{"id":1771,"date":"2017-02-21T10:50:46","date_gmt":"2017-02-21T08:50:46","guid":{"rendered":"https:\/\/www.fractioncollector.info\/?p=1771"},"modified":"2023-10-04T15:51:26","modified_gmt":"2023-10-04T13:51:26","slug":"izolace-a-charakterizace-alelopatickych-sloucenin","status":"publish","type":"post","link":"https:\/\/www.fractioncollector.info\/cs\/isolation-and-characterization-of-allelopathic-compounds\/","title":{"rendered":"Izolace, purifikace a charakterizace alelopatick\u00fdch slou\u010denin"},"content":{"rendered":"

Izolace a charakterizace alelopatick\u00fdch slou\u010denin pomoc\u00ed sloupcov\u00e9 chromatografie a frak\u010dn\u00edho sb\u011bra\u010de LAMBDA OMNICOLL<\/span><\/strong><\/h1>\n<\/div><\/div><\/div><\/div><\/div>

Izolace, objasn\u011bn\u00ed struktury a chemick\u00e1 synt\u00e9za alelopatick\u00fdch slou\u010denin<\/b> z rostlin, hub a dal\u0161\u00edch mikroorganism\u016f jsou velmi pracn\u00e9 a \u010dasov\u011b n\u00e1ro\u010dn\u00e9 studie. Identifikace vysoce \u00fa\u010dinn\u00fdch biologick\u00fdch struktur v\u0161ak (pou\u017e\u00edvaj\u00ed se jako p\u0159\u00edrodn\u00ed herbicidy, l\u00e9\u010diva, antioxidanty atd.)<\/i> stoj\u00ed za ten \u010das a \u00fasil\u00ed vynalo\u017een\u00e9 na to.<\/p>\n

Rutinn\u00ed studie alelochemick\u00e9 strukturn\u00ed anal\u00fdzy se st\u00e1v\u00e1 obt\u00ed\u017enou, pokud struktury nejsou v surov\u00e9m extraktu identifikov\u00e1ny, a to ani po n\u011bkolika p\u0159edb\u011b\u017en\u00fdch kroc\u00edch k dosa\u017een\u00ed frakcionace surov\u00fdch extrakt\u016f. (macerac\u00ed, extrakc\u00ed za pomoci ultrazvuku nebo mikrovln n\u00e1sledovanou dvouf\u00e1zovou vodnou extrakc\u00ed atd.)<\/i>.<\/p>\n

V\u00fdzkumn\u00edk pak z\u016fstane s velk\u00fdm objemem rozpou\u0161t\u011bdla, kter\u00e9 m\u016f\u017ee obsahovat \u010d\u00e1ste\u010dnou sm\u011bs struktur (s podobn\u00fdmi biologick\u00fdmi aktivitami nebo bez nich)<\/i>. Biologick\u00e1 aktivita<\/b> struktur lze identifikovat podle bio-guided studie a testy<\/b>. Tento velk\u00fd objem rozpou\u0161t\u011bdla by m\u011bl b\u00fdt rozd\u011blen na men\u0161\u00ed frakce pomoc\u00ed sm\u011bs\u00ed rozpou\u0161t\u011bdel s r\u016fzn\u00e9 polarity<\/b>. Sb\u011br frakc\u00ed pom\u016f\u017ee izolovat nejaktivn\u011bj\u0161\u00ed alelopatick\u00e9 molekuly pro po\u017eadovanou\/c\u00edlovou biologickou aktivitu.<\/p>\n

V\u011bt\u0161inou chromatografie na kapalinov\u00e9 kolon\u011b silikagelu<\/b> se pou\u017e\u00edv\u00e1 k frakcionaci extrakt\u016f. B\u011b\u017en\u00e9 je v\u0161ak frakcionovat a shroma\u017e\u010fovat elu\u00e1ty o objemu p\u0159ibli\u017en\u011b 65 - 200 frakc\u00ed<\/b>, aby se dos\u00e1hlo nej\u010dist\u0161\u00ed bioaktivn\u00ed struktura v ka\u017ed\u00e9m elu\u00e1tu<\/b>. Ru\u010dn\u00ed sb\u011br velk\u00e9ho po\u010dtu frakc\u00ed je \u010dasov\u011b a energeticky nejn\u00e1ro\u010dn\u011bj\u0161\u00ed f\u00e1z\u00ed v obecn\u00e9m v\u00fdzkumn\u00e9m projektu, a to kv\u016fli vysok\u00e9 cen\u011b spektrofotometrick\u00fdch sb\u011bra\u010d\u016f frakc\u00ed na trhu.<\/p>\n

Dal\u0161\u00ed kroky \u010di\u0161t\u011bn\u00ed zahrnuj\u00ed tenkovrstv\u00e1 chromatografie, bioaktivn\u00ed testy, preparativn\u00ed sloupcov\u00e1 chromatografie <\/b>atd. a nakonec seznam spektrometrick\u00e9 testy<\/b> pro vysoce aktivn\u00ed struktury, aby se v budoucnu objasnila a dos\u00e1hla jejich chemick\u00e9 synt\u00e9zy.<\/p>\n

Ten\/Ta\/To sb\u011br 200 a v\u00edce frakc\u00ed lze snadno automatizovat<\/b> co nejprovediteln\u011bj\u0161\u00edm zp\u016fsobem (pokud je spr\u00e1vn\u00e9 sb\u011bra\u010d frakc\u00ed<\/a> je vybr\u00e1n!)<\/em>.<\/p>\n<\/div><\/div><\/div>

\"\"<\/a>

Izolace a purifikace alelopatick\u00fdch slou\u010denin za pou\u017eit\u00ed sloupcov\u00e9 chromatografie na silikagelu, \u010dasov\u011b v\u00e1zan\u00fdch frakc\u00ed sb\u00edran\u00fdch pomoc\u00ed sb\u011bra\u010de frakc\u00ed OMNICOLL a tenkovrstv\u00e9 chromatografie (TLC). Purifikovan\u00e9 slou\u010deniny lze charakterizovat UV, IR, LC-MS a NMR spektroskopick\u00fdmi studiemi.<\/p><\/div><\/p>\n<\/div><\/div><\/div><\/div>

Frakcionace surov\u00e9ho extraktu<\/h2>\n

Nejprve se surov\u00e9 extrakty separuj\u00ed do r\u016fzn\u00fdch samostatn\u00fdch frakc\u00ed obsahuj\u00edc\u00edch slou\u010deniny s podobnou polaritou nebo molekul\u00e1rn\u00ed velikost\u00ed pomoc\u00ed sloupcov\u00e1 chromatografie<\/b>. V t\u00e9to chromatografick\u00e9 separaci asi Bylo odebr\u00e1no 65 frakc\u00ed o objemu 20 ml<\/b> (na z\u00e1klad\u011b \u010dasu) podle sb\u011bra\u010d frakc\u00ed<\/b> OMNICOLL.<\/p>\n<\/div><\/div><\/div><\/div>

Tenkovrstv\u00e1 chromatografie (TLC)<\/h2>\n

Alikvotn\u00ed pod\u00edl v\u0161ech shrom\u00e1\u017ed\u011bn\u00fdch frakc\u00ed byl nanesen na aktivovan\u00e9 silikagelov\u00e9 TLC desti\u010dky (20 x 20 cm). Frakce byly slou\u010deny na z\u00e1klad\u011b vzor skvrn a hodnoty Rf<\/b> na TLC desti\u010dce. Spojen\u00e9 frakce byly o\u010d\u00edslov\u00e1ny (Fr.1 - Fr.8).<\/p>\n<\/div><\/div><\/div><\/div>

Preparativn\u00ed TLC nebo HPLC sloupcov\u00e1 chromatografie<\/h2>\n

Frakce (Fr.1 - Fr.8) lze d\u00e1le \u010distit pomoc\u00ed preparativn\u00ed TLC a\/nebo HPLC sloupcov\u00e1 chromatografie<\/b>. Preparativn\u00ed HPLC pom\u016f\u017ee p\u0159esn\u011bji izolovat konkr\u00e9tn\u00ed slou\u010deninu.<\/p>\n<\/div><\/div><\/div><\/div>

Charakterizace alelopatick\u00fdch slou\u010denin<\/h2>\n

Izolovan\u00e9 \u010dist\u00e9 frakce se pot\u00e9 pou\u017eij\u00ed ke stanoven\u00ed struktury a tak\u00e9 k op\u011btovn\u00e9mu ov\u011b\u0159en\u00ed biologick\u00e9 aktivity.<\/p>\n

Charakterizace \u010dist\u00fdch alelopatick\u00fdch slou\u010denin se prov\u00e1d\u00ed pomoc\u00ed metody s vysok\u00fdm rozli\u0161en\u00edm. IR, MS, 1<\/sup>H a 13<\/sup>C NMR <\/b>spektra.<\/p>\n

IR spektra se pou\u017e\u00edvaj\u00ed ke zji\u0161t\u011bn\u00ed funk\u010dn\u00edch skupin izolovan\u00e9 \u010dist\u00e9 frakce. LC-MS ud\u00e1v\u00e1 bu\u010f empirickou, nebo p\u0159esnou (HRMS) molekulovou hmotnost a NMR spektra odhaluj\u00ed prost\u0159ed\u00ed C a H. LC-MS anal\u00fdza pom\u00e1h\u00e1 porovnat MS obrazce.<\/p>\n<\/div><\/div><\/div><\/div><\/div>","protected":false},"excerpt":{"rendered":"

Isolation and characterization of allelopathic compounds by column chromatography and fraction collector LAMBDA OMNICOLL Isolation, structural elucidation and chemical synthesis of allelopathic compounds from plants, fungi, and other microorganisms are a very labor-exhausting and time-consuming studies. However, the identification of highly potent biological structures (used as natural herbicides, medicaments, antioxidants, etc.) is worth the time […]<\/p>\n","protected":false},"author":2,"featured_media":1789,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"ai_generated_summary":"","footnotes":""},"categories":[132,4],"tags":[135,134,6,136,133],"class_list":["post-1771","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-post-name","category-omnicoll","tag-allelopathic-compounds","tag-chemical-synthesis","tag-fraction-collector","tag-liquid-column-chromatography","tag-structural-elucidation","post-with-thumbnail","post-with-thumbnail-icon"],"yoast_head":"\nIsolation, purification and characterization of allelopathic compounds – LAMBDA OMNICOLL | LAMBDA Laboratory Instruments<\/title>\n<meta name=\"description\" content=\"Isolation of allelopathic compounds with silica gel column 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