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2 .% TFA/acetonitrile TFA ph acetonitrile -propanol TFA Triart

3 YMC-Triart C8 / C8 YMC-Triart C8 / C8 C8 C8 ( ).9 µm 3 µm µm nm C8 % C8 7% ph - 7 for ph -7 for ph 7- YMC-Triart % of initial retention time % % 8% 6% % % acetonitrile/water/tfa(/9/, ph ), 7 % 6 8 Time (hr) YMC-Triart C8 ph (% TFA), 7 YMC-Triart C8 Silica based C8 Hybrid-silica based C8 Column : µm, X. mmi.d. Eluent : acetonitrile/water (6/) Flow rate :. ml/ Temperature : 37ºC Sample : butyl benzoate 3

4 Triart C8 Leu-Enkephalin MW 6 Oxytocin MW,7 γ-endorphin MW,89 β-endorphin MW 3,46 Amyloid β-protein(-) MW 4,33 Insulin MW,733 Cytochtome C MW, Lysozyme MW 4,3 β-lactoglobulin A MW 8, α-chymotrypsinogen A MW,7 4

5 Triart C8 HPLC Column Eluent Flow rate Detection Temperature : YMC-Triart C8 (.9 µm, nm), X. mmi.d. : A). % TFA or HCOOH in water B). % TFA or HCOOH in acetonitrile -9%B (- ) :.4 ml/ : nm :

6 Triart C8,7,7 8, 8, 4,3 4,3,, MW,733 4,33 MW,733 4,33 3,46 3,46,89,7 6.% TFA.% HCOOH,89, (% )() () α-chymotrypsinogen A (MW,7).% TFA.% HCOOH TFA % ().4 (). HCOOH D 97.D TFA 6

7 Triart C8 HPLC Column : YMC-Triart C8 (.9 µm, nm), X. mmi.d. Eluent : A) water/tfa (/.) B) acetonitrile/tfa (/.) -9%B (- ) Flow rate :.4 ml/ Detection : nm Temperature : or 7 7

8 Triart C8 MW,7 8, 4,3,,733 4,33 3,46,89, (% )() MW,7 8, 4,3,,733 4,33 3,46,89,7 6 () α-chymotrypsinogen A (MW,7) 7 % () () D 893.D

9 MW 6-8, MW 4,3-,7 9

10 . MW 6-8, Leu-Enkephalin MW 6 Oxytocin MW,7 β-endorphin MW 3,46 Insulin MW,733 β-lactoglobulin A MW 8, HPLC Column Eluent Flow rate Detection Temperature : YMC-Triart C8 (.9 µm, nm), X. mmi.d. : A) B) :.4 ml/ : nm :

11 . at.% TFA -8 %B (- ) slope : 4%B/ -8%B (- ) slope : 7%B/ -6%B (- ) slope :.%B/ 6 W/ () =.7 W/ () =.43 4, D 363.D_F33P 36.D_F38X 3 * % TFA 6 W/ () =.8 W/ () = D 3 6.% HCOOH * (peak 3) TFA peak W/ () =.8 W/ () = Oxytocin. Leu-Enkephalin 3. -Endorphin 64.D 3 * 4. Insulin. -Lactoglobulin A

12 . 3 Rs (3,4) =.78 W/ () =. W/ () =. 333.D_F33G Rs (3,4) = - W/ () =.9 W/ () = , 4 Column : YMC-Triart C8 (.9 µm, nm) X. mmi.d. Eluent : A) water/tfa (/.) B) acetonitrile/tfa (/.) -8%B (- ) Flow rate :.4 ml/ Detection: nm. Oxytocin. Leu-Enkephalin 3. -Endorphin 4. Insulin. -Lactoglobulin A 336.D_F33H Rs (3,4) =.97 W/ () =.7 W/ () = D_F33J peak Rs (3,4) = 3.47 W/ () =.6 W/ () = D_F33K 7 Rs (3,4) = 4.87 W/ () =. W/ () = D_F33L... 3

13 . MW 4,3-,7 Lysozyme MW 4,3 β-lactoglobulin A MW 8, α-chymotrypsinogen MW,7 HPLC Column : YMC-Triart C8 (.9 µm, nm), X. mmi.d. Eluent : A) water/tfa (/.) B) acetonitrile/-propanol/tfa (//.) 3-6%B (- ) Flow rate :.4 ml/ Detection : nm 3

14 . 7 Peak capacity = 67 Peak capacity = 7 6 Rs (-3) Rs (-3) F6B F6A * * Analytes MW (% ) () 7. Lysozyme. α-chymotrypsinogen 3. β-lactoglobulin A 4, , , Lysozyme. α-chymotrypsinogen 3. β-lactoglobulin A 7 peak 3 4

15 3. α-defensin- : A CYCRIPACIAGERRYGTCIYQGRLWAFCC MW 3,44 α-defensin- : CYCRIPACIAGERRYGTCIYQGRLWAFCC MW 3,37 α-defensin-3 : D CYCRIPACIAGERRYGTCIYQGRLWAFCC MW 3,486 Total 3 N or HPLC Column : YMC-Triart C8 (.9 µm, nm), X. mmi.d. Eluent : A) water/tfa (/.) B) acetonitrile/tfa (/.) -4%B (- ) Flow rate Detection :.4 ml/ : nm

16 3. 7. α-defensin-. α-defensin- 3. α-defensin-3 3,, 3 3, 3 68.D_F6A 6.D_F6E 7 peak,3 peak 6

17 3. at 7 6.% TFA, 3 Column Eluent Flow rate : YMC-Triart C8 (.9 µm, nm), X. mmi.d. : A) B) (. % HCOOH B.8%) :.4 ml/. α-defensin-. α-defensin- 3. α-defensin-3 Detection : nm Temperature : D_F46B 4 6.% TFA TFA 6 3 Rs (, ) =.6 Rs (, 3) =. 464.D_F46D %B (- ).% HCOOH slope :.%B/ -3%B (- ) slope :.%B/ 6 3 Rs (, ) =.44 Rs (, 3) =. 6 3 Rs (, ) =. Rs (, 3) = D F46C D_F46F 4 6 7

18 3. -propanol acetonitrile -3%B (- ) slope :.%B/ 6 Rs (, ) =. Rs (, 3) = D_F46F 3 4 -%B (- ) -propanol/acetonitrile (/) slope :.%B/. α-defensin-. α-defensin- 3. α-defensin-3 Column : YMC-Triart C8 (.9 µm, nm) 6 Rs (, ) =.86 Rs (, 3) =.47 3 Eluent Flow rate Detection X. mmi.d. : A).% formic acid in water B).8% formic acid in organic solvent :.4 ml/ : nm 3.D_FD Temperature : / peak 8

19 HPLC YMC-Triart C8 / C8 nm 9

Ł\”ƒ-2005

$Ł\”ƒ-2005$