LC/MS MS LC/MS LC LC (APCI) (ESI) FAB 2 LC (APCI) 4.1.1.1 LC LC/MS LC MS LC 3 ph APCI/MS LC (1) APCI ESI 100% 100% ESI 4.1.1-1 LC 1.00cp 100 34
a 4.1.1-1 LC b ( /mpas) n- 0.23 36 0.5 99 0.31 69 n- 0.92 174 1.0 81 0.47 50 0.37 46 0.97 77 p- 0.64 140 0.37 68 0.59 111 0.65 80 0.23 35 0.57 62 0.44 40 0.54 118 0.4 66 0.79 57 0.4 80 0.32 56 1.54 107 0.45 77 0.37 57 2.24 189 4.4 184 0.38 56 0.67 101 0.37 82 0.94 116 2.39 82 n- 2.2 97 0.61 78 1.19 64 19.9 198 1.22 118 1.00 100 a b 20 C (2)pH 3 LC ph ph LC/APCI/MS ph 35
ph LC/APCI-MS ph ph LC LC/APCI/MS ph ph [M+NH 4 ] + 4.1.1-2 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 7 11 1 2 0.05 2 6 4 6 6 7 8 10 4.4 6.4 APCI ph 100mM 36
ph ph APCI/MS 4.1.1-2 ph 4.2.2 (3) LC LC/APCI/MS ph LC/APCI-MS APCI-MS LC/APCI-MS 4.1.1-3 4.1.1-3 LC-APCI-MS LC-APCI-MS 1- CH 3 CH 2 CH 2 SO 3 146.14 Na 1- CH 3 (CH 2 ) 3 SO 3 N 160.16 a 1- CH 3 (CH 2 ) 4 SO 3 N 174.19 a 1- CH 3 (CH 2 ) 5 SO 3 N 188.22 a 1- CH 3 (CH 2 ) 7 SO 3 N 202.24 a [CH 3 (CH 2 ) 5 ] 3 N 269.15 [CH 3 (CH 2 ) 6 ] 3 N 311.6 [CH 3 (CH2) 7 ] 3 N 353.67 (CH 3 CH 2 ) 4 NOH 147.26 [CH 3 (CH 2 ) 3 ] 4 NO 259.47 H 4.1.1.2 APCI/MS 37
(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 + 182.1kcal/mol (1648kcal/mol) (202.3kcal/mol) A - + BH B - + AH BH A - A - BH A - BH 38
Linuron Diuron APCI 4.1.1-1 4.1.1-2 p-tert- ( 4.1.1-3 4.1.1-4) (C 60 ) APCI Linuron (200µg/ml) Scan ;100-400 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 3 4.1.1-1 Linuron APCI-MS Cl Cl N C N H O CH 3 CH 3 Diuron Diuron ( 0µg/ml) Scan ;100-400 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 4.1.1-2 Diuron APCI-MS 39
CH OH 3 HO 17β-Estradiol 17β-estradiol (2.0µg/ml) Scan ;100-400 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 4.1.1-3 17b-estradiol APCI-MS t-bu COOH 4-t-Butyl-benzoic acid p-t-butylbenzoic acid (5.0µg/ml) Scan ;100-400 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 4.1.1-4 p-t-butylbenzoic acid APCI-MS 40
(2) APCI ESI ph ph LC-MS 0.05% 1 4.1.1-5 ph 4-10 APCI/MS ph 4.1.1-4 ph 4.1.1-4 APCI/MS Hitachi M-1200 AP/ES 1.0ml/min 100-400 50amu/sec 70V 1,800V 3,000V 90V 48 180 400 120 20ml APCI ESI [M+NH 4 ] + ESI 18- -6- ESI APCI H 2 O + 4.1.1-6 41
4.1.1-5 ph Dibenzo-18-crown-6-ether (5.0µg/ml) Scan ;100-400 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 4.1.1-6 Dibenzo-18-crown-6-ether APCI-MS 42
(3) LC MS APCI ESI 4.1.1.3 (1) APCI MS MS LC ml/min 0.1-1.0ml/min LC/APCI/MS 1.0ml/min (2) CID(collision induced dissociation) CID SIM CID 4-50 166 4.1.1-7 90 166 122 4.1.1-8 972-100 150 ( 4.1.1-9) 100V 43
NO 2 4-Nitro-benzoic acid COOH 4-Nitrobenzoic acid (5.0µg/ml) Scan ;100-400 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 4.1.1-7 4-Nitrobenzoic acid APCI-MS (Fragmentor voltage ; 50V) 4.1.1-8 4-Nitrobenzoic acid APCI-MS (Fragmentor voltage ; 90V) α-cyclodextrin (1000µg/ml) Scan ;300-1000 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 4.1.1-9 a-cyclodextrin APCI-MS 44
(3) APCI 3000 4000V -2500-3500V 3000V -2500V (4) APCI APCI 4.1.1-10 / 4.1.1-10 1ml/min 45
(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) 4.1.1.4 APCI/MS 4.1.1-11 4.1.1-15 2,4- A, APCI/MS M-H ESI 2,3,3,4,5-pentachloro-4- biphenylol(10µg/ml) Scan ;100-450 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 4.1.1-11 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
2,2,3,4,5,5,6-heptachloro-4- biphenylol (10µg/ml) Scan ;100-450 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 4.1.1-12 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 ;100-400 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 4.1.1-13 2,4-Dichlorophenol APCI-MS 47
Bisphenol A (5.0µg/ml) Scan ;100-400 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 ; 4.1.1-14 CH 3 HO C OH CH 3 Bisphenol Bisphenol A APCI-MS Diethylstilbestrol (10µg/ml) Scan ;100-400 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 4.1.1-15 HO Diethylstilbestrol OH Diethylstilbestrol APCI-MS 48
(ESI) ESI ESI LC LC/MS/ESI ph MS 4.1.2.1 ESI ESI 4.1.2-1 4.1.2-1 ESI 1 1 cp Nm -1 1.0 0.073 100 0.61 0.022 78 0.37 0.029 82 1.19 0.022 64 2.20 0.024 97 2.39 0.021 82 2.95 0.025 83 0.32 0.024 56 1.80 0.038 101 1.22 0.028 118 0.44 0.028 40 2.24 0.043 189 (DMSO) 2 1 20 2 10% 49
4.1.2-1 ESI 4.1.2.2 ph LC ph LC LC/MS 4.1.2-2 4.1.2-3 35 NH 3 NH 4 + 50 CH 3 OH NH 4 + 68 CH 3 OH H 2 O NH 4 + 82 (CH 3 OH) 2 NH 4 + 36 H 2 O NH 4 + 54 (H 2 O ) 2 NH 4 + 77 (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 77 4.1.2-4 41 CH 3 CN + 59 + CH 3 CN NH 4 77 + CH 3 CN H 2 O NH 4 100 + (CH 3 CN) 2 NH 4 ESI ph H 3.2.3-2 50
4.1.2-2 4.1.2-4 ph ESI (10mM ) 4.1.2.3 (IPC) IPC IPC IPC ph ESI IPC 3.2.3-4 3.2.3-5 LC IPC LC/MS LC/MS IPC 4.1.2.4 ESI ESI ESI (1) 51
ESI ( ) ( ) N M+H M-H 4.1.2-1 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 4.1.2-1 0.2mm 52
0.1mm LC/MS LC 3.2.2 LC/MS ESI LC LC/MS LC/MS (3) LC MS 4.1.2-5 MS MS LC 4.1.2-2 2.0 1.0 PB APCI TSP ESI (ml/min) Frit-FAB or CF-FAB 4.1.2-2 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
4.1.2-5 LC MS ( ) ( cm 3 /min) column Conventional Semi-micro Micro mm 4.0~4.6 1.0~2.0 0.2~0.5 ml/min 1.0 0.2 0.005 Methanol 550 110 3 Water 1240 248 6 ESI 4.1.2-6 LC 4.1.2.1 (mm) 0.32 3-8 ESI 1.0 30-60 0.1mm 2.0 200-400 4.6 800-2000 l 200 l ESI 0.2ml/min ( 4.6mm 150mm) 1ml/min LC ESI Tee 4.1.2-6 LC (4) LC ESI Taylor cone ±3 5kV MS (5) ( ) 54 ( l/min)
( ) ( ) API CID(collision induced dissociation) ( ) CID (6) ESI LC LC ph HFBA TFA 4.1.2-4 50 M [M+Na] + ESI 4.1.2-3 Mixing Tee HPLC (UV) (API Needle) Pump 4.1.2-3 55
20 Signal Signal (%) 10 10 15 16 1.5 6 Formic Acetic Propionic Butyric Valeric Acid 4.1.2-4 20% HP 1998 LC/MSD ESI MS 4.1.2-5 0% 0.05% 0.1% 0.5% Signal ratio 2.0 1.0 benzoic acid o-toluic acid salicylic acid phenoxacetic acid 0% 0.05% 0.1% 0.5% 4.1.2-5 56
4.1.2.5 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 4.1.2-6 =82.5 =115.5 =173.3 m1:496 m1 496 m2:578.5 m2 578.5 m3:694.0 m3 694.0 m4:867.3 m4 m/z 500 600 700 800 900 4.1.2-6 n1=496 (578.5 496) M1=6(496-1) =6.012 = 2970.0 n2=578.5 (694.0 578.5) M2=5(578.5-1) =5.008 =2887.5 n3=694.0 (867.3 694.0) M3=4(694.0-1) =4.004 =2772 =M1 M2 M3 --- 57
4.1.2.6 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) 4.1.2.7 Paul Kebarle and Liang Tang ;AnalChem.,1993,65,972A-986A Robert D.Voyksner; Environ. Sci. Technol.,1994,28,118A-127A 15 LC/MS ( ) 58
CF(Frit)-FAB 4.1.3.1 LC Frit-FAB API Frit API LC Frit-FAB LC Frit-FAB (1) Frit-FAB FAB Frit- FAB 4.1.3.6 1) 4.1.3-1 (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] + 4.1.3-1 59
Positive Negative 4.1.3-1 4.1.3-1 JMS-700 Frit-FAB 0.5 ml/min FAB Xe kv 55 scan 80 SIM 10 kv 10-2 torr MS (Scan) m/z 45 500 1 sec/cycle MS (SIM) 100msec/ch 60
(2) Frit-FAB ph LC-Frit-FAB-MS K. Kamei, et al.(1988) ph HPLC APCI ESI FAB 4.1.3.2 (1) 8 Frit-FAB 4.1.3-2 4.1.3-2 20mM 0.2 % / (1:1) 100mg/L 20 L 4.1.3-1 FAB 4.1.3-4.1.3-10 APCI ESI a) [M-H] - [M+H] + M + FAB b) 61
1:1 m/z 208 Frit-FAB c) Frit-FAB 4.1.3-2 Frit-FAB [mg/ml] CAS 1 10380-28-6 0.04 0.4 2 137-26-8 0.006 0.06 0.06 3 3337-71-1 2 4 * 55335-06-3 0.06 5 (SAP) 741-58-2 1 0.1 6 (MCPP) 16484-77-8 0.05 7 13861-41-3 0.2 8 80844-07-1 0.08 * 62
4.1.3-2 63
H 2N SO2 + [M-H] - [M + H] + 4.1.3-3 FAB : : [M + H]+ [M-H] - 4.1.3-4 FAB : : CH3 + Cl O + H [M-H] - CH3 + Cl O-CH CH 3 M [M-H] [M-H] - - 4.1.3-5 FAB : : 64
N O Cu + H + N O - 4.1.3-6 FAB : : H 2NCH 2CH 2S 2P(OCH(CH 3) 2) 2 + H + S -SP(OCH(CH 3) 2) 2-4.1.3-7 FAB : : S -SCN(CH 3 ) 2-4.1.3-8 FAB : : 65
N + Cl CH 2 Cl N CH2 N NH 2 + N H [M-H] - [M + H] + 129 256 4.1.3-9 FAB : : CH3CH2O C(CH 3) 2 + 163 4.1.3-10 FAB : : 66
(2) 1 4.1.3-4 4.1.3-4 m/z 208 100 66 33 33 231 100 66 33 33 258 50 50 50 100 214 100 100 100 100 211 100 66 33 33 163 25 25 100 75 100 a) ph 20mM 0.2 % / (1:1) ph b) 67
c) FAB - ph HPLC MS (3) Frit-FAB m- (NBA) NBA 0.1% HPLC 0.2% 4.1.3.3 (1) HPLC MS 5 10 L/min (2) FAB Frit FAB 50 / (4:6) 50 68
80 FAB 4.1.3.4 LC-Frit-FAB-MS LC-Frit-FAB-MS LC Frit-FAB-MS LC MS (1) (2)LC (3) (4)LC-Frit-FAB-MS 4.1.3.5 FAB FAB Frit Frit-FAB Frit-FAB ppb Frit-FAB ppb 69
4.1.3.6 1) 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