JPH071263B2 - Molecular weight distribution analysis method of melanin - Google Patents
Molecular weight distribution analysis method of melaninInfo
- Publication number
- JPH071263B2 JPH071263B2 JP22794689A JP22794689A JPH071263B2 JP H071263 B2 JPH071263 B2 JP H071263B2 JP 22794689 A JP22794689 A JP 22794689A JP 22794689 A JP22794689 A JP 22794689A JP H071263 B2 JPH071263 B2 JP H071263B2
- Authority
- JP
- Japan
- Prior art keywords
- melanin
- molecular weight
- weight distribution
- dmso
- gpc
- 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.)
- Expired - Lifetime
Links
Landscapes
- Investigating Or Analysing Materials By Optical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Description
【発明の詳細な説明】 ［産業上の利用分野］ この発明はメラニンの分子量分布分析法に関する。さら
に詳しくは、ゲルパーミエーションクロマトグラフィを
用いた、メラニンの品質管理、性能評価等に好適な分子
量分布の分析法に関する。TECHNICAL FIELD The present invention relates to a molecular weight distribution analysis method for melanin. More specifically, it relates to a method for analyzing molecular weight distribution suitable for quality control, performance evaluation, etc. of melanin using gel permeation chromatography.
［従来の技術］ メラニンは、フェノール類がオキシダーゼにより酸化さ
れた褐色ないし黒色の高分子色素であり、通常タンパク
質と固く結合して存在し、水やほとんどの有機溶媒に不
溶の極めて扱いにくい物質である。以上の理由により、
メラニンは通常の液体クロマトグラフィでは分析不可能
である。したがって現状ではメラニンの品質管理は不十
分なままであった。[Prior Art] Melanin is a brown or black high-molecular pigment in which phenols are oxidized by oxidase, and is usually present in a state that it is firmly bound to proteins and is insoluble in water and most organic solvents and is a very difficult substance to handle. is there. For the above reasons
Melanin cannot be analyzed by conventional liquid chromatography. Therefore, quality control of melanin remains insufficient at present.
［発明が解決しようとする課題］ 前記従来技術の課題を解決するため、この発明はメラニ
ンの品質管理や性能評価が可能な分析法を提供しようと
するものである。[Problems to be Solved by the Invention] In order to solve the above-mentioned problems of the prior art, the present invention aims to provide an analytical method capable of quality control and performance evaluation of melanin.
すなわち、メラニンの希釈剤としてジメチルスルホキシ
ド（DMSO）を用い、ゲルパーミエーションクロマトグラ
フィ（以下GPCという）の移動相としてジメチルスルホ
キシド（DMSO）を用い、検出手段として紫外線分光光度
計を用いたGPCによって分析することにより、メラニン
の分子量分布の測定を効率よく、正確に行なう方法を提
供する。That is, dimethyl sulfoxide (DMSO) is used as a diluent for melanin, dimethyl sulfoxide (DMSO) is used as a mobile phase of gel permeation chromatography (hereinafter referred to as GPC), and analysis is performed by GPC using an ultraviolet spectrophotometer as a detection means. This provides a method for efficiently and accurately measuring the molecular weight distribution of melanin.
［課題を解決するための手段］ 前記課題を解決するため、本発明方法は、ゲルパーミエ
ーションクロマトグラフィ（GPC）によりメラニンの分
子量分布を測定する方法であって、メラニンをジメチル
スルホキシド（DMSO）で希釈し、ジメチルスルホキシド
（DMSO）を溶離液として用いたゲルパーミエーションク
ロマトグラフィ（GPC）に付して、紫外線分光光度計で
検出することにより、前記メラニンの分子量分布を測定
することを特徴とするメラニンの分子量分布分析法であ
る。[Means for Solving the Problems] In order to solve the above problems, the method of the present invention is a method of measuring the molecular weight distribution of melanin by gel permeation chromatography (GPC), in which melanin is diluted with dimethyl sulfoxide (DMSO). Then, it is subjected to gel permeation chromatography (GPC) using dimethylsulfoxide (DMSO) as an eluent, and the molecular weight distribution of melanin is measured by detecting with a UV spectrophotometer. It is a molecular weight distribution analysis method.
この発明において特徴的なことは、下記の４点を有機的
に組み合わせた点にある。A characteristic of this invention is that the following four points are organically combined.
GPCを用いる点 メラニンをジメチルスルホキシド（DMSO）で希釈す
る点 ジメチルスルホキシド（DMSO）を溶離液として用い
る点 GPCの検出手段として、紫外線分光光度計を用いる
点 GPCは通常ポリマーの分子量分布を測定するのに用いら
れる。すなわち溶液に溶解または分散させたポリマーを
移動相を用いて分離カラムで分離し、各分子量のポリマ
ーを検出器で検出して分子量分布を求めるのである。そ
してメラニン試料の希釈液についてのGPCからの溶離液
を分画して補集することにより、メラニンのゾル成分の
分子量分布を得るのである。A point using GPC A point diluting melanin with dimethylsulfoxide (DMSO) A point using dimethylsulfoxide (DMSO) as an eluent A point using an ultraviolet spectrophotometer as a means for detecting GPC GPC usually measures the molecular weight distribution of a polymer Used for. That is, the polymer dissolved or dispersed in the solution is separated by a separation column using a mobile phase, and the polymer of each molecular weight is detected by a detector to obtain the molecular weight distribution. The molecular weight distribution of the sol component of melanin is obtained by fractionating and collecting the eluate from GPC for the diluted melanin sample.
次に、メラニンの希釈剤としてジメチルスルホキシド
（DMSO）を用い、GPCの移動相としてジメチルスルホキ
シドを用いる。ジメチルスルホキシド（DMSO）は、メラ
ニンの分子量分布（粒度分布）に対応した溶離をしやす
いからである。Next, dimethyl sulfoxide (DMSO) is used as a diluent for melanin, and dimethyl sulfoxide is used as a mobile phase for GPC. This is because dimethyl sulfoxide (DMSO) easily elutes according to the molecular weight distribution (particle size distribution) of melanin.
次に、GPCの検出器として紫外線分光光度計を用いる。
メラニンの分子量分布の検出に優れているからである。Next, an ultraviolet spectrophotometer is used as a GPC detector.
This is because it is excellent in detecting the molecular weight distribution of melanin.
［作用］ この発明によれば、メラニンのDMSO溶液を、DMSOからな
る移動相中に導入してGPCに付すことにより、メラニン
を効率良く分離定量することができる。また定性分析も
可能である。[Operation] According to the present invention, melanin can be efficiently separated and quantified by introducing a DMSO solution of melanin into a mobile phase made of DMSO and subjecting it to GPC. Qualitative analysis is also possible.
［実施例］ この発明において、上記メラニンがDMSOに溶解されて注
入用溶液が調整された後、これがDMSOを移動相とするGP
Cに付される。この場合、メラニン溶解用のDMSOと移動
相のDMSOとは同一であることが装置の操作上好ましい。[Examples] In the present invention, after the above-mentioned melanin was dissolved in DMSO to prepare an injectable solution, this was a GP using DMSO as a mobile phase.
Attached to C. In this case, DMSO for dissolving melanin and DMSO for the mobile phase are preferably the same in terms of operation of the apparatus.
前記においてメラニンはDMSOに50ppm程度溶解・希釈さ
せる。In the above, melanin is dissolved / diluted in DMSO at about 50 ppm.
一方、GPCで用いる分離カラムとしては、汎用の高分子
ゲルを充填したものが使用可能であり、例えばポリスチ
レンゲルや、スチレンージビニルベンゼン共重合体ゲル
の充填カラムが好適な例として挙げられる。On the other hand, as a separation column used in GPC, a column packed with a general-purpose polymer gel can be used, and for example, a column packed with polystyrene gel or styrene-divinylbenzene copolymer gel can be mentioned as a suitable example.
またGPC自体は汎用のものを使用できる。The GPC itself can be general-purpose.
以下図面および測定例により本発明方法を詳細に説明す
る。なお本発明は下記の実施例に限定されない。The method of the present invention will be described in detail below with reference to the drawings and measurement examples. The present invention is not limited to the examples below.
第１図は、この発明の方法を実施する装置の一例の構成
説明図である。この図において、１は移動貯留槽、２は
送液ポンプ、３はマニュアルインジェクタ、４はカラ
ム、５はカラムオーブン、６は紫外線分光光度計、７は
データ処理装置である。移動相１からポンプ２によりあ
らかじめ溶離液を流し、定常状態を作る。次にマニュア
ルインジェクタ３から試料を注入する。注入された試料
は溶離液によりカラムオーブン５中のカラム４に送ら
れ、検出器６（紫外線分光光度計）により分子量分布が
検出される。このデータをデータ処理装置７で処理し、
分子量分布の計算を行う。FIG. 1 is a structural explanatory view of an example of an apparatus for carrying out the method of the present invention. In this figure, 1 is a moving storage tank, 2 is a liquid feed pump, 3 is a manual injector, 4 is a column, 5 is a column oven, 6 is an ultraviolet spectrophotometer, and 7 is a data processor. The eluent is made to flow in advance from the mobile phase 1 by the pump 2 to create a steady state. Next, the sample is injected from the manual injector 3. The injected sample is sent to the column 4 in the column oven 5 by the eluent, and the detector 6 (ultraviolet spectrophotometer) detects the molecular weight distribution. This data is processed by the data processing device 7,
Calculate the molecular weight distribution.
以上装置を用いて、下記の分析条件により分析を行っ
た。すなわち移動相１にDMSO、分析カラム４に“Shodex
GPC KD−80M"（昭和電工（株）製）、検出器６に紫外
可視分光光度計（SPD−6AV）：（株）島津製作所製）を
使用し、マニュアルインジェクタ３から試料を注入し
た。試料のメラニンはDMSOに50ppmの濃度で溶解・希釈
させた。そして実際の分析結果はデータ処理装置７で解
析し分子量分布の計算を行わせた。Using the above apparatus, analysis was performed under the following analysis conditions. That is, DMSO is used for mobile phase 1 and “Shodex” is used for analytical column 4.
GPC KD-80M "(manufactured by Showa Denko KK), an ultraviolet-visible spectrophotometer (SPD-6AV): Shimadzu Corp. was used as the detector 6, and the sample was injected from the manual injector 3. Was dissolved / diluted in DMSO at a concentration of 50 ppm, and the actual analysis results were analyzed by the data processor 7 to calculate the molecular weight distribution.
分析条件 カラム:Shodex GPC KD−80M（昭和電工（株）製）（8.0
mmI.D.×300mmL） 移動相：ジメチルスルホキシド（DMSO） 流 量:0.5ml/min 温 度:40℃ 検出器:SPD−6AV（紫外線分光光度計）（（株）島津製
作所製）、 スケール（scale）;0.32AUFS レスポンス（response）;STD 波長（wavelength）;261nm 試料の前処理 試料5mgを100mlのDMSOに溶解する。Analysis conditions Column: Shodex GPC KD-80M (manufactured by Showa Denko KK) (8.0
mmI.D. × 300mmL) Mobile phase: Dimethyl sulfoxide (DMSO) Flow rate: 0.5ml / min Temperature: 40 ℃ Detector: SPD-6AV (UV spectrophotometer) (Shimadzu Corporation) Scale ( scale); 0.32AUFS response; STD wavelength (wavelength); 261 nm Pretreatment of sample Dissolve 5 mg of sample in 100 ml of DMSO.
次に50μｌをGPCに注入する。Then inject 50 μl into the GPC.
上記分析条件で分析した結果を第２図に示す。この第２
図によりGPCを用いてメラニンが分析できることがわか
る。The results of analysis under the above analysis conditions are shown in FIG. This second
The figure shows that GPC can be used to analyze melanin.
なお第２図中の縦軸は信号強度、横軸は保持時間（分）
である。The vertical axis in FIG. 2 is the signal intensity, and the horizontal axis is the holding time (minutes).
Is.
［発明の効果］ 本発明は、メラニンの希釈剤としてジメチルスルホキシ
ド（DMSO）を用いて、ゲルパーミエーションクロマトグ
ラフィ（以下GPCという）の移動相としてジメチルスル
ホキシド（DMSO）を用い、検出手段として紫外線分光光
度計を用いたGPCによって分析することにより、メラニ
ンの分子量分布の測定を効率よく、正確に行なうことが
できた。EFFECTS OF THE INVENTION The present invention uses dimethyl sulfoxide (DMSO) as a diluent for melanin, dimethyl sulfoxide (DMSO) as a mobile phase for gel permeation chromatography (hereinafter referred to as GPC), and ultraviolet spectrophotometry as a detection means. By the GPC analysis using a meter, the molecular weight distribution of melanin could be measured efficiently and accurately.
また、この分子量分布をゲルパーミエーションクロマト
グラフィにより簡便に得ることができた。また、得られ
た分子量分布によりメラニンの品質管理、性能評価が簡
単に行えるという特別の効果を有する。Moreover, this molecular weight distribution could be easily obtained by gel permeation chromatography. In addition, the obtained molecular weight distribution has a special effect that melanin quality control and performance evaluation can be easily performed.
第１図は本発明の方法を実施する装置の一例の構成説明
図、第２図は本発明のメラニンの希釈試料液についての
分子量分布を表すグラフである。 1:移動相貯留槽 2:送液ポンプ 3:マニュアルインジェクタ 4:カラム 5:カラムオーブン 6:紫外線分光光度計 7:データ処理装置FIG. 1 is a configuration explanatory view of an example of an apparatus for carrying out the method of the present invention, and FIG. 2 is a graph showing a molecular weight distribution of a diluted sample solution of melanin of the present invention. 1: Mobile phase reservoir 2: Liquid feed pump 3: Manual injector 4: Column 5: Column oven 6: Ultraviolet spectrophotometer 7: Data processor
Claims (1)
よりメラニンの分子量分布を測定する方法であって、メ
ラニンをジメチルスルホキシドで希釈し、ジメチルスル
ホキシドを溶離液として用いたゲルパーミエーションク
ロマトグラフィに付して、紫外線分光光度計で検出する
ことにより、前記メラニンの分子量分布を測定すること
を特徴とするメラニンの分子量分布分析法。1. A method for measuring the molecular weight distribution of melanin by gel permeation chromatography, which comprises subjecting melanin to dilution with dimethyl sulfoxide and subjecting it to gel permeation chromatography using dimethyl sulfoxide as an eluent to obtain an ultraviolet spectrophotometer. A molecular weight distribution analysis method for melanin, which comprises measuring the molecular weight distribution of the melanin by detecting with a meter.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP22794689A JPH071263B2 (en) | 1989-09-01 | 1989-09-01 | Molecular weight distribution analysis method of melanin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP22794689A JPH071263B2 (en) | 1989-09-01 | 1989-09-01 | Molecular weight distribution analysis method of melanin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0390858A JPH0390858A (en) | 1991-04-16 |
| JPH071263B2 true JPH071263B2 (en) | 1995-01-11 |
Family
ID=16868754
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP22794689A Expired - Lifetime JPH071263B2 (en) | 1989-09-01 | 1989-09-01 | Molecular weight distribution analysis method of melanin |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH071263B2 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4921990B2 (en) | 2006-10-10 | 2012-04-25 | 本田技研工業株式会社 | Vehicle yaw moment control device |
| CN106324147A (en) * | 2016-11-02 | 2017-01-11 | 百奥森（江苏）食品安全科技有限公司 | Detection method for melamine in milk powder |
-
1989
- 1989-09-01 JP JP22794689A patent/JPH071263B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0390858A (en) | 1991-04-16 |
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