A Validated HPLC-MS/MS Method for Simultaneous Determination of Militarine and Its Three Metabolites in Rat Plasma: Application to a Pharmacokinetic Study

<table class="fixed-width table-group" id="tab6"><tr><td><table class="table"><colgroup><col style="width:5.76em"/><col style="width:5.10em"/><col style="width:6.84em"/><col style="width:7.94em"/><col style="width:9.50em"/><col style="width:6.41em"/></colgroup><tr><td class="thead-hr" colspan="6"><hr/></td></tr><tr class="thead"><td class="align_left" rowspan="2">Parameters</td><td class="align_center" rowspan="2">Unit</td><td class="align_center" colspan="4">Analytes</td></tr><tr class="thead"><td class="align_center">militarine</td><td class="align_center">gastrodin</td><td class="align_center"><i>α</i>-isobutylmalic acid</td><td class="align_center">gymnoside I</td></tr><tr><td class="thead-hr" colspan="6"><hr/></td></tr><tr><td class="align_left"><span style="width: 24.7431ptpx;"><svg height="12.0621pt" id="M31" style="vertical-align:-3.2957pt" version="1.1" viewbox="-0.0498162 -8.7664 24.7431 12.0621" width="24.7431pt" xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink"><g transform="matrix(.013,0,0,-0.013,0,0)"><path d="M592 498C586 556 582 629 581 675H560C545 656 537 650 510 650H115C87 650 74 652 60 675H40C38 620 33 555 28 495H57C69 539 77 568 89 585C102 607 122 616 205 616H267V123C267 44 259 34 168 28V0H454V28C360 34 352 44 352 123V616H422C498 616 514 608 531 584C542 568 552 543 562 495L592 498Z"></path></g><g transform="matrix(.0091,0,0,-0.0091,8.021,3.132)"><path d="M810 0V30C751 35 744 40 744 106V296C744 396 694 451 614 451C583 451 558 438 537 425C512 409 485 391 454 368C434 420 390 451 342 451C309 451 285 439 260 424C228 405 206 387 185 372V453C141 434 90 421 43 413V386C102 378 105 373 105 309V106C105 41 96 35 29 30V0H243V30C194 35 185 41 185 106V335C216 360 256 389 297 389C357 389 384 348 384 275V106C384 41 374 35 312 30V0H530V30C475 35 465 41 465 104V294C465 313 464 324 463 336C502 367 539 389 576 389C637 389 663 346 663 274V109C663 41 653 35 593 30V0H810Z"></path></g><g transform="matrix(.0091,0,0,-0.0091,15.583,3.132)"><path d="M442 39L431 68C420 63 406 57 394 57C377 57 359 71 359 118V301C359 353 349 393 314 424C291 443 261 451 230 451C171 438 107 401 79 381C59 366 46 355 46 340C46 316 72 295 91 295C106 295 117 303 122 319C131 350 139 371 152 384C163 397 178 404 197 404C246 404 281 365 281 293V278C260 262 186 232 124 213C69 194 41 161 41 112C41 47 92 -12 163 -12C195 -12 243 25 283 51C288 35 293 22 308 7C319 -4 339 -12 354 -12L442 39ZM281 87C262 68 226 51 199 51C169 51 129 74 129 126C129 162 151 181 190 198C211 208 259 229 281 240V87Z"></path></g><g transform="matrix(.0091,0,0,-0.0091,19.669,3.132)"><path d="M482 0V30C420 38 407 48 370 103L273 248C308 299 331 332 352 357C387 397 401 404 463 409V439H277V409C318 405 324 400 307 369C290 337 270 307 251 279L194 367C175 396 180 402 220 409V439H18V409C74 401 87 395 117 349L204 213C173 168 147 130 119 95C78 46 67 38 5 30V0H195V30C144 38 143 47 162 79C181 112 202 149 225 181L297 70C314 43 307 35 264 30V0H482Z"></path></g></svg></span></td><td class="align_center">h</td><td class="align_center">0.21 ± 0.08</td><td class="align_center">1.38 ± 0.25</td><td class="align_center">7.50 ± 1.00</td><td class="align_center">1.88 ± 0.25</td></tr><tr><td class="align_left"><span style="width: 25.3694ptpx;"><svg height="12.0621pt" id="M32" style="vertical-align:-3.2957pt" version="1.1" viewbox="-0.0498162 -8.7664 25.3694 12.0621" width="25.3694pt" xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink"><g transform="matrix(.013,0,0,-0.013,0,0)"><path d="M614 175C564 76 510 21 408 21C256 21 146 149 146 336C146 488 235 629 402 629C510 629 570 586 597 480L626 488C620 541 614 582 606 638C578 643 510 665 429 665C206 665 44 527 44 316C44 157 153 -15 402 -15C474 -15 558 5 586 11C604 45 629 119 643 165L614 175Z"></path></g><g transform="matrix(.0091,0,0,-0.0091,8.645,3.132)"><path d="M810 0V30C751 35 744 40 744 106V296C744 396 694 451 614 451C583 451 558 438 537 425C512 409 485 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240V87Z"></path></g><g transform="matrix(.0091,0,0,-0.0091,20.293,3.132)"><path d="M482 0V30C420 38 407 48 370 103L273 248C308 299 331 332 352 357C387 397 401 404 463 409V439H277V409C318 405 324 400 307 369C290 337 270 307 251 279L194 367C175 396 180 402 220 409V439H18V409C74 401 87 395 117 349L204 213C173 168 147 130 119 95C78 46 67 38 5 30V0H195V30C144 38 143 47 162 79C181 112 202 149 225 181L297 70C314 43 307 35 264 30V0H482Z"></path></g></svg></span></td><td class="align_center">ng/mL</td><td class="align_center">62.31 ± 10.01</td><td class="align_center">620.50 ± 75.98</td><td class="align_center">466.86 ± 96.49</td><td class="align_center">32.82 ± 5.92</td></tr><tr><td class="align_left"><span style="width: 37.6797ptpx;"><svg height="12.0392pt" id="M33" style="vertical-align:-3.272799pt" version="1.1" viewbox="-0.0498162 -8.7664 37.6797 12.0392" width="37.6797pt" xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink"><g 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452 212C400 220 373 229 322 255C373 281 400 290 452 298C463 316 471 339 471 357C456 366 431 371 410 370C377 329 356 310 308 279C311 336 317 364 336 413C326 432 310 451 294 459C279 451 262 432 252 413C271 364 277 336 280 279C232 310 211 329 178 370C157 371 132 367 117 357C117 340 125 316 136 298C188 290 215 281 266 255C215 229 188 220 136 212C125 194 117 171 117 153C132 144 157 139 178 140C211 181 232 200 280 231C277 174 271 146 252 97C262 78 278 59 294 51C309 59 326 78 336 97C317 146 311 174 308 231C356 200 377 181 410 140C431 139 456 143 471 153Z"></path></g></svg>ng/mL</td><td class="align_center">123.41 ± 9.95</td><td class="align_center">2077.40 ± 358.66</td><td class="align_center">6285.32 ± 1319.90</td><td class="align_center">122.27 ± 9.54</td></tr><tr><td class="align_left"><span style="width: 44.375ptpx;"><svg height="12.0575pt" id="M35" style="vertical-align:-3.2911pt" version="1.1" viewbox="-0.0498162 -8.7664 44.375 12.0575" width="44.375pt" 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height="6.01072pt" id="M36" style="vertical-align:-0.04980993pt" version="1.1" viewbox="-0.0498162 -5.96091 7.75925 6.01072" width="7.75925pt" xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink"><g transform="matrix(.013,0,0,-0.013,0,0)"><path d="M471 153C471 170 463 194 452 212C400 220 373 229 322 255C373 281 400 290 452 298C463 316 471 339 471 357C456 366 431 371 410 370C377 329 356 310 308 279C311 336 317 364 336 413C326 432 310 451 294 459C279 451 262 432 252 413C271 364 277 336 280 279C232 310 211 329 178 370C157 371 132 367 117 357C117 340 125 316 136 298C188 290 215 281 266 255C215 229 188 220 136 212C125 194 117 171 117 153C132 144 157 139 178 140C211 181 232 200 280 231C277 174 271 146 252 97C262 78 278 59 294 51C309 59 326 78 336 97C317 146 311 174 308 231C356 200 377 181 410 140C431 139 456 143 471 153Z"></path></g></svg>ng/mL</td><td class="align_center">140.34 ± 10.02</td><td class="align_center">2116.92 ± 384.37</td><td class="align_center">6316.32 ± 1327.54</td><td class="align_center">123.73 ± 9.81</td></tr><tr><td class="align_left"><span style="width: 38.6322ptpx;"><svg height="12.0392pt" id="M37" style="vertical-align:-3.272799pt" version="1.1" viewbox="-0.0498162 -8.7664 38.6322 12.0392" width="38.6322pt" xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink"><g transform="matrix(.013,0,0,-0.013,0,0)"><path d="M861 0V28C774 35 771 41 768 147L759 509C756 612 762 614 851 622V650H681L449 149L221 650H57V622C148 613 153 609 144 479L130 271C123 166 117 123 111 88C104 46 85 34 26 28V0H259V28C192 35 169 42 167 90C166 130 166 173 170 256L185 541H187L411 7H431L675 555H679L683 147C683 41 680 35 598 28V0H861Z"></path></g><g transform="matrix(.013,0,0,-0.013,11.583,0)"><path d="M631 18C609 24 585 35 559 65C534 91 514 117 478 169C448 214 406 281 389 313C462 346 516 399 516 485C516 545 490 590 449 616C412 641 363 650 290 650H42V622C120 615 128 612 128 527V125C128 40 120 34 38 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253Z"></path></g><g transform="matrix(.0091,0,0,-0.0091,35.22,3.132)"><path d="M298 36L289 62C276 55 253 45 228 45C202 45 169 60 169 141V397H276C289 405 292 426 282 437H169V574L155 576L90 509V437H45L17 408L21 397H90V107C90 28 125 -12 188 -12C198 -12 213 -8 230 1L298 36Z"></path></g></svg></span></td><td class="align_center">h</td><td class="align_center">3.29 ± 0.36</td><td class="align_center">2.66 ± 0.50</td><td class="align_center">8.71 ± 0.67</td><td class="align_center">3.28 ± 0.39</td></tr><tr><td class="align_left"><span style="width: 45.3275ptpx;"><svg height="12.0575pt" id="M38" style="vertical-align:-3.2911pt" version="1.1" viewbox="-0.0498162 -8.7664 45.3275 12.0575" width="45.3275pt" xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink"><g transform="matrix(.013,0,0,-0.013,0,0)"><path d="M861 0V28C774 35 771 41 768 147L759 509C756 612 762 614 851 622V650H681L449 149L221 650H57V622C148 613 153 609 144 479L130 271C123 166 117 123 111 88C104 46 85 34 26 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class="align_center">2.88±0.63</td><td class="align_center">8.87 ± 0.68</td><td class="align_center">3.41 ± 0.39</td></tr><tr class="table-tr"><td colspan="6"><hr class="tbody-hr"/></td></tr></table></td></tr></table>

<div>The pharmacokinetic parameters of militarine and its three metabolites after the single oral dose (60 mg/kg) of militarine to rats (n = 6, mean ± SD).</div>

Evidence-Based Complementary and Alternative Medicine

tab6

Table 6

Table 6: A Validated HPLC-MS/MS Method for Simultaneous Determination of Militarine and Its Three Metabolites in Rat Plasma: Application to a Pharmacokinetic Study 

Table 6 | A Validated HPLC-MS/MS Method for Simultaneous Determination of Militarine and Its Three Metabolites in Rat Plasma: Application to a Pharmacokinetic Study 