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Appendix: Common Steroid Identification—GC-MS Mass Spectra Gary Woodward, Francis Lam, and Gill Rumsby As described in Chap. 3, steroid profiling in the context of steroidogenic conditions is performed by GC-MS analysis following conjugate hydrolysis and derivatisation. Below are the mass spectra used for identification of urine steroids described throughout this book, and compiled during routine practice within our laboratory. Steroids are given in approximate order of their retention times. Androstanediol (internal standard) % 100 129 256 346 107 241 331 436 215 287 372 484 516 569 637 677 0 100 150 200 250 300 350 400 450 500 550 600 650 70 Pregnadienol % 129 50 243 267 372 173 357 0 421 494 525 556 620 665 7 100 150 200 250 300 350 400 450 500 550 600 650 70 G. Woodward · G. Rumsby Department of Clinical Biochemistry, University College London Hospitals, London, UK F. Lam Level 2-Chemistry, Health Services Laboratories, London, UK © Springer International Publishing AG, part of Springer Nature 2019 177 G. Rumsby, G. M. Woodward (eds.), Disorders of Steroidogenesis, https://doi.org/10.1007/978-3-319-96364-8 178 Appendix: Common Steroid Identification—GC-MS Mass Spectra Androsterone % 100 270 360 107 147 213 253 362 0 434 461 546 610 687 100 150 200 250 300 350 400 450 500 550 600 650 700 Aetiocholanolone % 100 270 360 105 131 213 422 0 255 362 460 506 564 644 679 100 150 200 250 300 350 400 450 500 550 600 650 700 Dehydroepiandrosterone % 100 129 268 260 358 105 374 0 211 432 459 523 592 642 682 7 100 150 200 250 300 350 400 450 500 550 600 650 700 Androstenediol % 100 129 239 119 215 344 305 434 0 389 464 531583 623 673 100 150 200 250 300 350 400 450 500 550 600 650 700 17-Hydroxypregnanolone (peak 1) % 100 476 100 156 188 386 255 296 364 507 0 443 574 609 694 100 150 200 250 300 350 400 450 500 550 600 650 700 11β-Hydroxyandrosterone % 100 448 358 105125 213 268 253 0 374 476 508 564 634 664 100 150 200 250 300 350 400 450 500 550 600 650 700 Appendix: Common Steroid Identification—GC-MS Mass Spectra 179 11β-Hydroxyaetiocholanolone % 100 448 268 125 213 358 105 253 388 0 464 537558 608 693 100 150 200 250 300 350 400 450 500 550 600 650 700 17-Hydroxypregnanolone (peak 2) % 476 100 100 156 188 364 386 255 296 443 507 574 609 694 0 100 150 200 250 300 350 400 450 500 550 600 650 700 16α-Hydroxydehydroepiandrosterone (two epimers) % 100 446 129 266 356 105 174 462 258 372 564 0 506 649 674 7 100 150 200 250 300 350 400 450 500 550 600 650 700 15,17-Dihydroxypregnanolone % 100 258 474 564 384 105 147188 294 506 0 317 433 610 688 719 100 150 200 250 300 350 400 450 500 550 600 650 700 750 Enterodiol % 100 410 180 231 103 147 500 0 294 354 409 575 648 693 100 150 200 250 300 350 400 450 500 550 600 650 700 Pregnane-3α,20α-diol % 117 10 269 147 175 347 449 0 374 474 552 644 674 704 100 150 200 250 300 350 400 450 500 550 600 650 700 180 Appendix: Common Steroid Identification—GC-MS Mass Spectra Enterolactone % 100 180 442 105 165 263 0 309 335 409 462 537 562 632 696 100 150 200 250 300 350 400 450 500 550 600 650 700 16β-Hydroxydehydroepiandrosterone (two epimers) % 100 446 129 266 356 105 462 174 239 372 0 529 559 642 674 100 150 200 250 300 350 400 450 500 550 600 650 700 Pregnanetriol % 100 117 255 435 147 345 173 308 552 0 403 464 608 672 100 150 200 250 300 350 400 450 500 550 600 650 700 5-Pregnene-3β,20α-diol % 117 50 418 129 460 211 238 282 328 372 0 550 610 694 100 150 200 250 300 350 400 450 500 550 600 650 700 16-Oxoandrostenetriol and 15β,16α-dihydroxydehydroepiandrosterone % 446 100 158 462 117 266 356 565 232 405 534 606 651 701 0 100 150 200 250 300 350 400 450 500 550 600 650 700 Androstenetriol % 129 239 432 100 117 329 402 213 303 522 567 607 653 0 100 150 200 250 300 350 400 450 500 550 600 650 700 Appendix: Common Steroid Identification—GC-MS Mass Spectra 181 Tetrahydro-11-deoxycortisol % 100 564 474 103 147 255 294 384 0 168 343 433 536 595 643676 100 150 200 250 300 350 400 450 500 550 600 650 700 Tetrahydrodeoxycorticosterone (THDOC) % 100 476 271 117 147 188 241 327 361 386 420 507 0 564 599 663 69 100 150 200 250 300 350 400 450 500 550 600 650 70 16,18-Dihydroxydehydroepiandrosterone (peak 1) % 100 103 444 129 283 534 252 388 564 183 342 504 0 624 654 70 100 150 200 250 300 350 400 450 500 550 600 650 70 5-Pregnene-3α,16α,20α-triol % 117 50 147 460 196 445 241 282 355 544 566 625 679 0 100 150 200 250 300 350 400 450 500 550 600 650 700 16,18-Dihydroxydehydroepiandrosterone (peak 2) % 100 103 444 129 283 534 252 388 564 183 342 504 0 624 654 70 100 150 200 250 300 350 400 450 500 550 600 650 700 Oestriol % 117 25.0 311 205 345 386 504 231 414 506 0.0 584 614 693 100 150 200 250 300 350 400 450 500 550 600 650 700 182 Appendix: Common Steroid Identification—GC-MS Mass Spectra 11-Oxopregnanetriol % 449 100 359 147 117 269 187 227 0 402 461 566 620 675 70 100 150 200 250 300 350 400 450 500 550 600 650 700 16α-Hydroxypregnenolone % 474 50 129 188 384 433 0 253 294 327 536 567 607 652 100 150 200 250 300 350 400 450 500 550 600 650 700 Hexahydro11-deoxycortisol (peak 1) % 100 255 243 435 117 345 488 537 578 0 191 304 398 609 640 686 100 150 200 250 300 350 400 450 500 550 600 650 700 5-Pregnene-3β,17α,20α-triol % 100 253 433 117 343 147 211 0 281 370 460 550 575 622 652 100 150 200 250 300 350 400 450 500 550 600 650 700 5-Androstene-3β, 15β,16α,17β-tetrol % 191 100 105 147 235 305325 405 479 520 595 666 0 100 150 200 250 300 350 400 450 500 550 600 650 700 6-Hydroxy-tetrahydro-11-deoxycortisol % 652 100 103 147 562 202 253276 343 382 418 472 521 624 741 0 100 150 200 250 300 350 400 450 500 550 600 650 700 750 Appendix: Common Steroid Identification—GC-MS Mass Spectra 183 3β,16α-Dihydroxypregnatrien-20-one % 188 290 100 235 129 340 380 119 411 501 533 567 607 69 0 100 150 200 250 300 350 400 450 500 550 600 650 700 3β,16α-Dihydroxy-5,8(9)-pregnadien-20-one % 100 188 398 235 308 119 156 325 488 0 415 544 574 624 679 699 100 150 200 250 300 350 400 450 500 550 600 650 700 3β,16α-Dihydroxy-5,7-pregnadien-20-one % 382 100 188 129 251 292 341 431 488 521 560 635 684 0 100 150 200 250 300 350 400 450 500 550 600 650 700 Hexahydro-11-deoxycortisol (peak 1) % 255 100 435 105 147 191 243 297 345 398 488 537 578 609 640 676 0 100 150 200 250 300 350 400 450 500 550 600 650 700 Tetrahydrocortisone (THE) % 578 100 488 103 398 580 147168 258 308 357 447 626 0 100 150 200 250 300 350 400 450 500 550 600 650 700 Hexahydro-11-deoxycortisol (peak 2) % 255 100 243 435 345 117 191 304 398 488 537 578 609 640 0 100 150 200 250 300 350 400 450 500 550 600 650 700 184 Appendix: Common Steroid Identification—GC-MS Mass Spectra 5-Androstene-3β, 15β, 16α,18-tetrol % 431 100 117 391 521 147 191 251269 341 472 562 638 0 100 150 200 250 300 350 400 450 500 550 600 650 700 Tetrahydro-11-dehydrocorticosterone (THA) % 188 100 103 400 431 147 241 490 521 310328 564 621 681 0 100 150 200 250 300 350 400 450 500 550 600 650 700 Tetrahydroaldosterone hemiacetal % 506 100 103 147 191 239 280 341 370 416 474 594 667 705 0 100 150 200 250 300 350 400 450 500 550 600 650 700 Tetrahydrocorticosterone (THB) % 188 100 103 474 143 312 384 564 228 356 448 506 638 683 0 100 150 200 250 300 350 400 450 500 550 600 650 700 Allo-tetrahydro-11-dehydrocorticosterone (alloTHA) % 188 100 490 105 147 241 282 359 400 418 521 560 606 650 0 100 150 200 250 300 350 400 450 500 550 600 650 700 Allo-tetrahydrocorticosterone (alloTHB) % 188 100 474 103 143 564 239 312 384 446 535 648 678 0 100 150 200 250 300 350 400 450 500 550 600 650 700 Appendix: Common Steroid Identification—GC-MS Mass Spectra 185 Tetrahydrocortisol (THF) % 652 100 103 562 147 246 276 382 472 202 343 432 534 638 0 100 150 200 250 300 350 400 450 500 550 600 650 700 5α-Tetrahydrocortisol (5α-THF) % 652 100 103 562 147 246 276 472 172 343 382 432 537 625 7 0 100 150 200 250 300 350 400 450 500 550 600 650 700 α-Cortolone % 100 449 359 269 103 147 191 243 551 0 371 461 654 705 100 150 200 250 300 350 400 450 500 550 600 650 700 18-Hydroxy-11-dehydrocorticosterone (18OH THA) % 103 100 267 578 147 188 457 253 339 398 429 506 609 652 0 714 740 100 150 200 250 300 350 400 450 500 550 600 650 700 750 β-Cortolone, β-cortol % 100 449 147 243 359 103 269 551 191 371 461 654 0 554 625 100 150 200 250 300 350 400 450 500 550 600 650 700 Hexahydro-11-dehydrocorticosterone (HHA 20α) % 100 283 373 103 147 175 265 343 463 0 433 566 602 651 100 150 200 250 300 350 400 450 500 550 600 650 700 186 Appendix: Common Steroid Identification—GC-MS Mass Spectra Hexahydro-11-hydroxycorticosterone (HHB) % 267 100 129 357 185 225 372 447 491 549 593 614 674 0 100 150 200 250 300 350 400 450 500 550 600 650 700 6α-Hydroxy THE % 666 100 103 147 576 195 267 396 486 243 355 431 506 607 698 0 100 150 200 250 300 350 400 450 500 550 600 650 700 750 6α-Hydroxytetrahydro-11-dehydrocorticosterone (6αOH THA) % 188 100 103 519 129 329 398 504 248 278 429 578 609 650 0 100 150 200 250 300 350 400 450 500 550 600 650 700 18-Hydroxy tetrahydrocorticosterone (18OH THB) % 103 100 147 652 472 531 562 188 253 342 382 444 292 609 740 0 100 150 200 250 300 350 400 450 500 550 600 650 700 750 Cholesterol % 100 129 105 329 368 458 519 188 255 578 609 0 650 100 150 200 250 300 350 400 450 500 550 600 650 700 Tetrahydroaldosterone % 506 100 103 147 207 241 267 341377 416 489 576 610 666 714 0 100 150 200 250 300 350 400 450 500 550 600 650 700 75 Appendix: Common Steroid Identification—GC-MS Mass Spectra 187 Hexahydro-tetrahydrocorticosterone (HHB) % 267 357 100 147 103 560 191 253 382 4475470 31 652 7327 0 100 150 200 250 300 350 400
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    Br J Clin Pharmacol 1998; 45: 107–114 Differential selectivity of cytochrome P450 inhibitors against probe substrates in human and rat liver microsomes Victoria A. Eagling, John F. Tjia & David J. Back Department of Pharmacology & Therapeutics, University of Liverpool, P.O. Box 147, Liverpool L69 3GE, UK Aims Chemical inhibitors of cytochrome P450 (CYP) are a useful tool in defining the role of individual CYPs involved in drug metabolism. The aim of the present study was to evaluate the selectivity and rank the order of potency of a range of isoform-selective CYP inhibitors and to compare directly the eVects of these inhibitors in human and rat hepatic microsomes. Methods Four chemical inhibitors of human cytochrome P450 isoforms, furafylline (CYP1A2), sulphaphenazole (CYP2C9), diethyldithiocarbamate (CYP2E1), and ketoconazole (CYP3A4) were screened for their inhibitory specificity towards CYP- mediated reactions in both human and rat liver microsomal preparations. Phenacetin O-deethylation, tolbutamide 4-hydroxylation, chlorzoxazone 6-hydroxylation and testosterone 6b-hydroxylation were monitored for enzyme activity. Results Furafylline was a potent, selective inhibitor of phenacetin O-deethylation = (CYP1A2-mediated) in human liver microsomes (IC50 0.48 mm), but inhibited both phenacetin O-deethylation and tolbutamide 4-hydroxylation (CYP2C9- mediated) at equimolar concentrations in rat liver microsomes (IC50=20.8 and 24.0 mm respectively). Sulphaphenazole demonstrated selective inhibition of tolbutamide hydroxylation in human liver microsomes but failed to inhibit this reaction in rat liver microsomes. DDC demonstrated a low level of selectivity as an inhibitory probe for chlorzoxazone 6-hydroxylation (CYP2E1-mediated). DDC also inhibited testosterone 6b-hydroxylation (CYP3A-mediated) in man and rat, and tolbutamide 4-hydroxylase activity in rat.
  • The Role of the Enterohepatic Circulation of Bile Salts and Nuclear Hormone Receptors in the Regulation of Cholesterol Homeostasis: Bile Salts As Ligands for Nuclear Hormone Receptors

    The Role of the Enterohepatic Circulation of Bile Salts and Nuclear Hormone Receptors in the Regulation of Cholesterol Homeostasis: Bile Salts As Ligands for Nuclear Hormone Receptors

    Redinger.qxd 3/28/03 11:21 AM Page 265 ORIGINAL ARTICLE The role of the enterohepatic circulation of bile salts and nuclear hormone receptors in the regulation of cholesterol homeostasis: Bile salts as ligands for nuclear hormone receptors Richard N Redinger MD RN Redinger. The role of the enterohepatic circulation of bile Le rôle de la circulation entérohépatique des salts and nuclear hormone receptors in the regulation of sels biliaires et des récepteurs de l’hormone cholesterol homeostasis: Bile salts as ligands for nuclear hormone receptors. Can J Gastroenterol 2003;17(4):265-271. nucléaire dans la régulation de l’homéostasie cholestérolique : Les sels biliaires comme The coordinated effect of lipid activated nuclear hormone receptors; ligands des récepteurs de l’hormone nucléaire liver X receptor (LXR), bound by oxysterol ligands and farnesoid X receptor (FXR), bound by bile acid ligands, act as genetic transcrip- L’effet coordonné des récepteurs nucléaires de l’hormone activée par les tion factors to cause feed-forward cholesterol catabolism to bile acids lipides, du récepteur X hépatique (RXH), lié par les ligands d’oxystérol et and feedback repression of bile acid synthesis, respectively. It is the le récepteur X famésoïde (RXF), liés par les ligands acidobiliaires, agit coordinated action of LXR and FXR, each dimerized to retinoid X comme des facteurs de transcription génétique afin de provoquer, respec- receptor, that signal nuclear DNA response elements to encode pro- tivement, un catabolisme cholestérolique non récurrent des sels biliaires et teins that prevent excessive cholesterol accumulation and bile salt une répression à rétroaction de la synthèse de l’acide biliaire.