{"id":133,"date":"2015-08-03T13:25:11","date_gmt":"2015-08-03T04:25:11","guid":{"rendered":"http:\/\/www-p.cc.ocha.ac.jp\/bio-shimada-lab\/?page_id=133"},"modified":"2015-10-02T18:09:38","modified_gmt":"2015-10-02T09:09:38","slug":"pr_03hplc","status":"publish","type":"page","link":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/ex_protocol\/pr_03hplc.html","title":{"rendered":"HPLC\u95a2\u4fc2"},"content":{"rendered":"

\u00a0HPLC\u30d7\u30ed\u30c8\u30b3\u30eb\uff082007.12\u6539\u8a02\u7248\uff09<\/h2>\n\n\n\n\n\n\n\n\n\n
Ion-pairing solution<\/th>\n\u7d14\u6c34 (100mL)\u306bammonium acetate(15.4g)\u5165\u308c\u3001tetrabutylammonium hydroxide\u3000(1.0 M solution) \u309210 ml\u5165\u308c\u7d14\u6c34\u3067\u6700\u7d42volume 200mL\u306b\u3057\u3001\u9162\u9178\u3067pH7.1\u306b\u3059\u308b<\/td>\n<\/tr>\n
SolventA<\/th>\nion-pairing solution-water-acetone-acetonitrile, 5:25:20:50ips (100mL)-\u7d14\u6c34(500mL)-acetone (400mL)-acetonitrile (1000mL)<\/td>\n<\/tr>\n
SolventB<\/th>\nacetone-ethyl acetate, 50:50acetone (500mL)-ethyl acetate (500mL)<\/td>\n<\/tr>\n
\u00a0<\/th>\n\u906e\u5149\u51b7\u6697\u6240\u4fdd\u5b58<\/td>\n<\/tr>\n
\u30ab\u30e9\u30e0\u30bb\u30c3\u30c8<\/th>\nPartisil-5 ODS-3 column (4\u00d7250mm; GL Science, tokyo, Japan)<\/td>\n<\/tr>\n
\u30bf\u30a4\u30e0\u30d7\u30ed\u30b0\u30e9\u30e0<\/th>\nlinear gradient from 100% solvent A to 100% solvent B (20min)\u3001100% solvent B (10 min)\u3001solvent A (15 min)<\/td>\n<\/tr>\n
detector\u306e\u691c\u51fa<\/th>\n400-440nm<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

HPLC\u6e96\u5099<\/h4>\n

\u96fb\u6e90\u3092\u5165\u308c\u308b\uff1a\u672c\u4f53\u5f8c\u308d\u3001\u30d7\u30ea\u30f3\u30bf\u30fc\u306e\u5f8c\u308d\u3001\u691c\u51fa\u5668\u306e\u30e9\u30f3\u30d7\u3001\u30d1\u30bd\u30b3\u30f3 \u30d7\u30ec\u30e9\u30f3\uff1asolventA(\u2460)\u3001solventB\uff08\u2461\uff09\u3092HPLC\u306b\u30bb\u30c3\u30c8\u3002\u30c9\u30ec\u30a4\u30f3\u30d0\u30eb\u30d6\u3092\u958b\u3051\uff08\u53cd\u6642\u8a08\u56de\u308a\uff09\u3001\u30b7\u30ea\u30f3\u30b8\u3067\u6eb6\u5a92\u3092\u672c\u4f53\u306b\u5faa\u74b0\u3055\u305b\u308b\u3002<\/p>\n

\"image5\"<\/a><\/p>\n

\u6ce2\u9577\u8a2d\u5b9a<\/h4>\n

\u672c\u4f53\u306e\u96fb\u6e90\u3092\u4f9b\u7d66\u3057\u3001\u30e9\u30f3\u30d7\u3092\u70b9\u706f\u3057\u3066\u30a4\u30cb\u30b7\u30e3\u30e9\u30a4\u30ba\u304c\u7d42\u4e86\u3057\u305f\u5f8c\u306b\u3001 \u672c\u4f53\u5168\u9762\u306e\u6570\u5b57\u30ad\u30fc\u3092\u4f7f\u7528\u3057\u3066\u3001\u30e2\u30cb\u30bf\u30ea\u30f3\u30b0\u3059\u308b\u6ce2\u9577\u3092\u5165\u529b\u3002 \u6ce2\u9577\u5909\u66f4\u3092\u53cd\u6620\u3055\u305b\u308b\u5834\u5408\u306f\u3001\u30a8\u30f3\u30bf\u30fc\u30ad\u30fc\u3092\u62bc\u3059\u3053\u3068\u306b\u3088\u308a\u78ba\u5b9a\u3002 \u30e2\u30cb\u30bf\u30ea\u30f3\u30b0\u3059\u308b\u6ce2\u9577\u57df\uff08\u4f8b\u3048\u3070400nm\u304b\u3089440nm\u306a\u3069\uff09\u3092\u8a2d\u5b9a\u3059\u308b\u3053\u3068\u306f\u3067\u304d\u306a\u3044\u3002<\/p>\n

\u9023\u65e5\u4f7f\u7528\u306e\u5834\u5408\u306f\u3053\u3053\u304b\u3089 \u30d1\u30bd\u30b3\u30f3\u753b\u9762\u3067\u3001Manual\u2192Mobile Time 0, Flow 10, %B 0\u3092\u5165\u529b\u3057Begin (2-3min)\u3001stop\u3002<\/p>\n

\u540c\u69d8\u306bTime 0, Flow 10, %B 100\u3092\u5165\u529b\u3057start run (2-3min)\u3001stop\u3002<\/p>\n

\u30c9\u30ec\u30a4\u30f3\u30d0\u30eb\u30d6\u3092\u9589\u3081\u3001 \u30ab\u30e9\u30e0\u306e\u4e21\u3075\u305f\u3092\u3068\u3063\u3066\u5de6\u53f3\u9593\u9055\u3048\u305a\u30e9\u30a4\u30f3\u3092\u5165\u308c\u308b\u3002 Time 0, Flow 1, %B 0\u300045min prerun\u3002<\/p>\n

\u30b5\u30f3\u30d7\u30eb\u6e96\u5099<\/h4>\n

\u85fb\u4f53\u306e\u30b3\u30f3\u30bf\u30df\u30c1\u30a7\u30c3\u30af\u2192\u30ad\u30e0\u30ef\u30a4\u30d7\u3067\u6c34\u6c17\u3092\u3068\u308b\u2192\u6c37\u4e0a\u3067\u51b7\u3084\u3057\u305f\u4e73\u9262\u306b\u85fb\u4f53\u3092\u9069\u5f53\u306b\u5c0f\u3055\u304f\u5207\u3063\u3066\u3044\u308c\u308b\u3002\u4e2d\u6027buffer\u3092\u6570ml\u5782\u3089\u3057\u3001\u305d\u306e\u4e0a\u304b\u3089\u3082\u3046\u4e00\u5ea6\u30ad\u30e0\u30ef\u30a4\u30d7\u3067\u6c34\u6c17\u3092\u3068\u308b\u2192\u30a2\u30bb\u30c8\u30f3\u3092\u6570ml\u5165\u308chomogenize\uff08\u5f31\u6697\u6240\uff09\u3002<\/p>\n

\u6eb6\u6db2\u30921.5ml\u30c1\u30e5\u30fc\u30d6\u306b\u79fb\u3057\u30c1\u30d3\u30bf\u30f3\u306730sec.\u2192milliporefilter\u306b\u304b\u3051\u6c37\u4e0a\u906e\u5149\u3067\u4fdd\u5b58<\/p>\n

INJECTION\u76f4\u524d\u306b3\/10\u306eIon-pairing Solution\u3092\u52a0\u3048\u308b\u3002<\/p>\n

\u30b5\u30f3\u30d7\u30eb\u306e\u30a4\u30f3\u30b8\u30a7\u30af\u30b7\u30e7\u30f3 Prerun\u7d42\u4e86\u5f8c\u3001\u300cRUN\u300d\u3092\u62bc\u3059\u3002<\/p>\n

HPLC\u5c02\u7528\u306e\u30b7\u30ea\u30f3\u30b8\u3092\u30e1\u30bf\u30ce\u30fc\u30eb\u3067\u6d17\u6d44\uff08\u91dd\u3092\u62ed\u3044\u3066\u304b\u3089\u51fa\u3057\u5165\u308c\uff09\u3002\u30b5\u30f3\u30d7\u30eb\u3092\u5438\u3044\u6570\u56de\u51fa\u3057\u5165\u308c\u3002\u30a4\u30f3\u30b8\u30a7\u30af\u30b7\u30e7\u30f3\u30d0\u30eb\u30d6\u3092Load\u306b\u3057\u3066\u3055\u3057\u3066\u3001\u6ce8\u5165\u3001\u91dd\u3092\u629c\u3044\u3066\u30d0\u30eb\u30d6\u3092inject\u306b\u623b\u3059\u3002 \u30b7\u30ea\u30f3\u30b8\u3092\u30e1\u30bf\u30ce\u30fc\u30eb\u3067\u6d17\u6d44\uff08\u91dd\u3092\u62ed\u3044\u3066\u304b\u3089\u51fa\u3057\u5165\u308c\uff09\u3002<\/p>\n

\u7d42\u4e86\u5f8c\u3001\u6b21\u306e\u30b5\u30f3\u30d7\u30eb\u3092\u30a4\u30f3\u30b8\u30a7\u30af\u30b7\u30e7\u30f3<\/p>\n

\u307e\u305f\u306f\u3001100\uff05\u30e1\u30bf\u30ce\u30fc\u30eb\u6d17\u6d44\uff0830-60min\uff09<\/p>\n

\u30ab\u30e9\u30e0\u3092\u306f\u305a\u3059\u3002\u30e9\u30a4\u30f3\u306f\u30d1\u30e9\u30d5\u30a3\u30eb\u30e0\u3067\u8986\u3046\u3002 \u96fb\u6e90\u3092\u5207\u308b\u3002\u30d1\u30bd\u30b3\u30f3\u56de\u53ce\u3002<\/p>\n

\u30c7\u30fc\u30bf\u89e3\u6790File, Data or Peprot Clip\u3067\u30b3\u30d4\u30fc<\/p>\n","protected":false},"excerpt":{"rendered":"

\u00a0HPLC\u30d7\u30ed\u30c8\u30b3\u30eb\uff082007.12\u6539\u8a02\u7248\uff09 Ion-pairing solution \u7d14\u6c34 (100mL)\u306bammonium acetate(15.4g)\u5165\u308c\u3001tetrabutylammonium hydroxide\u3000 … \u7d9a\u304d\u3092\u8aad\u3080 →<\/span><\/a><\/p>\n","protected":false},"author":18,"featured_media":0,"parent":94,"menu_order":3,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-133","page","type-page","status-publish","hentry"],"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/pages\/133","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/users\/18"}],"replies":[{"embeddable":true,"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/comments?post=133"}],"version-history":[{"count":3,"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/pages\/133\/revisions"}],"predecessor-version":[{"id":693,"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/pages\/133\/revisions\/693"}],"up":[{"embeddable":true,"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/pages\/94"}],"wp:attachment":[{"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/media?parent=133"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}