JPH0461809B2 - - Google Patents
Info
- Publication number
- JPH0461809B2 JPH0461809B2 JP60299160A JP29916085A JPH0461809B2 JP H0461809 B2 JPH0461809 B2 JP H0461809B2 JP 60299160 A JP60299160 A JP 60299160A JP 29916085 A JP29916085 A JP 29916085A JP H0461809 B2 JPH0461809 B2 JP H0461809B2
- Authority
- JP
- Japan
- Prior art keywords
- silica gel
- groups
- group
- hydrophobic
- sample
- 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
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 104
- 229910002027 silica gel Inorganic materials 0.000 claims description 97
- 239000000741 silica gel Substances 0.000 claims description 97
- 125000001165 hydrophobic group Chemical group 0.000 claims description 36
- 125000005372 silanol group Chemical group 0.000 claims description 32
- 239000011148 porous material Substances 0.000 claims description 28
- 238000009832 plasma treatment Methods 0.000 claims description 17
- 125000000217 alkyl group Chemical group 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 claims description 6
- 229960001866 silicon dioxide Drugs 0.000 description 90
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 27
- 239000000523 sample Substances 0.000 description 27
- 102000004169 proteins and genes Human genes 0.000 description 26
- 108090000623 proteins and genes Proteins 0.000 description 26
- -1 amino acids Chemical class 0.000 description 24
- ZUOUZKKEUPVFJK-UHFFFAOYSA-N diphenyl Chemical compound C1=CC=CC=C1C1=CC=CC=C1 ZUOUZKKEUPVFJK-UHFFFAOYSA-N 0.000 description 18
- 239000012488 sample solution Substances 0.000 description 15
- 125000000962 organic group Chemical group 0.000 description 14
- 238000004458 analytical method Methods 0.000 description 13
- 210000002966 serum Anatomy 0.000 description 10
- 239000004305 biphenyl Substances 0.000 description 9
- 235000010290 biphenyl Nutrition 0.000 description 9
- 125000004469 siloxy group Chemical group [SiH3]O* 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 238000000034 method Methods 0.000 description 8
- DDBREPKUVSBGFI-UHFFFAOYSA-N phenobarbital Chemical compound C=1C=CC=CC=1C1(CC)C(=O)NC(=O)NC1=O DDBREPKUVSBGFI-UHFFFAOYSA-N 0.000 description 8
- 229960002695 phenobarbital Drugs 0.000 description 8
- 230000002209 hydrophobic effect Effects 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 6
- 229940098773 bovine serum albumin Drugs 0.000 description 6
- 239000003480 eluent Substances 0.000 description 6
- 239000000945 filler Substances 0.000 description 6
- 239000000499 gel Substances 0.000 description 6
- 238000012856 packing Methods 0.000 description 6
- 238000000862 absorption spectrum Methods 0.000 description 5
- 239000003463 adsorbent Substances 0.000 description 5
- 239000003054 catalyst Substances 0.000 description 5
- 150000001875 compounds Chemical class 0.000 description 5
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 4
- 229910001882 dioxygen Inorganic materials 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- 239000000377 silicon dioxide Substances 0.000 description 4
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 230000007423 decrease Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000000921 elemental analysis Methods 0.000 description 3
- 238000004811 liquid chromatography Methods 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 239000002798 polar solvent Substances 0.000 description 3
- 238000004366 reverse phase liquid chromatography Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 230000005526 G1 to G0 transition Effects 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 239000001307 helium Substances 0.000 description 2
- 229910052734 helium Inorganic materials 0.000 description 2
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 125000005647 linker group Chemical group 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 125000001570 methylene group Chemical group [H]C([H])([*:1])[*:2] 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000010992 reflux Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000002211 ultraviolet spectrum Methods 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- 239000003570 air Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 125000004103 aminoalkyl group Chemical group 0.000 description 1
- 229910052786 argon Inorganic materials 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- YBQPKOVSNHDRKD-UHFFFAOYSA-N chloro-dimethyl-[3-(oxiran-2-ylmethoxy)propyl]silane Chemical compound C[Si](C)(Cl)CCCOCC1CO1 YBQPKOVSNHDRKD-UHFFFAOYSA-N 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000000975 co-precipitation Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 125000004966 cyanoalkyl group Chemical group 0.000 description 1
- 239000002274 desiccant Substances 0.000 description 1
- 238000003795 desorption Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- LRCFXGAMWKDGLA-UHFFFAOYSA-N dioxosilane;hydrate Chemical compound O.O=[Si]=O LRCFXGAMWKDGLA-UHFFFAOYSA-N 0.000 description 1
- KPUWHANPEXNPJT-UHFFFAOYSA-N disiloxane Chemical group [SiH3]O[SiH3] KPUWHANPEXNPJT-UHFFFAOYSA-N 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001879 gelation Methods 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 229910001872 inorganic gas Inorganic materials 0.000 description 1
- 150000008040 ionic compounds Chemical class 0.000 description 1
- FBNXYLDLGARYKQ-UHFFFAOYSA-N methoxy-dimethyl-[3-(oxiran-2-ylmethoxy)propyl]silane Chemical compound CO[Si](C)(C)CCCOCC1CO1 FBNXYLDLGARYKQ-UHFFFAOYSA-N 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 125000002347 octyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- SCPYDCQAZCOKTP-UHFFFAOYSA-N silanol Chemical compound [SiH3]O SCPYDCQAZCOKTP-UHFFFAOYSA-N 0.000 description 1
- 229960004029 silicic acid Drugs 0.000 description 1
- 125000003808 silyl group Chemical group [H][Si]([H])([H])[*] 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 125000004079 stearyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- DQZNLOXENNXVAD-UHFFFAOYSA-N trimethoxy-[2-(7-oxabicyclo[4.1.0]heptan-4-yl)ethyl]silane Chemical compound C1C(CC[Si](OC)(OC)OC)CCC2OC21 DQZNLOXENNXVAD-UHFFFAOYSA-N 0.000 description 1
- BPSIOYPQMFLKFR-UHFFFAOYSA-N trimethoxy-[3-(oxiran-2-ylmethoxy)propyl]silane Chemical compound CO[Si](OC)(OC)CCCOCC1CO1 BPSIOYPQMFLKFR-UHFFFAOYSA-N 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Description
産業上の利用分野
本発明はシリカゲルの細孔内面部が疎水化され
ていると共に、外表面部にシラノール基を有する
部分親水化シリカゲル及びその製造方法に関する
ものである。
従来の技術及び発明が解決しようとする問題点
従来より、シリカゲルは多孔性、表面における
活性なシラノール基の存在等の特性を利用して吸
着剤、乾燥剤、触媒担体など、各種用途に使用さ
れており、分析の分野では液体クロマトグラフイ
ーにおける担体或いは充填剤としての用途が知ら
れている。
この液体クロマトグラフイーにおいては、シリ
カゲルは主に順相クロマトグラフイーの担体とし
て広く使用されているが、シリカゲルは上述した
ように表面にシラノール基が存在しているため、
水、その他の極性溶媒、或いはアミノ酸等のイオ
ン性化合物を強く吸着し、従つてこのような極性
溶媒を使用したり、強いイオン性を有する化合物
を分離対象とする場合はシラノール基の保護が必
要となる。このために従来、シリカゲルのシラノ
ール基にアルキル基、特にオクタデシルシリル基
を導入することが行なわれており、この種のアル
キル基を導入したシリカゲルが極性溶媒を用いる
逆相クロマトグラフイーに多く使用されている。
しかし、この疎水性のアルキル基を導入したシリ
カゲルは、細孔内面部を含む全表面がアルキル基
の導入で疎水化されているもので、シリカゲルの
全表面の一部にのみ疎水基が導入されたもの或い
は一部に疎水基、残りの部分に親水性有機基が導
入されたものはない。
ところで、最近において、医学、薬学等の分野
で蛋白質と共存する微量成分を分析することが多
くなつているが、このような蛋白質と共存する成
分を逆相クロマトグラフイーで分析する場合、上
述したオクタデシルシリル基導入シリカゲルを用
い、これに蛋白質を含む試料を直接接触させる
と、シリカゲルの表面に蛋白質がゲル化するた
め、蛋白質を含む試料は最初に塩化アンモニウム
等を添加してPH調整することにより蛋白質を沈殿
させ、これを遠心分離により除去するという前処
理を行なう必要がある。しかしながら、このよう
な蛋白質を沈殿、遠心分離による除去はその操作
が面倒である上、時間もかかり、しかも分析すべ
き成分の共沈のおそれもあり、従つて蛋白質と共
存する成分のより簡単な分析法が望まれていた。
本発明の目的は、細孔内面部にオクタデシルシ
リル基等の疎水基が導入され、かつ表面部にシラ
ノール基を有し、このため疎水基と活性なシラノ
ール基とが共存しているので、それ自体吸着剤、
触媒や分析用の担体など、各種の用途を有し、ま
た蛋白質と共存する成分を逆相クロマトグラフイ
ーで分析する場合の充填剤を製造するのに有効に
使用される部分親水化シリカゲル及びその製造方
法を提供するものである。
問題点を解決するための手段及び作用
本発明者らは、シリカゲル、特にオクタデシル
シリル基等の疎水基が導入されたシリカゲルの特
性を改善することについて鋭意研究を行なつてい
るうち、オクタテジルシリル基等の疎水基を導入
したシリカゲルにプラズマ処理を施した場合、意
外にもシリカゲルからオクタデシルシリル基等の
疎水基が脱離し、シリカゲル表面のシラノール基
が顕出することを見い出すと共に、このような疎
水基の脱離はプラズマ処理された部分にのみ生
じ、疎水基導入シリカゲルのプラズマ処理により
その細孔内面部の疎水基は脱離せずにそのまま残
つて外表面部のみがシラノール化されることを知
見した。そして、このように全表面疎水基導入シ
リカゲルをプラズマ処理することにより得られた
細孔内面部に疎水基を有すると共に、外表面部に
シラノール基を有するシリカゲルは、細孔内が疎
水化しており、かつ外表面部に活性なシラノール
基を有するので、従来の全表面にシラノール基を
有するシリカゲル或いは細孔内面部を含む全表面
にオクタデシルシリル基等の疎水基が導入された
シリカゲルに比べて親水性と疎水性の両者の性質
を兼ね備えた特異な性状を有し、吸着剤、触媒や
分析用等の担体などとして有用であることを知得
すると共に、この部分親水化シリカゲルの外表面
部のシラノール基には親水性有機基を簡単に導入
することができること、そしてこのように細孔内
面部に疎水基を有し、かつ外表面部に親水性有機
基が導入されたシリカゲルは、意外なことは蛋白
質を吸着もゲル化させず、従つて蛋白質と共存す
る成分の分析にこの部分疎水化・親水性有機基導
入シリカゲルを充填剤としてカラムに充填して用
いると、分析対象成分がこのシリカゲルの疎水基
に吸着し、蛋白質はこのシリカゲルに吸着された
りゲル化されることなくそのままカラムから流出
するので蛋白質が確実に分離除去され、このため
分析対象成分を効果的に分析に供することができ
ることを知見して、本発明を完成するに至つたも
のである。
従つて、本発明はシリカゲルの細孔内面部に疎
水基が導入されていると共に、外表面部にシラノ
ール基を有することを特徴とする部分親水化シリ
カゲル及びこのような部分親水化シリカゲルを得
る方法として、細孔内面部を含む全表面に疎水基
が導入されたシリカゲルにプラズマ処理を施し
て、外表面部の疎水基を脱離し、シラノール基を
顕出させることを特徴とする部分親水化シリカゲ
ルの製造方法を提供するものである。
以下、本発明につき更に詳しく説明する。
本発明に係る部分親水化シリカゲルは、上述し
たようにシリカゲルの細孔内面部に疎水基を、外
表面部にシラノール基を有するものである。第１
図はこれを模型的に示したもので、ａがシリカゲ
ル、ｂは細孔、ｃがその内面部、ｄが外表面部で
あり、内面部ｃに疎水基ｅが導入されていると共
に、外表面部ｄにはシリカゲルａに由来するシラ
ノール基ｆが存在している。
ここで、疎水基としては、本発明シリカゲルの
使用目的等の応じて適宜選択される。例えば、液
体カラムクロマイトグラフイー用の充填剤等とし
て用いる場合は、プロピル、オクチル、オクタデ
シル基等のアルキル基、シアノプロピル基等のシ
アノアルキル基、アミノプロピル基等のアミノア
ルキル基などが挙げられる。
なお、本発明シリカゲルの粒子形状に制限はな
く、球状、破砕状等、適宜な形状のものが用いら
れ、また粒径、細孔の大きさ、表面積なども本発
明シリカゲルの使用目的等に応じて選定される。
本発明の上述した細孔内面部に疎水基が導入さ
れ、外表面部にシラノール基を有する部分親水化
シリカゲルは、細孔内面部を含む全表面に疎水基
が導入されたシリカゲルにプラズマ処理を施すこ
とにより効率よく得ることができるもので、この
プラズマ処理により外表面部の疎水基が脱離さ
れ、シリカゲルに由来する表面シラノール基が顕
われるものである。
この場合、プラズマ処理としては低温プラズマ
処理が好適に採用され、これにより全表面に疎水
基が導入されたシリカゲルに対する熱作用を殆ん
ど与えず、しかも外表面部のみを処理することが
できる。プラズマ処理条件としては、圧力0.1〜
2Torr、特に0.5Torr前後、出力10〜150W、特に
80W前後、処理時間10〜120分、特に60分前後の
条件を採用することができ、また封入ガスとして
は酸素、水蒸気、空気、アルゴン、ヘリウム、炭
酸ガス等の無機ガスが使用され、特に酸素ガスが
好適に用いられる。
なお、全表面に疎水基が導入されたシリカゲル
は従来公知の方法で製造でき、各種市販品を用い
ることもできる。
本発明の部分親水化シリカゲルは、細孔内が疎
水化され、外表面部に活性なシラノール基を有す
るため、それ自体吸着剤、触媒担体、分析用担体
などとして有効に使用できるほか、シラノール基
に親水性有機基を導入した新規シリカゲルの製造
にも使用される。
即ち、細孔内が疎水化され、外表面部にシラノ
ール基を有するシリカゲルに対し、シラノールと
反応する化学結合基を有すると共に、親水性有機
基又は加水分解等により親水性有機基に変わり得
る有機基を有する化合物を反応させ、必要により
反応後有機基を親水性化することにより、親水性
有機基を導入し得るものである。
この場合、このようなシラノール基との反応性
化学結合基を有し、かつ親水性有機基もしくは親
水化可能な有機基を有する化合物としては、特に
限定するものではないが、β−（３，４−エポキ
シシクロヘキシル）エチルトリメトキシシラン、
γ−グリシドキシプロピルトリメトキシシラン、
γ−グリシドプロピルメチルジエトキシシラン、
γ−グリシドキシプロピルジメチルメトキシシラ
ン、γ−グリシドキシプロピルジメチルクロルシ
ランなどが挙げられる。
なお、これらの化合物と疎水基及びシラノール
基を有するシリカゲルとの反応を行なう場合の条
件、或いは必要により行なわれる親水化の条件
は、使用する化合物の種類等に応じて選択され
る。
このシラノール基に親水性有機基を導入するこ
とによつて得られた部分親水化シリカゲルも、細
孔内面部に疎水基が導入され、外表面部に親水性
有機基が導入され、疎水基と親水基とが共存して
いるので、吸着剤、触媒担体、分析用担体等とし
て有効に使用されるが、特に蛋白質と共存する成
分を分析する場合の分離剤或いは濃縮剤等として
好適に用いられる。即ち、この疎水基を細孔内面
部に有し、親水性有機基を外表面部に有するシリ
カゲルには蛋白質が全く作用せず、吸着やゲル化
がないので、蛋白質と分析対象成分とを導入する
と分析対象成分はこのシリカゲルの疎水基には吸
着されるが、蛋白質はそのままこのシリカゲル外
に排出される。従つて、蛋白質中の微量成分を分
析する場合、従来のように蛋白質を沈殿し、遠心
分離により除去するという面倒で時間のかかる前
処理を必要とせず、親水性有機基導入シリカゲル
の充填したカラムに試料に通すだけで蛋白質が確
実に分離溶出され、微量成分は保持されるのでこ
れを簡単に分析できる。また、このシリカゲル
は、このような性質を利用して蛋白質の精糖にも
好適に用いられる。
発明の効果
以上説明したように、本発明の部分親水化シリ
カゲルは、各種用途に好適に使用することがで
き、また本発明の製造方法より、かかる部分親水
化シリカゲルを効率よく確実に得ることができ
る。
以下、実施例と参考例を示し、本発明を具体的
に説明するが、本発明は下記の実施例に制限され
るものではない。
実施例 １
細孔内面部を含む全表面にオクタデシルシリル
基が導入された平均粒径5μｍのシリカゲル約0.8
ｇを容量50mlのナス型フラスコに入れ、このナス
型フラスコ内の真空ポンプで約10-2Torrに減圧
した後、酸素ガスを導入して0.45Torrに調整し
た。次いで、このナス型フラスコを互いに所定間
隔を存して離間対向させた２枚の銅製電極板の間
に位置するように配置してナス型フラスコを回転
させ、同時に電極板に周波数13.56MHz、実効出
力80Wの高周波を印加し、ナス型フラスコ内に酸
素プラズマを発生持続させ、酸素プラズマにより
上記シリカゲルを60分間処理した。
上述したプラズマ処理によるシリカゲルの外表
面時の経時変化を赤外線吸収スペクトル
（KBrdisk法）で測定したところ、第２図に示す
結果が得られた。また、プラズマ処理前後のシリ
カゲルの元素分析を行なつたところ、第１表に示
す結果が得られた。
INDUSTRIAL APPLICATION FIELD The present invention relates to a partially hydrophilized silica gel in which the inner surface of the pores of silica gel is made hydrophobic and has silanol groups on the outer surface, and a method for producing the same. Conventional technology and problems to be solved by the invention Silica gel has traditionally been used for various purposes such as adsorbents, desiccants, and catalyst supports, taking advantage of its characteristics such as porosity and the presence of active silanol groups on the surface. In the field of analysis, it is known to be used as a carrier or filler in liquid chromatography. In this liquid chromatography, silica gel is mainly used as a carrier for normal phase chromatography, but as mentioned above, silica gel has silanol groups on its surface, so
It strongly adsorbs water, other polar solvents, or ionic compounds such as amino acids, so protection of the silanol group is necessary when using such polar solvents or when separating compounds with strong ionicity. becomes. For this purpose, alkyl groups, especially octadecylsilyl groups, have been introduced into the silanol groups of silica gel, and silica gels with this type of alkyl group are often used in reversed-phase chromatography using polar solvents. ing.
However, in silica gel into which hydrophobic alkyl groups have been introduced, the entire surface, including the inner surface of the pores, has been made hydrophobic by introducing the alkyl groups, and the hydrophobic groups have been introduced into only a portion of the entire surface of the silica gel. There are no products in which a hydrophobic group is introduced into a part or a hydrophilic organic group is introduced into the remaining part. By the way, in recent years, it has become common to analyze trace components that coexist with proteins in fields such as medicine and pharmacy.When analyzing such components that coexist with proteins using reversed phase chromatography, When octadecylsilyl group-introduced silica gel is used and a sample containing proteins is brought into direct contact with it, the protein gels on the surface of the silica gel. It is necessary to perform pretreatment to precipitate proteins and remove them by centrifugation. However, removal of such proteins by precipitation and centrifugation is not only cumbersome and time-consuming, but also involves the risk of co-precipitation of the components to be analyzed. An analytical method was desired. The object of the present invention is to introduce a hydrophobic group such as an octadecylsilyl group into the inner surface of the pore and a silanol group on the surface, so that the hydrophobic group and the active silanol group coexist. Adsorbent itself,
Partially hydrophilized silica gel, which has various uses such as catalysts and carriers for analysis, and is effectively used to produce packing materials for analyzing components coexisting with proteins by reversed-phase chromatography. A manufacturing method is provided. Means and Effects for Solving the Problems The present inventors have conducted intensive research on improving the properties of silica gel, particularly silica gel into which hydrophobic groups such as octadecylsilyl groups have been introduced, and found that octadecylsilyl They discovered that when plasma treatment is applied to silica gel into which hydrophobic groups such as silyl groups have been introduced, the hydrophobic groups such as octadecylsilyl groups are unexpectedly removed from the silica gel, and silanol groups on the surface of the silica gel are exposed. Desorption of hydrophobic groups occurs only in the plasma-treated area, and when plasma treatment of hydrophobic group-introduced silica gel occurs, the hydrophobic groups on the inner surface of the pores do not detach and remain as they are, and only the outer surface becomes silanolized. I found out. Silica gel that has hydrophobic groups on the inner surface of the pores and silanol groups on the outer surface obtained by plasma treating silica gel with hydrophobic groups introduced on its entire surface has hydrophobicized inside of the pores. , and has active silanol groups on the outer surface, so it is more hydrophilic than conventional silica gels that have silanol groups on the entire surface or silica gels that have hydrophobic groups such as octadecylsilyl groups introduced on the entire surface including the inner surface of the pores. This partially hydrophilic silica gel has a unique property that combines both hydrophilic and hydrophobic properties, and has been found to be useful as an adsorbent, a catalyst, and a carrier for analysis. It is surprising that hydrophilic organic groups can be easily introduced into the silica gel, which has hydrophobic groups on the inner surface of the pores and hydrophilic organic groups on the outer surface. does not adsorb or gel proteins; therefore, when this partially hydrophobicized, hydrophilic organic group-introduced silica gel is used as a packing material in a column to analyze components that coexist with proteins, the target components are The protein is adsorbed to the hydrophobic group and flows out of the column as it is without being adsorbed to the silica gel or gelated, so the protein is reliably separated and removed, and the target component can therefore be effectively analyzed. These findings led to the completion of the present invention. Therefore, the present invention provides a partially hydrophilic silica gel characterized in that a hydrophobic group is introduced into the inner surface of the pores of the silica gel and a silanol group is present on the outer surface, and a method for obtaining such a partially hydrophilized silica gel. Partially hydrophilic silica gel is characterized in that silica gel having hydrophobic groups introduced into its entire surface including the inner surface of pores is subjected to plasma treatment to remove the hydrophobic groups on the outer surface and expose silanol groups. The present invention provides a method for manufacturing. The present invention will be explained in more detail below. As described above, the partially hydrophilized silica gel according to the present invention has hydrophobic groups on the inner surface of the pores of the silica gel and silanol groups on the outer surface. 1st
The figure shows this schematically, where a is a silica gel, b is a pore, c is an inner surface, and d is an outer surface.A hydrophobic group e is introduced into the inner surface c, and the outer Silanol groups f derived from silica gel a are present in the surface portion d. Here, the hydrophobic group is appropriately selected depending on the intended use of the silica gel of the present invention. For example, when used as a packing material for liquid column chromatography, examples thereof include alkyl groups such as propyl, octyl, and octadecyl groups, cyanoalkyl groups such as cyanopropyl groups, and aminoalkyl groups such as aminopropyl groups. Note that there is no restriction on the particle shape of the silica gel of the present invention, and any suitable shape such as spherical or crushed shape may be used, and the particle size, pore size, surface area, etc. may also be determined depending on the purpose of use of the silica gel of the present invention. Selected based on The above-mentioned partially hydrophilized silica gel of the present invention having hydrophobic groups introduced into the inner surface of the pores and silanol groups on the outer surface is obtained by plasma treatment of the silica gel having hydrophobic groups introduced into the entire surface including the inner surface of the pores. This plasma treatment removes hydrophobic groups on the outer surface and exposes surface silanol groups derived from silica gel. In this case, low-temperature plasma treatment is suitably employed as the plasma treatment, whereby only the outer surface can be treated with almost no thermal effect on the silica gel into which hydrophobic groups have been introduced over the entire surface. The plasma processing conditions are pressure 0.1~
2Torr, especially around 0.5Torr, output 10~150W, especially
Conditions of around 80W and processing time of 10 to 120 minutes, especially around 60 minutes, can be adopted, and inorganic gases such as oxygen, water vapor, air, argon, helium, carbon dioxide, etc. are used as the sealed gas, especially oxygen Gas is preferably used. Note that silica gel having hydrophobic groups introduced onto its entire surface can be produced by a conventionally known method, and various commercially available products can also be used. The partially hydrophilic silica gel of the present invention has pores made hydrophobic and has active silanol groups on the outer surface, so it can be effectively used as an adsorbent, catalyst carrier, analytical carrier, etc. It is also used to produce new silica gels with hydrophilic organic groups introduced into them. In other words, silica gel whose pores are hydrophobic and has silanol groups on its outer surface has a chemical bonding group that reacts with silanol, and a hydrophilic organic group or an organic group that can be converted into a hydrophilic organic group by hydrolysis, etc. A hydrophilic organic group can be introduced by reacting a compound having a group and, if necessary, making the organic group hydrophilic after the reaction. In this case, the compound having a chemical bonding group reactive with such a silanol group and having a hydrophilic organic group or a hydrophilic organic group is not particularly limited, but β-(3, 4-epoxycyclohexyl)ethyltrimethoxysilane,
γ-glycidoxypropyltrimethoxysilane,
γ-glycidopropylmethyldiethoxysilane,
Examples include γ-glycidoxypropyldimethylmethoxysilane and γ-glycidoxypropyldimethylchlorosilane. The conditions for reacting these compounds with silica gel having a hydrophobic group and a silanol group, or the conditions for hydrophilization if necessary, are selected depending on the type of compound used, etc. Partially hydrophilic silica gel obtained by introducing a hydrophilic organic group into the silanol group also has a hydrophobic group introduced into the inner surface of the pores and a hydrophilic organic group introduced into the outer surface. Since it coexists with hydrophilic groups, it can be effectively used as an adsorbent, catalyst carrier, analytical carrier, etc., but it is particularly suitable for use as a separating agent or concentrating agent when analyzing components that coexist with proteins. . In other words, proteins do not act on the silica gel, which has hydrophobic groups on the inner surface of the pores and hydrophilic organic groups on the outer surface, and there is no adsorption or gelation. Then, the target component to be analyzed is adsorbed to the hydrophobic groups of this silica gel, but the protein is directly discharged from the silica gel. Therefore, when analyzing trace components in proteins, it is possible to use columns packed with hydrophilic organic group-introduced silica gel without the need for the conventional pretreatment of precipitating proteins and removing them by centrifugation. By simply passing the sample through the sample, proteins are reliably separated and eluted, and trace components are retained, making them easy to analyze. Moreover, this silica gel is suitably used for protein sugar refining by utilizing such properties. Effects of the Invention As explained above, the partially hydrophilized silica gel of the present invention can be suitably used for various purposes, and the production method of the present invention makes it possible to efficiently and reliably obtain such partially hydrophilized silica gel. can. EXAMPLES Hereinafter, the present invention will be specifically explained with reference to Examples and Reference Examples, but the present invention is not limited to the Examples below. Example 1 Silica gel with an average particle size of 5 μm and approximately 0.8 μm in which octadecylsilyl groups were introduced on the entire surface including the inner surface of the pores.
g was placed in a 50 ml eggplant-shaped flask, and after reducing the pressure to about 10 -2 Torr with a vacuum pump inside the eggplant-shaped flask, oxygen gas was introduced to adjust the pressure to 0.45 Torr. Next, this eggplant-shaped flask was placed between two copper electrode plates facing each other with a predetermined distance between them, and the eggplant-shaped flask was rotated. At the same time, a frequency of 13.56 MHz and an effective output of 80 W were applied to the electrode plates. A high frequency wave was applied to generate and sustain oxygen plasma in the eggplant-shaped flask, and the silica gel was treated with the oxygen plasma for 60 minutes. When the change over time of the outer surface of the silica gel due to the plasma treatment described above was measured by infrared absorption spectroscopy (KBrdisk method), the results shown in FIG. 2 were obtained. Further, elemental analysis of the silica gel before and after plasma treatment was conducted, and the results shown in Table 1 were obtained.
【表】
第２図の結果は、全表面にオクタデシルシリル
基を導入したシリカゲルに対するプラズマ処理の
経時に従い、赤外線吸収スペクトルの2900cm-1近
傍のメチレン基の吸収が減少し、逆に3400cm-1近
傍の水親基の吸収が増加していることを示し、ま
た第１表の結果はプラズマ処理後の炭素及び水素
の減少を示すものであり、双方の結果からシリカ
ゲルの外表面部のオクタデシルシリル基がシラノ
ール基に変化していると判断された。
また、酸素プラズマ処理されたシリカゲル及び
未処理のシリカゲルをそれぞれ充填剤として4.6
mmφ×30mmのカラムに充填して固定相を形成し、
また移動相として75％CH3OH水溶液からなる溶
離液を流速0.5ml／minにて移動させ、それぞれ
にベンゼン1000ppm、ビフエニル20ppmを含有す
るメタノール試料液を5μ注入して固定相を通
過した液のUVスペクトルを測定した。なお、
UVスペクトルの測定はUV検出波長254nｍ、感
度0.32AUFSで行なつた。
上述したプラズマ処理を施したシリカゲルを充
填剤として用いた場合の測定結果を第３図に、未
処理のシリカゲルを充填剤として用いた場合の測
定結果を第４図に示す。なお、第３図、第４図に
おいてははベンゼンの検出ピーク、はビフエ
ニルの検出ピークを示すものである。また、第３
図、第４図のベンゼン及びビフエニルの検出ピー
クから得られたベンゼンとビフエニルの分離性能
を第２表に示す。[Table] The results shown in Figure 2 show that as the plasma treatment of silica gel with octadecylsilyl groups introduced into its entire surface increases over time, the absorption of methylene groups near 2900 cm -1 in the infrared absorption spectrum decreases, and conversely, the absorption of methylene groups near 3400 cm -1 decreases. This shows that the absorption of hydrophilic groups in the silica gel increases, and the results in Table 1 show a decrease in carbon and hydrogen after plasma treatment. was determined to have changed to a silanol group. In addition, oxygen plasma treated silica gel and untreated silica gel were used as fillers at 4.6
Fill a mmφ x 30mm column to form a stationary phase.
In addition, an eluent consisting of a 75% CH 3 OH aqueous solution was moved at a flow rate of 0.5 ml/min as a mobile phase, and 5μ of a methanol sample solution containing 1000 ppm of benzene and 20 ppm of biphenyl was injected into each of the eluents that had passed through the stationary phase. UV spectra were measured. In addition,
The UV spectrum was measured at a UV detection wavelength of 254 nm and a sensitivity of 0.32 AUFS. FIG. 3 shows the measurement results when the plasma-treated silica gel described above was used as the filler, and FIG. 4 shows the measurement results when the untreated silica gel was used as the filler. In addition, in FIGS. 3 and 4, the detected peak of benzene indicates the detected peak of biphenyl. Also, the third
Table 2 shows the separation performance of benzene and biphenyl obtained from the detection peaks of benzene and biphenyl shown in FIGS.
【表】
第３図及び第４図の結果から、プラズマ処理さ
れたシリカゲルは、未処理の全表面にオクタデシ
ルシリル基が導入されたシリカゲルに比したベン
ゼンとビフエニルとの分離性能にやや劣るものの
非極性のベンゼンとビフエニルを分離し得、また
全表面にシラノール基を有するシリカゲルを液体
クロマトグラフ用充填剤として用いた場合には、
非極性のベンゼンやビフエニルは吸着分離しない
ことから、プラズマ処理されたオクタデシルシリ
ル基導入シリカゲルが部分親水化されていること
は明らかである。
実施例 ２
実施例１の方法において、酸素ガスに代えて水
蒸気を導入してナス型フラスコ内を0.3Torrに調
整し、また、高周波の実効出力を50W、プラズマ
処理時間30分としたほかは実施例１と同様にして
細孔内面部を含む全表面にオクタデシルシリル基
が導入されたシリカゲルをプラズマ処理（水蒸気
プラズマ処理）した。
処理されたシリカゲルにつき実施例１と同様に
して赤外線吸収スペクトルを測定すると共に、元
素分析を行なつた結果、実施例１と同様にシリカ
ゲルの外表面部がシラノール基に変化しているこ
とが認められた。
実施例 ３
実施例１の方法において酸素ガスに代えてヘリ
ウムガスを導入してナス型フラスコ内を0.6Torr
に調整し、また、高周波の実効出力を70W、プラ
ズマ処理時間40分としたほかは実施例１と同様に
して細孔内面部を含む全表面にオクタデシルシリ
ル基が導入されたシリカゲルをプラズマ処理し
た。
処理されたシリカゲルにつき実施例１と同様に
して赤外線吸収スペクトルを測定すると共に、元
素分析を行なつた結果、実施例１と同様にシリカ
ゲルの外表面部がシラノール基に変化しているこ
とが認められた。
参考例
実施例１により得られた細孔内面部にオクタデ
シルシリル基が導入され、外表面部にシラノール
基を有する部分親水化シリカゲル0.8ｇをトリエ
チルアミン３mgを含むトルエン溶液５mlに懸濁さ
せた後、水10mg、γ−グリシドキシトリメトキシ
シラン0.37ｇを加え、還流下に17時間反応させ
た。次いで、吸引過して固体相を分離回収し、
これをトルエン及びアセトンで順次洗浄し、乾燥
した後、約10-4規定硫酸水溶液中に懸濁させ、還
流下で２時間加水分解反応を行なつた。処理物を
分離、洗浄後、80℃で乾燥して、下記式に示すよ
うに前記シラノール基のOH基を置換シロキシ基
に変えた、細孔内面面にオクタデシルシリル基、
外表面部に置換シロキシ基が導入された部分親水
化シリカゲルを得た。
次に、上記方法で得られた置換シロキシ基導入
部分親水化シリカゲルを濃縮カラム用充填剤とし
て使用した下記実験を行なつた。
実験１
第５図に示した装置を使用し、人血清中のフエ
ノバルビタールの分析を行なつた。
ここで、第５図中１は試料ループ２が接族され
た第１六方バルブ、３は濃縮カラム４及び分析カ
ラム５がそれぞれ接続された第２六方バルブで、
まず試料を注入する場合は、第１六方バルブ１を
図の実線で示す流路状態とし、試料を第１六方バ
ルブ１の試料溶液注入口６から試料溶液を導入す
る。これにより試料溶液は第１六方バルブ１の第
１流路口１ａから試料ループ２を通つて所定量の
試料溶液が試料ループ２に採取され、余剰の試料
溶液は第４流路口１ｄ、第３流路口１ｃを順次経
てドレン７に排出される。次いで、試料ループ２
に採取された試料溶液を濃縮カラム４に濃縮する
場合は、第１六方バルブ１を点線で示す流路状
態、第２六方バルブ３を実線で示す流路状態と
し、ポンプ８を作動させて濃縮用キヤリヤー容器
９内の濃縮用キヤリヤーの第１六方バルブの第５
流路口１ｅから導入する。これにより、キヤリヤ
ーは第４流路口１ｄを通り、試料ループ２内の試
料溶液を押し出し、第１流路口１ａ、第６流路１
ｆを順次経て、第六方バルブ３の第１流路口３ａ
に導入され、更に第６流路口３ｆから濃縮カラム
４に導入されて試料溶液がこの濃縮カラム４で濃
縮されると共に、このカラム４を通過した液は第
３流路口３ｃ、第２流路口３ｂを順次経てドレン
１０に排出される。そして、前記濃縮カラム４内
の試料を分析カラム５に送り、分析を行なう場合
は、第２六方バルブ３を点線に示す流路状態と
し、ポンプ１１を作動させて溶離液容器１２内の
溶離液を第二六方バルブ３の第４流路口３ｄに導
入するもので、これにより溶離液は第３流路口３
ｃ、濃縮カラム４、第６流路口３ｆ、第５流路口
３ｅを順次経て分析カラム５に流入し、溶離液に
より溶離せしめられた試料はこの分析カラム６か
ら検出器１３に送られ、分析される。
なお、第５図中１４は分流用抵抗ループ、１
５，１６はそれぞれ三方ジヨイントであり、試料
濃縮時において濃縮用キヤリヤーはその一部が一
方の三方ジヨイント１５から分流抵抗ループ１４
に分流され、他方の三方ジヨイント１６におい
て、第１六方バルブ１の第６流路口１ｆから第２
六方バルブ３の第１流路口３ａに向けて流れる試
料ループ２からの試料溶液と合流し、これにより
試料が希釈されるものである。
而して、上記した装置において、試料溶液とし
て、６倍に希釈した人血清にフエノバルビタール
を53ppm添加した試料Ａ、水にフエノバルビター
ルを53ppm溶解した試料Ｂ、人血清を５倍に希釈
した試料Ｃをそれぞれ使用し（注入量50μ）、
濃縮用キヤリヤーとして水、溶離液として50％メ
タノール水溶液を使用し（いずれも流速１ml／
min）、上述した方法により試料の濃縮、分析を
行なつた。この場合、濃縮カラムは4.6mmφ×30
mmの大きさと、前記置換シロキシ基導入部分親水
化シリカゲルを充填し、分析カラムは4.6mmφ×
250mmの大きさとして、ZorbaxODS（デユポン社
製）を充填した。また、濃縮時において、一方の
三方ジヨイント１５における余流比（希釈比）は
１：3.84（試料ループ導ん入量：分流用抵抗ルー
プ導入量）とし、分析は波長254nｍ、感度
0.32AUFSで行なつた。
以上の結果を第６図（試料Ａ）、第７図（試料
Ｂ）、第８図（試料Ｃ）に示す。
第６，８図のクロマトグラムを第７のクロマト
グラムと比較してみると、第６，８図の試料は人
血清を含むにもかかわらず、第７図の人血清を含
まない試料と同様にクロマトグラムに人血清に由
来する蛋白質のピークが現われていないことが認
められる。このことは、濃縮の段階で試料溶液を
濃縮カラムに導入した時に試料溶液中の人血清が
濃縮カラムに充填された置換シロキン基導入部分
類水化シリカゲルに吸着したりゲル化したりする
ことなく、濃縮用キヤリヤーと共に濃縮カラムか
ら排出されたことを示すもので、実際、ドレン１
０に排出された溶液から人血清の存在が確認され
た。また、このことから、第６，７図のクロマト
グラフを比較すれば明らかなように、蛋白質（人
血清）中のフエノバルビタールが蛋白質に全く影
響されることなく確実に分析し得ることが認めら
れ、置換シロキシ基導入部分親水化シリカゲルを
濃縮用充填剤として用いることにより、フエノバ
ルビタールと蛋白質とが混合されていても、フエ
ノバルビタールと蛋白質とを確実に分離し、フエ
ノバルビタールのみを置換シロキシ基導入部分親
水化シリカゲルに吸着し得ることが認められた。
更に、蛋白質除去能を検討するために下記の実
験を行なつた。
実験２
上記した外表面部に置換シロキシ基を有する部
分親水化シリカゲル及び全表面にオクタデシルシ
リル基を有するシリカゲルをカラムに充填し、こ
のカラムに50〜500ppmの牛血清アルブミンを通
してこのカラムを通過する牛血清アルブミン量を
測定した。結果を第９図に示す。なお、図中は
外表面部に置換シロキシ基を有する部分親水化シ
リカゲル、は全表面にオクタデシルシリル基を
有するシリカゲルの結果である。また、分析条件
は下記の通りである。
分析条件：カラム 4.6mmφ×30mm
溶離液 リン酸緩衝液（PH＝７）
測定波長 280nｍ
感 度 0.04AUFS
注入量 10μ
第９図の結果より、外表面部に置換シロキシ基
を有する部分親水化シリカゲルは、牛血清アルブ
ミン量に比例してピーク高さが高くなり、従つて
このことから牛血清アルブミン（蛋白質）はこの
部分親水化シリカゲルに補足されることなく通過
していることが知見される。これに対し、オクタ
デシルシリル基を全表面に有するシリカゲルは牛
血清アルブミンに吸着され、カラムを殆んど通過
しないものであつた。[Table] From the results shown in Figures 3 and 4, the plasma-treated silica gel has slightly inferior benzene and biphenyl separation performance compared to the untreated silica gel with octadecylsilyl groups introduced on its entire surface, but it is still acceptable. When silica gel that can separate polar benzene and biphenyl and has silanol groups on its entire surface is used as a packing material for liquid chromatography,
Since nonpolar benzene and biphenyl are not adsorbed and separated, it is clear that the plasma-treated octadecylsilyl group-introduced silica gel is partially hydrophilized. Example 2 The method of Example 1 was carried out except that water vapor was introduced instead of oxygen gas and the inside of the eggplant-shaped flask was adjusted to 0.3 Torr, and the effective output of high frequency was 50 W and the plasma treatment time was 30 minutes. In the same manner as in Example 1, silica gel in which octadecylsilyl groups were introduced into the entire surface including the inner surface of the pores was subjected to plasma treatment (steam plasma treatment). As a result of measuring the infrared absorption spectrum of the treated silica gel in the same manner as in Example 1 and conducting elemental analysis, it was found that the outer surface of the silica gel had changed to silanol groups as in Example 1. It was done. Example 3 In the method of Example 1, helium gas was introduced instead of oxygen gas, and the inside of the eggplant-shaped flask was heated to 0.6 Torr.
In addition, silica gel in which octadecylsilyl groups were introduced into the entire surface including the inner surface of the pores was plasma treated in the same manner as in Example 1, except that the effective power of the high frequency was 70 W and the plasma treatment time was 40 minutes. . As a result of measuring the infrared absorption spectrum of the treated silica gel in the same manner as in Example 1 and conducting elemental analysis, it was found that the outer surface of the silica gel had changed to silanol groups as in Example 1. It was done. Reference Example After suspending 0.8 g of partially hydrophilized silica gel obtained in Example 1, in which octadecylsilyl groups were introduced into the inner surface of the pores and silanol groups were introduced into the outer surface, in 5 ml of toluene solution containing 3 mg of triethylamine, 10 mg of water and 0.37 g of γ-glycidoxytrimethoxysilane were added, and the mixture was reacted under reflux for 17 hours. Next, the solid phase is separated and collected by suction,
This was washed successively with toluene and acetone, dried, suspended in an aqueous solution of about 10 -4 N sulfuric acid, and subjected to a hydrolysis reaction under reflux for 2 hours. After separating and washing the treated material, it is dried at 80°C, and the OH group of the silanol group is changed to a substituted siloxy group, and an octadecylsilyl group is formed on the inner surface of the pore, as shown in the formula below.
Partially hydrophilic silica gel with substituted siloxy groups introduced onto the outer surface was obtained. Next, the following experiment was conducted using the substituted siloxy group-introduced partially hydrophilized silica gel obtained by the above method as a packing material for a concentration column. Experiment 1 Using the apparatus shown in Figure 5, phenobarbital in human serum was analyzed. Here, in FIG. 5, 1 is a first six-way valve to which a sample loop 2 is connected, 3 is a second six-way valve to which a concentration column 4 and an analysis column 5 are connected, respectively.
First, when injecting a sample, the first six-way valve 1 is placed in the flow path state shown by the solid line in the figure, and the sample solution is introduced from the sample solution inlet 6 of the first six-way valve 1. As a result, the sample solution passes through the sample loop 2 from the first flow path port 1a of the first hexagonal valve 1, and a predetermined amount of the sample solution is collected into the sample loop 2, and the excess sample solution passes through the fourth flow path port 1d and the third flow path. The water is discharged to the drain 7 through the outlet 1c in sequence. Next, sample loop 2
When concentrating the sample solution collected in the concentration column 4, the first six-way valve 1 is set to the flow path shown by the dotted line, the second six-way valve 3 is set to the flow path shown by the solid line, and the pump 8 is operated to perform the concentration. The fifth of the first six-way valve of the carrier for concentration in the carrier container 9 for
It is introduced from the channel opening 1e. As a result, the carrier passes through the fourth channel port 1d, pushes out the sample solution in the sample loop 2, and pushes out the sample solution in the sample loop 2 through the first channel port 1a and the sixth channel 1.
f sequentially, and then the first flow path port 3a of the six-way valve 3.
The sample solution is introduced into the concentration column 4 from the sixth flow path port 3f, and the sample solution is concentrated in this concentration column 4, and the liquid that has passed through this column 4 is introduced into the third flow path port 3c and the second flow path port 3b The water is sequentially discharged to the drain 10. When the sample in the concentration column 4 is sent to the analysis column 5 for analysis, the second hexagonal valve 3 is set to the flow path shown by the dotted line, and the pump 11 is operated to remove the eluent in the eluate container 12. is introduced into the fourth flow path port 3d of the second six-way valve 3, whereby the eluent is introduced into the third flow path port 3d.
c. The sample flows into the analysis column 5 through the concentration column 4, the sixth flow path port 3f, and the fifth flow path port 3e in order, and is eluted with the eluent.The sample is sent from the analysis column 6 to the detector 13 and analyzed. Ru. In addition, 14 in FIG. 5 is a resistor loop for shunt, 1
5 and 16 are three-way joints, and during sample concentration, a part of the carrier for concentration flows from one three-way joint 15 to the shunt resistance loop 14.
At the other three-way joint 16, the flow is diverted from the sixth flow path opening 1f of the first six-way valve 1 to the second
It joins with the sample solution from the sample loop 2 flowing toward the first channel port 3a of the six-way valve 3, thereby diluting the sample. In the above-mentioned apparatus, the sample solutions were Sample A, in which 53 ppm of phenobarbital was added to human serum diluted 6 times, Sample B, in which 53 ppm of phenobarbital was dissolved in water, and Human serum diluted 5 times. Using each sample C (injection amount 50μ),
Water was used as the carrier for concentration, and 50% methanol aqueous solution was used as the eluent (flow rate of 1 ml/ml in both cases).
min), the sample was concentrated and analyzed using the method described above. In this case, the concentration column is 4.6mmφ×30
The analytical column is 4.6 mmφ×
ZorbaxODS (manufactured by Dupont) was filled to a size of 250 mm. In addition, during concentration, the afterflow ratio (dilution ratio) in one three-way joint 15 is 1:3.84 (amount introduced into the sample loop: amount introduced into the diversion resistance loop), and the analysis is performed at a wavelength of 254 nm and a sensitivity of
This was done at 0.32 AUFS. The above results are shown in FIG. 6 (sample A), FIG. 7 (sample B), and FIG. 8 (sample C). Comparing the chromatograms in Figures 6 and 8 with the chromatogram in Figure 7, although the samples in Figures 6 and 8 contain human serum, they are similar to the samples in Figure 7 that do not contain human serum. It is observed that the peak of proteins derived from human serum does not appear in the chromatogram. This means that when the sample solution is introduced into the concentration column during the concentration stage, the human serum in the sample solution does not adsorb or gelatinize on the substituted siloxane group-introduced hydrated silica gel packed in the concentration column. This indicates that it was discharged from the concentration column together with the concentration carrier, and in fact, drain 1
The presence of human serum was confirmed from the solution discharged. In addition, from this, as is clear from the comparison of the chromatographs in Figures 6 and 7, it is recognized that phenobarbital in proteins (human serum) can be reliably analyzed without being affected by proteins at all. By using hydrophilized silica gel with a substituted siloxy group introduced as a packing material for concentration, even if phenobarbital and protein are mixed, phenobarbital and protein can be reliably separated and only phenobarbital can be extracted. It was found that the substituted siloxy group-introduced moiety can be adsorbed on hydrophilized silica gel. Furthermore, the following experiment was conducted to examine protein removal ability. Experiment 2 A column is filled with the above-mentioned partially hydrophilized silica gel having substituted siloxy groups on the outer surface and silica gel having octadecylsilyl groups on the entire surface, and 50 to 500 ppm of bovine serum albumin is passed through the column. Serum albumin level was measured. The results are shown in Figure 9. In the figure, the partially hydrophilized silica gel having substituted siloxy groups on the outer surface is the result of silica gel having octadecylsilyl groups on the entire surface. Moreover, the analysis conditions are as follows. Analysis conditions: Column 4.6mmφ The peak height increases in proportion to the amount of bovine serum albumin, which indicates that bovine serum albumin (protein) passes through the partially hydrophilized silica gel without being captured. In contrast, silica gel having octadecylsilyl groups on its entire surface was adsorbed to bovine serum albumin and hardly passed through the column.
第１図は本発明の部分親水化シリカゲルを模型
的に説明する部分断面図、第２図は全表面にオク
タデシルシリル基が導入されたシリカゲルに低温
プラズマ処理を施した場合における経時的な外表
面部変化を示す赤外線吸収スペクトル、第３図は
本発明の部分親水化シリカゲルを充填剤として用
いてベンゼン及びビフエニル混合溶液を通過させ
た場合の流出液の紫外線吸収スペクトル、第４図
は全表面にオクタデシルシリル基が導入されたシ
リカゲルを充填剤として用いてベンゼン及びビフ
エニル混合溶液を通過させた場合の流出液の紫外
線吸収スペクトル、第５図は置換シロキシ基導入
部分親水化シリカゲルの蛋白質分離特性を調べる
ために用いた装置のフローチヤート、第６図乃至
第８図は同装置を用いて試料分析を行なつた場合
のクロマトグラムで、第６図は試料として人血清
中にフエノバルビタノールを添加したものを用い
た場合のクロマトグラム、第７図は水にフエノバ
ルビタールを添加した試料のクロマトグラム、第
８図は人血清のクロマトグラム、第９図は牛血清
アルブミンを外表面部に置換シロキシ基を有する
部分親水化シリカゲル及び全表面にオクタデシル
シリル基を有するシリカゲルを充填したカラムに
通した場合の牛血清アルブミンの通過量を示すグ
ラフである。
ａ……シリカゲル、ｂ……細孔、ｃ……その内
面部、ｄ……外表面部、ｅ……疎水基、ｆ……シ
ラノール基。
Figure 1 is a partial cross-sectional view schematically illustrating the partially hydrophilized silica gel of the present invention, and Figure 2 is the outer surface over time of a silica gel with octadecylsilyl groups introduced into its entire surface that is subjected to low-temperature plasma treatment. Figure 3 shows the ultraviolet absorption spectrum of the effluent when a mixed solution of benzene and biphenyl is passed through it using the partially hydrophilized silica gel of the present invention as a filler. The ultraviolet absorption spectrum of the effluent when a mixed solution of benzene and biphenyl is passed through using silica gel into which an octadecylsilyl group has been introduced as a filler. Figure 5 shows the protein separation characteristics of partially hydrophilized silica gel into which a substituted siloxy group has been introduced. Figures 6 to 8 are flowcharts of the equipment used for this purpose, and chromatograms when sample analysis is performed using the same equipment. Figure 7 is a chromatogram of a sample prepared by adding phenobarbital to water, Figure 8 is a chromatogram of human serum, and Figure 9 is a sample in which bovine serum albumin is substituted on the outer surface. It is a graph showing the amount of bovine serum albumin that passes through a column filled with partially hydrophilized silica gel having siloxy groups and silica gel having octadecylsilyl groups on the entire surface. a... Silica gel, b... Pore, c... Inner surface portion, d... Outer surface portion, e... Hydrophobic group, f... Silanol group.
Claims (1)
ていると共に、外表面部にシラノール基を有する
ことを特徴とする部分親水化シリカゲル。 ２ 疎水基がアルキル基である特許請求の範囲第
１項記載の部分親水化シリカゲル。 ３ 細孔内面部を含む全表面に疎水基が導入され
たシリカゲルにプラズマ処理を施して、外表面部
の疎水基を脱離し、シラノール基を顕出させるこ
とを特徴とするシリカゲルの細孔内面部に疎水基
が導入されていると共に、外表面部にシラノール
基を有する部分親水化シリカゲルの製造方法。 ４ 疎水基がアルキル基である特許請求の範囲第
３項記載の製造方法。[Scope of Claims] 1. A partially hydrophilized silica gel characterized by having a hydrophobic group introduced into the inner surface of the pores of the silica gel and having a silanol group on the outer surface. 2. The partially hydrophilized silica gel according to claim 1, wherein the hydrophobic group is an alkyl group. 3. The inner surface of the pores of silica gel, which is characterized in that silica gel having hydrophobic groups introduced into the entire surface including the inner surface of the pores is subjected to plasma treatment to remove the hydrophobic groups on the outer surface and expose silanol groups. A method for producing a partially hydrophilized silica gel having a hydrophobic group introduced therein and a silanol group on its outer surface. 4. The manufacturing method according to claim 3, wherein the hydrophobic group is an alkyl group.
Priority Applications (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP29916085A JPS62158112A (en) | 1985-12-27 | 1985-12-27 | Partially hydrophilized silica gel and its production |
| US06/946,078 US4694092A (en) | 1985-12-27 | 1986-12-24 | Partially hydrophilicized silica gel and process for producing the same |
| EP86118093A EP0228090B1 (en) | 1985-12-27 | 1986-12-29 | Partially hydrophilicized silica sel and process for producing the same |
| DE8686118093T DE3682886D1 (en) | 1985-12-27 | 1986-12-29 | PARTLY HYDROPHYLIZED SILICONE GEL AND METHOD FOR THE PRODUCTION THEREOF. |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP29916085A JPS62158112A (en) | 1985-12-27 | 1985-12-27 | Partially hydrophilized silica gel and its production |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS62158112A JPS62158112A (en) | 1987-07-14 |
| JPH0461809B2 true JPH0461809B2 (en) | 1992-10-02 |
Family
ID=17868906
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP29916085A Granted JPS62158112A (en) | 1985-12-27 | 1985-12-27 | Partially hydrophilized silica gel and its production |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS62158112A (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1019188B1 (en) * | 1997-09-29 | 2005-07-13 | MERCK PATENT GmbH | Chemically-modified porous materials with an electroneutral hydrophilic outer surface |
| JP2002256169A (en) * | 2001-02-28 | 2002-09-11 | Mitsubishi Materials Corp | Silica powder and its manufacturing method |
| US6900157B2 (en) * | 2001-09-10 | 2005-05-31 | Tosoh Corporation | Process for production of partially hydrophilized porous adsorbents |
| KR100613685B1 (en) | 2004-10-20 | 2006-08-21 | 한국화학연구원 | Method for modifying surface of silica |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4544485A (en) * | 1984-08-31 | 1985-10-01 | Purdue Research Foundation | Chromatographic method and means |
-
1985
- 1985-12-27 JP JP29916085A patent/JPS62158112A/en active Granted
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4544485A (en) * | 1984-08-31 | 1985-10-01 | Purdue Research Foundation | Chromatographic method and means |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS62158112A (en) | 1987-07-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US4131544A (en) | Macroencapsulated sorbent element and process for using the same | |
| Pu et al. | 2-Mercaptobenzothiazole-bonded silica gel as selective adsorbent for preconcentration of gold, platinum and palladium prior to their simultaneous inductively coupled plasma optical emission spectrometric determination | |
| Falter et al. | Determination of mercury species in natural waters at picogram level with on-line RP C18 preconcentration and HPLC-UV-PCO-CVAAS | |
| EP0228090B1 (en) | Partially hydrophilicized silica sel and process for producing the same | |
| CN110240705B (en) | Preparation method and application of nitrogen-rich covalent organic framework material | |
| EP1175933A1 (en) | Gas separation apparatus | |
| Liu et al. | Amino-functionalized hypercrosslinked polymer as sorbent for effective extraction of nitroimidazoles from water, drink and honey samples | |
| Gambero et al. | Use of chemically modified silica with β-diketoamine groups for separation of α-lactoalbumin from bovine milk whey by affinity chromatography | |
| JPH0461809B2 (en) | ||
| JPH0774797B2 (en) | Packing agent for liquid chromatography and method for producing and using the same | |
| JPH0468244B2 (en) | ||
| JPH0338891B2 (en) | ||
| Rykowska et al. | Extraction of copper ions using silica gel with chemically modified surface | |
| Kasturi et al. | Reordering/resonation studies of alkylamide phases | |
| Nazario et al. | Analysis of fluoxetine and norfluoxetine in human plasma by HPLC-UV using a high purity C18 silica-based SPE sorbent | |
| Wei et al. | In‐situ preparation of porous monolithic polymer inside hollow fiber as a micro‐solid phase extraction device for glucocorticoids in cosmetics | |
| JP2004003897A (en) | Carrier for chromatography, carrier for pretreatment, and kit | |
| Masters et al. | Ligand-exchange chromatography of amino sugars and amino acids on copper-loaded silylated controlled-pore glass | |
| JP2792038B2 (en) | Analysis method and pretreatment method for sample in which water-soluble polymer substance and low-molecular component coexist, and filler for chromatography | |
| CN105675773B (en) | A kind of method of prealbumin in solid extracting agent enrichment human plasma | |
| JPS58223061A (en) | Assay of non-protein material | |
| CN109942686A (en) | A kind of refining methd of pitressin acetylation impurity | |
| Faria et al. | Development of a polymer‐coated stationary phase with improved chemical stability in alkaline mobile phases | |
| Cole et al. | Cyclohexylamine additives for enhanced peptide separations in reversed phase liquid chromatography | |
| Huber | Biopolymer chromatography |