JP2019045165A - Method for measuring interaction of hair protein and water - Google Patents
Method for measuring interaction of hair protein and water Download PDFInfo
- Publication number
- JP2019045165A JP2019045165A JP2017165038A JP2017165038A JP2019045165A JP 2019045165 A JP2019045165 A JP 2019045165A JP 2017165038 A JP2017165038 A JP 2017165038A JP 2017165038 A JP2017165038 A JP 2017165038A JP 2019045165 A JP2019045165 A JP 2019045165A
- Authority
- JP
- Japan
- Prior art keywords
- water
- hair
- infrared absorption
- measuring
- measured
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 53
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 34
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 34
- 238000000034 method Methods 0.000 title claims abstract description 30
- 230000003993 interaction Effects 0.000 title claims abstract description 22
- XLYOFNOQVPJJNP-ZSJDYOACSA-N Heavy water Chemical compound [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims abstract description 83
- 150000001408 amides Chemical class 0.000 claims abstract description 31
- 238000000862 absorption spectrum Methods 0.000 claims abstract description 26
- 238000004566 IR spectroscopy Methods 0.000 claims abstract description 23
- 238000009792 diffusion process Methods 0.000 claims abstract description 19
- 239000000126 substance Substances 0.000 claims description 20
- YZCKVEUIGOORGS-OUBTZVSYSA-N Deuterium Chemical group [2H] YZCKVEUIGOORGS-OUBTZVSYSA-N 0.000 claims description 13
- 229910052805 deuterium Inorganic materials 0.000 claims description 13
- 239000010432 diamond Substances 0.000 claims description 6
- 229910003460 diamond Inorganic materials 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 3
- 230000003595 spectral effect Effects 0.000 abstract description 2
- 239000013076 target substance Substances 0.000 abstract 2
- 238000005259 measurement Methods 0.000 description 19
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 4
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- 239000001257 hydrogen Substances 0.000 description 4
- 238000000691 measurement method Methods 0.000 description 4
- 230000035515 penetration Effects 0.000 description 3
- 239000011347 resin Substances 0.000 description 3
- 229920005989 resin Polymers 0.000 description 3
- 230000010354 integration Effects 0.000 description 2
- 230000004807 localization Effects 0.000 description 2
- 238000011158 quantitative evaluation Methods 0.000 description 2
- 238000003848 UV Light-Curing Methods 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000003705 background correction Methods 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- WUKWITHWXAAZEY-UHFFFAOYSA-L calcium difluoride Chemical compound [F-].[F-].[Ca+2] WUKWITHWXAAZEY-UHFFFAOYSA-L 0.000 description 1
- 229910001634 calcium fluoride Inorganic materials 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 238000003795 desorption Methods 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 238000010603 microCT Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 230000006916 protein interaction Effects 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 229940057950 sodium laureth sulfate Drugs 0.000 description 1
- SXHLENDCVBIJFO-UHFFFAOYSA-M sodium;2-[2-(2-dodecoxyethoxy)ethoxy]ethyl sulfate Chemical compound [Na+].CCCCCCCCCCCCOCCOCCOCCOS([O-])(=O)=O SXHLENDCVBIJFO-UHFFFAOYSA-M 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000036962 time dependent Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Landscapes
- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
Description
本発明は毛髪タンパク質の水との相互作用測定方法に関し、詳しくは、赤外吸収分光法を用いた毛髪タンパク質の水との相互作用測定方法に関する。 The present invention relates to a method for measuring the interaction of hair proteins with water, and more particularly to a method for measuring the interaction of hair proteins with water using infrared absorption spectroscopy.
水はキューティクルから毛髪内部へと浸透し、毛髪の物性や官能に影響を及ぼす。従来の毛髪の水分量測定法は重量測定や、膨潤度測定およびＸ線マイクロＣＴを用いた形態観察があるが、いずれも直接水を評価する方法ではなく、リアルタイムでの水浸透の測定は実現されていない（特許文献１、非特許文献１）。また、これまでに発明者らは、毛髪に重水を接触させることで、毛髪内部に存在する水と毛髪外部からの水とを判別できることを見出している（特許文献２）。 Water penetrates from the cuticle into the hair and affects the physical properties and sensory characteristics of the hair. Conventional methods for measuring the water content of hair include weight measurement, swelling measurement, and morphological observation using X-ray micro-CT, but it is not a method to directly evaluate water, but measurement of water permeation in real time is realized It is not done (patent documents 1, non-patent documents 1). In addition, the inventors have found that by contacting heavy water with hair, it is possible to distinguish between water present inside the hair and water from the outside of the hair (Patent Document 2).
顕微ＦＴ−ＩＲを用いて、キューティクルから毛髪内部へ拡散する重水によって引き起こされる毛髪タンパク質のスペクトル変化を連続的に評価することで、毛髪内部への水拡散による毛髪タンパク質の水との相互作用を可視化する方法を見出そうとするものである。 Visualize the hair protein interaction with water by water diffusion inside the hair by continuously evaluating the spectral change of the hair protein caused by heavy water that diffuses from the cuticle into the hair interior using micro-FT-IR Trying to find a way to
本発明者は、これらの従来の問題点を解決するために鋭意検討した結果、毛髪の横断面切片を赤外光に透明なセルで挟んだ後に重水を接触させ、赤外吸収分光法によって測定するアミドII及びＯＤ伸縮振動の信号強度を用いることにより、これまでの問題点を解決できることを見出し、本発明を完成するに至った。 As a result of intensive investigations to solve these conventional problems, the inventor of the present invention measured the infrared cross-section of the cross section of the hair with infrared light and then contacted with heavy water to measure by infrared absorption spectroscopy. It has been found that the above problems can be solved by using the signal strengths of the amide II and OD stretching vibrations to complete the present invention.
本願第一の発明は、毛髪の横断面切片を赤外光に透明なセルで挟んだ後に赤外吸収分光法によって前記被測定物質の赤外吸収スペクトルを測定する工程Ｉと、前記被測定物質に重水を接触させた後に赤外吸収分光法によって測定した赤外吸収スペクトルのうち、１４４０ｃｍ−１付近の重水素置換されたアミドIIの信号強度Ａｘを測定する工程と、前記Ａｘを得た波数における工程Ｉの信号強度Ａ０を求める工程と、Ａｘ−Ａ０を計算する工程とを含むことを特徴とする水拡散による毛髪タンパク質の水との相互作用測定方法である。 The first invention of the present application is a process I of measuring the infrared absorption spectrum of the substance to be measured by infrared absorption spectroscopy after sandwiching a cross section of the hair with a transparent cell to infrared light, and the substance to be measured Measuring the signal intensity Ax of the deuterium-substituted amide II in the vicinity of 1440 cm −1 in the infrared absorption spectrum measured by infrared absorption spectroscopy after bringing heavy water into contact with A method for measuring the interaction of hair proteins with water by water diffusion, comprising the steps of: determining the signal intensity A0 of step I in step 1; and calculating Ax-A0.
本願第二の発明は、毛髪の横断面切片を赤外光に透明なセルで挟んだ後に赤外吸収分光法によって前記被測定物質の赤外吸収スペクトルを測定する工程Ｉと、前記被測定物質に重水を接触させた後に赤外吸収分光法によって測定した赤外吸収スペクトルのうち、１４４０ｃｍ−１付近の重水素置換されたアミドIIの信号強度Ａｘ及び２５２０ｃｍ−１付近のＯＤ伸縮振動の信号強度Ｂｘを測定する工程と、前記Ａｘ及びＢｘを得た波数における工程Ｉの信号強度Ａ０及びＢ０を求める工程と、（Ａｘ−Ａ０）／（Ｂｘ−Ｂ０）を計算する工程とを含むことを特徴とする水拡散による毛髪タンパク質の水との相互作用測定方法
である。
The second invention of the present application is a process I of measuring the infrared absorption spectrum of the substance to be measured by infrared absorption spectroscopy after sandwiching a cross section of the hair with a transparent cell to infrared light, and the substance to be measured Of deuterium-substituted amide II in the vicinity of 1440 cm −1 and signal intensity of OD stretching vibration in the vicinity of 2520 cm −1 among infrared absorption spectra measured by infrared absorption spectroscopy after contacting heavy water with A step of measuring Bx, a step of obtaining signal intensities A0 and B0 of step I at a wave number at which Ax and Bx are obtained, and a step of calculating (Ax-A0) / (Bx-B0) This is a method for measuring the interaction of hair proteins with water by water diffusion.
本願第三の発明は、毛髪の横断面切片を赤外光に透明なセルで挟んだ後に赤外吸収分光法によって前記被測定物質の赤外吸収スペクトルを測定する工程Ｉと、前記被測定物質に重水を接触させた後に赤外吸収分光法によって測定した赤外吸収スペクトルのうち、１４４０ｃｍ−１付近の重水素置換されたアミドIIの信号強度Ａｘを測定する工程と、前記Ａｘを得た波数における工程Ｉの信号強度Ａ０及び１５４０ｃｍ−１付近のアミドIIの信号強度Ｃ０を求める工程と、（Ａｘ−Ａ０）／Ｃ０を計算する工程とを含むことを特徴とする水拡散による毛髪タンパク質の水との相互作用測定方法である。 The third invention of the present application relates to a process I of measuring an infrared absorption spectrum of the substance to be measured by infrared absorption spectroscopy after sandwiching a cross section of the hair between cells transparent to infrared light, and the substance to be measured Measuring the signal intensity Ax of the deuterium-substituted amide II in the vicinity of 1440 cm −1 in the infrared absorption spectrum measured by infrared absorption spectroscopy after bringing heavy water into contact with Determining the signal intensity A0 of the step I at step I and the signal intensity C0 of the amide II in the vicinity of 1540 cm -1, and calculating (Ax-A0) / C0; And its interaction measurement method.
本願第四の発明は、毛髪の横断面切片を赤外光に透明なセルで挟んだ後に赤外吸収分光法によって前記被測定物質の赤外吸収スペクトルを測定する工程Ｉと、前記被測定物質に重水を接触させた後に赤外吸収分光法によって測定した赤外吸収スペクトルのうち、１５４０ｃｍ−１付近のアミドIIの信号強度Ｃｘを測定する工程と、前記Ｃｘを得た波数における工程Ｉの信号強度Ｃ０を求める工程と、（Ｃ０−Ｃｘ）／Ｃ０を計算する工程とを含むことを特徴とする水拡散による毛髪タンパク質の水との相互作用測定方法である。 The fourth invention of the present application relates to a process I of measuring an infrared absorption spectrum of the substance to be measured by infrared absorption spectroscopy after sandwiching a cross section of the hair between cells transparent to infrared light, and the substance to be measured Measuring the signal intensity Cx of the amide II in the vicinity of 1540 cm −1 in the infrared absorption spectrum measured by infrared absorption spectroscopy after bringing heavy water into contact with the above, and the signal of the step I at the wave number at which Cx was obtained A method of measuring the interaction of hair proteins with water by water diffusion, comprising the steps of determining the strength C0 and calculating (C0-Cx) / C0.
本願第五の発明は、本願第一の発明〜本願第四の発明に記載の赤外光に透明なセルがダイヤモンドセルであることを特徴とする水拡散による毛髪タンパク質の水との相互作用測定方法である。 In the fifth invention of the present application, the interaction between the hair protein and the water by water diffusion is characterized in that the cell transparent to infrared light according to the first invention to the fourth invention of the present application is a diamond cell. It is a method.
本願第六の発明は、本願第一の発明〜本願第五の発明に記載の重水を接触させる工程がセル間に液体の重水を注水することを特徴とする水拡散による毛髪タンパク質の水との相互作用測定方法である。 A sixth invention of the present application relates to a method for water dispersion of hair protein by water diffusion, wherein the step of bringing heavy water into contact according to the first invention to the fifth invention of the present application injects liquid heavy water between cells. It is an interaction measurement method.
本発明によれば、顕微ＦＴ−ＩＲを用いて、キューティクルから毛髪内部へ拡散する重水によって引き起こされる毛髪タンパク質のスペクトル変化を連続的に評価することで、毛髪内部への水拡散による毛髪タンパク質の水との相互作用を可視化できる。 According to the present invention, water of hair protein by water diffusion inside hair is continuously evaluated by using FT-IR to continuously evaluate the spectrum change of hair protein caused by heavy water that diffuses from cuticle into hair inside. You can visualize the interaction with
以下、本発明を詳細に説明する。 Hereinafter, the present invention will be described in detail.
本発明の水拡散による毛髪タンパク質の水との相互作用測定方法は、毛髪の横断面切片を赤外光に透明なセルで挟んだ後に赤外吸収分光法によって前記被測定物質の赤外吸収スペクトルを測定する工程Ｉを含む。ここで、毛髪の横断面切片は、毛髪を樹脂包埋、中空樹脂内への挿入固定などの固定処理を行った後にミクロトームなどで作成することができ、その切片の厚さは０．５〜１５μｍ、望ましくは４〜８μｍである。０．５μｍより薄い場合には赤外吸収スペクトルの信号強度が弱くなったり測定時の積算時間が長くなったりノイズが強くなりやすく望ましくない。１５μｍより厚い場合には赤外吸収スペクトルの信号強度が強くなりすぎ定量性に問題が生じやすい。毛髪の横断面切片の両断面が平行になるように作成することがセルと毛髪の横断面との密着を適切に行う上で望ましく、そのためにはミクロトームを用いて作成することが望ましい。 The method for measuring the interaction of hair protein with water by water diffusion according to the present invention comprises the infrared absorption spectrum of the substance to be measured by infrared absorption spectroscopy after the cross section of the hair is sandwiched between cells transparent to infrared light. And measuring I. Here, the cross section of the hair can be prepared with a microtome etc. after fixing the hair into resin, embedding in hollow resin, etc., and the thickness of the section is 0.5 to 0.5. 15 μm, preferably 4 to 8 μm. If the thickness is smaller than 0.5 μm, the signal strength of the infrared absorption spectrum may be weak, the integration time at measurement may be long, or the noise may be strong, which is not desirable. When the thickness is more than 15 μm, the signal intensity of the infrared absorption spectrum becomes too strong, which tends to cause a problem in quantitativeness. It is desirable to make the cross sections of the cross section of the hair parallel to each other in order to properly adhere the cell to the cross section of the hair, and for that purpose, it is desirable to use a microtome.
また、赤外光に透明なセルとしては、ダイヤモンドセル、石英セルやフッ化カルシウムセルなどがある。ダイヤモンドセルが広く普及しており取り扱いにも優れていることから望ましい。 Moreover, as a cell transparent to infrared light, there are a diamond cell, a quartz cell, a calcium fluoride cell and the like. Diamond cells are desirable because they are widely used and excellent in handling.
前記被測定物質に重水を接触させた後に赤外吸収分光法による赤外吸収スペクトルを測定する工程を含む。毛髪と重水との接触は、毛髪の横断面切片の横断面をセルに均一に接するように挟んだ後に、そのセル間に液体の重水を注入する方法、重水蒸気下に毛髪を放置する方法などがあり、キューティクル面から選択的に重水を拡散させることができる。さらに、そのセル間を水蒸気濃度既知の空気とする方法もあり、重水と接触した後の毛髪の横断面切片の飽和重水蒸気圧よりも低い水蒸気圧の空気とすることで、毛髪内部からキューティクル面に向かって重水を拡散させることができる。 And a step of measuring an infrared absorption spectrum by infrared absorption spectroscopy after bringing the heavy water into contact with the substance to be measured. The contact between the hair and heavy water is carried out by sandwiching the cross section of the cross section of the hair in a uniform contact with the cells and then injecting the liquid heavy water between the cells, leaving the hair under heavy water vapor, etc. Can selectively diffuse heavy water from the cuticle surface. Furthermore, there is also a method in which air between the cells is known to have a known water vapor concentration, and by using air with a water vapor pressure lower than the saturated heavy water vapor pressure of the cross section of the hair after contact with heavy water Can diffuse heavy water towards the
上記工程を経て毛髪の横断面切片について測定した赤外吸収スペクトルのうち、１４４０ｃｍ−１付近の重水素置換されたアミドIIの信号強度Ａｘを測定する。赤外吸収分光法としては、透過法により測定する方法が好ましい。装置としては顕微ＦＴ−ＩＲが一般的に良く用いられる。 Among the infrared absorption spectra measured for the cross-sectional section of the hair through the above steps, the signal intensity Ax of the deuterium-substituted amide II in the vicinity of 1440 cm −1 is measured. As infrared absorption spectroscopy, a method of measurement by transmission is preferred. As an apparatus, microscopic FT-IR is generally and often used.
更に、前記Ａｘを得た波数における工程Ｉの信号強度Ａ０を求める工程と、Ａｘ−Ａ０を計算する工程とを含む。この工程により、１４４０ｃｍ−１付近の重水素置換されたアミドIIの信号強度に含まれるバックグラウンドを補正することができる。 The method further includes the steps of determining the signal intensity A0 of step I at the wave number at which Ax is obtained, and calculating Ax-A0. By this process, it is possible to correct the background contained in the signal intensity of deuterium substituted amide II around 1440 cm −1 .
本発明の水拡散による毛髪タンパク質の水との相互作用測定方法によれば、毛髪が重水に接触した直後から経時的な測定が可能であり、また、同一毛髪について重水を接触させた場合の変化、重水浸透後に重水を含まない環境に接触させた場合の重水脱離変化など繰り返し処理した時についても測定することが可能である。 According to the method for measuring the interaction of hair proteins with water by water diffusion of the present invention, it is possible to measure with time immediately after the hair comes in contact with heavy water, and changes when heavy water is brought into contact with the same hair. It is also possible to measure when repeatedly treated such as heavy water desorption change when brought into contact with an environment not containing heavy water after heavy water penetration.
上記工程を経て毛髪の横断面切片について測定した赤外吸収スペクトルのうち、１４４０ｃｍ−１付近の重水素置換されたアミドIIの信号強度Ａｘ及び２５２０ｃｍ−１付近のＯＤ伸縮振動の信号強度Ｂｘを測定する工程と、前記Ａｘ及びＢｘを得た波数における工程Ｉの信号強度Ａ０及びＢ０を求める工程と、（Ａｘ−Ａ０）／（Ｂｘ−Ｂ０）を計算する工程とを含む。このことにより、重水の拡散と重水素置換が起こった毛髪タンパク質の関係を指標化することができる。 From the infrared absorption spectrum of the cross-sectional sections of hair through the above steps, measure the signal strength Bx of OD stretching vibration in the vicinity of the signal strength Ax and 2520cm -1 Deuterium substituted amides II near 1440cm -1 And determining the signal intensities A0 and B0 of step I at the wavenumber at which Ax and Bx are obtained, and calculating (Ax-A0) / (Bx-B0). This makes it possible to index the relationship between the diffusion of heavy water and the hair protein in which deuterium substitution has occurred.
また、上記工程を経て毛髪の横断面切片について測定した赤外吸収スペクトルのうち、１４４０ｃｍ−１付近の重水素置換されたアミドIIの信号強度Ａｘを測定する工程と、前記Ａｘを得た波数における工程Ｉの信号強度Ａ０及び１５４０ｃｍ−１付近のアミドIIの信号強度Ｃ０を求める工程と、（Ａｘ−Ａ０）／Ｃ０を計算する工程とを含む。 In the infrared absorption spectrum measured for the cross section of the hair through the above steps, the step of measuring the signal intensity Ax of deuterium substituted amide II in the vicinity of 1440 cm −1 and the wave number at which Ax is obtained Step of determining signal intensity A0 of Step I and signal intensity C0 of amide II near 1540 cm −1, and calculating (Ax−A0) / C0.
毛髪が重水と接触する前にその毛髪タンパク質の代表的なアミドIIの信号強度Ｃ０を測定することができ、その後に前記被測定物質に重水を接触させ水素・重水素交換されることにより新たに得られる１４４０ｃｍ−１付近の重水素置換されたアミドIIの信号強度Ａｘを測定し、前記同様にバックグラウンド補正した後に、（Ａｘ−Ａ０）／Ｃ０を算出することで、被測定毛髪のタンパク質の量を基準とした水素・重水素交換後のタンパク質の量を評価することができる。 Before the hair comes into contact with heavy water, the signal intensity C0 of a representative amide II of the hair protein can be measured, and then the above-mentioned substance to be measured is brought into contact with heavy water to exchange it with hydrogen and deuterium. The signal intensity Ax of the deuterium-substituted amide II in the vicinity of 1440 cm −1 obtained is measured, and after background correction as described above, (Ax−A0) / C0 is calculated to obtain the protein of the hair to be measured. The amount of protein after hydrogen and deuterium exchange can be evaluated based on the amount.
毛髪の横断面切片の厚みの不均一さを相殺した水浸透による毛髪タンパク質の水分との相互作用の量的評価に適している。また、毛髪の横断面切片中のタンパク質の局在を考慮した水拡散による毛髪タンパク質の水との相互作用の量的評価に適している。 It is suitable for the quantitative evaluation of the interaction of the hair proteins with the water by water penetration, which compensates for the thickness unevenness of the cross-sectional section of the hair. In addition, it is suitable for quantitative evaluation of the interaction of the hair protein with water by water diffusion in consideration of the localization of the protein in the cross section of the hair.
また、上記工程を経て毛髪の横断面切片について測定した赤外吸収スペクトルのうち、１５４０ｃｍ−１付近のアミドIIの信号強度Ｃｘを測定する工程と、前記Ｃｘを得た波数における工程Ｉの信号強度Ｃ０を求める工程と、（Ｃ０−Ｃｘ）／Ｃ０を計算する工程とを含む。 In the infrared absorption spectrum measured for the cross section of the hair through the above steps, the step of measuring the signal intensity Cx of the amide II in the vicinity of 1540 cm −1 , and the signal intensity of the step I at the wave number at which Cx was obtained The steps of determining C0 and calculating (C0-Cx) / C0 are included.
毛髪が重水と接触する前にその毛髪タンパク質の代表的なアミドIIの信号強度Ｃ０を測定することができ、その後に前記被測定物質に重水を接触させ後でも水素・重水素交換されない１５４０ｃｍ−１付近のアミドIIの信号強度Ｃｘを測定し、（Ｃ０−Ｃｘ）／Ｃ０を算出することで、被測定毛髪のタンパク質の量を基準とした水素・重水素交換の起こったタンパク質の量を評価することができる。 Before the hair comes into contact with heavy water, the signal strength C0 of the representative amide II of the hair protein can be measured, and then after the heavy water is brought into contact with the substance to be measured, 1540 cm −1 which is not hydrogen / deuterium exchanged. By measuring the signal intensity Cx of the nearby amide II and calculating (C0-Cx) / C0, the amount of protein in which hydrogen and deuterium exchange have occurred is evaluated based on the amount of protein in the hair to be measured be able to.
以下に、実施例により本発明をより詳細に説明するが、本発明はこれによってなんら限定されるものではない。 Hereinafter, the present invention will be described in more detail by way of examples, but the present invention is not limited thereto.
健康な人毛黒髪の毛束（３０ｃｍ，２ｇ）を８質量％ラウレス硫酸ナトリウム水溶液で洗浄後、任意に選んだ毛髪についてカール半径を求め、その半径が２ｃｍ以下の毛髪をうねり毛として試験に供した。 After washing a healthy human dark brown hair bundle (30 cm, 2 g) with an 8% by weight aqueous solution of sodium laureth sulfate, the curl radius was determined for arbitrarily selected hair, and the hair having a radius of 2 cm or less was subjected to a test as a curly hair. .
次に、上記うねり毛をアクリル系ＵＶ硬化樹脂にて包埋し、工業用回転式ミクロトームを用いて厚さ６．０μｍの平滑な毛髪の横断面切片を作成した。 Next, the above-mentioned umbilical hair was embedded with an acrylic UV curing resin, and a cross section of smooth hair having a thickness of 6.0 μm was formed using an industrial rotary microtome.
上記うねり毛の横断面切片を、ダイヤモンドセル上に設置し、実体顕微鏡で確認しながら、ダイヤモンドセル間に挟み込んだ。さらにそのセル間に重水を注入した後、この毛髪切片を顕微ＦＴ−ＩＲ（サーモサイエンティフィック社製のＮｉｃｏｌｅｔ ｉＮ１０）を用い次の条件で、キューティクル最表面より２０μｍ内側の任意の２箇所（Ｓ及びＴ）における重水接触前後の１４４０ｃｍ−１付近の重水素置換されたアミドIIの信号強度及び２５２０ｃｍ−１付近のＯＤ伸縮振動の信号強度を測定した。
A cross-sectional section of the above-mentioned umbilical hair was placed on a diamond cell and sandwiched between the diamond cells while checking with a stereomicroscope. Furthermore, after injecting heavy water between the cells, this hair section is subjected to micro FT-IR (Nicolet iN10 manufactured by Thermo Scientific Co., Ltd.) under the following conditions at any two
＜測定条件＞
測定手法；ポイントマッピング、波数；４０００〜８００ｃｍ−１、積算回数；５１２、分解能；８ｃｍ−１、エリア；キューティクル最表面より２０μｍ内側の１０μｍ×１０μｍのエリア、アパーチャー幅；１０μｍ
<Measurement conditions>
Measurement Method: point mapping, wave number; 4000～800Cm -1, the number of integrations: 512, resolution; 8 cm -1, area; cuticle 20μm inner 10 [mu] m × 10 [mu] m from the outermost surface area, aperture width; 10 [mu] m
その結果のうち、キューティクル最表面より２０μｍ内側の１０μｍ×１０μｍのエリア２箇所（Ｓ及びＴ）における、重水接触後の１４４０ｃｍ−１付近の重水素置換されたアミドIIの各計測時における信号強度Ａｘと、計測開始時における１４４０ｃｍ−１付近の重水素置換されたアミドIIの信号強度Ａ０の差 Ａｘ−Ａ０の変化を表１に例示する。
Among the results, the signal intensity Ax at each measurement of deuterium-substituted amide II in the vicinity of 1440 cm −1 after contact with heavy water at two areas (S and T) of 10 μm × 10
また、上記２箇所において重水接触前後の１４４０ｃｍ−１付近の重水素置換されたアミドIIの各計測時における信号強度Ａｘと計測開始時における１４４０ｃｍ−１付近の重水素置換されたアミドIIの信号強度Ａ０、および２５２０ｃｍ−１付近のＯＤ伸縮振動の各計測時における信号強度Ｂｘと計測開始時における２５２０ｃｍ−１付近のＯＤ伸縮振動の信号強度Ｂ０から、各計測時間における（Ａｘ−Ａ０）／（Ｂｘ−Ｂ０）を算出し、表２に例示する。 In addition, the signal intensity Ax at each measurement of deuterium-substituted amide II in the vicinity of 1440 cm -1 before and after heavy water contact and the signal intensity of deuterium-substituted amide II in the vicinity of 1440 cm -1 at the start of measurement A0, and from 2520cm -1 signal strength of OD stretching vibration in the vicinity of 2520cm -1 in signal strength Bx and measurement start time at each measurement of the OD stretching vibration in the vicinity of B0, at each measurement time (Ax-A0) / (Bx Calculate -B0) and illustrate in Table 2.
水の浸透が進行しているにもかかわらず、場所によって毛髪タンパク質が水から受ける影響に差が存在することも明らかにすることができた。 It was also possible to show that there is a difference in the influence of the hair protein on water depending on the location despite the progress of water penetration.
重水接触後の１４４０ｃｍ−１付近の重水素置換されたアミドIIの各計測時における信号強度Ａｘと計測開始時における１４４０ｃｍ−１付近の重水素置換されたアミドIIの信号強度Ａ０、および１５４０ｃｍ−１付近のアミドIIの各計測時における信号強度Ｃｘと計測開始時における１５４０ｃｍ−１付近のアミドIIの信号強度Ｃ０から、各計測時間における（Ａｘ−Ａ０）／Ｃ０および（Ｃ０−Ｃｘ）／Ｃ０を算出し、表３に例示する。
Signal intensity Ax at each measurement of deuterium-substituted amide II in the vicinity of 1440 cm −1 after heavy water contact and
本発明の毛髪の水分浸透測定方法により、毛髪の横断面切片中のタンパク質の局在を考慮した水の影響の量的な比較が可能となった。 The moisture permeation measurement method of the hair of the present invention has made it possible to quantitatively compare the effects of water in consideration of the localization of proteins in cross-sectional sections of the hair.
Claims (6)
A method for measuring the interaction of hair proteins with water by water diffusion, wherein the step of bringing heavy water into contact according to any one of claims 1 to 5 comprises pouring liquid heavy water between cells.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2017165038A JP7058857B2 (en) | 2017-08-30 | 2017-08-30 | Method for measuring the interaction of hair protein with water |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2017165038A JP7058857B2 (en) | 2017-08-30 | 2017-08-30 | Method for measuring the interaction of hair protein with water |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2019045165A true JP2019045165A (en) | 2019-03-22 |
| JP7058857B2 JP7058857B2 (en) | 2022-04-25 |
Family
ID=65814017
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2017165038A Active JP7058857B2 (en) | 2017-08-30 | 2017-08-30 | Method for measuring the interaction of hair protein with water |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP7058857B2 (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004170142A (en) * | 2002-11-18 | 2004-06-17 | Mandom Corp | Method for measuring penetrance of reducing agent to hair |
| JP2006170915A (en) * | 2004-12-17 | 2006-06-29 | Kao Corp | Image processing method |
| JP2007010351A (en) * | 2005-06-28 | 2007-01-18 | Canon Inc | Sample for infrared absorption spectrum analysis, its preparing method and its preparing device |
| US20090105260A1 (en) * | 2005-02-23 | 2009-04-23 | Samir Mitragotri | Molecules to Enhance Percutaneous Delivery and Methods for Discovery Therefor |
| JP2010223609A (en) * | 2009-03-19 | 2010-10-07 | Kao Corp | Measuring method of ingredient amount in skin |
| JP2010256104A (en) * | 2009-04-23 | 2010-11-11 | Kao Corp | Evaluation method |
| JP2013533217A (en) * | 2010-05-17 | 2013-08-22 | セビックス・インコーポレイテッド | PEGylated C-peptide |
| JP2014528571A (en) * | 2011-09-29 | 2014-10-27 | アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル | Method for measuring biological values of the liver |
| JP2016224025A (en) * | 2015-05-26 | 2016-12-28 | 花王株式会社 | Method for evaluating moisture retention state of horny layer |
-
2017
- 2017-08-30 JP JP2017165038A patent/JP7058857B2/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004170142A (en) * | 2002-11-18 | 2004-06-17 | Mandom Corp | Method for measuring penetrance of reducing agent to hair |
| JP2006170915A (en) * | 2004-12-17 | 2006-06-29 | Kao Corp | Image processing method |
| US20090105260A1 (en) * | 2005-02-23 | 2009-04-23 | Samir Mitragotri | Molecules to Enhance Percutaneous Delivery and Methods for Discovery Therefor |
| JP2007010351A (en) * | 2005-06-28 | 2007-01-18 | Canon Inc | Sample for infrared absorption spectrum analysis, its preparing method and its preparing device |
| JP2010223609A (en) * | 2009-03-19 | 2010-10-07 | Kao Corp | Measuring method of ingredient amount in skin |
| JP2010256104A (en) * | 2009-04-23 | 2010-11-11 | Kao Corp | Evaluation method |
| JP2013533217A (en) * | 2010-05-17 | 2013-08-22 | セビックス・インコーポレイテッド | PEGylated C-peptide |
| JP2014528571A (en) * | 2011-09-29 | 2014-10-27 | アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル | Method for measuring biological values of the liver |
| JP2016224025A (en) * | 2015-05-26 | 2016-12-28 | 花王株式会社 | Method for evaluating moisture retention state of horny layer |
Non-Patent Citations (1)
| Title |
|---|
| 和田徹・松井とも子・田中誠之: "変成アルブミン二次構造のアミドI,II赤外吸収帯による解析", 高分子論文集, vol. 45, no. 7, JPN6021021354, July 1985 (1985-07-01), pages 461 - 465, ISSN: 0004672072 * |
Also Published As
| Publication number | Publication date |
|---|---|
| JP7058857B2 (en) | 2022-04-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Hajjarian et al. | Evaluating the viscoelastic properties of tissue from laser speckle fluctuations | |
| Luo et al. | Ultrasensitive U-shaped fiber optic LSPR cytosensing for label-free and in situ evaluation of cell surface N-glycan expression | |
| DE50212550D1 (en) | METHOD AND DEVICE FOR DETERMINING A LIGHT TRANSPORT PARAMETER AND AN ANALYTE IN A BIOLOGICAL MATRIX | |
| ATE347852T1 (en) | METHOD AND DEVICE FOR DETECTING SUBSTANCES IN BODY FLUID USING RAMAN SPECTROSCOPY | |
| DE602005025214D1 (en) | DEVICE FOR MEASURING AN ANALYTE IN A BODY LIQUID | |
| Zhang et al. | Molecularly imprinted carbon based electrodes for tetrahydrocannabinol sensing | |
| Tohmyoh et al. | Accurate determination of the structural elasticity of human hair by a small-scale bending test | |
| JP2019045165A (en) | Method for measuring interaction of hair protein and water | |
| JP6913944B2 (en) | Method for measuring the interaction of hair protein with water | |
| JP2019045162A (en) | Method for measuring interaction of hair protein and water | |
| Büyüköztürk et al. | Near infrared for non-destructive testing of articular cartilage | |
| JP6802458B2 (en) | Hair moisture penetration measurement method | |
| CN107121362A (en) | A kind of method for studying molecular migration motion | |
| JP3902999B2 (en) | Optical scattering characteristic estimation apparatus and operation method thereof | |
| Jain et al. | Polarimetric imaging in backscattering for the structural characterization of strongly scattering birefringent fibrous media | |
| Mohamad et al. | Conceptual design of near infrared spectroscopy instrumentation for skin moisture measurement | |
| JP2007313286A (en) | Method and apparatus for measurement of optical organism information | |
| You et al. | Preparation of SERS base membrane with cellulose compound dopamine and determination of hypochlorite | |
| Kleshnin et al. | A technique for measuring oxygen saturation in biological tissues based on diffuse optical spectroscopy | |
| Esposito et al. | A new method for depth profiling reconstruction in confocal microscopy | |
| Buijs | Simpler, faster, and softer: Towards broad application of laser speckle imaging in art conservation and soft matter | |
| JP5288912B2 (en) | Method for measuring skin moisture content | |
| KR102046838B1 (en) | Method of measuring degree of hair elasticity using the shape retaining pressure | |
| Lourenço et al. | Brief descriptions of the principles of prominent methods used to study the penetration of materials into human hair and a review of examples of their use | |
| RU2148257C1 (en) | Method for examining blood |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20200722 |
|
| A977 | Report on retrieval |
Free format text: JAPANESE INTERMEDIATE CODE: A971007 Effective date: 20210519 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20210608 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20210803 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20220104 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20220106 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20220405 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20220406 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 7058857 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313111 |
|
| R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |