JPS61196162A - Fractioning column for chromatography - Google Patents
Fractioning column for chromatographyInfo
- Publication number
- JPS61196162A JPS61196162A JP60038528A JP3852885A JPS61196162A JP S61196162 A JPS61196162 A JP S61196162A JP 60038528 A JP60038528 A JP 60038528A JP 3852885 A JP3852885 A JP 3852885A JP S61196162 A JPS61196162 A JP S61196162A
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- JP
- Japan
- Prior art keywords
- column
- crystals
- paths
- sectional area
- cross
- 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.)
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/60—Construction of the column
- G01N30/6034—Construction of the column joining multiple columns
- G01N30/6039—Construction of the column joining multiple columns in series
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/50—Conditioning of the sorbent material or stationary liquid
- G01N30/52—Physical parameters
- G01N2030/524—Physical parameters structural properties
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/50—Conditioning of the sorbent material or stationary liquid
- G01N30/56—Packing methods or coating methods
- G01N2030/562—Packing methods or coating methods packing
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/80—Fraction collectors
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
Description
【発明の詳細な説明】
イ１発明の目的
〔産業上の利用分野〕
本発明は、ハイドロキシアパタイト結晶を充填剤とする
クロマトグラフィ分取用カラムに関する。DETAILED DESCRIPTION OF THE INVENTION (1) Object of the Invention [Field of Industrial Application] The present invention relates to a chromatography preparative column using hydroxyapatite crystals as a packing material.
ハイドロキシアパタイト（以下、ＨＡと略記する）は通
常、化学組成ＣａｔＯ（ＰＯ４） ６ （ＯＨ）２ 、
六方晶系単位格子：ａΔｂ−１２０°、ａＡｃ＝ｂＡｃ
＝８０°、 １ａｌ＝ Ｉｂ１＝ ９．４２Ａ 、 １
ｃｌ＝ ８．８８Ａ ニよッテ特徴づけられるリン酸カ
ルシウムの一種であり、板状ないしは鱗片状結晶体であ
る。Hydroxyapatite (hereinafter abbreviated as HA) usually has the chemical composition CatO(PO4) 6 (OH)2,
Hexagonal unit cell: aΔb-120°, aAc=bAc
=80°, 1al=Ib1=9.42A, 1
cl = 8.88A Niyotte It is a type of calcium phosphate characterized by plate-like or scaly crystals.
而して、該ＨＡ結晶はクロマトグラフィ分取用カラムの
充填剤として優れた物質分子分離精製能を有し、従来困
難とされていた微細な構造差を有する物質量の分析・分
離、例えば分子量１０４〜１０９ダルトンの生態関連高
分子（免疫グロブリン・インターフェロン・酵素類等の
蛋白質類、ＲＮＡ＠ＤＮＡ・プラスミド等の核酸類、ウ
ィルス類など）についても鋭敏・高精度にクロマト分離
することが可能であり、従ってＨＡ結晶充填カラム（以
下、ＨＡカラムと略記する）を用いたクロマトグラフィ
は１例えばバイオテクノロジー分野での遺伝子組換え、
細胞融合、細胞大量培養法等により合成された各種の目
的有用物質の優れた分離精製操作手段等として大いに注
目されている。Therefore, the HA crystal has excellent ability to separate and purify substance molecules as a packing material for chromatography preparative columns, and can be used for analysis and separation of substances with minute structural differences, for example, molecular weight 104, which was previously considered difficult. It is possible to perform sensitive and highly accurate chromatography separation of ecologically relevant macromolecules (proteins such as immunoglobulins, interferons, enzymes, nucleic acids such as RNA@DNA and plasmids, viruses, etc.) of ~109 daltons. , Therefore, chromatography using a column packed with HA crystals (hereinafter abbreviated as HA column) can be used for example, genetic recombination in the biotechnology field,
It has attracted much attention as an excellent means for separating and purifying various purpose-useful substances synthesized by cell fusion, cell mass culturing, etc.
第３図はＨＡカラムの一例として直管形カラムＡの縦断
面図を示したものである。ｌは直管形カラム容体、２・
３はそのカラム容体の上端側と下端側に取付けた試料流
体導入口部材と同排出口部材、４・５はカラム容体１内
の上端側と下端側に設けたフィルタ部材、６はその上下
のフィルタ部材４・５間のカラム容体内空に充填された
ＨＡ結晶である０部材ｌ・２−３の材質はステンレスス
チール・セラミック・プラスチック等である。FIG. 3 shows a longitudinal sectional view of a straight column column A as an example of an HA column. l is a straight column column container, 2.
3 is a sample fluid inlet member and an outlet member attached to the upper and lower ends of the column container, 4 and 5 are filter members installed at the upper and lower ends of the column container 1, and 6 is a sample fluid inlet member and an outlet member attached to the upper and lower ends of the column container 1, respectively. The material of the 0 members 1 and 2-3, which are HA crystals filled in the column space between the filter members 4 and 5, is stainless steel, ceramic, plastic, etc.
フィルタ部材４・５は例えば硬質或は半硬質の連続微気
孔性のプラスチックポーラス体等である。The filter members 4 and 5 are, for example, hard or semi-hard continuous microporous plastic porous bodies.
カラム容体ｌの下端側の口に下側フィルタ部材５を嵌込
み、試料流体排出口部材３を取付け、カラム容体ｌの上
端側の口からカラム容体内にＨＡ結晶６を充填操作し、
上側フィルタ部材４を嵌込み、試料流体導入口部材２を
取付けることにより上記構造のＨＡカラムＡを得ること
ができる。Fitting the lower filter member 5 into the opening at the lower end of the column container 1, attaching the sample fluid outlet member 3, and filling the column with HA crystals 6 from the opening at the upper end of the column container 1;
By fitting the upper filter member 4 and attaching the sample fluid inlet member 2, the HA column A having the above structure can be obtained.
試料流体導入口部材２の口２ａから試料流体を導入する
。導入試料流体はカラムＡ内を上側フィルタ部材４→）
ＩＡ結晶充填部６→下側フィルタ部材５の経路で流れて
試料流体排出口部材３の口３ａから流出する。而して試
料流体のＨＡ結晶充填部流れ過程でＨＡ結晶の物質分子
捕捉作用により試料流体中に含まれる各種物質分子間の
所謂クロマト分離が起きる。クロマト分離後、ＨＡカラ
ムＡ内に特定ｐＨ値の緩衝液（バッファー）を導入“す
ることにより、クロマト分離されている各種の物質分子
のうち特定の物質分子のみがＨＡ結晶から離脱して口３
ａからカラム外へ排出される。即ち試料流体に含まれる
各種物質のうち目的とする特定物質を精製分取すること
ができる。A sample fluid is introduced from the port 2a of the sample fluid inlet member 2. The introduced sample fluid passes through column A through the upper filter member 4→)
The fluid flows along a path from the IA crystal filling part 6 to the lower filter member 5 and flows out from the opening 3a of the sample fluid outlet member 3. During the flow of the sample fluid through the HA crystal-packed section, so-called chromatographic separation occurs between the various substance molecules contained in the sample fluid due to the substance molecule trapping action of the HA crystals. After chromatographic separation, by introducing a buffer solution (buffer) with a specific pH value into HA column A, only specific substance molecules out of the various substance molecules being chromatographically separated are separated from the HA crystals.
It is discharged to the outside of the column from a. That is, it is possible to purify and separate a target specific substance from among various substances contained in the sample fluid.
ところでＨＡ結晶６は前述したように板状もしくは鱗片
状結晶体で、方向性がある。従って。By the way, as mentioned above, the HA crystal 6 is a plate-like or scale-like crystal body, and has directionality. Therefore.
（ａ）該ＨＡ結晶を個々の結晶体の方向性は考慮せずに
カラム容体ｌ内に互いにランダムな配向で充填した場合
と（ランダム充填）、
（ｂ）個々の結晶６の平板面が第４図に模形的に示した
ように、各結晶６の平板面をカラム容体軸線方向０−Ｏ
に実質的に略並行に配向させて充填した場合と（配向充
填）。(a) A case in which the HA crystals are packed in a random orientation relative to each other in a column container 1 without considering the orientation of each crystal (random packing), and (b) a case in which the flat plate surface of each crystal 6 is As schematically shown in Figure 4, the flat plate surface of each crystal 6 is aligned in the axial direction of the column 0-O.
(Oriented filling).
では、カラム内導入流体の流通性は上記後者（ｂ）の配
向充填の場合の方が格段によく、カラム内に試料流体を
低圧でしかも高流速に流すことができ、迅速なりロマト
グラフィ操作を実行することができる。The flowability of the fluid introduced into the column is much better in the case of the latter (b) oriented packing, which allows the sample fluid to flow into the column at low pressure and at a high flow rate, allowing for rapid chromatography operations. can be executed.
カラム容体１内に対するＨＡ結晶６の配向充填は１次の
ような要領で充填操作す、ればよい、即ち、カラム容体
ｌの下端側の口に下側フィルタ部材５を嵌込み、試料流
体排出口部材３を取付け、カラム容体の上端側の口から
ＨＡ結晶の適当濃度の懸濁液（ディスパージョン）を供
給し、その供給懸濁液がカラム容体ｌ内を層流状態で流
れる流速になるように、排出口部材３の口３ａから母液
性を自然流排出、或は吸引流排出、或は加圧流排出状態
にする。そうするとカラム容体内に於けるＨＡ結晶懸濁
液の層流流れ過程で個々の懸濁ＨＡ結晶６は母液の層流
流れによりその結晶平板面がカラム容体軸線方向０−０
に実質的に略並行の配向姿勢状態となる。そしてその配
向姿勢がほぼ保たれた状態でカラム容体ｌ内に下側フィ
ルタ部材５側から順次に堆積充填していく、即ちカラム
容体ｌ内へのＨＡ結晶６の配向充填がなされていく、所
要量の充填が終ったら上側フィルタ部材４を嵌込み、試
料流体導入口部材２を取付ける。The oriented filling of the HA crystals 6 into the column container 1 can be carried out in the following manner. That is, the lower filter member 5 is fitted into the opening at the lower end of the column container 1, and the sample fluid is drained. Attach the outlet member 3 and supply a suspension (dispersion) of HA crystals at an appropriate concentration from the opening on the upper end side of the column container, and the flow rate is such that the supplied suspension flows in a laminar flow state within the column container 1. Thus, the mother liquor is discharged from the opening 3a of the discharge port member 3 in a natural flow, suction flow, or pressurized flow state. Then, during the laminar flow process of the HA crystal suspension in the column container, each suspended HA crystal 6 has a crystal flat surface that is 0-0 in the axial direction of the column container due to the laminar flow of the mother liquor.
The orientation state is substantially parallel to the direction of the direction shown in FIG. Then, the HA crystals 6 are deposited and filled sequentially into the column container 1 from the lower filter member 5 side while the orientation is almost maintained, that is, the HA crystals 6 are filled in the column container 1 with the desired orientation. Once the amount of filling is completed, the upper filter member 4 is fitted and the sample fluid inlet member 2 is attached.
ただし、ＨＡ結晶を上記のような要領で充填操作してＨ
Ａ結晶がほぼ理想的な配向状態で充填されたＨＡカラム
Ａを得ることができるのは実際上はカラム容体ｌについ
てその内空横断面積が７〜５０１履２程度（丸パイプカ
ラム容体でその内径が３〜１０ｍｍ程・度）のかなり小
口径のものである場合に限られる。内空横断面積が７ｍ
１Ｉｚ以下の場合はカラム内壁面によるＨＡ結晶懸濁液
流通抵抗が大きく関与してカラム容体内でのＨＡ結晶懸
濁液の良好な層流流れ状態を形成させ難い、 ５Ｑｍｍ
２以上の場合はカラム容体内のＨＡ結晶順次堆積部面近
傍でＨＡ結晶懸濁液に乱流現象を生じるようになり、そ
の結果堆積ＨＡ結晶の配向が実質的にランダムなものと
なる。However, if you fill the HA crystals as described above,
In practice, it is possible to obtain an HA column A packed with A crystals in an almost ideal orientation state when the internal cross-sectional area of the column is approximately 7 to 501 cm2 (in a round pipe column, its inner diameter This is limited to cases where the diameter is quite small (approximately 3 to 10 mm/degree). Internal cross-sectional area is 7m
If it is less than 1 Iz, the flow resistance of the HA crystal suspension due to the inner wall surface of the column will be greatly involved, making it difficult to form a good laminar flow state of the HA crystal suspension within the column.
If the number is 2 or more, a turbulent flow phenomenon will occur in the HA crystal suspension near the surface of the sequentially deposited HA crystals in the column, and as a result, the orientation of the deposited HA crystals will become substantially random.
ＨＡ結晶を配向充填した。横断面積７〜５０ｍｍ２程度
の小口径ＨＡカラムＡは実験室的には十分に活用するこ
とができるが、工業的規模のものとしてはそれよりもは
るかに大口径（大横断面積）、例えば口径１２０ｇ＋■
φとか３００層■φとかの大口径ＨＡカラムが望まれる
。しかしそのような大口径カラムについてはその内空全
体にＨＡ結晶を実質的に配向充填させたものを得ること
は極めて難しいものであった。HA crystals were oriented and packed. A small-diameter HA column A with a cross-sectional area of about 7 to 50 mm2 can be fully utilized in the laboratory, but it can be used on an industrial scale with a much larger diameter (large cross-sectional area), such as a diameter of 120 g+. ■
A large diameter HA column such as φ or 300 layers ■φ is desired. However, it has been extremely difficult to obtain such a large-diameter column in which the entire interior space is substantially oriented and packed with HA crystals.
本発明は上記に鑑みて開発−提案されたもので、工業的
規模の大口径カラムについてもその内空全体に充填剤と
してのＨＡ結晶をほぼ理想的に配向充填させたものを容
易に得ることができるように工夫したものである。The present invention was developed and proposed in view of the above, and it is an object of the present invention to easily obtain an industrial-scale large-diameter column in which HA crystals as a filler are packed in an almost ideal orientation throughout the interior space of the column. It has been devised so that it can be done.
口、発明の構成
〔問題点を解決するための手段〕
本発明は、ハイドロキシアパタイト結晶を充填剤とする
クロマトグラフィ分取用カラムに於て。Summary: Structure of the Invention [Means for Solving the Problems] The present invention relates to a chromatography preparative column using hydroxyapatite crystals as a packing material.
カラム容体内空を、互いにカラム容体軸線方向に略並行
で、個々の内空横断面積が７〜５０■１１２の小横断面
積通路の集合化状態に分割区画し、その各小横断面積通
路内にハイドロキシアパタイト結晶を、個々の結晶の平
板面を通路方向に実質的に略並行に配向させた状態で充
填処置した、ことを特徴とするクロマトグラフィ分取用
カラムを要旨とする。The interior space of the column is divided into a collection of small cross-sectional area passages, each having an internal cross-sectional area of 7 to 50 x 112, which are approximately parallel to each other in the axial direction of the column, and each of the small cross-sectional area passages has a The object of the present invention is to provide a column for preparative chromatography, characterized in that hydroxyapatite crystals are packed with the flat surfaces of the individual crystals oriented substantially parallel to the path direction.
即ち、横断面積７〜５０厘鳳２程度の小口径カラム容体
内へのＨＡ結晶の配向充填は前述の要領で十分に可能で
ある。そこで本発明は横断面３５０鳳腸２以上の大口径
カラムの場合にそのカラム内空をＨＡ結晶の配向充填処
理の可能な横断面７〜５０１■２、就中２０〜４０層層
２ （横断面円形として約５〜“７層層φ）の範囲の小
横断面積通路の集合化状態に分割区分する。而る後その
各小横断面積通路内にＨＡ結晶を配向充填処置する。こ
れによりカラムがいくら大口径のものであってもその内
空は、ＨＡ結晶が夫々配向充填している各小横断面積通
路の集合により全体的にＨＡ結晶がほぼ理想的に配向充
填した状態のものとなる。That is, oriented packing of HA crystals into a small-diameter column having a cross-sectional area of about 7 to 50 mm is fully possible in the above-described manner. Therefore, in the case of a large-diameter column with a cross section of 350 mm or more, the present invention aims to fill the column space with a cross section of 7 to 501 cm2, particularly 20 to 40 layers (cross section It is divided into clusters of small cross-sectional area passages with a circular surface and approximately 5 to 7 layers φ).Then, HA crystals are oriented and packed in each of the small cross-sectional area passages.As a result, the column No matter how large the diameter of the HA crystal is, its inner space will be filled with HA crystals in an almost ideal orientation due to the collection of small cross-sectional area passages each filled with HA crystals in an oriented manner. .
カラム容体内空を、互いにカラム容体軸線方向に略並行
で、個々の内空横断面積が７〜５０ｍｍ２の小横断面積
通路の集合化状態に分割区画する手段としては、第１図
（ａ）〜（ｄ）に各種例示するようにカラム容体ｌ内に
上記のような小横断面積通路７ａの集合構造物７を挿入
すればよい、第２図は第１図 （ａ）の■−■線断面を
示すものである。そのような集合構造物７はポリエチレ
ン樹脂などプラスチックの押出成形法、プラスチック製
膜ポール構造物の丸め加工等により得ることができる。As a means for dividing the interior space of the column into a collection of small cross-sectional area passages that are substantially parallel to each other in the axial direction of the column and each having a cross-sectional area of 7 to 50 mm2, the method shown in FIG. 1(a) to As shown in FIG. 1(d), the collective structure 7 of the small cross-sectional area passages 7a as described above may be inserted into the column body l. This shows that. Such an aggregate structure 7 can be obtained by extrusion molding of a plastic such as polyethylene resin, or by rolling a plastic membrane pole structure.
或はカラム容体ｌ内体がプラスチック等の押出し成形で
製造されるときはそのカラム容体の押出しと同時にその
内空に小横断面積通路７ａの集合構造部７を押出し造成
することもできる。Alternatively, when the inner body of the column container 1 is manufactured by extrusion molding of plastic or the like, the collective structure 7 of the small cross-sectional area passages 7a can be formed in the inner space by extrusion simultaneously with extrusion of the column container.
（＋）内径１２０１φ・実質長さくＨＡ結結晶充填長さ
寸法）５０■層の横断面円形のステンレススチール製プ
レカラム容体と、内径 １２０層謄φ・実質長さ １０
０１の横断面円形のステンレススチール製本カラム容体
とに、夫々その内空に個々の小通路７ａの横断面積が約
２８■腸２ （直径約６ｍｍの円形断面通路に相当）の
第１図（ａ）のような横断面蜂巣型の小通路集合構造物
（ポリエチレン樹脂製、各通路７ａの隔壁肉厚的０．３
膳腸）７を嵌入することによリカラム内空を小横断面積
通路の集合化状態に分割区画した。(+) Inner diameter 1201φ, actual length HA crystal packed length dimension) Stainless steel pre-column container with circular cross section of 50 layers, inner diameter 120 layers φ, actual length 10
A stainless steel book-bound column container with a circular cross section of 01 has a cross-sectional area of about 28 cm intestine 2 (corresponding to a circular cross-section passage with a diameter of about 6 mm) of each small passage 7a in its inner space. ) with a honeycomb-shaped cross section (made of polyethylene resin, the partition wall thickness of each passage 7a is 0.3
By inserting the recolumn 7, the space inside the recolumn was divided into a collection of small cross-sectional area passages.
そして上記プレカラムと木カラムの各小横断面積通路７
ａ内にＨＡ結晶を該ＨＡ結晶の懸濁液を層流として導入
することにより配向充填処理し、本発明に従うＨＡＡ晶
配向充填のプレカラムと木カラムを作成した。and each small cross-sectional area passage 7 of the pre-column and wooden column.
A precolumn and a wood column packed with oriented HAA crystals according to the present invention were prepared by introducing HA crystals in a laminar flow as a laminar flow.
充填するＨＡ結晶は、ストークス等価球直径で１０４ｍ
以下の小径のもの、或はそのような小径のものを多く含
んだものは好ましくなく、１０ｇｍ以下の小径のものは
含まないか、含んでもその混入率が１重量％以下であり
、平均粒径がストークス等価球直径で２０〜１００ ｇ
ｍ、就中３０〜６０ＩＬｍのものを用いるのが好ましい
。The HA crystal to be filled has a Stokes equivalent sphere diameter of 104 m.
It is not preferable to use particles with the following small diameters or those that contain a large amount of such small particles; particles with small diameters of 10 gm or less should not be included, or even if they are included, the contamination rate should be 1% by weight or less, and the average particle size should be is the Stokes equivalent sphere diameter of 20 to 100 g.
m, particularly preferably 30 to 60 ILm.
又番手横断面積通路７ａ内へのＨＡＡ晶配向充填の際の
ＨＡＡ晶懸濁液の通路内流速は、木実施例の場合は２Ｇ
＋ｎＪ１／分（１ｃｍの直径当り換算）程度に調節して
ＨＡ結晶を良好に配向充填させることができた。In addition, the flow velocity of the HAA crystal suspension in the channel when filling the cross-sectional area channel 7a with oriented HAA crystals is 2G in the case of the wood embodiment.
By adjusting the flow rate to about +nJ1/min (converted per 1 cm diameter), the HA crystals could be well oriented and packed.
而して上記のプレカラムと木カラムとを上下に連通パイ
プで連結し、ペプシンについてのクロマト分離操作を行
った。Then, the above precolumn and the wooden column were connected vertically with a communicating pipe, and a chromatographic separation operation for pepsin was performed.
この操作は試料流体（ペプシンを含む燐酸緩衝液）のカ
ラム内最大流速約２１ｍ ｌ ／分（直径ＩＣ１当り換
算、以下同じ）、最大圧力２．２ Ｋｇｆ／ｃ■２以内
で安定に運転可能であった。This operation can be operated stably at a maximum flow rate of sample fluid (phosphate buffer containing pepsin) in the column of approximately 21 ml/min (calculated per diameter IC, hereinafter the same) and a maximum pressure of 2.2 Kgf/c2. there were.
（２）これに対して上記のプレカラム及び木カラムの何
れも、その内空を小横断面積通路の集合化状態に分割区
画処置せずにＨＡ結晶を充填したもの、従ってＨＡ結晶
がランダム充填状態にあるものを用いた場合にはカラム
内最大流速４．１８ｍ ｌ ／分、最大圧力８．２ Ｋ
ｇｆ／ｃ■２以内での運転が可能であるに過ぎなかった
。(2) On the other hand, both the precolumn and the wooden column described above are filled with HA crystals without dividing the inner space into a cluster of small cross-sectional area passages, and therefore the HA crystals are packed randomly. When using a column with a maximum flow rate of 4.18 ml/min and a maximum pressure of 8.2 K.
It was only possible to operate within gf/c 2.
即ち本発明に従う （１）のカラムの場合は （２）の
場合に比べて、カラム内への流体の導入圧力は低圧力で
、しかも高流速で流体をカラム内に流すことができ、迅
速な・大容量のクロマトグラフィ操作を実行することが
可能であった。That is, in the case of the column (1) according to the present invention, compared to the case (2), the pressure for introducing the fluid into the column is lower, and the fluid can be flowed into the column at a higher flow rate, resulting in rapid flow. - It was possible to perform large volume chromatography operations.
ハ、発明の効果
以上のように本発明に依れば、工業的規模の大口径ＨＡ
カラムについてもその内空全体的に充填剤としてのＨＡ
結晶をほぼ理想的に配向充填させたもの、従って流体の
カラム内流通性がよく、多量の流体をカラム内に低圧力
でしかも高流速で流して短時間で大量の目的物質の分取
操作が可能なものを容易に実現することができる。又装
置内圧が低圧力ですむことは設備費の低減化ができると
共に、装置の安全運転を確保する上に大きな利点となる
。C. Effects of the invention As described above, according to the present invention, an industrial-scale large-diameter HA
The column also has HA as a packing material throughout its interior space.
The crystals are packed in an almost ideal orientation, so the flow of fluid in the column is good, and a large amount of fluid can be passed through the column at low pressure and high flow rate, allowing for preparative separation of a large amount of the target substance in a short time. What is possible can be easily realized. Furthermore, the fact that the internal pressure of the apparatus is low can reduce equipment costs and is a great advantage in ensuring safe operation of the apparatus.
第１図（ａ）乃至同図（ｄ）は小横断面積通路の集合構
造物の各種の例の横断面図、第２図は第１図（ａ）の■
−■線断面図、第３図は直管形ＨＡカラムの一例の縦断
面図、第４図はＨＡ結晶の配向模型図である。
ＡはＨＡカラム、１はカラム容体、６は充填ＨＡ結晶、
７は小横断面積通路７ａの集合構造物。Figures 1(a) to 1(d) are cross-sectional views of various examples of collective structures with small cross-sectional area passages, and Figure 2 is the cross section of Figure 1(a).
3 is a longitudinal sectional view of an example of a straight pipe type HA column, and FIG. 4 is a schematic diagram of the orientation of HA crystals. A is the HA column, 1 is the column container, 6 is the packed HA crystal,
7 is a collection structure of small cross-sectional area passages 7a.
Claims (1)
マトグラフィ分取用カラムに於て、 カラム容体内空を、互いにカラム容体軸線方向に略並行
で、個々の内空横断面積が７〜５０ｍｍ＾２の小横断面
積通路の集合化状態に分割区画し、その各小横断面積通
路内にハイドロキシアパタイト結晶を、個々の結晶の平
板面を通路方向に実質的に略並行に配向させた状態で充
填処置した、ことを特徴とするクロマトグラフィ分取用
カラム。(1) In a chromatography preparative column using hydroxyapatite crystals as a packing material, the hollow space of the column is approximately parallel to each other in the axial direction of the column space, and the cross-sectional area of each hollow space is small with a cross-sectional area of 7 to 50 mm^2. It is divided into a group of cross-sectional area passages, and each of the small cross-sectional area passages is filled with hydroxyapatite crystals with the flat plate surfaces of the individual crystals oriented substantially parallel to the passage direction. A chromatography preparative column characterized by:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60038528A JPS61196162A (en) | 1985-02-27 | 1985-02-27 | Fractioning column for chromatography |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60038528A JPS61196162A (en) | 1985-02-27 | 1985-02-27 | Fractioning column for chromatography |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS61196162A true JPS61196162A (en) | 1986-08-30 |
| JPH0462337B2 JPH0462337B2 (en) | 1992-10-06 |
Family
ID=12527777
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP60038528A Granted JPS61196162A (en) | 1985-02-27 | 1985-02-27 | Fractioning column for chromatography |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS61196162A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2017051950A (en) * | 2010-12-23 | 2017-03-16 | エボニック コーポレイションＥｖｏｎｉｋ Ｃｏｒｐｏｒａｔｉｏｎ | Device for and method of producing emulsion |
-
1985
- 1985-02-27 JP JP60038528A patent/JPS61196162A/en active Granted
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2017051950A (en) * | 2010-12-23 | 2017-03-16 | エボニック コーポレイションＥｖｏｎｉｋ Ｃｏｒｐｏｒａｔｉｏｎ | Device for and method of producing emulsion |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0462337B2 (en) | 1992-10-06 |
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