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高性能液相色谱介绍——学员手册-Agilent_504页.pdf

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18、y?j(xy?z0?1xy?x?(x?x?(xy4?hijklm(?0hijklm(?0hijklm(?0hijklm(?0 15?!?$/0?!?$/0?!?$/0?!?$/0 31HPLC?&?&?&?&?0?0?0?0?!?f?f?f?f?#$?%&(?)*+,?-./01234*hijklm Zx?hx?(ddx?79N(?T?(xyi?hijklm7?0?O(?=qPHt?O(?0P?hijklm(?0hijklm(?0hijklm(?0hijklm(?0 16?(HPLC)?(HPLC)9?t?x9?t?x9?t?x9?t?x0?0?0?0?18 12?3?12?3?12?3?12

19、?3?04567045670456704567 2?HPLC?.?HPLC?.?(HPLC)?19 89:;89:;89:;89:;3?OPQV?Q?TURSQ8-#$%!%8-?W?OPQ?TU?TU?sQhijklm?$?q?Z?“fi?R)?(jklm(?(?0S?T%S?T(!d?&?dUV&/&Q?xy%&Q?A0?&S?T d?W“0ht?0X67N(T%?hixy2x?u?0=0?W“0?(HPLC)?20?9?9?9?9 4?10 m?!?#$2%0.5 m?&?()*?+?&,-./?k5YE(?67(?j?9?k?Z5A0+?d 10 m?k5YE(?“?6?5YE(?-?

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22、5(#j?#?F#j?(0?_?(HPLC)h?j?h?j?h?j?h?j?25?FC?GHIJK?FC?GHIJK?FC?GHIJK?FC?GHIJK 9?q?q?q?q?56789:;7?7A7?A0(q?|?/?%&?s?0(?fl?X?%?d?9Xuevh?Z?A0?/?%&?H?Yd0%&?sV“,?-j?j?a?%?(HPLC)h?j?h?j?h?j?h?j?26 10?A?B?AB?st?“,?k%&?lm“?qt“,(ktkr 180Cq“,?:(o?6GZo?Jst“,?k?lm“?st“,?kM?(l?1090?a?“,?(HPLC)h?j?h?j?h?j?h?j?27 1

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27、A0?6?A0?st?(?_?st0+?(?T?5?T0(?eF?9?5?A0?(?(?D?-A0?(?;I?_(?9?qJ?T5(l?_fl?t?O?x?%?T?*lm“(?(?T5(?lm“9?5?T?p#?Vs?x?-?s?x?(?/?Z?9?-?TX?7?“?“A0?4?(HPLC)?37 21OPQV?Q?TURSQ8-1234?1234?1234?1234?7hij?7hij?n?)?-?(t?#?9?5?T?fllm“0?LM?(?W(?_(t?TZd?(s?x?#fllm“(?_ev9?t?p#?T h?wxx?(f9rt?5?-dwx?T?_(?Xu?8?(HPLC)?38

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31、enzo(e)pyrenePeryleneBenzo(k)fluorantheneBenzo(a)pyreneBenzo(ghi)peryleneIndeno(123-cd)pyrene?/fi?0?HPLCs?Fn UV-VIS fi?#?Xu?(fi?s?Fn UV-VIS fi?(?-fl?fi?9?P(?5A0 UV fi?7fi?%?6?0fl?fi?(HPLC)HPLC fi?44 28FGHIAJ?FGHIAJ?FGHIAJ?FGHIAJ?BCKLMBCKLMBCKLMBCKLMFlecainide in SerumTherapeutic concentration:1.8mg/

32、l,20ul injectedUV and fluorescence signalFL signalUV signal?LM?V?(?Q5fi?&Q?cfi?(?rs/?&Q?V?(?Q5fiF(7d?flx?&?&Q?,?(fi?_?UV fi?(?flx?&QM5?kO%?fi?_?DA0?4F?(fi?fl?fi?JA0lmxy4?7?flx?&Qx?X flx?&?XuA0fl?Y?(?E?6?0?-?F?F?(HPLC)fifififi?7?7?7?7?49 35?-?678(LOD)9:;?ABCDDE?)67FGH?+8(LOQ)9I?JK?)IL?MNO?./?=?MQLMDL

33、?e.g.,RSD 20e.g.,S/N 3?5?)fi?(?t?u(7?LODLOQ?U-?fib LOD?cd?y(2*3 LOD(6?st?*fi?T*?&?K?7?Y?X67?(?&(u?(?LOD 6?Z*100 pgfl?-?fi?7fi*9Z(?%*HPLC?(Qmfi?(LODw?A0(?E?(?6?fi*?fi?s?0?500 ng(P?LOQ?b?cd?y(10*20UVfi?6?0?YZh?500 pg(?fi?,?6?0?U-?U?U?6:6?(fi?=?M?(Y?-Y?(?D?U7:?(fi?=?(?Q?U?09?tU?,?=?T*s?t?Us?10 x?1(,?(U

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35、?st?NC&(?9?(?U?x?&(?6?(?(HPLC)UV-VIS fi?51 37UVW?XYUVW?XYUVW?XYUVW?XY?!#$%$&$-NH-C=C-C=O-COOR-CHO-COOH-NO195190195205210200-210310202,255220,275max(nm)22-()*UV-Visfi?(fi?210 nm*850 nmA0?|?-?(?Xu(7?kJ?180 nm(?/?(?fi?z0(H?Nx?67?M5?-?H?Nx?x?5(?-?k?H?=%?-?(?&s?fi?0(M5rt?7?(?D?UH9?P(?5?(HPLC)UV-VIS fi?52

36、 38UV-VIS?-Z?Z?Z?Z?OPQRSTUVQRSTWXY,Z_QRabUV 55556767676789:89:89:89:;?=?=?=?=?ABCABCABCABCDEFG6HDEFG6HDEFG6HDEFG6HIJIJIJIJCCCC2CCCC 16W?fi?9?(6W?fi?(?(?l?P?-P?*|l?(?|l?(?U(?5F(?x?5(?QM5?(M?fi?Ni?(?9?7dT?(?.-i?(?.T*(?fi?6?6?stlmx?5(ht?d)fi?st?&(?mXuux?k(?ufi?(&Qef9?5?t?U?(HPLC)UV-VIS fi?53 39?UV-VIS?

37、PQRST UV-VIS 67FUVWX7+Y=$Z./190-900 nm$ZM 1 nm?_a$?bc$Zde&()*+,+-./#$?!0%123?+45+67UV8Vis8 7dTHfi?7?|?g?fi?/fi?7d?x?8?m?m?-:(=?P?1100 7dTH(G?(?-?7:190*850 nm(?st?!W(?!?xZ|?(?ht?7dTst?(?f(?Dh 12#$?st?(?m:=?8?9?t?5H?1024 t7dTht7dTfi?st?(?f(?t?(U?fi?.(WT*)M5?m?6?d?1*16 nm?%&T*Yh(?6?8?m?T*(7dTHfi?(-8?H?

38、m(7AT6?Z?m?6?0?5r/?&(?stlmU?(?m6?(HPLC)UV-VIS fi?55 39?_a?bc.?_a?bc.?_a?bc.?_a?bc.nrstuvwDojnojxykz|ojD?Dn?x?*9?(?55?8?)m?(?6?+,?lm?9lm?(?5?m?lm(ht?mj-?T*(?m?9?m?-?m?(rs?J?lm(m?5?&?lm9?(?mk?67?(G?Q67?:?HM5=%?-95?.?(?x0?5(H(?J?/67?(HPLC)UV-VIS fi?56 40de.fde.fde.fde.f?GC?6?5.*stBM?(?BM?)stM?-?(?/?/?_

39、6?+,x?*?rs4(Y?fi?g|t#9?“Zs?0s?0(4?fl?(?GZ)?(?d=?fi(67eF)fi*?(fiy(Y?(S/N?1x?10(?x?E?fl?(?fl?ATx?6?M5s?U(?M5?*?t?2?l?(x?3ls?fl?x?7?M5(7?M5?(?h?M5?9?W?(?Ifl?.?qF?+?T*)?m?(HPLC)fl?fi?fl?fi?fl?fi?fl?fi?58 42hihihihi-ijijijijUV?DAD-Tfi?d)%fib-?x?*LM)vux?&(fl?Q6?(?-?fi*Y?(fib-?s?D?T*(fl?m67?4(?w?A0(?-(9?(4

40、?-fl?mk?(UV?m5?(?mKA?TY?T*?m?ht?&?t?LMu9A0fl?x?g9 HPLC(u?q?V(s?ht?&jXux5?s?ht?&T*s?-?m$?t?56?J?x?7?H?(?I80?/4?(HPLC)fl?fi?fl?fi?fl?fi?fl?fi?59 43g?g?g?g?fl?4Q?G?Q?G?QP?s9:;?9?HU?(Y?.:?(?$?9fi?(?x?C?(A0A6?fi*1t?:;?zM6?fi*?(?st!?d|l?:(?l?x?Xu(?*?:Y?d)?fi?s?F(?#*9?“Z?0(4?Hst!?d|l?qt|l?j96?U?Y?(?“T?fi?Z

41、.fl?(Y?$?;?Bh?;?(.(rsst?7dT(?fi?(.?fi?=?(?$?Y?h?280 nmstCx?m?5?;1xZ?t?(?*?|l?(?+?(Cx?m-s?(E?(20 nm)UV)?(-?stE?6D(?m-?(?bX(SE?(HPLC)fl?fi?fl?fi?fl?fi?fl?fi?60?-|l?7?9=?-?m?9=?*st.?(t?tt?Xu(?2st9?(fi?$?b?(?j?9st?-?T*?/?)YZ(fib?m?0?T*?=?g?m=!;?(?#?g|l?(?(0?(Ffi?z?A(?2 PC h?s?0(Ffi?zs?J?G*fist-?(t?:?st;

42、?;?v&?2#$?(?H(?14#$?|l?I?m?(?9?(?UVfi?(HPLC)fl?fi?fl?fi?fl?fi?fl?fi?61 46?90?!?#!?$%&?()*+,-./0/0/0/0?12&?345 O2 6.7?8?#?9:?;FLD!?=?6ABABABABCD?D?D?D?EF?GH?!?!?!?!?fl?fi(?0U?tx?&6?“0?fl?fi?1?x?g5(0?7?“?(?st(“?qb?7777?-NH2 o-Phthaladehyde?7?-NHR 9-Fluorenylmethychloroformate 9-?-COOH p-Bromophenylacy

43、l bromidep-?J?,2-Naphtacyl bromide2-KJ?-OH Phenylisocyanate(?sL)-CHO,=C=O 2,4-Dinitrophenylhydrazine(2,4-7M?N)-CO-COOH 2,4-Dinitrophenylhydrazine(2,4-7M?N)?(HPLC)?fi?fi?fi?fi 62 6YZ6YZ6YZ6YZ 47?,-?./?0?123456?37&89:;NH(x)SH-?AB23456 C3DEFA B+e-(23)?fi?9K?5(?K?5x?5x?T*?K?9?d?d(?P?zG?&?K?#?(?BO?d-?Q(?

44、j(PrA?7.4?(K?P-/7(C?6?EK?-d?(?(?H?(?H?PD?x?&(?(?(HPLC)?fi?fi?fi?fi 63 46klm?klm?klm?klm?Zcde;fg?hei:jklei=min#cde;feihg?:jklei=oi?E2E1/2E1#”V?b?9fi?5A0td?d?9?d?d-?d?Zd b?d?k-?d(Pr?d?jE(W?dQ?R*d?5?d(?Iev?(Pr fi?(,?$?(g?-?&(=?:?0Jl?(?z6?fi?Sfl?x?&:(kJZ(?XT?fi?7fi?t_?/(&Q9x?V?(?/Tb?Zds?%?=dh)fi(?d)w?Y%

45、(?dP?hst?&jXuUfi?,?-b?P?(C?b(?-?(CV)?9PZ?E1?=db?(7?(?mK?7P“?*E1/2?vsh9d?P(x?Y?,?Xust?Th?E2(P?sP?(?ddb?(limiting current)?(s?h?(“?VDU,fi?(HPLC)?fi?fi?fi?fi 64 47klm?klm?klm?klm?*#?2?*#?2?*#?2?*#?2?*#?4?*#?4?*#?4?*#?44444“?*#?o?*#?o?*#?o?*#?o?*#?f?*#?f?*#?f?*#?ffifoflffifoflffifoflffifoflf?SW5?O?X)?(?

46、Y?6?d/?u?s?(thin-layer design)Z?(wall-jet design)-?+?(porous flow-through design)?+?-?q/?4(rs?/?eF?fi?5d?P(?:Ed 100%opqr!#nopqr!#nopqr!#nopqr!#stn?stn?stn?stn?;uv*wx;uv*wx;uv*wx;uv*wx?*Y?z”?EdC?0D0?Nx?DY?(stRAT?tfi?6?0?Nx?fghi5jkl?(x?-”5(Nfi 9fi?5Sfl?5?A0?d(x?A0?o#?x?(?6?h?XT6?4?&5hs/?&(m?Fw?Y%Pn?t%

47、?6?A0bn?“?ZHPLC?9st“?(PUq$P5xP-“?s?H(?9%?5?9?eF9qs?x?evk(?o?(HPLC)?fi?fi?fi?fi 66 49LC-MSt?u?GC/MS?/Z8 V?(?Zd LC x?5(stCLp?&?6?0A07dTH?z(UV M5fi?F?J?mrs?/4?.sfl?Bfi?z6?UV fi?h(.s?fi?(?&?t?z?7?qclG(A0Qmfi?6?r?t?(x?&/s?GC/MS?x?O?st?t(?z?9 LC/MS 9%uv?ww?(st?_?GC(x?0?t(?(?&?D?&?(_?h(d?0?_-#0?Ey?&?S?T%S?

48、T?|?y?-h?x?Jy?y?S?T?xy?1?xy?1?y?i?(?06?5(?|0-?x?xy?2?y?9y?*fi?d?h?5&Q(?03d?&?d?(y?s?v?5&Q?x?3d?*9?d?y?(7?x6(?0?(HPLC)?fi?fi?fi?fi 69?d?Q?h?stQ(y?h7“fi?(HED)?y?g?=?s?y?g?Qm?k?(?9?(st?S?5?(HPLC)?fi?fi?fi?fi 70 523504004505005506006507002000400060008000m/z-Abundance439467502684.653504004505005506006507

49、002000400060008000m/z-Abundance411439467495684.65Fragmentor=175 voltsFragmentor=75 voltsloss of C14 fatty acidloss of C12 fatty acidpqrsn?NMOPn?NMOPn?NMOPn?NMOP JA0?|?y?zx?x?P?qQm5?u?Q?(x?y?5?e?(phenylbutazone)(?cQ?308 k?Qm?Y?(?x?y?M+H+Qd 309?(:(?x?y?Sfl(?%P?uw?k?-?-y?q?(y?M+Na+=M+23+M+K+=M+39+M+NH4

50、+=M+18+M+X+%?Xd?#j?y?2M+H+h?q?70 M+H+S+?&?(HPLC)?fi?fi?fi?fi 71 My?q?M H-$M+X-%?Xd?#j?y?M H+S-?&?(e8?k-?j(U,Y?(?st4?X?f?t=?_?yt?(hx?&(:?(?(?y?st?Q9 q?5(?1-O1 mfj6?Q?bx?(QLM9?d?Qm?(Q?U?API?s/k-(y?z?:st?x?y?k?yv?y?5|0 x?k?DD?v(?x?-?x?j?60(?x?(?x?(?=!89?Cy?H?6?h?.CID 6?y?S?T-xy?1(y?7?F!W?Wst9!W(5?d?v?(

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