stored in the storage unit 13 in advance. The age of the user to be measured is stored in the storage unit 13 in advance, or is input by the operation unit 15 at the time of measurement.

  The determination unit 12 reads the reference fluorescence intensity corresponding to the age of the measurement target user from the storage unit 13 and compares it with the actually measured fluorescence intensity. For example, when the measured fluorescence intensity is higher than the reference fluorescence intensity, the determination unit 12 determines that the user's skin condition is bad.

  When the determination of the skin condition is finished, the determination unit 12 outputs the determination result to the display unit 14 and displays the determination result on the display unit 14 (S4).

(Example of changing the evaluation method)
The determination unit 12 calculates a ratio between the fluorescence intensity derived from collagen that is easily affected by aging and the fluorescence intensity derived from tryptophan AGEs, and based on the result of comparing the calculated ratio value with the reference fluorescence intensity, the skin state The degree of deterioration may be determined. The reference fluorescence intensity in this case is a statistical value (for example, an average value) of the ratio between the fluorescence intensity of collagen and the fluorescence intensity of tryptophan AGEs obtained for each age group.

[Experimental example]
Next, the fact that tryptophan AGEs can be used as an index of deterioration of the skin condition will be described using actual measurement results.

<Fluorescence measurement examples for different measurement subjects>
Using a commercially available fluorometer (SkinSkan, JY Horiba) as the measuring apparatus 1, a fluorescence spectrum was obtained noninvasively from the forearm skin of the measurement subject. The result is shown in FIG. FIG. 3 is a diagram showing excitation spectra at a fluorescence wavelength of 400 nm obtained from different measurement subjects.

  In the measurement example shown in FIG. 3, the fluorescence wavelength is set to 400 nm, and the excitation light wavelength is scanned from 270 nm to 370 nm.

  In subjects A (42 years old) and B (55 years old), a peak of excitation light derived from AGE collagen was observed at around 340 nm, and there was almost no difference between subjects in the peak intensity.

  On the other hand, in subject B, a peak was observed near 290 nm, but in subject A, it was not observed.

  In order to examine what the peak at 290 nm of the excitation light originates from, fluorescence spectra of artificially synthesized AGEs derived from various amino acids were measured. A commercially available fluorometer (Hitachi High-Technologies Corporation, Horiba, Ltd.) was used for this measurement.

  As amino acids, 20 kinds of amino acids existing in the living body were used. In the synthesis of amino acid-derived AGEs, an amino acid (4.5 g / dl) and ribose (0.5 M) were mixed in a 1/15 M phosphate buffer and reacted in an incubator set at 40 ° C. for 60 days.

  The AGEs derived from various amino acids after the reaction were irradiated with an excitation wavelength of 365 nm, and the peak wavelength of the obtained fluorescence was measured. FIG. 4 is a diagram showing fluorescence peak wavelengths of AGEs derived from various amino acids.

  As shown in FIG. 4, the peak wavelength of fluorescence obtained when irradiated with an excitation wavelength of 365 nm was observed only around 390 nm for tryptophan AGEs, and other 19 amino acid-derived AGEs were observed around 430 nm to 460 nm.

  Therefore, in the measurement example of FIG. 3, it is estimated that the peak seen only in the subject B that emits fluorescence of 400 nm when irradiated with excitation light of 290 nm is a peak derived from tryptophan AGEs. In order to further verify this, the following experiments were conducted.

(Production and quantification experiment of tryptophan AGEs)
Tryptophan AGEs were artificially synthesized, and the fluorescence of the synthesized tryptophan AGEs was measured using a fluorometer. A commercially available fluorometer (Hitachi High-Technologies Corporation, HORIBA, Ltd.) was used for the measurement.

  For the synthesis of tryptophan AGEs, tryptophan (4.5 g / dl) and ribose (0.5 M) were mixed in a 1/15 M phosphate buffer and reacted in an incubator set at 40 ° C.

  FIG. 5 shows the relationship between the fluorescence intensity derived from tryptophan AGEs (vertical axis) and saccharification time (horizontal axis). The fluorescence intensity on the vertical axis is the intensity of the fluorescence wavelength around 390 nm obtained when the excitation wavelength of 365 nm is irradiated.

  As shown in FIG. 5, the fluorescence intensity increased in proportion to the increase in saccharification time (days). From this result, it was found that the amount of tryptophan AGEs produced increased in proportion to the saccharification reaction time of tryptophan, and the amount produced could be determined quantitatively by measuring the fluorescence intensity.

  Therefore, it was shown that changes in the amount of tryptophan AGEs accumulated in the skin can be quantitatively measured by measuring the fluorescence intensity.

(Measurement of fluorescence spectrum of tryptophan AGEs)
Next, the fluorescence spectrum of tryptophan AGEs synthesized as described above was measured. FIG. 6 shows the fluorescence spectrum of tryptophan (4.5 g / dl) and the fluorescence spectrum of tryptophan AGEs incubated for 159 days as described above.

  The fluorescence spectrum of tryptophan is obtained by setting the excitation light wavelength to 310 nm and scanning the fluorescence wavelength from 320 nm to 550 nm. The fluorescence spectrum of tryptophan AGEs is obtained by setting the excitation light wavelength to 290 nm and scanning the fluorescence wavelength from 300 nm to 550 nm.

  As shown in FIG. 6, a tryptophan-derived fluorescence peak was detected in the vicinity of 350 nm, and a tryptophan AGEs-derived fluorescence peak was detected in the vicinity of 390 nm.

  Moreover, the excitation spectrum of tryptophan AGEs incubated for 159 days as described above is shown in FIG. This excitation spectrum is obtained by setting the fluorescence wavelength to 390 nm and scanning the excitation light wavelength from 250 nm to 370 nm. As shown in FIG. 7, a peak of excitation light derived from tryptophan AGEs was detected near 290 nm.

  From these results, in the measurement example of FIG. 3, it is considered that the peak seen only in the subject B that emits fluorescence of 400 nm when irradiated with excitation light of 290 nm is a peak derived from tryptophan AGEs.

<Verification of availability of tryptophan AGEs as markers>
The peak of tryptophan AGEs was not detected in the 42-year-old subject A, but was detected in the older 55-year-old subject B. Therefore, it is possible that tryptophan AGEs may be an indicator (marker) of deterioration of the skin condition. It was.

  In order to verify the possibility that tryptophan AGEs could be a marker for deterioration of skin condition, the following experiment was conducted.

  In order to realize reaction conditions that are as close as possible to the in vivo conditions, tryptophan was saccharified using glucose, which is less reactive than ribose. For saccharification, tryptophan (4.5 g / dl) and glucose (5.6 mM) were mixed in a 1/15 M phosphate buffer and reacted in an incubator set at 37 ° C. The glucose concentration was adjusted so as to fall within the normal blood glucose concentration (80-100 mg / dl) range of a healthy person.

  A commercially available fluorometer (Hitachi High-Technologies Corporation, HORIBA, Ltd.) was used for the fluorescence measurement.

  FIG. 8 shows the fluorescence spectrum of the sample incubated for 134 days as described above. This fluorescence spectrum is obtained by setting the excitation light wavelength to 290 nm and scanning the fluorescence wavelength from 300 nm to 550 nm.

  As shown in FIG. 8, only a tryptophan-derived fluorescence peak around 350 nm was detected, and if tryptophan AGEs were generated, a tryptophan AGEs-derived fluorescence peak that should be seen around 390 nm was not detected.

  From this result, it was found that tryptophan AGEs are difficult to be produced in the presence of glucose at a normal blood sugar concentration of a healthy person. On the other hand, as shown in FIG. 6, tryptophan AGEs are produced by reacting a high concentration of sugar with tryptophan.

  For this reason, it has been shown that tryptophan AGEs may be generated by an insidious diet and chronic hyperglycemia.

  From the above results, it can be seen that, among amino acid-derived AGEs, tryptophan AGEs can be suitably used as a marker for deterioration of skin condition, particularly because tryptophan AGEs sensitively reflects deterioration of skin condition due to long-term dietary disturbance. It was.

  This example does not deny the use of amino acid-derived AGEs other than tryptophan AGEs as an indicator of skin condition, but shows that tryptophan AGEs are particularly preferred as an indicator of skin condition among amino acid-derived AGEs. It is.

  In the measurement method of the present invention, only the tryptophan AGEs are monitored, so the excitation light source 2 provided in the measurement device 1 may be a single light source. Therefore, the specification of the measuring device can be made compact. Furthermore, since correction for each wavelength is not necessary in the fluorescence measurement, an effect such as shortening the measurement time can be obtained.

  In the above-described fluorescence measurement, in order to measure the fluorescence intensity more accurately, it is preferable to irradiate a specific reference substance with excitation light and correct the fluorescence intensity using the fluorescence as a reference (reference).

  That is, in the measurement method of the present invention, the intensity of the fluorescence received in the light receiving process is determined by using the standard fluorescence receiving process for receiving the fluorescence that is the standard for measuring the fluorescence intensity and the intensity of the fluorescence received in the standard fluorescence receiving process. And a correction step of correcting. By performing such correction of the fluorescence intensity, the reliability of the measured value can be increased.

  As an example of calculation in the case of performing correction using a reference, for example, taking a ratio of a measured value and a reference can be mentioned.

  Further, as the reference material, it is preferable to select a material in which the fluorescence intensity does not easily decrease even when the excitation light is irradiated for a long time. As such a reference substance, it is preferable to use a nanoparticle phosphor using nanometer-size particles. Nanoparticle phosphors are less likely to reduce the intensity of fluorescence generated even when irradiated with excitation light continuously. When fluorescent beads are used as the reference substance, there is a problem that the fluorescence intensity gradually decreases when excitation light is continuously irradiated.

  In addition, a nanoparticle fluorescent substance can adjust a fluorescence wavelength by adjusting the size of a nanoparticle.

(Other)
The present invention is not limited to the above-described embodiments, and various modifications are possible within the scope shown in the claims, and embodiments obtained by appropriately combining technical means disclosed in the embodiments. Is also included in the technical scope of the present invention.

  Moreover, each block of the determination apparatus 10 mentioned above, especially the determination part 12, may be comprised by hardware logic, and may be implement | achieved by software using CPU as follows.

  That is, the determination apparatus 10 includes a CPU (central processing unit) that executes instructions of a control program that realizes each function, a ROM (read only memory) that stores the program, a RAM (random access memory) that expands the program, A storage device (recording medium) such as a memory for storing the program and various data is provided. The object of the present invention is to record the program code (execution format program, intermediate code program, source program) of the control program (determination program) of the determination apparatus 10 which is software that realizes the above-described functions in a computer-readable manner. This can also be achieved by supplying the recording medium to the determination apparatus 10 and reading and executing the program code recorded on the recording medium by the computer (or CPU or MPU).

  Examples of the recording medium include a tape system such as a magnetic tape and a cassette tape, a magnetic disk such as a floppy (registered trademark) disk / hard disk, and an optical disk such as a CD-ROM / MO / MD / DVD / CD-R. Card system such as IC card, IC card (including memory card) / optical card, or semiconductor memory system such as mask ROM / EPROM / EEPROM / flash ROM.

  The determination device 10 may be configured to be connectable to a communication network, and the program code may be supplied via the communication network. The communication network is not particularly limited. For example, the Internet, intranet, extranet, LAN, ISDN, VAN, CATV communication network, virtual private network, telephone line network, mobile communication network, satellite communication. A net or the like is available. Further, the transmission medium constituting the communication network is not particularly limited. For example, even in the case of wired such as IEEE 1394, USB, power line carrier, cable TV line, telephone line, ADSL line, etc., infrared rays such as IrDA and remote control, Bluetooth ( (Registered trademark), 802.11 wireless, HDR (high data rate), mobile phone network, satellite line, terrestrial digital network, and the like can also be used. The present invention can also be realized in the form of a computer data signal embedded in a carrier wave in which the program code is embodied by electronic transmission.

  The present invention can also be expressed as follows.

  That is, the measurement method of the present invention is characterized by evaluating pathological aging of the skin using tryptophan AGEs contained in the skin.

  Further, in the above measurement method, it is possible to evaluate pathological aging of the skin by irradiating a specific site or specific position of the skin with excitation light, observing the fluorescence of tryptophan AGEs, and measuring the fluorescence intensity. preferable.

  The excitation light preferably has a wavelength range suitable for measuring tryptophan AGEs.

  The wavelength of the excitation light is preferably within a wavelength range from about 280 nm to about 400 nm.

  Moreover, it is preferable that the fluorescence wavelength of the tryptophan AGEs detected by the excitation is in a wavelength range from about 370 nm to about 410 nm.

  In the measurement method described above, it is preferable to correct the fluorescence intensity of tryptophan AGEs using the fluorescence intensity as a standard for measuring the fluorescence intensity.

  When tryptophan AGEs are used as skin aging markers, the health condition of the skin can be determined with high accuracy. Therefore, the present invention can be used in many aspects such as selection of cosmetics / pharmaceuticals such as anti-aging and anti-glycation effects suitable for the skin condition, skin counseling, and evaluation / monitoring of the effectiveness of cosmetics / pharmaceuticals.

1 Measuring device (measurement unit)
2 Excitation light source 3 Detector 10 Determination device (determination unit)
12 determination unit 20 skin condition determination system (skin condition determination device)
30 Measurement target part

Claims (13)

A measurement unit for measuring the fluorescence intensity of tryptophan AGEs (Advanced Glycation Endproducts) in the skin of an individual to be measured;
A skin condition determination apparatus comprising: a determination unit that determines the skin condition using the fluorescence intensity measured by the measurement unit.   The skin state determination device according to claim 1, wherein the determination unit determines the skin state based on a result of comparing the fluorescence intensity measured by the measurement unit with a predetermined reference value.   3. The determination unit according to claim 2, wherein when the fluorescence intensity measured by the measurement unit is higher than the predetermined reference value, the determination unit determines that the skin state is out of a normal state. Skin condition judging device.   The skin condition determination device according to claim 2 or 3, wherein the predetermined reference value is a value based on a statistical value of fluorescence intensity of tryptophan AGEs in an age group to which the individual belongs.   The said measurement part irradiates excitation light with respect to the said skin, and measures the intensity | strength of the fluorescence generate | occur | produced by irradiation of the said excitation light, The skin state of any one of Claims 1-4 characterized by the above-mentioned. Judgment device.   6. The skin condition determination apparatus according to claim 5, wherein the excitation light has a wavelength suitable for detecting fluorescence of tryptophan AGEs.   The skin condition determination apparatus according to claim 6, wherein the peak wavelength of the excitation light is included in a wavelength range of 280 nm to 400 nm.   The skin condition determination device according to any one of claims 1 to 7, wherein the measurement unit detects fluorescence having a peak wavelength within a wavelength range of 370 nm to 410 nm as fluorescence of tryptophan AGEs. .   The skin state determination device according to claim 1, wherein the measurement unit and the determination unit are physically separable and can be connected to be communicable.   The skin condition determination program for functioning a computer as a determination part with which the skin condition determination apparatus of any one of Claims 1-9 is provided.   The computer-readable recording medium which recorded the skin condition determination program of Claim 10. A measurement step of measuring fluorescence intensity of tryptophan AGEs (Advanced Glycation Endproducts) in the skin;
And a determination step of determining the skin state using the fluorescence intensity measured in the measurement step.   The skin condition determination method according to claim 12, further comprising a correction step of correcting the fluorescence intensity of tryptophan AGEs using the fluorescence intensity as a standard for measuring the fluorescence intensity.

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