JP4657175B2 - Measuring method of protein fiber retention - Google Patents

Description

  The present invention relates to a method for measuring the amount of protein fiber retentate, a method for measuring the permeation distribution of protein fiber solution using the same, a method for measuring the swelling rate of protein fibers, and a method for measuring the amount of agent permeation using the same. About.

  In recent years, many agents used for hair such as shampoos, rinses, conditioners, etc., to which water-retaining components, moisturizing components and the like are added have been provided. In the development of hair dyes and bleaching agents, the effect on the water retention and moisture retention of the hair after use is also considered. Under such circumstances, in the development of agents used for hair such as shampoos, rinses, conditioners, hair dyes, bleaching agents, etc., it is necessary to grasp the amount of water retention and the degree of water penetration in the local area of the hair. It is considered to be extremely effective in selecting the above and grasping the penetration of these agents into the hair.

  Various techniques have been proposed so far for techniques for measuring the water retention amount and water penetration distribution (see Patent Document 1 below). However, since these techniques determine the average moisture content of the entire hair, It was difficult to grasp the water retention amount of the typical hair.

  On the other hand, as in the techniques described in Non-Patent Documents 1 and 2 below, a technique has been proposed in which the Raman spectrum of the skin is measured by confocal Raman spectroscopy and the moisture content in the depth direction is measured. In the technique of Non-Patent Document 1, the skin is pressed against a plate having a pinhole, and the skin surface exposed through the pinhole is measured by confocal Raman measurement. In the technique of Non-Patent Document 2, the skin is pressed against the transparent substrate, and the measurement of the transparent substrate is performed on the formed skin plane. None of the techniques can acquire chemical composition information in the local area of the skin with the skin surface immersed in the agent, so even if these techniques are applied as they are, the hair remains immersed in the agent. It was difficult to grasp the amount of water retained locally.

JP 2003-344279 A L. Chrit et al .: "In vivo chemical investigation of human skin using a confocal Raman fiber optic microprobe", Journal of Biomedical Optics 10 (4), 044007 (July / August 2005) PJ Caspers et al .: "In vivo confocal Raman microscopy of the skin: noninvasive determination of molecular concentration profile", Journal of Investigative Dermatology 116 (3), 434 (2001)

  The present invention has been made in view of the above-described problems, and is suitable for the amount of liquid retention, the liquid permeation distribution, the method for measuring the swelling rate, and the permeation amount of the agent in the region of the protein fiber immersed in the agent. A method for measuring the amount of protein retentate locally, a method for measuring the permeation distribution of protein fiber liquid, a method for measuring the swelling rate of protein fibers, and a method for measuring the amount of agent permeation into protein fibers. The purpose is to provide.

  The present invention measures the Raman spectrum of each known reference protein having a different liquid retention amount by confocal Raman spectroscopy, creates a calibration curve of the liquid retention amount of the reference protein based on the measurement result, and becomes a measurement object A method for measuring a protein fiber retentive amount by measuring a Raman spectrum of a protein fiber by confocal Raman spectroscopy, and using the measurement result of the protein fiber and the calibration curve to determine a retentive amount of the protein fiber locally. By providing the above, the above object is achieved.

  The present invention also provides a method for measuring the amount of protein fiber retained in the present invention, wherein the Raman spectrum of the protein fiber is measured by confocal Raman spectroscopy while changing the depth from the surface. A method for measuring the permeation distribution of a protein fiber liquid is provided.

  Moreover, the present invention measures the amount of the protein fiber locally retained by the method for measuring the amount of protein fiber retained according to the present invention, and based on the measurement result, the density of water and the hair in a non-swelled state is measured. The present invention provides a method for measuring the swelling rate of protein fibers to obtain the local swelling rate in a protein solution.

  In addition, the present invention measures the Raman spectrum of each known reference protein having a different liquid retention amount and each solution of a known agent having a different concentration by confocal Raman spectroscopy, and the reference protein is based on the measurement results. A calibration curve of the amount of the agent held in is prepared, the Raman spectrum of the protein fiber to be measured is measured by confocal Raman spectroscopy, and the protein fiber locally in the protein fiber using the measurement result and the calibration curve The present invention provides a method for measuring the amount of penetration of an agent into a protein fiber for obtaining the amount of the agent to be retained.

  ADVANTAGE OF THE INVENTION According to this invention, the liquid retention amount in the local of protein fibers, such as hair, the penetration distribution of a liquid, a swelling rate, and the penetration amount of an agent can be measured suitably.

The present invention will be described below based on preferred embodiments thereof.
First, a preferred embodiment in which the method for measuring the permeation distribution of protein fiber liquid and the method for measuring the amount of protein fiber retention of the present invention is applied to the method of measuring the water penetration distribution of hair and the method of measuring the water retention amount of hair. Based on

  The reference protein used in this embodiment is selected according to the protein fiber to be measured. As in this embodiment, when the protein fiber to be measured is hair, preferred reference proteins include water-soluble proteins such as gelatin, albumin, and casein. Among these, gelatin is preferable from the viewpoint of high water solubility, handleability, availability, and the like.

  In the measurement method of this embodiment, a Raman spectrum for each known reference protein having a different water retention amount is measured by confocal Raman spectroscopy, and a calibration curve for the water retention amount of the reference protein is created based on the measurement result. Here, the water retention amount is expressed as water [mass%] / reference protein [mass%]. Then, the Raman spectrum of the hair to be measured is measured by confocal Raman spectroscopy, and the water retention amount in the local area of the hair is obtained using the measurement result and the calibration curve. Hereinafter, the measurement method of this embodiment will be described in more detail. In the following description, the term “Raman spectrum” refers to a Raman spectrum measured by confocal Raman spectroscopy.

  As described above, when measuring the Raman spectrum of known reference proteins with different water retention amounts, water and reference proteins are uniformly mixed at a fixed ratio, and the mixed sample is irradiated with laser light. The Raman scattered light generated from the unit is collected and introduced into a spectrometer equipped with a detector. Laser irradiation and condensing of the mixed sample may be performed through a window material such as glass, or a water immersion type objective lens may be brought into direct contact with the sample.

Next, in the Raman spectrum obtained as a result of the measurement, the Raman spectrum IM1 (ω) of the reference protein whose water retention amount is known has α and β as parameters and does not contain water as shown in the following formula (1). It is represented by the Raman spectrum IN1 (ω) of the reference protein (absolutely dry state: after drying at 25 ° C. for 72 hours and the same applies below) and the Raman spectrum IN2 (ω) of water, and IM1 (ω), IN1 ( Parameters α and β, or β / α are obtained from ω) and IN2 (ω). Here, ω is the wave number of the Raman shift. Then, a calibration curve is created from the relationship between the obtained parameters α and β, or β / α, and the water retention amount of the reference protein (water [mass%] / reference protein [mass%]).
IM1 (ω) = αIN1 (ω) + βIN2 (ω) (1)

Specifically, for example, a Raman spectrum is measured only for a reference protein with a different amount of water retention, a reference protein that does not contain water, and water. Then, the above formula (1) is applied to the wave number region including the peak derived from the CH stretching vibration of hair near 2900 cm ⁻¹ and the peak derived from the OH stretching vibration of water near 3300 cm ⁻¹ in each Raman spectrum. As shown in FIG. 1, a calibration curve expressed by (β / α) vs water [mass%] / reference protein [mass%] is obtained.

  Next, the Raman spectrum of the hair to be measured is measured. In this measurement, when acquiring the spectrum of the hair not containing water, the measurement is performed by irradiating laser on the hair placed on the window material or in the air. In the case of an inverted confocal Raman spectroscope, the hair and the agent are placed in a container with a window material attached to the bottom, and laser is irradiated to the hair through the window material. To measure the spectrum. In the case of an erecting confocal Raman spectrometer, the hair and the agent are placed in a container, the tip of a water immersion lens is inserted into the agent, and the spectrum is measured by irradiating the hair with laser.

Next, in the Raman spectrum obtained as a result of the measurement, the Raman spectrum IM2 (ω) of the hair whose amount of water retention is unknown is represented by the following formula (2). It is expressed by a Raman spectrum IN1 (ω) in a completely dry state and a Raman spectrum IN2 (ω) of water, and their parameters α and β, or β / α are obtained. And the water retention amount Hw of hair is calculated | required from the calculated | required parameter (alpha) and beta, or (beta) / (alpha), and the calibration curve about the water retention amount of a reference | standard protein.
IM2 (ω) = αIN1 (ω) + βIN2 (ω) (2)
Next, the hair is approximated with two components of protein and water, and the water content WW [mass%] in the hair is obtained from the following formula (3).
WW = {HW / (HW + 1)} × 100 (3)

Specifically, for example, the Raman spectrum is measured only for hair whose water retention amount is unknown, hair not containing water, and water. Then, in the wave number region including the peak area derived from the CH stretching vibration of hair near 2900 cm ⁻¹ and the peak derived from the OH stretching vibration of water near 3300 cm ⁻¹ in each Raman spectrum, the above formula (2) (Β / α) is obtained, and a water retention amount HW (water [mass%] / protein [mass%]) in the local area of the hair is obtained from a calibration curve (FIG. 1) for the water retention amount of the reference protein. Next, the water content Ww [mass%] in hair is calculated | required by applying HW to said Formula (3). Here, the local means a part of hair that becomes a focal part when the laser beam is irradiated, and depends on the irradiation area (beam diameter) of the laser beam and the pinhole of the confocal optical system of the confocal Raman spectrometer. A fixed range.

  Next, the Raman spectrum of the hair as described above is measured while changing the depth from the surface, and the permeation distribution of water locally in the hair is obtained. The narrower the measurement depth, the more accurate the results are obtained. However, considering that the thickness of the hair cuticle is about 0.5 μm, the measurement is performed at intervals of 0.25 to 0.5 μm. Thus, a desired measurement result can be obtained.

  As described above, according to the method for measuring the water retention amount of the hair and the method for measuring the water penetration distribution of the hair according to this embodiment, the local water retention amount and the water penetration distribution of the hair can be easily measured. . Therefore, as will be described later, the local swelling rate of hair and the penetration amount of the agent can be easily measured. In this embodiment, the water retention amount of the hair is measured with respect to the hair in water, and the water penetration amount and water penetration distribution are measured. Can be grasped.

  Next, the method for measuring the swelling rate of the protein fiber of the present invention will be described based on a preferred embodiment applied to the method for measuring the swelling rate by retaining the hair.

In the method for measuring the swelling rate of this embodiment, first, the Raman spectrum of hair is measured as in the method for measuring the water retention amount of the hair of the above embodiment. Then, the amount of water in the local area of the hair is measured, and the measurement result (HW), the density of the water (DW), and the density of the hair in the non-swelled state (absolutely dry state) (DH) are swollen by the following formula (4). Find the rate. However, water and hair are ideally mixed with water retention, and there is no volume shrinkage associated with mixing.
Swelling ratio [%] = [(HW / DW) + {(1-HW) / DH}] / {(1-HW) / DH} × 100 (4)

  Thus, according to the method for measuring the swelling rate of the present embodiment, the local swelling rate of hair accompanying water retention can be suitably measured. According to the method for measuring the swelling rate of the present embodiment, since the change in the local swelling rate of the hair can be grasped, the influence of the damage caused by the distortion of the hair internal structure due to the local swelling of the hair is grasped. can do.

  Next, the method for measuring the penetration amount of the agent in the protein fiber of the present invention will be described based on a preferred embodiment applied to the method for measuring the penetration amount of hydrogen peroxide in the aqueous bleaching agent in the hair.

  In the measurement method of the present embodiment, the Raman spectrum is measured for each known reference protein having a different water retention amount and each aqueous solution of a known bleaching agent having a different concentration, and the bleaching agent for the reference protein is determined based on the measurement results. Create a calibration curve for the amount of protein in it, measure the Raman spectrum of the protein fiber to be measured, and determine the amount of bleaching agent retained locally in the protein fiber using the measurement result and the calibration curve, The amount of hydrogen peroxide is determined from the weight fraction of hydrogen peroxide in the bleaching agent. Hereinafter, the measurement method of this embodiment will be described in more detail.

  First, in the same manner as the method for measuring the water retention amount of the hair in the above embodiment, the Raman spectrum of a reference protein with a known water retention amount is measured, and the obtained measurement results are obtained with parameters α and β, or β / α, A calibration curve is created from the relationship with the water content of the reference protein. For example, a calibration curve represented by (β / α) vs (water [mass%] / reference protein [mass%]) as shown in FIG. 1 is created. Also in this embodiment, it is preferable that the measurement of the Raman spectrum of hair is performed in water, except for the measurement of hair not containing water, as in the above embodiment.

  Next, the Raman spectrum of an aqueous solution of bleaching agent having a known concentration is measured. This measurement method is basically the same as the above-described method for measuring hair in water. However, bleaching agent generates hydrogen peroxide after mixing one agent and two agents, so one agent and two agents. It is preferable to start the hair treatment immediately after mixing.

Next, in the Raman spectrum obtained as a result of the measurement, the Raman spectrum IM4 (ω) of the aqueous solution of bleaching agent having a known concentration is represented by the following equation (5), with β and γ as parameters, The parameters β and γ, or γ / β are determined by the Raman spectrum IN2 (ω) and the Raman spectrum IN3 (ω) of the bleaching agent only. Then, a calibration curve is created from the relationship between the obtained parameters β and γ, or γ / β, and bleaching agent [mass%] / water [mass%]).
IM4 (ω) = βIN2 (ω) + γIN3 (ω) (5)

Specifically, for example, a Raman spectrum is measured for an aqueous solution of bleaching agent with different concentrations, only the bleaching agent, and only water. For each Raman spectrum, the wave number region including the peak derived from the OH stretching vibration of water near 3300 cm ⁻¹ and the OO stretching vibration of hydrogen peroxide in the bleaching agent near 870 cm ^{−1 is} expressed by the above formula (5). Is applied to obtain a calibration curve represented by (γ / β) vs (bleaching agent [mass%] / water [mass%]).

  Then, the obtained slopes of the calibration curves relating to the above formulas (1) and (5) were integrated, and (γ / α) vs (bleaching agent [mass%] / reference protein [mass%]). The slope coefficient of the calibration curve of the amount of bleaching agent held in the reference protein represented by

Next, when immersed in a bleaching agent aqueous solution of a predetermined concentration, the Raman spectrum of hair (the Raman spectrum of hair with unknown bleaching agent retention amount) IM5 (ω) is expressed as α, It is assumed that γ is a parameter and is represented by a Raman spectrum IN1 (ω) of hair not containing water (absolutely dry state) and a Raman spectrum IN3 (ω) of a bleaching agent, and those parameters α and γ, or γ / α Ask for. From the combination of the obtained parameters α and γ, or γ / α, and the above-mentioned calibration curve for the water retention amount and the concentration of the bleaching agent, the bleaching agent holding amount HB of the hair bleaching agent unknown Ask.
IM5 (ω) = αIN1 (ω) + γIN3 (ω) (6)
Next, when the hair is approximated by three components of protein, water, and bleaching agent, the bleaching agent amount WB [% by mass] in the hair is represented by the following formula (7).
WB = {HB / (HB + HW + 1)} × 100 (7)
Further, the hydrogen peroxide amount WD [mass%] in the hair is obtained from the weight fraction WBD [mass%] of the hydrogen peroxide in the bleaching agent by the following formula (8).
WD = (WB × WBD) / 100 (8)

Specifically, for example, the Raman spectrum is measured only for hair with unknown amount of bleaching agent, hair without water, and bleaching agent only. Then, for each Raman spectrum, the above-mentioned formula is used for the wave number region including the peak derived from the CH stretching vibration of hair near 2900 cm ⁻¹ and the peak derived from the OO stretching vibration of hydrogen peroxide in the bleaching agent near 870 cm ^−1. (6) is applied to obtain (γ / α), and the amount of bleaching agent in the local area of the hair where the amount of bleaching agent is unknown HB from the combination of calibration curves for the water retention amount of the reference protein and the concentration of bleaching agent HB (Hydrogen peroxide [mass%] / reference protein [mass%]) is determined. Next, HB is applied to the formula (7) to determine the bleaching agent amount WBD [mass%] in the hair. Further, WBD [mass%] is applied to the formula (8) to determine the amount of hydrogen peroxide WB [mass%] in the hair. Also in this embodiment, it is preferable to measure the Raman spectrum in a state where the hair is immersed in an aqueous solution of a bleaching agent.

  Thus, according to the method for measuring the amount of penetration of the hair agent of this embodiment, the local bleaching agent for hair and the penetration amount (retention amount) of hydrogen peroxide in the bleaching agent can be easily measured. it can. Moreover, in this embodiment, since the penetration amount can be measured in a state where the hair is immersed in the bleaching agent (an aqueous solution thereof), it is possible to obtain a measurement result of the penetration amount in accordance with the use form of the bleaching agent.

  The present invention is not limited to the embodiment. In the present invention, hair is suitable as the protein fiber to be measured as in the above embodiment, but the protein fiber to be measured in the present invention is not particularly limited as long as it is made of protein fiber. Preferred protein fibers to be measured include natural protein fibers such as animal hair and silk and various artificial fibers in addition to human hair.

In the present invention, the agent for measuring the amount of penetration is not limited to the bleaching agent as in the above embodiment. The agent to be measured for the amount of penetration to which the present invention is applied has a Raman spectrum peak that does not overlap with the expression position (cm ⁻¹ ) of the Raman spectrum peak of the protein fiber to be measured and water. Agents are preferred. Preferable agents that serve as measurement materials include shampoos, rinses, conditioners, hair dyes, hair straighteners, and the like.

  Hereinafter, the present invention will be described more specifically with reference to examples. The scope of the present invention is not limited to such examples.

[Example 1]
Using gelatin as a reference protein, a plurality of samples with different water retention amounts were prepared as described below, and Raman spectra were measured under the following measurement conditions. Moreover, the Raman spectrum of only water was also measured. A commercially available confocal Raman spectrometer (manufactured by Tokyo Instruments Inc., Nano Finder) was used for the measurement of the Raman spectrum. FIG. 2 shows the measurement results. CH stretching vibration of the hair near 2900 cm ^-1, as shown in FIG. 2, the Raman spectrum having a peak derived from OH stretching vibration of water is obtained in the vicinity of 3300 cm ^-1. And the said Formula (1) was applied about the wavenumber area | region containing these two peaks, and the calibration curve about the water retention amount of a reference | standard protein as shown in FIG. 3 was calculated | required.

<Reference protein sample>
Properties: Uniform mixture of water and gelatin powder Water retention amount: 19, 9.0, 5.7, 4.0

<Measurement conditions>
A reference protein sample or water was placed in a glass bottom container. The bottom surface of the container was brought into close contact with the lens via oil immersion oil, and laser light having a wavelength of 633 nm was incident to measure the Raman spectrum.

Next, the Raman spectrum of the hair to be measured was measured by changing the depth from the hair surface under the following measurement conditions. FIG. 4 shows the measurement results. Hair CH stretching vibration near 2900 cm ^-1, as shown in FIG. 4, a Raman spectrum having a peak derived from OH stretching vibration of water is obtained in the vicinity of 3300 cm ^-1.

<Hair>
Dimensions / shape: Hair cut to a length suitable for the container. Moreover, the hair (white hair) which does not contain a melanin pigment is preferable.

<Measurement conditions>
The hair was immersed in water in a container with a glass bottom. The bottom surface of the container was brought into close contact with the lens via oil immersion oil, a laser beam having a wavelength of 633 nm was incident, and the focal position was changed in the depth direction to measure the Raman spectrum.

  Obtain the area of the above two peaks of the obtained Raman spectrum, apply it to the equation (2), obtain (β / α), and determine the local water retention amount of hair (water penetration at each depth) from the calibration curve in FIG. Distribution). Furthermore, the hair was approximated with two components of water and protein, and the depth direction distribution of the water content [% by mass] was determined based on the formula (3). The result is shown in FIG.

  As shown in FIG. 5, it was confirmed that the water retention amount decreased as the surface depth increased.

[Example 2]
Based on the water retention amount obtained under the above conditions, the swelling rate was obtained by the above formula (4). The result is shown in FIG.

  As shown in FIG. 6, it has been confirmed that the swelling rate decreases as the depth of the surface increases.

Example 3
A Raman spectrum of a reference protein with a known water retention amount was measured in the same manner as in [Example 2], and a calibration curve for the water retention amount of the reference protein as shown in FIG. 3 was obtained.

Bleaching agent aqueous solutions with different concentrations were prepared, and Raman spectra were measured in the same manner as in [Example 2]. Moreover, the Raman spectrum of only water was also measured. A Raman spectrum was obtained in which a peak derived from the OH stretching vibration of water expressed in the vicinity of 3300 cm ⁻¹ and the OO stretching vibration of hydrogen peroxide in the bleaching agent expressed in the vicinity of 870 cm ⁻¹ was obtained. And the said Formula (5) was applied about the wavenumber area | region containing these two peaks, and the calibration curve about the quantity of a bleaching agent was calculated | required.

  These calibration curves were combined to create a calibration curve for the amount of bleaching agent in the reference protein.

The Raman spectrum was measured only for hair with unknown amount of bleaching agent, hair without water and bleaching agent. Peak derived from CH stretching vibration of hair around 2900 cm ^−1, peak derived from OH stretching vibration of water near 3300 cm ⁻¹ , and OO stretching vibration of hydrogen peroxide in bleaching agent around 870 cm ⁻¹ , respectively. A Raman spectrum in which a peak appears was obtained.

  Applying to the equation (6) to obtain (γ / α), and holding the bleaching agent in the local area of the hair where the amount of the bleaching agent is unknown based on the combination of the calibration curve for the water retention amount of the reference protein and the concentration of the bleaching agent The amount (retention amount [mass%] / reference protein amount [mass%]) was determined. Next, this result was applied to the formula (7) to determine the amount of bleaching agent WBD [mass%] in the hair. Furthermore, WBD [mass%] was applied to the formula (8) to determine the amount of hydrogen peroxide [mass%] in the hair. The result is shown in FIG.

  As shown in FIG. 7, it was confirmed that the amount of hydrogen peroxide [mass%] decreased as the surface depth increased.

It is the schematic of the calibration curve of the water retention amount by the reference | standard protein used in the measuring method of the water retention amount and permeation distribution of the protein fiber of this invention. It is a figure which shows an example of the Raman spectrum of water, the reference | standard protein, and the reference | standard protein with known water retention amount in the Example of this invention. It is a figure which shows the calibration curve of the amount of water retention by the reference | standard protein obtained by the Example of this invention. It is a figure which shows the Raman spectrum in each depth of the hair whose water retention amount is unknown in the Example of this invention. It is a figure which shows the local moisture content (water penetration distribution) of the hair in each depth by the Example of this invention. It is a figure which shows the local swelling rate of the hair in each depth by the Example of this invention. It is a figure which shows the local hydrogen peroxide amount (permeation distribution of hydrogen peroxide) of the hair in each depth by the Example of this invention.

Claims (8)

  The Raman spectrum of each known reference protein with different liquid retention amount is measured by confocal Raman spectroscopy, a calibration curve of the liquid retention amount of the reference protein based on the measurement result is created, and the Raman of the protein fiber to be measured A method for measuring a retained amount of protein fiber, wherein a spectrum is measured by confocal Raman spectroscopy, and a measured amount of the protein fiber and a calibration curve are used to obtain a locally retained amount of the protein fiber.   The method for measuring the protein fiber retention amount according to claim 1, wherein the Raman spectrum is measured in a state where the protein fiber is immersed in a liquid.   The method for measuring a protein fiber retention amount according to claim 1 or 2, wherein the protein fiber is hair.   The method for measuring the amount of protein fiber retentate according to any one of claims 1 to 3, wherein the Raman spectrum of the protein fiber is measured by confocal Raman spectroscopy while changing the depth from the surface. A method for measuring the permeation distribution of protein fiber liquid to determine the local liquid permeation distribution.   The method for measuring the protein fiber retentive amount according to any one of claims 1 to 3, wherein the protein fiber local retentate amount is measured, and based on the measurement results, the water and the hair in a non-swelled state are measured. A method for measuring the swelling ratio of a protein fiber to obtain a local swelling ratio in the protein fiber from the density.   The Raman spectrum of each known reference protein having a different liquid retention amount and each solution of a known agent having a different concentration are measured by confocal Raman spectroscopy, and the amount of the agent retained in the reference protein based on the measurement results. A calibration curve of the protein fiber is prepared, the Raman spectrum of the protein fiber to be measured is measured by confocal Raman spectroscopy, and the amount of the agent held locally in the protein fiber is determined using the measurement result and the calibration curve. A method for measuring the amount of an agent penetrating into a protein fiber.   The method according to claim 6, wherein the Raman spectrum is measured in a state where the protein fiber is immersed in a liquid.   The method for measuring the amount of penetration of an agent in a protein fiber according to claim 6 or 7, wherein the protein fiber is hair.

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