Chinese Journal of Chromatography ›› 2020, Vol. 38 ›› Issue (8): 900-905.DOI: 10.3724/SP.J.1123.2019.12025
Previous Articles Next Articles
DAI Jing1, GAO Lehong2, PENG Fangda1, YU Lijia1, WANG Chaodong2, WANG Yuping2, DING Chunguang1,*()
Received:
2019-12-18
Online:
2020-08-08
Published:
2020-12-11
Contact:
DING Chunguang
Supported by:
DAI Jing, GAO Lehong, PENG Fangda, YU Lijia, WANG Chaodong, WANG Yuping, DING Chunguang. Simultaneous determination of twelve antiepileptic drugs in serum by ultra high performance liquid chromatography-tandem mass spectrometry[J]. Chinese Journal of Chromatography, 2020, 38(8): 900-905.
No. | Compound | Retention time/min | Parent ion (m/z) | Product ion (m/z) | Collision energy/eV | Cone voltage/V | Ionization mode | IS |
1 | pregabalin | 2.51 | 159.97 | 97.06 | 14 | 26 | [M+H]+ | phenacetin |
2 | gabapentin | 2.55 | 172.13 | 154.1 | 10 | 24 | [M+H]+ | phenacetin |
3 | levetiracetam | 2.77 | 171.11 | 126.1 | 15 | 10 | [M+H]+ | phenacetin |
4 | lacosamide | 3.90 | 251.10 | 108.1 | 8 | 8 | [M+H]+ | phenacetin |
5 | lamotrigine | 4.10 | 255.98 | 108.9 | 50 | 70 | [M+H]+ | phenacetin |
6 | phenobarbital | 4.33 | 230.99 | 188.1 | 8 | 20 | [M+H]- | chlorzoxazone |
7 | topiramate | 4.51 | 338.09 | 77.90 | 27 | 50 | [M+H]- | chlorzoxazone |
8 | oxcarbazepine | 4.77 | 253.06 | 180.0 | 32 | 52 | [M+H]+ | phenacetin |
9 | phenytoin sodium | 5.09 | 253.20 | 182.2 | 15 | 45 | [M+H]+ | phenacetin |
10 | carbamazepine | 5.29 | 237.06 | 194.1 | 17 | 58 | [M-H]+ | phenacetin |
11 | clonazepam | 5.34 | 316.07 | 270.1 | 24 | 45 | [M-H]+ | phenacetin |
12 | sodium valproic | 5.56 | 143.10 | 143.1 | 3 | 30 | [M-H]- | chlorzoxazone |
13 | phenacetin | 4.35 | 179.94 | 110.0 | 18 | 48 | [M+H]+ | |
14 | chlorzoxazone | 5.34 | 168.02 | 132.0 | 18 | 44 | [M-H]- |
Table 1 MS parameters of the 12 antiepileptic drugs and internal standards
No. | Compound | Retention time/min | Parent ion (m/z) | Product ion (m/z) | Collision energy/eV | Cone voltage/V | Ionization mode | IS |
1 | pregabalin | 2.51 | 159.97 | 97.06 | 14 | 26 | [M+H]+ | phenacetin |
2 | gabapentin | 2.55 | 172.13 | 154.1 | 10 | 24 | [M+H]+ | phenacetin |
3 | levetiracetam | 2.77 | 171.11 | 126.1 | 15 | 10 | [M+H]+ | phenacetin |
4 | lacosamide | 3.90 | 251.10 | 108.1 | 8 | 8 | [M+H]+ | phenacetin |
5 | lamotrigine | 4.10 | 255.98 | 108.9 | 50 | 70 | [M+H]+ | phenacetin |
6 | phenobarbital | 4.33 | 230.99 | 188.1 | 8 | 20 | [M+H]- | chlorzoxazone |
7 | topiramate | 4.51 | 338.09 | 77.90 | 27 | 50 | [M+H]- | chlorzoxazone |
8 | oxcarbazepine | 4.77 | 253.06 | 180.0 | 32 | 52 | [M+H]+ | phenacetin |
9 | phenytoin sodium | 5.09 | 253.20 | 182.2 | 15 | 45 | [M+H]+ | phenacetin |
10 | carbamazepine | 5.29 | 237.06 | 194.1 | 17 | 58 | [M-H]+ | phenacetin |
11 | clonazepam | 5.34 | 316.07 | 270.1 | 24 | 45 | [M-H]+ | phenacetin |
12 | sodium valproic | 5.56 | 143.10 | 143.1 | 3 | 30 | [M-H]- | chlorzoxazone |
13 | phenacetin | 4.35 | 179.94 | 110.0 | 18 | 48 | [M+H]+ | |
14 | chlorzoxazone | 5.34 | 168.02 | 132.0 | 18 | 44 | [M-H]- |
Fig. 1 Total ion chromatogram of the 12 antiepileptic drugs and two internal standards 1. pregabalin; 2. gabapentin; 3. levetiracetam; 4. lacosamide; 5. lamotrigine; 6. phenobarbital; 7. phenacetin (IS); 8. topiramate; 9. oxcarbazepine; 10. chlorzoxazone (IS); 11. phenytoin sodium; 12. carbamazepine; 13. clonazepam; 14. sodium valproic.
No. | Compound | Linear range/(μg/L) | Linear equation | r2 | LOQ/(μg/L) |
Y: peak area ratio of the compound to internal standard; X: mass concentration, μg/L. | |||||
1 | pregabalin | 25.0-2500 | Y=0.1508X+0.0008 | 0.992 | 16.5 |
2 | gabapentin | 12.5-2500 | Y=0.3420X+0.0022 | 0.992 | 10.0 |
3 | levetiracetam | 12.5-2500 | Y=1.9180X+0.0078 | 0.995 | 0.750 |
4 | lacosamide | 12.5-2500 | Y=4.3480X+0.0160 | 0.997 | 0.210 |
5 | lamotrigine | 12.5-2500 | Y=0.2405X+0.0008 | 0.996 | 0.500 |
6 | phenobarbital | 25.0-2500 | Y=0.0769X+0.0020 | 0.997 | 21.0 |
7 | topiramate | 12.5-2500 | Y=0.7476X+0.0078 | 0.998 | 5.30 |
8 | oxcarbazepine | 12.5-2500 | Y=1.3070X+0.0045 | 0.996 | 0.140 |
9 | phenytoin sodium | 12.5-2500 | Y=0.3903X+0.0018 | 0.995 | 2.30 |
10 | carbamazepine | 12.5-2500 | Y=8.5390X+0.0460 | 0.995 | 0.150 |
11 | clonazepam | 1.25-250 | Y=1.2230X+0.0006 | 0.995 | 0.330 |
12 | sodium valproic | 125-12500 | Y=0.9634X+0.0810 | 0.997 | 103 |
Table 2 Linear ranges, linear equations, correlation coefficients (r2), limits of quantification (LOQs) of the 12 antiepileptic drugs
No. | Compound | Linear range/(μg/L) | Linear equation | r2 | LOQ/(μg/L) |
Y: peak area ratio of the compound to internal standard; X: mass concentration, μg/L. | |||||
1 | pregabalin | 25.0-2500 | Y=0.1508X+0.0008 | 0.992 | 16.5 |
2 | gabapentin | 12.5-2500 | Y=0.3420X+0.0022 | 0.992 | 10.0 |
3 | levetiracetam | 12.5-2500 | Y=1.9180X+0.0078 | 0.995 | 0.750 |
4 | lacosamide | 12.5-2500 | Y=4.3480X+0.0160 | 0.997 | 0.210 |
5 | lamotrigine | 12.5-2500 | Y=0.2405X+0.0008 | 0.996 | 0.500 |
6 | phenobarbital | 25.0-2500 | Y=0.0769X+0.0020 | 0.997 | 21.0 |
7 | topiramate | 12.5-2500 | Y=0.7476X+0.0078 | 0.998 | 5.30 |
8 | oxcarbazepine | 12.5-2500 | Y=1.3070X+0.0045 | 0.996 | 0.140 |
9 | phenytoin sodium | 12.5-2500 | Y=0.3903X+0.0018 | 0.995 | 2.30 |
10 | carbamazepine | 12.5-2500 | Y=8.5390X+0.0460 | 0.995 | 0.150 |
11 | clonazepam | 1.25-250 | Y=1.2230X+0.0006 | 0.995 | 0.330 |
12 | sodium valproic | 125-12500 | Y=0.9634X+0.0810 | 0.997 | 103 |
No. | Compound | Added/(mg/L) | Recovery/% | RSDs/% | |
Intra-assay (n=6) | Inter-assay (n=3) | ||||
1 | pregabalin | 0.05 | 95.30 | 5.99 | 14.2 |
0.25 | 105.0 | 5.85 | 5.99 | ||
1.25 | 98.10 | 4.75 | 9.07 | ||
2 | gabapentin | 0.05 | 93.30 | 5.53 | 14.1 |
0.25 | 104.0 | 5.30 | 6.60 | ||
1.25 | 99.70 | 6.03 | 9.43 | ||
3 | levetiracetam | 0.05 | 97.70 | 4.01 | 12.8 |
0.25 | 102.7 | 7.45 | 6.52 | ||
1.25 | 96.50 | 2.58 | 9.21 | ||
4 | lacosamide | 0.05 | 94.00 | 4.46 | 14.8 |
0.25 | 95.70 | 5.08 | 5.43 | ||
1.25 | 93.90 | 3.68 | 9.69 | ||
5 | lamotrigine | 0.05 | 111.3 | 4.63 | 10.5 |
0.25 | 110.3 | 4.46 | 5.12 | ||
1.25 | 108.6 | 2.63 | 9.37 | ||
6 | phenobarbital | 0.05 | 102.0 | 4.84 | 12.5 |
0.25 | 101.9 | 4.85 | 5.08 | ||
1.25 | 103.6 | 5.14 | 11.0 | ||
7 | topiramate | 0.05 | 109.3 | 4.72 | 14.5 |
0.25 | 102.7 | 6.31 | 7.49 | ||
1.25 | 106.0 | 4.93 | 10.8 | ||
8 | oxcarbazepine | 0.05 | 112.0 | 4.78 | 13.5 |
0.25 | 112.5 | 4.98 | 12.9 | ||
1.25 | 109.0 | 2.23 | 10.8 | ||
9 | phenytoin sodium | 0.05 | 96.70 | 13.2 | 14.6 |
0.25 | 105.2 | 6.93 | 6.29 | ||
1.25 | 101.9 | 3.31 | 9.03 | ||
10 | carbamazepine | 0.05 | 91.70 | 10.7 | 11.7 |
0.25 | 95.90 | 1.88 | 5.48 | ||
1.25 | 90.80 | 2.39 | 9.37 | ||
11 | clonazepam | 0.005 | 110.0 | 10.5 | 10.4 |
0.025 | 114.0 | 5.56 | 5.40 | ||
0.125 | 113.1 | 2.61 | 6.89 | ||
12 | sodium valproic | 0.25 | 107.6 | 9.34 | 7.32 |
1.25 | 110.0 | 2.75 | 4.12 | ||
6.25 | 108.7 | 1.99 | 11.1 |
Table 3 Spiked recoveries and RSDs of the 12 antiepileptic drugs
No. | Compound | Added/(mg/L) | Recovery/% | RSDs/% | |
Intra-assay (n=6) | Inter-assay (n=3) | ||||
1 | pregabalin | 0.05 | 95.30 | 5.99 | 14.2 |
0.25 | 105.0 | 5.85 | 5.99 | ||
1.25 | 98.10 | 4.75 | 9.07 | ||
2 | gabapentin | 0.05 | 93.30 | 5.53 | 14.1 |
0.25 | 104.0 | 5.30 | 6.60 | ||
1.25 | 99.70 | 6.03 | 9.43 | ||
3 | levetiracetam | 0.05 | 97.70 | 4.01 | 12.8 |
0.25 | 102.7 | 7.45 | 6.52 | ||
1.25 | 96.50 | 2.58 | 9.21 | ||
4 | lacosamide | 0.05 | 94.00 | 4.46 | 14.8 |
0.25 | 95.70 | 5.08 | 5.43 | ||
1.25 | 93.90 | 3.68 | 9.69 | ||
5 | lamotrigine | 0.05 | 111.3 | 4.63 | 10.5 |
0.25 | 110.3 | 4.46 | 5.12 | ||
1.25 | 108.6 | 2.63 | 9.37 | ||
6 | phenobarbital | 0.05 | 102.0 | 4.84 | 12.5 |
0.25 | 101.9 | 4.85 | 5.08 | ||
1.25 | 103.6 | 5.14 | 11.0 | ||
7 | topiramate | 0.05 | 109.3 | 4.72 | 14.5 |
0.25 | 102.7 | 6.31 | 7.49 | ||
1.25 | 106.0 | 4.93 | 10.8 | ||
8 | oxcarbazepine | 0.05 | 112.0 | 4.78 | 13.5 |
0.25 | 112.5 | 4.98 | 12.9 | ||
1.25 | 109.0 | 2.23 | 10.8 | ||
9 | phenytoin sodium | 0.05 | 96.70 | 13.2 | 14.6 |
0.25 | 105.2 | 6.93 | 6.29 | ||
1.25 | 101.9 | 3.31 | 9.03 | ||
10 | carbamazepine | 0.05 | 91.70 | 10.7 | 11.7 |
0.25 | 95.90 | 1.88 | 5.48 | ||
1.25 | 90.80 | 2.39 | 9.37 | ||
11 | clonazepam | 0.005 | 110.0 | 10.5 | 10.4 |
0.025 | 114.0 | 5.56 | 5.40 | ||
0.125 | 113.1 | 2.61 | 6.89 | ||
12 | sodium valproic | 0.25 | 107.6 | 9.34 | 7.32 |
1.25 | 110.0 | 2.75 | 4.12 | ||
6.25 | 108.7 | 1.99 | 11.1 |
No. | Compound | Effective blood concentrations/(mg/L)[ | Contents/(mg/L) | |||||||||
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | |||
/: not detected; * below the effective concentration; # beyond the effective concentration. | ||||||||||||
1 | pregabalin | 2-8 | / | / | / | / | / | / | / | / | / | / |
2 | gabapentin | 2-20 | / | / | / | / | / | / | / | / | / | / |
3 | levetiracetam | 12-46 | / | 28.4 | 6.54* | 7.57* | 9.30* | 16.1 | / | 25.5 | 7.86* | / |
4 | lacosamide | 10-20 | / | / | / | / | / | / | / | / | / | / |
5 | lamotrigine | 2.5-15 | / | 13.0 | / | 1.95* | / | / | / | 9.16 | / | / |
6 | phenobarbital | 10-40 | 12.3 | / | / | / | / | / | / | / | / | 0.0200* |
7 | topiramate | 5-20 | 4.06* | / | 8.67 | / | / | / | / | / | / | 13.3 |
8 | oxcarbazepine | 3-35 | / | / | / | / | / | / | / | / | 0.120* | / |
9 | phenytoin sodium | 10-20 | / | / | / | / | / | / | / | / | / | / |
10 | carbamazepine | 4-12 | / | / | / | / | / | / | / | / | / | / |
11 | clonazepam | 0.02-0.07 | / | 0.0150* | / | / | / | / | / | / | / | / |
12 | sodium valproic | 50-100 | 96.2 | 92.4 | / | / | / | / | 82.3 | 110# | / | / |
Table 4 Determination results of the 12 antiepileptic drugs in real samples
No. | Compound | Effective blood concentrations/(mg/L)[ | Contents/(mg/L) | |||||||||
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | |||
/: not detected; * below the effective concentration; # beyond the effective concentration. | ||||||||||||
1 | pregabalin | 2-8 | / | / | / | / | / | / | / | / | / | / |
2 | gabapentin | 2-20 | / | / | / | / | / | / | / | / | / | / |
3 | levetiracetam | 12-46 | / | 28.4 | 6.54* | 7.57* | 9.30* | 16.1 | / | 25.5 | 7.86* | / |
4 | lacosamide | 10-20 | / | / | / | / | / | / | / | / | / | / |
5 | lamotrigine | 2.5-15 | / | 13.0 | / | 1.95* | / | / | / | 9.16 | / | / |
6 | phenobarbital | 10-40 | 12.3 | / | / | / | / | / | / | / | / | 0.0200* |
7 | topiramate | 5-20 | 4.06* | / | 8.67 | / | / | / | / | / | / | 13.3 |
8 | oxcarbazepine | 3-35 | / | / | / | / | / | / | / | / | 0.120* | / |
9 | phenytoin sodium | 10-20 | / | / | / | / | / | / | / | / | / | / |
10 | carbamazepine | 4-12 | / | / | / | / | / | / | / | / | / | / |
11 | clonazepam | 0.02-0.07 | / | 0.0150* | / | / | / | / | / | / | / | / |
12 | sodium valproic | 50-100 | 96.2 | 92.4 | / | / | / | / | 82.3 | 110# | / | / |
|
Viewed | ||||||||||||||||||||||||||||||||||||||||||||||||||
Full text 110
|
|
|||||||||||||||||||||||||||||||||||||||||||||||||
Abstract 184
|
|
|||||||||||||||||||||||||||||||||||||||||||||||||