橡マニュアルv4c.PDF

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1 LC/MS MS LC/MS LC LC (APCI) (ESI) FAB 2 LC (APCI) LC LC/MS LC MS LC 3 ph APCI/MS LC (1) APCI ESI 100% 100% ESI LC 1.00cp

2 a LC b ( /mpas) n n p n a b 20 C (2)pH 3 LC ph ph LC/APCI/MS ph 35

3 ph LC/APCI-MS ph ph LC LC/APCI/MS ph ph [M+NH 4 ] ph ph APCI-MS ph LC-APCI-MS H 3 PO 4 /KH 2 PO 4 /K 2 HPO 4 /KOH 2 11 / 3 6 / /NH 4 OH 5 9 / 8 10 /NH 4 OH / /NaOH 9 11 H 3 BO 3 /Na 3 BO 3 /NaOH APCI ph 100mM 36

4 ph ph APCI/MS ph (3) LC LC/APCI/MS ph LC/APCI-MS APCI-MS LC/APCI-MS LC-APCI-MS LC-APCI-MS 1- CH 3 CH 2 CH 2 SO Na 1- CH 3 (CH 2 ) 3 SO 3 N a 1- CH 3 (CH 2 ) 4 SO 3 N a 1- CH 3 (CH 2 ) 5 SO 3 N a 1- CH 3 (CH 2 ) 7 SO 3 N a [CH 3 (CH 2 ) 5 ] 3 N [CH 3 (CH 2 ) 6 ] 3 N [CH 3 (CH2) 7 ] 3 N (CH 3 CH 2 ) 4 NOH [CH 3 (CH 2 ) 3 ] 4 NO H APCI/MS 37

5 (1) APCI APCI AH + + B BH + + A ( G<0 A,B A B AH + B BH + APCI AH + (CH 3 OH)H + ((CH 3 OH) 2 H kcal/mol (1648kcal/mol) (202.3kcal/mol) A - + BH B - + AH BH A - A - BH A - BH 38

6 Linuron Diuron APCI p-tert- ( ) (C 60 ) APCI Linuron (200µg/ml) Scan ; Cap.Vol ; 3500V Corona current ; 4.0µA Drying gas temp. ; 350 Vaporizer temp.; 473 Drying gas flow ; 6.0 l/min Neb. Pressure ; 60psig positive mode Mobile phase ; MeOH:H 2 O=1:1 Flow rate ; 0.5ml/min Fragmentor voltage ; 80V Injection volume ; 5µl Instrument ; HP1100MSD Cl Cl N C N H O Linuron OCH 3 CH Linuron APCI-MS Cl Cl N C N H O CH 3 CH 3 Diuron Diuron ( 0µg/ml) Scan ; Cap.Vol ; 3500V Corona current ; 4.0µA Drying gas temp. ; 350 Drying gas flow ; 6.0 l/min Vaporizer temp.; 473 Neb. Pressure ; 60psig positive mode Mobile phase ; MeOH:H 2 O=1:1 Flow rate ; 0.5ml/min Fragmentor voltage ; 50V Injection volume ; 5µl Instrument ; HP1100MSD Diuron APCI-MS 39

7 CH OH 3 HO 17β-Estradiol 17β-estradiol (2.0µg/ml) Scan ; Cap.Vol ; 3500V Corona current ; 4.0µA Drying gas temp. ; 350 Vaporizer temp.; 473 Drying gas flow ; 6.0 l/min Neb. Pressure ; 60psig negative mode Mobile phase ; MeOH:H 2 O=1:1 Flow rate ; 0.5 ml/min Fragmentor voltage ;150V Injection volume ; 5µl Instrument ; HP1100MSD b-estradiol APCI-MS t-bu COOH 4-t-Butyl-benzoic acid p-t-butylbenzoic acid (5.0µg/ml) Scan ; Cap.Vol ; 3500V Corona current ; 4.0µA Drying gas temp. ; 350 Vaporizer temp.; 473 Drying gas flow ; 6.0 l/min Neb. Pressure ; 60psig negative mode Mobile phase ; MeOH:H 2 O=1:1 Flow rate ; 0.5 ml/min Fragmentor voltage ;70V Injection volume ; 5µl Instrument ; HP1100MSD p-t-butylbenzoic acid APCI-MS 40

8 (2) APCI ESI ph ph LC-MS 0.05% ph 4-10 APCI/MS ph ph APCI/MS Hitachi M-1200 AP/ES 1.0ml/min amu/sec 70V 1,800V 3,000V 90V ml APCI ESI [M+NH 4 ] + ESI ESI APCI H 2 O

9 ph Dibenzo-18-crown-6-ether (5.0µg/ml) Scan ; Cap.Vol ; 3500V Corona current ; 4.0µA Drying gas temp. ; 350 Vaporizer temp.; 473 Drying gas flow ; 6.0 l/min Neb. Pressure ; 60psig positive mode Mobile phase ; MeOH:H 2 O=1:1 Flow rate ; 0.5 ml/min Fragmentor voltage ;50V Injection volume ; 5µl Instrument ; HP1100MSD O O O H + O O O O O O H 3 O + O O O Dibenzo-18-crown-6-ether APCI-MS 42

10 (3) LC MS APCI ESI (1) APCI MS MS LC ml/min ml/min LC/APCI/MS 1.0ml/min (2) CID(collision induced dissociation) CID SIM CID ( ) 100V 43

11 NO 2 4-Nitro-benzoic acid COOH 4-Nitrobenzoic acid (5.0µg/ml) Scan ; Cap.Vol ; 3500V Corona current ; 4.0µA Drying gas temp. ; 350 Vaporizer temp.; 473 Drying gas flow ; 6.0 l/min Neb. Pressure ; 60psig negative mode Mobile phase ; MeOH:H 2 O=1:1 Flow rate ; 0.5 ml/min Injection volume ; 5µl Instrument ; HP1100MSD Nitrobenzoic acid APCI-MS (Fragmentor voltage ; 50V) Nitrobenzoic acid APCI-MS (Fragmentor voltage ; 90V) α-cyclodextrin (1000µg/ml) Scan ; Cap.Vol ; 3500V Corona current ; 4.0µA Drying gas temp. ; 350 Vaporizer temp.; 473 Drying gas flow ; 6.0 l/min Neb. Pressure ; 60psig negative mode Mobile phase ; MeOH:H 2 O=1:1 Flow rate ; 0.5 ml/min Fragmentor voltage ;150V Injection volume ; 5µl Instrument ; HP1100MSD a-cyclodextrin APCI-MS 44

12 (3) APCI V V 3000V -2500V (4) APCI APCI / ml/min 45

13 (5)LC/APCI-MS LC/APCI-MS LC APCI-MS LC MS a, : =1:1 : =1:1 b, g/ml SCAN c, d, LC a, b, ph a, LC-APCI-MS SCAN SIM) APCI/MS ,4- A, APCI/MS M-H ESI 2,3,3,4,5-pentachloro-4- biphenylol(10µg/ml) Scan ; Cap.Vol ; 3500V Corona current ; 4.0µA Drying gas temp. ; 350 Vaporizer temp.; 473 Drying gas flow ; 6.0 l/min Neb. Pressure ; 60psig negative mode Mobile phase ; MeOH:H 2 O=1:1 Flow rate ; 0.5 ml/min Fragmentor voltage ;130V Injection volume ; 5µl Instrument ; HP1100MSD Cl Cl Cl 2,3,3,4,5-pentachloro-4-biphenylol APCI-MS 46 Cl OH Cl 2,3,3',4',5-Pentachlorobipheniyl-4-ol

14 2,2,3,4,5,5,6-heptachloro-4- biphenylol (10µg/ml) Scan ; Cap.Vol ; 3500V Corona current ; 4.0µA Drying gas temp. ; 350 Vaporizer temp.; 473 Drying gas flow ; 6.0 l/min Neb. Pressure ; 60psig negative mode Mobile phase ; MeOH:H 2 O=1:1 Flow rate ; 0.5 ml/min Fragmentor voltage ;130V Injection volume ; 5µl Instrument ; HP1100MSD Cl Cl Cl Cl OH Cl Cl Cl 2,2',3,4',5,5',6-Heptachlorobipheniyl-4-ol 2,2,3,4,5,5,6-heptachloro-4-biphenylol APCI-MS 2,4-Dichlorophenol (5.0µg/ml) Scan ; Cap.Vol ; 3500V Corona current ; 4.0µA Drying gas temp. ; 350 Vaporizer temp.; 473 Drying gas flow ; 6.0 l/min Neb. Pressure ; 60psig negative mode Mobile phase ; MeOH:H 2 O=1:1 Flow rate ; 0.5 ml/min Fragmentor voltage ;90V Injection volume ; 5µl Instrument ; HP1100MSD OH Cl Cl 2,4-DIchlorophenol ,4-Dichlorophenol APCI-MS 47

15 Bisphenol A (5.0µg/ml) Scan ; Cap.Vol ; 3500V Corona current ; 4.0µA Drying gas temp. ; Vaporizer temp.; 473 Drying gas flow ; 6.0 Neb. Pressure ; negative mode Mobile phase ; MeO :H 2 O=1:1 Flow rate ; 0.5 Fragmentor Injection volume ;µl Instrument ; CH 3 HO C OH CH 3 Bisphenol Bisphenol A APCI-MS Diethylstilbestrol (10µg/ml) Scan ; Cap.Vol ; 3500V Corona current ; 4.0µA Drying gas temp. ; 350 Vaporizer temp.; 473 Drying gas flow ; 6.0 l/min Neb. Pressure ; 60psig negative mode Mobile phase ; MeOH:H 2 O=1:1 Flow rate ; 0.5 ml/min Fragmentor voltage ;130V Injection volume ; 5µl Instrument ; HP1100MSD HO Diethylstilbestrol OH Diethylstilbestrol APCI-MS 48

16 (ESI) ESI ESI LC LC/MS/ESI ph MS ESI ESI ESI 1 1 cp Nm (DMSO) % 49

17 ESI ph LC ph LC LC/MS NH 3 NH CH 3 OH NH CH 3 OH H 2 O NH (CH 3 OH) 2 NH H 2 O NH (H 2 O ) 2 NH (CH 3 COOH NH 3 NH 4-18) + 78 CH 3 COOH NH 4 + Negative (CH 3 COOH) 2-H 119 CH 3 COOH H 2 O-H CH 3 CN CH 3 CN NH CH 3 CN H 2 O NH (CH 3 CN) 2 NH 4 ESI ph H

18 ph ESI (10mM ) (IPC) IPC IPC IPC ph ESI IPC LC IPC LC/MS LC/MS IPC ESI ESI ESI (1) 51

19 ESI ( ) ( ) N M+H M-H N R HA R 1 R 1 N R A R 3 R 3 Base Acid Sample (M+H) O O R C OH B R C O H B Acid Base Sample (M-H) (2) (+) R-NH 2 R-NH-R ESI R 2 -PH R 2 -PH ( ) R-OH R-SH R-OR R-C=O ( RCOOH ) RSO H RPO H ( ) ESI ±3 5kV mm 52

20 0.1mm LC/MS LC LC/MS ESI LC LC/MS LC/MS (3) LC MS MS MS LC PB APCI TSP ESI (ml/min) Frit-FAB or CF-FAB LC/MS APCI Atomospheric pressure chemical ionization ESI Electrospray ionization PB Particle beam TSP Themospray Frit-FAB Frit-Fast atom bombardment 53 CF-FAB Continuous flow-fast atom bombardment

21 LC MS ( ) ( cm 3 /min) column Conventional Semi-micro Micro mm 4.0~ ~ ~0.5 ml/min Methanol Water ESI LC (mm) ESI mm l 200 l ESI 0.2ml/min ( 4.6mm 150mm) 1ml/min LC ESI Tee LC (4) LC ESI Taylor cone ±3 5kV MS (5) ( ) 54 ( l/min)

22 ( ) ( ) API CID(collision induced dissociation) ( ) CID (6) ESI LC LC ph HFBA TFA M [M+Na] + ESI Mixing Tee HPLC (UV) (API Needle) Pump

23 20 Signal Signal (%) Formic Acetic Propionic Butyric Valeric Acid % HP 1998 LC/MSD ESI MS % 0.05% 0.1% 0.5% Signal ratio benzoic acid o-toluic acid salicylic acid phenoxacetic acid 0% 0.05% 0.1% 0.5%

24 ESI (M+nH) n+ (M-nH) n- QMS QMS n2=n1+1 n n1< n2 n1=m1/(m2-m1) M1=n1(m1- ) mn m1< m2 Mn MW=ave(M1,M2,M3.) MW =82.5 =115.5 =173.3 m1:496 m1 496 m2:578.5 m m3:694.0 m m4:867.3 m4 m/z n1=496 ( ) M1=6(496-1) =6.012 = n2=578.5 ( ) M2=5( ) =5.008 = n3=694.0 ( ) M3=4( ) =4.004 =2772 =M1 M2 M

25 LC/ESI/MS LC/ESI/MS LC ESI/MS a ( LC ) b SCAN g/ml c ( ) LC a b ph ( LC ) a --- (LC/MS ) LC LC-ESI-MS (SCAN SIM) Paul Kebarle and Liang Tang ;AnalChem.,1993,65,972A-986A Robert D.Voyksner; Environ. Sci. Technol.,1994,28,118A-127A 15 LC/MS ( ) 58

26 CF(Frit)-FAB LC Frit-FAB API Frit API LC Frit-FAB LC Frit-FAB (1) Frit-FAB FAB Frit- FAB ) (1:1) ph 20mM 1v/v% m/z 59 CH 3 COO - m/z 151 [Gly + CH 3 COO] - m/z 91 [Gly-H] - m/z 93 [Gly+H] + m/z 185 [2Gly+H]

27 Positive Negative JMS-700 Frit-FAB 0.5 ml/min FAB Xe kv 55 scan 80 SIM 10 kv 10-2 torr MS (Scan) m/z sec/cycle MS (SIM) 100msec/ch 60

28 (2) Frit-FAB ph LC-Frit-FAB-MS K. Kamei, et al.(1988) ph HPLC APCI ESI FAB (1) 8 Frit-FAB mM 0.2 % / (1:1) 100mg/L 20 L FAB APCI ESI a) [M-H] - [M+H] + M + FAB b) 61

29 1:1 m/z 208 Frit-FAB c) Frit-FAB Frit-FAB [mg/ml] CAS * (SAP) (MCPP) * 62

30

31 H 2N SO2 + [M-H] - [M + H] FAB : : [M + H]+ [M-H] FAB : : CH3 + Cl O + H [M-H] - CH3 + Cl O-CH CH 3 M [M-H] [M-H] FAB : : 64

32 N O Cu + H + N O FAB : : H 2NCH 2CH 2S 2P(OCH(CH 3) 2) 2 + H + S -SP(OCH(CH 3) 2) FAB : : S -SCN(CH 3 ) FAB : : 65

33 N + Cl CH 2 Cl N CH2 N NH 2 + N H [M-H] - [M + H] FAB : : CH3CH2O C(CH 3) FAB : : 66

34 (2) m/z a) ph 20mM 0.2 % / (1:1) ph b) 67

35 c) FAB - ph HPLC MS (3) Frit-FAB m- (NBA) NBA 0.1% HPLC 0.2% (1) HPLC MS 5 10 L/min (2) FAB Frit FAB 50 / (4:6) 50 68

36 80 FAB LC-Frit-FAB-MS LC-Frit-FAB-MS LC Frit-FAB-MS LC MS (1) (2)LC (3) (4)LC-Frit-FAB-MS FAB FAB Frit Frit-FAB Frit-FAB ppb Frit-FAB ppb 69

37 ) Application Note, MS42, MS43, MS55, MS62, MS63, MS72, MS101, MS109, MS131, MS144, MS159 2)K. Kamei, K. Kitahara, A. Momose, K. Matsuura, H. Yuki : Mass Spectrosc. (Japan), vol.36, 115(1988) 70

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