LAAN-J-MS-E141

GC-MS Gas Chromatograph Mass Spectrometer Analysis of Nootropics Using GC-MS(/MS) -Part1 141

Nootropics is the general term for substances intended to improve cognitive function or abilities, such as certain drugs and supplements used to improve concentration or memory. Such nootropics are sometimes sold via the Internet without proper licensing, and there is concern that they could be harmful if used without the prescription of a physician because nootropics also include therapeutic drugs intended for epilepsy, attention-deficit hyperactivity disorder (ADHD), or depression. Due to the relative newness of such substances, there are cases in which they are not registered in NIST, Wiley, or other mass spectral libraries commonly used for GC/MS analysis. This Application Data Sheet describes an example of measuring 19 nootropics measurable by GC/MS.

Experimental

Standards of each component were dissolved in methanol to prepare a 10 g/mL standard mixture solution. For components that cannot be measured without derivatization, 500 L of the 10 g/mL standard mixture solution was obtained, dried under a stream of nitrogen gas, and dissolved in 500 L of BSTFA. Then the solutions were heated for 30 minutes at 60 C for TMS derivatization. Target compounds are listed in Table 1 and analytical conditions in Table 2.

Table 1: Target Compounds Compound Name Compound Name Adrafinil Vinpocetine Atomoxetine Pramiracetam Aniracetam Pregnenolone Oxiracetam Levetiracetam Dihydroergotoxine Etiracetam Dehydroepiandrosterone (DHEA) Atenolol Nicergoline Nadolol Nimodipine Idebenone Nefiracetam Furosemide Piracetam Tianeptine

Table 2: Analytical Conditions

GC-MS: GCMS-QP2020 Column: SH-RxiTM-5Sil MS (30 m long, 0.25 mm I.D., df = 0.25 μm) (Shimadzu, P/N: 221-75940-30) Glass Liner: Deactivated splitless liner with wool (PN:221-48876-03)

MS GC Interface Temp.: 280 C Injection Temp.: 260 C Ion Source Temp.: 200 C Column Oven Temp.: 60 C(2min) (10 C/min)  320 C (15 min) Solvent Cut Time: 2 min Carrier Gas: Helium Acquisition Time: 6 to 43 min Flow Control: Linear velocity (45.6 cm/sec) Acquisition Mode: Scan Injection Mode: Splitless Mass Range: m/z 40 to 600 High Pressure Injection: 250 kPa (1.5 min) Event Time: 0.3 sec 141

Analytical Results

The total ion current chromatogram (TIC) obtained by measuring the underivatized 10 g/mL standard mixture solution is shown in Fig. 1 and the TIC obtained by measuring the TMS derivatized 10 g/mL standard mixture solution is shown in Fig. 2. Decomposition products were detected for nicergoline, adrafinil, and dihydroergotoxine. The decomposition products for adrafinil had the same retention time and mass spectral pattern as the decomposition products for modafinil. Because levetiracetam and etiracetam are optical isomers, they were detected at the same retention time and mass spectral pattern. (10,000,000) TIC (1.00) 13 2.75 11 14 2.50

2.25 10 9 2.00 15 8 1.75

1.50 4 7 16 1.25 1.00 2,3 6 17 5 12 0.75 0.50 1

10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5

Retention Retention Retention Retention ID Compound Name ID Compound Name Time Index Time Index 1 Nicergoline deg.1 10.707 1349 10 Nefiracetam 20.310 2230 2 Oxiracetam 12.603 1493 11 Dehydroepiandrosterone 22.950 2551 3 Piracetam 12.611 1493 12 Nicergoline deg. 2 24.202 2718 4 Levetiracetam/Etiracetam 13.458 1563 13 Pregnenolone 24.344 2738 5 Adrafinil deg. 1 14.348 1638 14 Vinpocetine 25.516 2904 6 Adrafinil deg. 2 17.683 1949 15 Nimodipine 26.074 2988 7 Atomoxetine 17.785 1959 16 Dihydroergotoxine deg. 26.585 3066 8 Aniracetam 18.608 2044 17 Nicergoline 32.430 3813 9 Pramiracetam 19.686 2160 Fig. 1: Total Ion Current Chromatogram (TIC) of 10 g/mL Standard Solution (Underivatized)

(10,000,000) 1.75 TIC (1.00) 3 1.50

1.25 1

1.00 2 6 0.75 4 5 0.50

0.25

15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5

ID Compound Name Retention Time Retention Index 1 Oxiracetam-2TMS 16.535 1836 2 Atenolol-2TMS 21.849 2412 3 Nadolol-3TMS 22.380 2478 4 Idebenone-TMS 24.214 2720 5 Furosemide-2TMS 25.539 2908 6 Tianeptine-TMS 29.959 3567 Fig. 2: Total Ion Current Chromatogram (TIC) of 10 g/mL Standard Solution (TMS Derivatives) 141

Mass Spectra

The mass spectra for typical nootropics obtained by measuring the underivatized standard mixture are shown in Fig. 3 and the mass spectra obtained by measuring the TMS derivatized standard are shown in Fig. 4.

Underivatized ID:1 Nicergoline deg.1 ID:2 Oxiracetam % %

100 156 100 41 184 123 96 75 75 50 76 136 53 68 50 215 50

25 25 108 131 44 67 140 53 65 171 58 7882 0 93 117 197 0 50 100 150 200 50.0 75.0 100.0 125.0

ID:3 Piracetam ID:4 Levetiracetam/Etiracetam % %

100 98 100 126

75 75 70 84 50 41 50

25 142 25 41 69 44 59 58 98 55 97 125 44 84 112 170 0 67 87 114 0 59 99 142 50.0 75.0 100.0 125.0 50.0 75.0 100.0 125.0 150.0 175.0

ID:5 Adrafinil deg.2 (Same as Modafinil deg.) ID:6 Adrafinil deg.1 (Same as Modafinil deg.) % %

100 105 100 165

75 75 105 184 50 121 214 50 77 77 198 25 25 82 185 51 165 51 63 98 115 139 152 63 76 115 152 164 171 200 0 53 89 128 139 0 50.0 75.0 100.0 125.0 150.0 175.0 50 100 150 200

ID:7 Atomoxetine ID:8 Aniracetam % %

100 44 100 135

75 75

50 50 25 25 77 107 148 92 191 219 65 7791 104 255 51 64 104 134 0 51 132 0 50 100 150 200 250 50 100 150 200

ID:9 Pramiracetam ID:10 Nefiracetam % %

100 114 100 98

75 75 121 50 50 72 70 25 25 98 254 126 148 43 56 70 86 169 226 65 79 106 228 246 0 126 0 51 163 50 100 150 200 250 50 100 150 200 250

Fig. 3-1: Mass Spectra of Underivatized Typical Nootropics 141

ID:11 Dehydroepiandrosterone (DHEA) ID:12 Nicergoline deg.2 % %

100 91 105 100 181 79 75 159 75 203 119 145 255 50 270 288 50 270 67 183 55 174 237 198 25 199 227 25 237 242 100115 144 167 285300 58 77 254 0 0 45 50 100 150 200 250 50 100 150 200 250 300

ID:13 Pregnenolone ID:14 Vinpocetine % %

100 43 100 321

75 75 280 316 50 91105 231 50 145 298 350 161 187 213 55 67 119 25 255 25 252 199 301 206 123 175 249 292 0 0 42 77 352 50 100 150 200 250 300 50 100 150 200 250 300 350

ID:15 Nimodipine ID:16 Dihydroergotoxine deg. % %

100 296 100 269

75 75 254 196 154 50 50 223 25 25 127 182 150 273 44 102 209 43 59 106 197 227 317 359 418 72 251 0 77 0 415 100 200 300 400 100 200 300 400

ID:17 Nicergoline %

100 181

75

50 168 198 453 25 251 44 125 221 485 70 283 375 405 436 0 329 100 200 300 400 500

Fig. 3-2: Mass Spectra of Underivatized Typical Nootropics 141

TMS Derivatives

ID:1 Oxiracetam-2TMS ID:2 Atenolol-2TMS % %

100 187 100 72 97 75 75 73 172 50 50 116 25 25 130 287 45 59 212 76 102 197 56 101188 223 294 395 0 245 0 321 50 100 150 200 250 100 200 300 400

ID:3 Nadolol-3TMS ID:4 Idebenone-TMS % %

100 86 100 197 410 75 75 73

50 50 196 239 269 103 25 25 380 73 41 81 153 320 147 409 510 267 413 484 0 41 116 200 0 100 200 300 400 500 100 200 300 400

ID:5 Furosemide-2TMS ID:6 Tianeptine-TMS % %

100 81 100 292

75 75

50 50 294 25 474 25 216 53 73 356 73 126 165 193 257 231 313 384 459 45 377 429 0 103 140 168 204 281 0 321 493 100 200 300 400 100 200 300 400 500

Fig. 4: Mass Spectra of TMS Derivatized Typical Nootropics

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First Edition: March 2018

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