杞嚜 鐜颁唬绉戝浠櫒缃?/p>

锛堜竴锛夐叾瀵归鍝佸姞宸ョ殑褰卞搷

閰舵槸鐢熺墿娲荤粏鑳炰骇鐢熺殑涓€绫诲叿鏈夊偓鍖栧姛鑳界殑铔嬬櫧璐ㄣ€傞叾闄や簡鍏锋湁楂樼殑鍌寲鏁堢巼鐗规€у锛岃繕鍏锋湁寰堥珮鐨勪笓涓€鎬э紝鍒╃敤瀹冭兘閫夋嫨鎬у湴灏嗕釜鍒鍝佺粍鍒嗘敼鎬э紝鑰屼笉褰卞搷鍒板叾瀹冪粍鍒嗐€傚洜姝わ紝閰跺湪椋熷搧绉戝涓浉褰撻噸瑕侊紝閫氳繃閰剁殑浣滅敤鑳藉紩璧烽鍝佸師鏂欑殑鍝佽川鍙戠敓鍙樺寲锛屼篃鑳藉湪姣旇緝娓╁拰鐨勬潯浠朵笅鍔犲伐鍜屾敼鑹鍝併€?/p>

1. 鏈夊埄褰卞搷

灏嗘灉鑳堕叾銆佺氦缁寸礌閰剁瓑閰跺埗鍓傚簲鐢ㄤ簬鏋滈厭銆佹灉姹佺殑鐢熶骇锛屾湁鍒╀簬姒ㄦ眮銆佹緞娓呭拰杩囨护绛変綔涓氥€?/p>

椋熷搧涓殑澶у垎瀛愮墿璐紝濡傝剛绫汇€佽泲鐧借川銆佹牳閰哥瓑锛屽湪閰剁殑浣滅敤涓嬪彲鐢熸垚椋庡懗鐗╄川銆?/p>

涔崇硸閰堕€氳繃姘磋В涔崇硸鏉ユ彁楂樹钩鍒跺搧鐨勫彲娑堝寲鎬э紝鏈夊埄浜庣紦瑙ｄ钩绯栦笉鑰愮棁銆?/p>

鏋滃疄鐨勫悗鐔熶綔鐢ㄦ槸鍦ㄥ悇绉嶉叾鐨勫弬涓庝笅杩涜鐨勬瀬鍏跺鏉傜殑鐢熺悊鐢熷寲杩囩▼銆傚娣€绮夊垎瑙ｄ负绯栵紝鏋滃疄鍙樼敎锛涘彲婧舵€у崟瀹佸嚌鍥猴紝鏋滃疄娑╁懗娑堝け锛涘師鏋滆兌姘磋В涓烘灉鑳讹紝鏋滃疄鍙樿蒋锛涘悓鏃舵灉瀹炶壊娉藉姞娣憋紝棣欏懗澧炲姞銆?/p>

2. 涓嶅埄褰卞搷

鍦ㄧ綈澶村姞宸ヤ腑锛岃嫻鏋溿€佹ⅷ鍜岄┈閾冭柉绛夋按鏋溿€佽敩鑿滃湪鍓婄毊鍒囧紑鍚庯紝鐢变簬缁勭粐鍐呭閰氭哀鍖栭叾鐨勪綔鐢紝浼氬彂鐢熼叾淇冭鍙橈紝涓ラ噸褰卞搷浜у搧鐨勫瑙傚拰鍝佽川銆?/p>

鏈変簺寰敓鐗╁垎娉岀殑鍚勭閰跺彲灏嗛鍝佷腑铔嬬櫧璐ㄦ按瑙ｆ垚澶氳偨鍜屾皑鍩洪吀锛屽苟鑳借繘涓€姝ュ皢姘ㄥ熀閰稿垎瑙ｇ敓鎴愭皑銆佺～鍖栨阿绛夌墿璐紝鑰屽紩璧烽鍝佺殑鑵愯触鍙樿川銆?/p>

閰跺湪椋熷搧鍔犲伐涓殑浣滅敤灏卞儚涓€鎶婂弻鍒冨墤锛屾垜浠瓒嬪埄閬垮銆傞叾鐨勭Н鏋佷綔鐢ㄦ垜浠鍔犲己锛屽湪椋熷搧鍔犲伐杩囩▼涓坊鍔犻叾鍒跺墏锛屼娇鍏朵綔鐢ㄥ厖鍒嗗彂鎸ワ紱娑堟瀬浣滅敤鎴戜滑瑕佸敖閲忛伩鍏嶏紝鍙互閫氳繃鍔犵儹绛夋柟娉曞皢閰剁伃娲伙紝娑堥櫎鍏朵笉鍒╁奖鍝嶃€?/p>

涓轰簡灏嗛叾鏇村ソ鍦板簲鐢ㄤ簬椋熷搧鍔犲伐锛岀爺绌堕叾鐨勬€ц川鏄崄鍒嗗繀瑕佺殑銆傝€岀传澶?鍙鍒嗗厜鍏夊害娉曟槸鐮旂┒閰舵€ц川鐨勯噸瑕佹柟娉曚箣涓€銆備笅闈粙缁嶇敤-鍙鍒嗗厜鍏夊害璁℃祴瀹氶叾娲荤殑鍏蜂綋鏂规硶銆?/p>

锛堜簩锛夌传澶?鍙鍒嗗厜鍏夊害娉曟祴瀹氶叾娲?/p>

1. β涓€鍗婁钩绯栬嫹閰?

β涓€鍗婁钩绯栬嫹閰讹紝鍙堢О涔崇硸閰?Lactase)銆傝兘姘磋В涔崇硸鏉ラ檷浣庝钩鍒跺搧鐨勪钩绯栧惈閲忥紝浠庤€屾彁楂樹钩鍒跺搧鐨勫彲娑堝寲鎬э紝鐢ㄤ簬浣庝钩绯栫墰濂跺拰闈炵粨鏅跺瀷娴撶缉鐗涘ザ鐨勭敓浜у強濂堕叒椋庡懗鐨勬敼鍙橈紝鍚屾椂杩樺彲鐢ㄤ簬鐢熶骇浣庤仛鍗婁钩绯栥€?/p>

銆愰叾娲绘祴瀹氥€戜互ONPG涓哄簳鐗╂祴瀹?beta;-鍗婁钩绯栬嫹閰舵椿鍔涖€?/p>

銆愰叾娲诲畾涔夈€戜互ONPG涓哄簳鐗╋紝37鈩冧繚娓╅叾瑙ｏ紝姣忓垎閽熼噴鏀緇μmol锛廘閭荤鍩洪厷鐨勯叾閲忥紝瀹氫箟涓?涓叾娲诲姏鍗曚綅銆?/p>

2. 瓒呮哀鍖栫墿姝у寲閰?/p>

瓒呮哀鍖栫墿姝у寲閰?Superoxide Dismutase锛岀畝绉癝OD)鏄竴绉嶅崄鍒嗛噸瑕佺殑鐢熺墿浣撻槻姝㈡哀鍖栨崯浼ょ殑閰剁被锛屾槸鐢熺墿浣撳唴瓒呮哀闃寸瀛愭竻闄ゅ墏锛屼繚鎶ょ粏鑳炲厤鍙楁崯浼ゃ€係OD骞挎硾瀛樺湪浜庡悇绫荤敓鐗╀綋鍐咃紝鎵€鏈夊ソ姘у井鐢熺墿缁嗚優涓兘鍚湁SOD銆傝嚜1969骞碝ccord绛変汉棣栨鍙戠幇浜哠OD鐢熺墿娲绘€у悗锛屽尰瀛︾晫瀵瑰叾鍖荤枟浣滅敤鍋氫簡璁稿鐮旂┒锛岃瘉鏄庡畠鍏锋湁鎶楄“鑰併€佹姉鑲跨槫銆佹姉杈愬皠銆佹姉缂鸿銆佹彁楂樹汉浣撳厤鐤姏绛変綔鐢紝琚笓瀹剁О涓?1涓栫邯鏈€鏈夊墠閫旂殑鑽敤閰躲€傛缇庡浗瀹跺凡寮€濮嬪皢鍏跺簲鐢ㄤ簬鍖荤枟銆侀鍝併€佷繚鍋ャ€佸寲濡嗗搧绛夐鍩熴€?/p>

銆愰叾娲绘祴瀹氥€戝湪25鈩?4.5ml 50mmol锛廘 pH8.3鐨凨2HPO4- KH2PO4缂撳啿娑蹭腑鍔犲叆寰呮祴SOD鏍锋恫锛屽啀鍔犲叆10ul 50mmol锛廘鐨勮繛鑻笁閰氾紝杩呴€熸憞鍖€锛屽€掍汉鍏夊緞lcm 鐨勬瘮鑹叉澂锛屽湪325nm娉㈤暱涓嬫瘡闅?0s娴嬩竴娆鍊笺€傚悓鏃舵祴瀹歋igma鍏徃鐨凷OD鏍囧噯鍝侊紝瀵规墍娴嬫牱鍝丼OD娲绘€ц繘琛屾牎姝ｃ€?/p>

銆愰叾娲诲畾涔夈€戝湪lml鍙嶅簲娑蹭腑锛屾瘡鍒嗛挓鎶戝埗杩炶嫰涓夐厷鑷哀鍖栭€熺巼杈?0锛呮椂鐨勯叾閲忎负涓€涓叾娲诲崟浣嶏紝 鍗?25nm 0.035OD锛弇in涓轰竴涓叾娲诲崟浣嶃€?/p>

3. 澶氶厷姘у寲閰?/p>

澶氶厷姘у寲閰?PPO)鏄竴绫荤敱鏍稿熀鍥犵紪鐮佸湪缁嗚優璐ㄤ腑鍚堟垚鐨勫惈閾滆川浣撶殑閲戝睘閰讹紝鍏朵綔鐢ㄦ満鐞嗗湪浜庨摐鐨勬哀鍖?杩樺師浣滅敤銆傚ぇ楹︿腑鐨勫閰氱墿璐ㄧ粡杩嘝PO鐨勫偓鍖栨哀鍖栧悗锛屽叿鏈夊崟瀹佹€ц川锛屾槗鍜岃泲鐧借川璧蜂氦鑱斾綔鐢ㄨ€屾矇娣€鍑烘潵锛岃繘鑰屽奖鍝嶅暏閰掔殑鑹叉辰銆佹场娌€侀鍛冲拰闈炵敓鐗╃ǔ瀹氭€с€傚ぇ楹︿腑閮ㄥ垎PPO浠?ldquo;娼滀紡”鐘舵€佸瓨鍦紝鍦ㄦ蹈楹﹁繃绋嬩腑锛屽彲閫氳繃鍘婚櫎涓€浜涢叾鎶戝埗鍓傘€佽泲鐧介叾浣滅敤鎴栧叾浠栫殑涓€浜涙柟寮忎娇鍏舵椿鎬у湪娴搁害杩囩▼涓彁楂樸€傚湪澶ч害鍙戣娊杩囩▼涓紝PPO鐨勬椿鍔涜緝楂橈紝褰卞搷鐫€楹﹁娊鐨勮川閲忋€傚洜姝わ紝鍦ㄥ埗楹﹁繃绋嬩腑搴旇闄嶄綆PPO娲绘€с€?/p>

銆愰叾娲绘祴瀹氥€?.1mL閰舵恫涓巐mL 0.1 mol锛廘閭昏嫰浜岄厷(鐢╬H 8.4鏌犳閰?纾烽吀缂撳啿娑查厤鍒?80oC鍙嶅簲3min锛岀敤2mL 20锛匱CA缁堟鍙嶅簲锛屼簬娉㈤暱450nm澶勭敤鍒嗗厜鍏夊害璁℃祴瀹氬惛鏀跺€笺€?/p>

銆愰叾娲诲畾涔夈€戜笂杩版潯浠朵笅锛屽皢姣忓垎閽烵D鍊煎鍔?.01瀹氫箟涓?涓叾娲诲姏鍗曚綅銆?/p>

4. 钁¤悇绯栨哀鍖栭叾

涓婁笘绾?0骞翠唬浠ュ墠锛岃憽钀勭硸姘у寲閰朵富瑕佺敤浜庤泲鍝佸姞宸ヤ笟锛岄櫎鍘昏憽钀勭硸锛岄槻姝㈠彂鐢烳aillard鍙嶅簲锛岀ǔ瀹氫骇鍝佽川閲忥紝寤堕暱淇濊川鏈熴€?0骞翠唬鏈紝鍙堥檰缁紑鍙戜簡鍘婚櫎鍟ら厭婧惰В姘у拰鐡堕姘ф敼鍠勯鍛冲欢闀夸繚璐ㄦ湡銆佹灉姹佹灉閰掗櫎姘ф姢鑹茬ǔ瀹氳川閲忋€侀潰绮夊搧璐ㄦ敼鍠勩€佹捣浜у搧鍜岃倝绫讳繚椴溿€侀ゲ鏂欐坊鍔犲幓姘у钩琛＄暅绂借儍鑲犱唬璋㈢瓑鐢ㄩ€旓紝骞跺缓绔嬩簡钁¤悇绯栨哀鍖栭叾涓撲笟鐢熶骇杞﹂棿銆?/p>

銆愰叾娲绘祴瀹氥€戦噰鐢?鑱?浜?鑼撮鑳哄垎鍏夊厜搴︽硶銆傚湪鏈夋哀瀛樺湪鏉′欢涓嬶紝钁¤悇绯栨哀鍖栭叾鍌寲钁¤悇绯栬劚姘骇鐢烪2O2锛屽湪杩囨哀鍖栫墿閰?POD)浣滅敤涓嬶紝姘т緵浣撳嵆閭讳竴鑱?浜?鑼撮鑳?DH2)琚哀鍖栨垚妫曡壊浜х墿銆備簬436nm澶勬祴閲忓惛鍏夌巼鐨勫彉鍖栵紝鍗冲彲鎹㈢畻涓鸿憽钀勭硸姘у寲閰剁殑娲绘€с€?/p>

銆愰叾娲诲畾涔夈€戜竴涓叾娲绘€у崟浣嶅畾涔変负鍦?6鈩冧笅姣忓垎鍌寲閲婃斁涓€寰厠鍒嗗瓙H2O2鎵€闇€鐨勯叾閲忋€?/p>

5. 鏋滆兌閰?/p>

鏋滆兌閰跺彲浠ユ敼鍠勮憽钀勯厭鐨勮壊娉姐€佸鍔犻厭棣欙紝骞舵彁楂樿憽钀勯厭鍑烘眮鐜囩瓑锛屽鎻愰珮绾㈣憽钀勯厭鐨勫搧璐ㄦ湁閲嶈浣滅敤銆?/p>

銆愰叾娲绘祴瀹氥€戝悜鍏峰璇曠涓姞鍏?.8mL 0.30锛?鑱氬崐涔崇硸閱涢吀婧舵恫(鐢?.05 mol锛廘 Na2CO3锛廚aHCO3缂撳啿婧舵恫閰嶅埗锛岃皟鑺俻H 10.0)锛屼簬55鈩冩按娴翠笅淇濇俯5 min锛岀劧鍚庡姞鍏ヤ竴瀹氱█閲婂€嶆暟鐨勯叾娑?.2 mL鍙嶅簲15 min锛屽姞鍏NS璇曞墏1.5 mL锛屾贩鍖€锛屽湪娌告按涓姞鐑?0min锛岀珛鍗崇敤鑷潵姘村喎鍗达紝鍐嶅姞鍏?1.5 mL钂搁姘达紝娣峰寑锛屼簬鍒嗗厜鍏夊害璁′笂520 nm娴嬪畾鍚稿厜搴︺€傜┖鐧界敤鐓哺澶辨椿鐨勯叾娑蹭唬鏇裤€?/p>

銆愰叾娲诲畾涔夈€戝湪涓婅堪璇曢獙鏉′欢涓嬶紝浠ユ瘡鍒嗛挓鍒嗚В搴曠墿浜х敓1μmol鍗婁钩绯栭啗閰告墍闇€鐨勯叾閲忎负1涓叾娲诲姏鍗曚綅銆?/p>

6. 鑳?alpha;-娣€绮夐叾

銆愰叾娲绘祴瀹氥€戠敤绉绘恫绠″皢0锛? mL閰舵憾娑茬Щ鍏ヨ瘯绠★紝骞跺湪姘存荡涓皟娓╄嚦25鈩冦€傚姞0锛? mL宸插悓鏍烽鐑埌25鈩冪殑1锛呮穩绮夋憾娑层€? min鍚庡姞1锛? mL DNS鏄捐壊鍓傘€傜疆璇曠浜庢哺姘翠腑5 min锛屽喎鍗村悗鐢?0 mL钂搁姘寸█閲娿€傜敤鍒嗗厜鍏夊害璁′簬540 nm澶勪互绌虹櫧浣滃鐓ф祴瀹氬惛鍏夊害銆備互0锛? mL纾烽吀鐩愮紦鍐叉恫浠ｆ浛閰舵恫娴嬪畾绌虹櫧鍊笺€?/p>

7. 妞嶉吀閰?/p>

銆愰叾娲绘祴瀹氥€?浠ユ閰搁挔涓哄簳鐗╋紝閲囩敤閽掗捈閰搁摰娉曟祴瀹氭閰搁叾鐨勯叾娲汇€傚惛鍙? mmol锛弆鐨勬閰搁挔(閰朵綔鐢ㄥ簳鐗╋紝鐢?锛?5 mol锛弆銆乸H鍊?锛?0閱嬮吀-閱嬮吀閽犵紦鍐叉恫閰嶅埗)1锛? ml锛?7 oC棰勭儹5 min鍚庯紝鍑嗙‘鍔犲叆閫傚綋绋€閲婄殑閰舵恫0锛? ml(瀵圭収缁勫厛鍔犲弽搴旂粓姝㈡恫)锛?7 oC姘存荡鍙嶅簲30min鍚庯紝鍐嶅姞鍏? ml鍙嶅簲缁堟娑?鐜扮敤鐜伴厤)鍐峰嵈鑷冲娓╋紝鍦?15 nm澶勬祴瀹歄D鍊硷紝鍒╃敤鏃犳満纾锋爣鍑嗘洸绾胯绠楀嚭閰舵椿銆?/p>

銆愰叾娲诲畾涔夈€戝湪37鈩冿紝姣忓垎閽熶粠1 mmol锛弆妞嶉吀閽犳憾娑蹭腑(鐢╬H鍊?锛?鐨勯唻閰?閱嬮吀閽犵紦鍐叉恫閰嶅埗)閲婃斁鍑? nmol鏃犳満纾锋墍闇€瑕佺殑閰堕噺涓轰竴涓叾娲诲崟浣嶃€?/p>

8. 缇х敳鍩虹氦缁寸礌閰?/p>

銆愰叾娲绘祴瀹氥€戝湪0.5mL閫傚綋绋€閲婄殑閰舵恫涓姞鍏ヨ川閲忓垎鏁颁负0.625%鐨勭晶鐢插熀绾ょ淮绱犻挔婧舵恫锛屼簬50鈩冩按娴翠腑淇濇俯30min锛屽姞2.0 mL璐ㄩ噺鍒嗘暟涓?0%鐨凬aOH婧舵恫缁堟鍙嶅簲锛屽啀鍔犲叆2.5mLDNS璇曞墏锛屼簬娌告按娴翠腑鐓哺15min锛屽喎鍗达紝鍦?50 nm娉㈤暱澶勬瘮鑹诧紝瑕佹眰鍏夊瘑搴﹁鏁板敖鍙兘鍦ㄦ爣鍑嗘洸绾跨殑0. 5 mg钁¤悇绯栧銆傛瘡娆℃祴瀹氬潎璁剧珛绌虹櫧瀵圭収锛屽鐓х鍏堝姞鍏?.5mL閰舵恫锛岀劧鍚庡姞鍏?. 0 mL璐ㄩ噺鍒嗘暟涓?0%鐨凬aOH婧舵恫灏嗛叾鐏椿锛屽啀渚濇鍔犲叆璐ㄩ噺鍒嗘暟涓?.625%鐨勫簳鐗?. 0 mL锛屼笌寰呮祴绠′竴璧蜂繚娓╋紝鍔犲叆DNS锛岀叜娌搞€佸喎鍗淬€佺█閲婃瘮鑹层€佷笌鏍囧噯鏇茬嚎瀵圭収銆?/p>

銆愰叾娲诲畾涔夈€戝湪涓婅堪鏉′欢涓嬶紝姣?0min浜х敓0.5mg杩樺師绯栦负1涓狢x娲诲姏鍗曚綅锛屽嵆姣? h浜х敓1.0mg杩樺師绯栦负1涓狢x娲诲姏鍗曚綅銆?/p>

9. 鏈ㄨ仛绯栭叾

     鏈ㄨ仛绯栭叾鑳芥按瑙ｄ笉鍙憾鎴婅仛绯?闃挎媺浼湪鑱氱硸)锛屼娇鍏舵垚涓烘按婧舵€х殑锛屾按瑙ｇ巼杈?5锛呫€傞潰绮変腑澧炲姞姘存憾鎬ч樋鎷変集鏈ㄨ仛绯栵紝鑳芥彁楂橀潰鍥㈢殑鎸佹按鎬у拰鏈烘寮哄害锛屼娇闈㈠洟鍏锋湁鏇村ソ鐨勬寔姘旇兘鍔涘拰鎿嶄綔鑰愬姏锛岃繘鑰屾彁楂橀澶村拰闈㈠寘鐨勫搧璐ㄣ€?/p>

銆愰叾娲绘祴瀹氥€戜互鏈ㄨ仛绯栦负搴曠墿锛岀敤DNS娉曟祴瀹氳繕鍘熺硸鐨勭敓鎴愰噺銆傚彇绋€閲婇叾娑?.1ml锛屽姞1%鏈ㄨ仛绯栨恫0.9ml锛岀疆50鈩冩按娴翠繚娓?0min鍚庯紝鍔燚NS 2ml浜庢哺姘存荡涓樉鑹?min锛岀敤鑷潵姘村喎鍗村悗锛屽姞钂搁姘村畾瀹?5ml锛屾祴O.D鍊笺€?/p>

銆愰叾娲诲畾涔夈€戞瘡姣崌閰舵恫姣忓垎閽熸按瑙ｆ湪鑱氱硸鐢熸垚1μmol鏈ㄧ硸鎵€鐩稿綋鐨勯叾閲忋€?/p>

10. 铔嬬櫧閰?/p>

銆愭祴瀹氬師鐞嗐€戜腑鎬ц泲鐧介叾鍦ㄤ竴瀹氱殑娓╁害鍜宲H鏉′欢涓?姘磋В閰礌搴曠墿浜х敓鍚湁閰氬熀鐨勬皑鍩洪吀,鍦ㄧ⒈鎬ф潯浠朵笅,灏嗙鏋楄瘯鍓傝繕鍘?鐢熸垚閽艰摑涓庨挩钃?鍏堕鑹叉繁娴呬笌閰氬熀姘ㄥ熀閰稿惈閲忔垚姝ｆ瘮銆傞€氳繃鍦?60 nm娴嬪畾鍏跺惛鍏夊害,鍙緱鍒伴叾瑙ｄ骇鐢熺殑閰氬熀姘ㄥ熀閰哥殑閲?璁＄畻鍑鸿泲鐧介叾娲诲姏,浠ユ浠ｈ〃铔嬬櫧閰剁殑鎬婚叾娲诲姏銆?/p>

銆愰叾娲绘祴瀹氥€戝彇3鏀瘯绠?涓€鏀┖鐧界,涓ゆ敮鏍峰搧绠?,鍒嗗埆鍚?鏀瘯绠′腑鍑嗙‘鍔犲叆绋€閲婂ソ鐨勫緟娴嬮叾娑?.0 mL,灏?鏀瘯绠℃斁鍏?0鈩冩按娴翠腑棰勭儹5 min,鍚屾椂灏嗘祴瀹氬簳鐗?1%閰礌)缃悓涓€娓╁害姘存荡涓鐑? min銆傚垎鍒悜涓ゅ彧鏍峰搧绠′腑鍔犲叆1%閰礌婧舵恫1.0 mL,鍑嗙‘璁℃椂,鍙嶅簲10 min鍙栧嚭銆傝繀閫熷噯纭悜涓ゅ彧鏍峰搧绠″姞鍏?.4 mol/L涓夋隘涔欓吀2mL,浜庣┖鐧界涓姞鍏?.4 mol/L涓夋隘涔欓吀2 mL鍜?.0 mL 1%閰礌婧舵恫,鎽囧寑銆?鏀瘯绠′腑鍙嶅簲娑茬敤婊ょ焊杩囨护銆傚彟鍙?鏀瘯绠?鍒嗗埆鍚稿彇涓婅堪鐩稿簲婊ゅ嚭娑?.0 mL,0.4 mol/L纰抽吀閽犳憾娑?.0 mL鍜岀█绂忔灄璇曞墏1.0 mL,鎽囧寑,缃?0鈩冩按娴翠腑鏀剧疆20 min(鏄捐壊),鍙栧嚭,杩呴€熷喎鍗寸疆瀹ゆ俯銆備互绌虹櫧绠¤皟浠櫒闆剁偣,鍦ㄥ垎鍏夊厜搴﹁娉㈤暱660 nm澶勫垎鍒祴涓ゆ敮鏍峰搧绠″惛鍏夊害,鍙栧钩鍧囧€笺€傞€氳繃鏌ユ爣鍑嗘洸绾挎眰鍑虹敓鎴愰叒姘ㄩ吀鐨勫惈閲忋€?/p>

11. α-娣€绮夐叾

α-娣€绮夐叾鏄涓€涓簲鐢ㄤ簬闈㈠寘鍒朵綔鐨勫井鐢熺墿閰讹紝瀹冨彇浠ｄ簡楹﹁娊鏄敱浜庨害鑺戒腑鐨勬穩绮夐叾鍚噺涓嶇ǔ瀹氳€屼笖鍚湁铔嬬櫧姘磋В閰躲€?/p>

銆愰叾娲绘祴瀹氥€戝湪10 mL 璇曠涓緷娆″姞鍏?. 5%鍙憾鎬ф穩绮夋憾娑? mL , pH 4. 2鐨勭７閰告阿浜岄挔-鏌犳閰哥紦鍐叉恫2. 5 mL, 40 鈩冮鐑?0 min銆傜劧鍚庡姞鍏ュ鍊嶇█閲婄殑0. 5 mL 閰舵恫, 40 鈩冧繚娓? min鍚?鍔犲叆0. 1 mol/L H2 SO4 1 mL 缁堟鍙嶅簲銆傚啀鍙? mL 鍙嶅簲娑蹭笌0. 5% KI - I2 婧舵恫0. 5 mL鏄捐壊,娴嬪畾OD620銆備笌娣€绮夋搴︽爣鍑嗘洸绾垮鐓у緱鍙嶅簲娑叉穩绮夋祿搴﹀悗,璁＄畻閰舵椿鍔涖€?/p>

銆愰叾娲诲畾涔夈€戝湪pH 4. 2銆佹俯搴?0 鈩冧笅, 5 min姘磋В鍙憾鎬ф穩绮? mg鎵€闇€鐨勯叾閲忎负涓€涓叾娲诲姏鍗曚綅 銆?/p>

12. 鑴傝偑閰?/p>

鍒╃敤鑴傝偑閰朵綔鐢ㄥ悗閲婃斁鍑虹殑閾捐緝鐭殑鑴傝偑閰革紝澧炲姞鍜屾敼杩涢鍝佺殑椋庡懗鍜岄鍛筹紱鍒╃敤鑴傝偑閰跺偓鍖栫殑閱囪В鍜岄叝鍖栧弽搴旀潵鐢熶骇鍚勭棣欑簿閰紝浣滆皟鏂欏墏锛涚敤鏈変笓涓€鎬х殑寰敓鐗╄剛鑲叾鎻愰珮椋熺敤娌硅惀鍏讳环鍊煎拰澧炲姞椋熺敤娌圭殑鑺辫壊鍝佺锛屽埗澶囦唬鍙彲閰瓑楂樻。娌硅剛锛涘彟澶栵紝鑴傝偑閰惰繕鍙互鐢ㄤ簬閰掔被鐨勫幓娴婇櫎娓ｃ€佹敼鍠勯潰鍖呰川閲忋€佹敼鍠勮泲鐧界殑鍙戞场绛夋柟闈€?/p>

銆愰叾娲诲畾涔夈€戝湪40鈩冦€乸H 7锛?锛屼互姣忓垎閽熶粠姗勬娌逛腑閲婃斁lμmol鑴傝偑閰哥殑閰堕噺瀹氫箟涓轰竴涓叾娲诲姏鍗曚綅銆?/p>

13. 婕嗛叾

銆愰叾娲绘祴瀹氥€?.0mL鍙嶅簲娑蹭腑鍚?0 mmol/LABTS 0.7mL锛岄叾娑?.lmL鍜?5 mmol/L鐞ョ弨閰哥紦鍐叉恫(pH4.5)锛屽湪25鈩冧笅鍙嶅簲2min鍚庣珛鍗崇敤鍐版荡缁堟鍙嶅簲锛屾祴瀹?36nm澶勫惛鍏夊害鐨勫閲忋€?/p>

14. 鏈ㄨ川绱犺繃姘у寲鐗╅叾

銆愰叾娲绘祴瀹氥€?0鈩冩潯浠朵笅鍙嶅簲浣撶郴涓惈閰舵恫锛屾祿搴︿负0.2mol/L锛屼笖pH鍊间负2.4鐨勯厭鐭抽吀缂撳啿娑插強娴撳害涓?.008mol/L鐨勯粠鑺﹂唶鍚刲mL锛屽姞浜烘祿搴︿负0.016mol/L鐨凥2O2 lmL 鍚姩鍙嶅簲锛屾祴瀹氬弽搴旀渶鍒?min鍐呭湪310nm澶勭殑鍚稿厜搴︼紝浠ョ伃杩囬叾娲荤殑閰舵恫浣滃鐓с€?/p>

15. 閿拌繃姘у寲鐗╅叾

銆愰叾娲绘祴瀹氥€戝娓╀笅鍙嶅簲浣撶郴涓惈鏈?0mmol/L pH4.5鐨勪钩閰搁挔缂撳啿娑?.4mL锛?.6mmol/L鐨凪nSO4婧舵恫0.lrnL锛岄叾娑?.4rnL锛屽姞浜?.6 mmol/L鐨凥2O2 0.lmL鍚姩鍙嶅簲銆傛祴瀹氬弽搴旀渶鍒?min鍐呭湪240nm澶勭殑鍚稿厜搴︺€?/p>

銆愰叾娲诲畾涔夈€戞瘡鍒嗛挓浣?μmol鐨凪n2+杞寲鎴怣n3+鎵€闇€瑕佺殑閰堕噺涓轰竴涓叾娲诲崟浣嶃€?/p>

锛堜笁锛変华鍣ㄩ厤缃?/p>

Thermo鍏徃Genesys绯诲垪銆丠elios绯诲垪鍜孍volution绯诲垪绱-鍙鍒嗗厜鍏夊害璁″潎鍙繘琛岄鍝侀叾鍒嗘瀽銆侲nzLab杞欢鏄疶hermo鍏徃鐮斿埗鐨勪笓涓氶鍝侀叾鍒嗘瀽杞欢锛屽叿鏈夋櫤鑳芥彁绀哄姛鑳斤紝鍙互涓€姝ユ寮曞鎿嶄綔鍛樺畬鎴愰叾娲绘祴瀹氥€傚嵆浣夸笉鏄叾鍒嗘瀽涓撳锛屼篃鑳借幏寰楀噯纭€佸彲闈犵殑鏁版嵁銆傝€屼笖璇ヨ蒋浠舵敮鎸佹墜鍔ㄨ緭鍏ユā寮忥紝鍗充娇瀹為獙瀹や娇鐢ㄧ殑鏄叾瀹冨搧鐗岀殑鍒嗗厜鍏夊害璁★紝涔熻兘浜彈EnzLab杞欢甯︽潵鐨勬櫤鑳藉拰渚挎嵎銆?/p>

鍙厤锛?/p>

l    鏍峰搧姹?/p>

l    鍗曟睜娑蹭綋鏍峰搧鏋讹紙鏃?鍚俯鎺э級

l    澶氭睜鏃嬭浆鏀灦锛堟棤/鍚俯鎺э級

l    鍏夌氦闄勪欢