2,4- (2,4-Dichlorophenoxy) acetic acid 2,4-D 2,4-PA OH O O C A S 94-75-7 C 8 H 6 Cl 2 O 3 Cl Cl ( ) 221. (219.9694) 1) (g/cm 3 ) (25 C) 2) ( C) (.4 mm Hg) 2) ( C) 2) (mmhg) (2 C) 2) (mg/l) (25 C) log P ow 2) 1.6 16 14.5 8.25 1-5 677 2.81 3) LD 5 375 mg/kg() ACGIH- TWA: 1 mg/m 3 ACGIH- A4 HCl PRTR 1) SPARC Performs Automated Reasoning in Chemistry v4.5 <http://sparc.chem.uga.edu/sparc/> 21.11.15 2) The Physical Properties Database (Phys Prop), Syracuse Research Corporation, USA 3) Kis-net 371
1 (1).5 mol/l / LC/MS/MS-SRM (ESI - negative) (2) 2,4- (2,4-D) (>98.) 2,4--d 5 (>99.) PCB 5 Waters OASIS-MAX 6 cc (15 mg) Cat.No.186369 2,4-D 1. mg 1 ml 1 μg/ml 1.4. ng/ml 1.2 ng/ml 2,4-D-d 5 5. ng/ml 2,4-D-d 5 1. mg 1 ml 1 μg/ml 1 ng/ml Zymark 1 372
Sep-Pak Concentrator 13 mm GD/X 1. μm 2.5 ml (3) 21 3 2. g 1. g 2 1 ng/ml 5 μl2,4-d-d 5 5. ng)1 mol/l.5 ml 3 ml 1 3 rpm 1 3 ml 3 3 ml 5 ml 1 ml OASIS-MAX 3 1 ml/min 2 ml 5 mmol/l 5 ml 5 ml.5 mol/l / 5 ml 1 ml 4 LC/MS LC Waters ACQUITY UPLC Atlantis dc18 (15 cm 2.1 mm, 3 μm) (Waters) 373
A: 1 mmol/l B: 7 min A: 6 1 B: 4 99 linear gradient 7 1 min A: 1 B: 99 1 1.1 min A: 1 6 B: 99 4linear gradient 1.1 15 min A: 6 B: 4.2 ml/min 4 C 5 μl MS Waters XEVO TQ MS 1 kv 15 C 4 C 5 L/Hr 8 L/Hr ESI - negative 2 V 14 ev SRM (m/z) 2,4-D219 > 1612,4-D-d 5 224 > 164 5 μl LC/MS 5 μl LC/MS C = (ng/g-dry) C = RQ/V R: Q: (ng) = (ng/μl) (μl) V: (g-dry) 374
Q: 5. (ng) = (.1 ng/μl) (5. μl) V: 1. (g-dry) C = R.5 (ng/g-dry) (IDL) IDL 5 IDL (ng/ml) 1 IDL (g-dry) (ml) IDL (μg/kg-dry) 2,4-D.17 1. 1.17 (MDL) (MQL) MDL MQL 5 (g-dry) 2 MDL MQL (ml) MDL (ng/g-dry) MQL (μg/kg-dry) 2,4-D 1. 1.63.16 1 2 21 3 3, 2 3OASIS-MAX 5 ml 1 ml 4 5IDL 21 3 3 375
3 IDL 2,4-D (g-dry) 1. (ml) 1 (ng/ml) 1. (μl) 5. 1 (ng/ml).928 2 (ng/ml).866 3 (ng/ml).988 4 (ng/ml).927 5 (ng/ml).898 6 (ng/ml).87 7 (ng/ml).957 (ng/ml).9191.447 IDL (ng/ml) *1.17 IDL (ng/g-dry).17 S/N *2 1 CV () 4.9 *1: IDL = t (n-1,.5) σ n-1 2 t (n-1,.5) = 1.9432 *2: 24D native 1ng d 5ng 24D_11614_33 1 2,4-D-d 5 224>164 7.42 224 > 164 (d-24d) 8.9e3 2. 4. 6. 8. 1. 12. 24D_11614_33 1 7.42 219 > 161 (24D) 1.75e3 2,4-D 219>161 7.77 2.17 4.84 5.15 3.66 4.32 6.5 8.69 9.69 Time 2. 4. 6. 8. 1. 12. 1 IDL 1. ng/ml 376
6MDLMQL 21 3 4 4 MDL MQL 2,4-D - (g-dry) 1. - (ng) 2. - (ng/g-dry).2 - (ml) 1 - (ng/ml) 2. - (μl) 5. - (ng/g) *1 ND 14.3 (ng/g) *2 ND 83.2 1 ng/gdry.211 89.1 2 (ng/g-dry).196 85.1 3 (ng/g-dry).228 81.2 4 (ng/g-dry).228 89.3 5 (ng/g-dry).239 86. 6 (ng/g-dry).242 97. 7 (ng/g-dry).219 92.5 (ng/g-dry).2234 88.6 (ng/g-dry).161 MDL (ng/g-dry) *3.63 MQL (ng/g-dry) *4.16 S/N *5 14 CV () 7.2 *1 (n = 2) *2 MDL (n = 2 ) *3 MDL= t (n-1,.5)σ n-1 2 t (n-1,.5) = 1.9432 *4 MQL= σ n-1 1 *5 377
24D MDL2 native 2ng d 5ng 11612 24D_11614_58 1 2,4-D-d 5 224>164 7.33 224 > 164 (d-24d) 7.66e3 2. 4. 6. 8. 1. 12. 24D_11614_58 7.37 1 219 > 161 (24D) 3.31e3 2,4-D 219>161 5.23 7.68 2.17 7.9 4.8 5.58 5.89 Time 2. 4. 6. 8. 1. 12. 2 MDL 378
2 3 2. g 1. g (2,4-D-d 5 5. ng) 1 mol/l.5 ml 3 ml 1 min 1 min 3 rpm 1 min 3 3 ml 65 ml OASIS-MAX 5 mmol/l 1 ml 1 ml/min 2 ml 5 ml 5 ml 5 ml LC/MS/MS SRM.5 mol/l / 5 ml 1 ml ESI-negative 3 4 5-15-2 (ng/ml) (1) (2) (3) (4) (ng/ml) (5) (1) (15) (2) 4 1.4. ng/ml 1.2 ng/ml 379
5-1 : ng/ml (A s ) (A is ) (A s /A is ) (C s ) 2,4-D 2,4-D-d 5 1. 2. 5. 369.86 718.38 274.337 2147.42 259.849 211.389.172.349.987 1. 3796.399 265.118 1.838 2. 8416.428 2247.472 3.745 4. 16771.979 2237.952 7.494 *: : 5 ng/ml (C is ) 5-2 (A s ) : ng/ml 2,4-D (A is ) (A s /A is ) (C s ) 2,4-D-d 5 1. 3754.475 2139.73 1.755 2. 8312.93 2397.18 3.468 5. 18249.355 2233.71 8.17 1 36353.289 2169.937 16.753 2 7438.414 279.623 33.871 *: : 5 ng/ml (C is ) 38
5 6 21166_24D_MS_SCAN 1 (.17) 1 a) XEVO-TQMS#VBA588 7-Jun-211 15:5:1 MS2 ES- 219 3.65e6 221 161 91 91 113 163 97 16 196 198 62 62 62 69 69 81 81 81 89 93 13 15 119 131 155 121 137 145 164 2 18 182 185 28 223 218 224 229 m/z 5 6 7 8 9 1 11 12 13 14 15 16 17 18 19 2 21 22 23 21166_24D_MSMS_SCAN 1 (.17) 1 b) XEVO-TQMS#VBA588 161 7-Jun-211 15:8:36 Daughters of 219ES- 3.33e6 125 175 219 m/z 5 6 7 8 9 1 11 12 13 14 15 16 17 18 19 2 21 22 23 5 2,4-D a) b) 21166_D-24D_MS_SCAN 1 (.17) 1 c) XEVO-TQMS#VBA588 7-Jun-211 15:18:28 MS2 ES- 224 3.68e6 226 224 91 228 164 166 81 58626262 6969 75 113 163 81 89 9797 1315 117 131 135 198 223 129 145 147 166173 196 18 189 26 28 228 m/z 5 6 7 8 9 1 11 12 13 14 15 16 17 18 19 2 21 22 23 21166_D-24D_MSMS_SCAN 2 (.34) 1 d) XEVO-TQMS#VBA588 164 7-Jun-211 15:2:39 Daughters of 224ES- 4.51e6 127 163 166 13 128 18 224 m/z 5 6 7 8 9 1 11 12 13 14 15 16 17 18 19 2 21 22 23 6 2,4-D-d 5, c) d) 381
2,4-D 6 8 (g-dry) 1 6 (ng) (ng/g-dry) () () () 無 添 加 2 ND - - 8 5 5.54 18 5.1 72 2 g 1 g 5 ng1 mol/l.5 ml 1 4 ml2 2 ml3 2 ml 2 5 ml 1 1 ml 23 5 ml 1 ml 2,4-D-d 5 5 ng 7 7 (n=1) () () 1 74.4 2 8.2 87.1 3 4.5 1 69. 2 5. 74. 3 1 73.2 2 6.4 79.6 3 1 78.2-2 /= 1 : 1 1.1 3 88.3 382
OASIS-MAX ml1 ml5 ml5 ml 1 ml ml 2 ng 1 mol/l.5 ml 1 mol/l.5 ml OASIS-MAX 5 mmol/l.5 mol/l / 5 ml 1 ml 2,4-D-d 5 5 ng 8 1 5 3 8 OASIS-MAX (n=2) (ml) 1 (ml) (ng) () 5.3 5 2 9.4 1. 5.4 5 2 8.5 1 41.4 121.3 5 2 117.5 1 11.9 113.5 5 2 117.5 1 48.2 383
2,4-D MDL <.63 ng/g-dry 5 ng 7 8 24D Rive sed d 5ng 11612 24D_11614_53 1 2,4-D-d 5 224>164 7.37 224 > 164 (d-24d) 7.23e3 5.36 2. 4. 6. 8. 1. 12. 24D_11614_53 7.37 1 219 > 161 (24D) 459 2,4-D 219>161 2.26 7.64 2.87 3.66 4.32 6.26.15 7.11 5.2 7.2 7.94 8.69 9.3 11. 9.87 Time 2. 4. 6. 8. 1. 12. 24D MDL2 native 2ng d 5ng 11612 24D_11614_58 1 2,4-D-d 5 224>164 7.33 5 ng 224 > 164 (d-24d) 7.66e3 2. 4. 6. 8. 1. 12. 24D_11614_58 1 7.37 219 > 161 (24D) 3.31e3 2,4-D 219>161 5.23 7.68 2.17 7.9 4.8 5.58 5.89 Time 2. 4. 6. 8. 1. 12. 7 5 ng 384
24D Sea sed d 5ng 11612 24D_11614_54 1 2,4-D-d 5 224>164 7.33 224 > 164 (d-24d) 6.86e3 5.28 2. 4. 6. 8. 1. 12. 24D_11614_54 7.42 1 219 > 161 (24D) 48 2,4-D 219>161 2.17 7.68 2.48 2.7 3.1 3.88 4.674.93 5.54 6.24 7.86 8.77 8.9 9.95 1.39 11.18 11.92 Time 2. 4. 6. 8. 1. 12. 24D Rec river sed native 5ng d 5ng 11612 24D_11614_66 7.29 1 2,4-D-d 5 224>164 5 ng 224 > 164 (d-24d) 7.56e3 5.23 2. 4. 6. 8. 1. 12. 24D_11614_66 7.37 1 219 > 161 (24D) 8.54e3 2,4-D 219>161 Time 2. 4. 6. 8. 1. 12. 8 5 ng 385
MDL 4 C 6 14 1 9 6 14 14 18 ()( n = 7) 2,4-2,4-D 14 ng/ml R 2 >.999 (1 2 ng/ml) R 2 >.999 MDL.63 ng/g-drymql.16 ng/g-dry 1 g-dry 2,4-D 5 ng (1511.63 ng/g-dry 2,4-D 18 p.827-8412,4-d 654-37 3-1-27 TEL78-735-6918 FAX78-735-7817 E-mail Azusa_Oita@pref.hyogo.lg.jp, Shuusuke_Takemine@pref.hyogo.lg.jp Masahiro_Tsurukawa@pref.hyogo.lg.jp, Chisato_Matsumura@pref.hyogo.lg.jp 386
(2,4-Dichlorophenoxy) acetic acid 2,4-D, 2,4-PA This method is suitable for the determination of (2,4-Dichlorophenoxy) acetic acid in sediment by liquid chromatography-tandem mass spectrometry (LC/MS/MS). Twenty grams-wet of the sediment sample is placed into a centrifugal sedimentation tube, and 5. ng of 2,4-D-d 5 as a surrogate,.5 ml of 1 mol/l formic acid and 3 ml methanol are added. Thereafter, the sample is extracted by sonication and shaking (1 minute each), centrifuged at 3 rpm. The extract is subsequently separated off, another 3 ml methanol is added to the residue, and the preceding process is repeated another two times. The combined extract is concentrated to 3 ml. A concentrated solution is placed into a beaker, and diluted with 65 ml of distilled water. The mixed solution is loaded on a OASIS-MAX cartridge. The cartridge is washed with 5 ml of 5 mmol/l ammonium acetate in distilled water and 5 ml of methanol that rinsed the beaker. Thereafter, the cartridge is eluted with 5 ml of.5 mol/l formic acid in methanol, and the elution is concentrated to 1 ml. The solution is analyzed by the LC/MS/MS-SRM (ESI negative). The method detection limits (MDL) and the method quantification limit (MQL) is.63 and.16 ng/g-dry, respectively. The average of recoveries (n=5) from 2. g-wet of sediment sample spiked with 5 ng (2,4-Dichlorophenoxy) acetic acid was 18, and the relative standard deviation was 5.1. 3 times Sediment sample 2. g-wet (Corresponding to 1. g-dry) surrogate (2,4-D-d 5 5. ng) 1 mol/l formic acid.5 ml Ultrasonic extraction Methanol 3 ml 1 min Shaking extraction 1 min Centrifugation 3 rpm, 1 min Concentration N 2 gas, to 3 ml Dilution Purified water 65 ml Rinse: Methanol 5 ml Solid phase extraction OASIS MAX 1 ml/min Rinse beaker:methanol 5 ml Dry Air 2 ml Wash 5 mmol/l Ammonium acetate 5 ml Beaker-rinsed-methanol 5 ml Elution.5 mol/l Formic acid methanol 5 ml Concentration N 2 gas Make up volume Methanol to 1 ml LC/MS/MS-SRM ESI-Negative 387
2,4-2,4-D 2,4-PA 2. g 1. g (2,4-D-d 5 5. ng) 1mol/L.5 ml 3 ml 1 1 LC/MS/MS-SRM -ESI-.63 ng/g-dry 3 rpm, 1 3 OASIS MAX 1 ml/min 2 ml 3 ml 65 ml 5 ml 5 mmol/l 5 ml 5 ml 分 析 条 件 : 機 器 LC:Waters ACQUITY UPLC MS:Waters XEVO TQ MS カラム Atlantis dc18 15 cm 2.1 mm, 3 μm (Waters).5 mol/l / 5 ml 1 ml LC/MS/ MS-SRM ESI-Negative 388