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 0 100 150 200 250 300 350 400 450 500 550 600 650 700 750

α-Cortol

% 343 100 253 117 147 433523 191 279 369 4606576 625 50 0 100 150 200 250 300 350 400 450 500 550 600 650 700

6α-Hydroxy-20α-cortolone

% 537 100 147 267 103 243 357 447 639 191 369 563 666 74 0 100 150 200 250 300 350 400 450 500 550 600 650 700750

6α-Hydroxy-20β-cortolone

% 100 243 537 147 639 103 191 267 357 447 74 0 369 459 562 652 100 150 200 250 300 350 400 450 500 550 600 650 700750

1β-Hydroxy-20β-cortolone

% 103 147 196 50 217 445 271 652 355 492 535 562 605 685 752 0 100 150 200 250 300 350 400 450 500 550 600 650 700 750

Stigmasterol

% 100 129 255 394 119 355 484 173 282 431 529 576 650 0 607 100 150 200 250 300 350 400 450 500 550 600 650 700 188 Appendix: Common Steroid Identification—GC-MS Mass Spectra

3β,16α,17α-Trihydroxy-5α pregnane-7,20-dione (‘607’)

% 100 607

117 147 217 258 296476 517 0 352 3865427 77 650 100150 200 250 300 350 400450 500550 600650 700

Prednisolone

% 100.0 262

75.0 352 603 50.0 103 513 149 175 293 423 25.0 391 482 573 0.0 662 726 770 840 878 922 995 100 200 300 400 500 600 700 800 900 1000

6-Hydroxyprednisolone

% 100.0 632

75.0 219 50.0 380 103 25.0 133 350 542 691 191 289 421 470 601 707 774 808 874 998 0.0 100 200 300 400 500 600 700 800 900 1000

Cortisol (two peaks)

% 100 605 103 402 515 147 191 245 296 361 425 0 575 683 713 7 100 150 200 250 300 350 400 450 500 550 600 650 700 7

18-Hydroxycortisol

% 693 100 103 513 603 147 246 423 191 279 344 384 482 575 724 0 783 100 150 200 250 300 350 400 450 500 550 600 650 700 750 8 Appendix: Common Steroid Identification—GC-MS Mass Spectra 189

6β-Hydroxycortisol

% 693 100 103 513 603 147 246 423 191 279 336 384 458 575 724 0 100 150 200 250 300 350 400450 500550 600650 700

Cholesteryl butyrate (internal standard)

% 368 100 147 105 260 353 213 274 412 4746516 574605 96 0 100 150 200 250 300 350 400450 500550 600650 700 Index

A primary adrenal insufficiency, Adrenoleukodystrophy protein 167–169 (ALDP), 169 secondary adrenal insufficiency, 170 Acquired 11β-HSD deficiency steroid profile, 173–175 Cushing’s syndrome, 123 tertiary adrenal insufficiency, 170 itraconazole, 124 treatment, 175 liquorice, 123 type 1 diabetic, 173 posaconazole, 124 Adrenocortical carcinoma, pregnancy, 124 24, 25, 150 renal and liver disease, 123 Adrenocortical pathophysiology triazole antifungal drugs, 124 adrenal adenoma, incidentaloma Acquired adrenal hyperplasia, 147 and adrenocortical carcinoma, Addison’s disease, 97, 170–172 24, 25 Adolescent period, 11 adrenal androgen excess, 24 Adrenal adenoma, 24, 25, 150 glucocorticoid excess, 23 Adrenal androgen, 4, 10, 19, 19, 19, 22, 75, steroids and homeostasis, 18 125, 127, 164 adrenal androgens, 19 Adrenal androgen excess, 24 aldosterone, 18 Adrenal carcinomas, 148 exogenous steroids, 20 Adrenal cortex, 3, 5, 10, 17 glucocorticoids, 19 Zona reticularis, 3, 18, 19, 67, 76, 131 mineralocorticoid excess, Adrenal deficiency 23, 24 CAH, 21, 22 types of, 20 pan-adrenal hypofunction, 22 congenital adrenal hyperplasia, Adrenal glands, 3, 8, 9, 17, 88, 137, 147, 20, 22 168, 172 inborn errors, effect of, 21 adolescent period, 11 pan-adrenal hypofunction, 22 adult adrenal, structure and function of, 3, 4 Adrenocorticotropic hormone (ACTH) fetal adrenal, 8–10 deficiency, 4, 41, 167, 170 neonatal period, 10, 11 Adrenocorticotropic hormone stimulation Adrenal hypoplasia, 10, 173 3βHSD deficiency, 69 Adrenal incidentaloma, 24, 151 11β-hydroxylase deficiency, 60, 61 Adrenal insufficiency Adrenogonadal primordium (AGP), 9 diagnosis, 171–173 Adrenoleukodystrophy, 169 genetics, 171 Adult adrenal, structure and function of, negative feedback, 167 3, 4 presentation, 171 Adult adrenal cortex, 17

AKR1C3, see 17βHSD type 5 Cholesterol 20-22 lyase, 86 Aldo-keto reductase (AKR1C) family, 14 Cholesterol biosynthesis Aldosterone, 3, 17, 18 cytochrome P450 oxidoreductase insufficiency, 53 diagnosis, 141 Aldosterone synthase deficiency, 7 genetics, 141 vs. adrenal steroid biosynthesis, 93 impaired androgen synthesis, 140 biochemical features, 94 presentation, 140 biosynthesis, 93 steroid profile, 144, 145 clinical and biochemical features, 96 treatment, 146 coding sequence, 93 esterification, 137 corticosterone metabolites, 99 estimated frequency, 138 corticosterone methyl oxidase type 1 and hydrolyse cholesteryl esters, 137 type 2, 94 low-density lipoprotein, 137 diagnosis, 96–97 multistep process, 137 genetics, 95–96 pathway, 137, 138 metabolites, 99 Smith-Lemli-Opitz syndrome mineralocorticoid effect, 93 clinical variability, 139 presentation, 94–95 diagnosis, 141 renin-angiotensin-aldosterone system, 94, 95 genetics, 141 treatment, 101 in utero diagnosis, 143 urine steroid profile, 97, 100, 101 incidence, 139 Allgrove syndrome, see Triple A deiciency inherited defects, 139 Alpha-blockers, 151 presentation, 140 Amberlite XAD-2 resin, 29 severity, 139 Amenorrhoea, 82, 89, 117 steroid profile, 142, 143 Androstenedione, 4, 77 treatment, 145–146 Anorexia nervosa, 115 sterol response element-binding Anti-Müllerian hormone (AMH), 104, 105, proteins, 137 111, 114 sterol synthesis, 137 Antley-Bixler syndrome (ABS), 139 Wolman’s disease, 139 Apparent cortisone reductase deficiency, 122 Cholesterol side-chain cleavage enzyme Apparent mineralocorticoid deficiency (CYP11A1), 5 (AME), 123 Chronic hypercortisolism, 115 Arylsulphatase C, see Steroid sulphatase (STS) Combined pituitary hormone deficiencies ATP-binding cassette subfamily D member 1 (CPHD), 170 (ABCD1) gene, 169 Compound B, 6 Autoimmune polyglandular syndromes Compound S, 6 (APS), 168 Congenital adrenal hyperplasia (CAH), 20, Autoimmune regulator protein (AIRE), 168 22, 41, 106 Autonomous aldosterone excess, 151 17-hydroxylase, 21 Autosomal recessive inheritance, 113 21-hydroxylase, 21 3β-hydroxysteroid dehydrogenase, 21 20-22 lyase, 21 B StAR deficiencies, 21 Backdoor pathway, 43 CYP11B1 deficiency, 55 Bartter and Gitelman syndromes, 24 salt-losing form of, 70 Beckwith-Wiedemann syndrome, 151 Non-salt-losing forms of, 67 Benzyloxime derivatives (BO-TMS), 30 Conjugates, hydrolysis of, 29, 30 Conjugation, 15 Conn syndrome, 23 C Corticosterone methyl oxidase (CMO) type 1 Calcium channel blockers, 151 and type 2, 94 cAMP/protein kinase A pathway, 150 Corticotrophic-releasing hormone Cholesterol, 86 (CRH), 4 Index 193

Cortisol, 4, 19 E Cortisol-producing adenomas, 150 Electrolyte disturbances, 56 Cushing’s syndrome, 23, 115, Exogenous steroids, 20 123, 147 Expression of masculinity score causes, 159 (EMS), 112 clinical and diagnostic features, 160 cortisol excess, 160 diagnosis, 162–163 F endogenous glucocorticoid excess, 159 Familial adenomatous polyposis, 151 genetic defects, 161 Familial glucocorticoid deficiency, 169 hypothalamic corticotrophin-releasing Fetal adrenal zone, 17 hormone, 159 Fibroblast growth factor receptor 2 negative feedback system, 160 (FGFR2), 139 pituitary adrenocorticotrophic Finasteride, 112, 119, 166 hormone, 159 Fluconazole, 62 presentation, 161 Fludrocortisone, 53, 89, 90 screening guidelines, 161 Fetal adrenal, 8–10, 66 steroid profile, 164, 165 Fetal neocortex, 76 treatment, 165, 166 CYP11A1 deficiency, 86, 88, 89 CYP11B1, 23 G CYP11B1-B2 hybrid construct, 96 Gas chromatography/mass spectrometry CYP11B2 deficiency, 101 (GC/MS), 31, 32 CYP17A1 gene, 78 Glucocorticoid(s), 2, 19, 20, 53, 72, 84, 87, CYP21, 41 122–124, 140, 159, 173 CYP21A2, 45, 46 Glucocorticoid excess, 23 CYP3A family, 15 Glucocorticoid remedial hyperaldosteronism Cytochrome P450 oxidoreductase (POR) (GRA), 57, 96 diagnosis, 141 β-Glucuronidase, 29 genetics, 141 Glycerol kinase deficiency, 171 impaired androgen synthesis, 140 Gonadal steroid synthesis, 76 presentation, 140 Gonadectomy, 115 steroid profile, 144, 145 Gonadotrophin analogues, 156 treatment, 146 Gonane, 1

D H DAX1, see Nuclear receptor 0B1 HSD3B2 gene, 67 (NROB1) Human chorionic gonadotrophin Dehydroepiandrosterone (DHEA), (hCG), 79 4, 42, 75 Human testicular testosterone Dehydroepiandrosterone sulphate (DHEAS), synthesis, 105 hydrolysis of, 29 Hydrocortisone, 51, 53 Deoxycorticosterone (DOC), 22, 56, 77 Hydroxime (HO), 30 Derivatisation, 30, 31 Hydroxylation, 2, 5, 6, 14, 15, 50, 86 Dexamethasone, 90 11βHSD1, 14 46XY disorder of sex development 11β-Hydroxylase deficiency, 7, 55 (DSD), 67, 77, 104 11-deoxycortisol, conversion of, 56 Drug inhibition, of 11β-hydroxylase ACTH stimulation test, 60, 61 deficiency, 62 CAH, 55 Duchenne muscular dystrophy, 171 diagnosis, 58, 59 Dutasteride, steroid profile, 119 drug inhibition of, 62 194 Index

11β-Hydroxylase deficiency cont.( ) Hydroxysteroid dehydrogenases (HSDs), genetics, 57, 58 7, 9 metabolic block causes, 57 3β-Hydroxysteroid dehydrogenase/ presentation, 56 isomerase (3βHSD) deficiency, steroid profiling, 60–62 18, 35, 65, 67 treatment, 63 diagnosis, 68, 69 urine steroid profile, 60 differential diagnosis, 70 17α-Hydroxylase deficiency, 75 genetic testing, 71 diagnosis, 78–80 genetics, 67 differential diagnosis, 79 inhibitors, 72 functions, enzyme, 75–77 metabolites, 72 genetics, 78 presentation, 67 inhibitors, 84 steroid profiling, 70–72 presentation, 77, 78 treatment, 72 primary amenorrhoea and pubertal 11β-Hydroxysteroid dehydrogenase (11β- delay, 82 HSD) deficiency steroid profile, 80, 82, 83 acquired treatment, 84 Cushing’s syndrome, 123 urine steroid profile, 81 itraconazole, 124 11β-Hydroxyandrostenedione, 43 liquorice, 123 17β-hydroxysteroid dehydrogenase (17βHSD) posaconazole, 124 deficiency pregnancy, 124 diagnosis, 106–107 renal and liver disease, 123 disorders of sex development, 104 triazole antifungal drugs, 124 enzyme deficiency, 104 apparent cortisone reductase genetics, 106 deficiency, 122 in human steroid hormone biosynthesis, apparent mineralocorticoid deficiency, 123 103, 104 cortisol clearance, 121 in human testicular testosterone diagnosis, 125 synthesis, 105 genetics, 124–125 metabolites, 109 mineralocorticoid receptor, 122 NR5A1, 104 presentation, 124 presentation, 105–106 reversible interconversion, 121, 122 with primary amenorrhoea, 107 steroid profile, 126–128 retention times, 109 treatment, 128 treatment, 109 Hyperaldosteronism, 57, 149, 151 urine steroid profile, 108, 109 Hyperandrogenism, 84, 133, 148, 156, 173 Wolffian duct development, 105 Hypercortisolism, 23, 148, 150, 163 17β-hydroxysteroid dehydrogenase (17βHSD) Hyperreninaemic hypoaldosteronism, 90, 97 type 5, 103 Hypertension, 56, 60, 77, 84 21-Hydroxylase deficiency, 6, 41, 173 Hyponatraemia, 87–89, 97 17OHP, response to ACTH Hypothalamic-pituitary-adrenal (HPA) stimulation, 47 axis, 121 androgens, source of, 42, 43 causes, 41 diagnosis, 46, 47 I genetics, 45, 46 Immunoassays, 20, 28, 79, 114, 163 presentation, 45 Inborn error of metabolism, 22 steroid profile, 48–52 Incidental adrenal mass, 151 treatment, 51, 53 Incidentaloma, 24, 25, 147 urinary metabolites, orgin, 44 Internal standards (IS), 32, 33, 51 Index 195

L P Li-Fraumeni (TP53 mutations), 151 P450 oxidoreductase (POR) deficiencies, Lipoid congenital adrenal hyperplasia 41, 77 (LCAH), 86 Pan-adrenal hypofunction, 22 amenorrhoea, 89 PAPSS2 deficiency, see Steroid clinical presentation, 87, 88 sulphotransferases diagnosis, 89 Peripheral hormone, 12 genetics, 88 Phaeochromocytoma, 152 steroid profile, 89, 90 Pituitary macroadenoma, 170 treatment, 90 Polycystic ovarian syndrome, 115 Liquid chromatography/MS (LC–MS/ Posaconazole, 62 MS), 28 Prednisolone, 50, 51, 53, 90, 169, 175 Liver disease, 37–39 Pregnancy, 36, 87, 116, 124 Low-density lipoproteins (LDL), 85 Pregnanediol, 36, 37, 144 Lynch syndrome, 151 Pregnenolone, 5, 10, 65, 79, 82 Primary adrenal insufficiency (PAI), 167–169 M Primary aldosteronism (PA), 23 Maternal urinary oestriol, 132 Progesterone, 4–6, 42, 53, 77, 87, 124, 153 Medroxyprogesterone, 72 Prostatic hyperplasia, 112 Menstrual cycle, 36 Pseudohyperaldosteronisms (PHA), 24 Methoxyamine and N-trimethylsilylimidazole (MO–TMS derivatives), 30 Methylene units (MU), 31 Q Metyrapone, 59 Quad test, 134 Mifepristone, 156 Mineralocorticoid, 17, 18 Mineralocorticoid effect, 93, 95 R Mineralocorticoid excess, 23, 24 Renin, 96 Mineralocorticoid receptors (MR), 18 Renin–angiotensin–aldosterone system Mineralocorticoid replacement (RAAS), 18, 94, 95 therapy, 101 Mitotane, 119, 155, 156 Modified Weiss system, 149 S Multiple endocrine neoplasia type 1 Salt wasting, 46, 67, 89 (MENIN), 151 Secondary adrenal insufficiency, 170 Sephadex LH-20, 29 Smith-Lemli-Opitz syndrome (SLOS) N clinical variability, 139 Neonatal period, 10, 11 diagnosis, 141 Nicotinamide nucleotide transhydrogenase genetics, 141 (NNT), 169 in utero diagnosis, 143 Non-classical LCAH, 88 incidence, 139 Nuclear receptor 0B1 (NROB1), 10, 169 inherited defects, 139 presentation, 140 severity, 139 O steroid profile, 142, 143 Oestrogens, 36 treatment, 145–146 17-hydroxyprogesterone (17OHP), 5, 48, 48 Sodium-dependant organic anion transporter Organic acid transporter polypeptides (SOAT), 131 (OATPs), 130 Solid packing materials, 29 196 Index

Solid-phase extraction (SPE), 28 aldosterone synthase deficiency, 100, 101 Sonic hedgehog (SHH) protein, 139 Cushing’s syndrome, 164, 165 Spironolactone, 128, 156 GC/MS, 31, 32 Sterol response element-binding proteins LC–MS/MS, 28 (SREBPs), 137 mitotane, 156 Sterane, 1 physiology and pathophysiology, 27 Steroids StAR, 89, 90 HSDs, 7, 9 steroid sulphotransferases, 135 metabolism and excretion, 12, 13 steroid-producing tumours, 153–155 conjugation, 15 STS hydroxylation, 14, 15 postnatal findings, 135 oxidation and reduction, 14 prenatal findings, 134 reduction of the Δ4 bond, 12 Interpretation, 32, 33 steroid centre, oxidation and Steroid sulphatase (STS), 134, 135 reduction, 14 case study, 134 terminals, oxidation and reduction, 14 diagnosis, 133 structures and nomenclature, 1–3 fetal deficiency, 132 synthesis, 4–7 fetoplacental unit, 131 Steroid 5-alpha-reductase deficiency formylglycine residue, 131 anorexia nervosa, 115 prenatal diagnostic test, 132 chronic hypercortisolism, 115 prevalence, 132 definitive diagnosis, 114 steroid profile genetics, 113 postnatal findings, 135 gonadectomy, 115 prenatal findings, 134 impaired androgen synthesis and Steroid sulphotransferases (SULT) action, 113 case study, 134 in normal individuals, 115 diagnosis, 133 late presentation, 117–118 expression of, 131 polycystic ovarian syndrome, 115 fetoplacental unit, 131 presentation, 112–113 OATPs, 130 prostatic hyperplasia, 112 postnatal findings, 135 reaction catalyse, 111, 112 presentation, 132 testicular Leydig cells, 111 primary substrates, 130 testicular-derived testosterone, 115 types, 130 thyroxine, 115 Steroidogenic acute regulatory protein (StAR) treatment, 119 clinical presentation, 87, 88 type 1 and 2 enzymes, 111 diagnosis, 88, 89 urine steroid profile, 114, 117–119 genetics, 88 Steroid hormone replacement therapy, 20 steroid profile, 89, 90 Steroid profile treatment, 90 changes in profile, 33–35 Steroidogenic acute regulatory protein with age, 33 (StAR)-related lipid transfer liver disease, 37–39 (START) domain, 85 pregnancy and menstrual cycle, 36 Steroidogenic factor 1 (SF1), 22 conjugates, hydrolysis of, 29, 30 Steroid-producing tumours derivatisation, 30, 31 acquired adrenal hyperplasia, 147 3βHSD deficiency, 70–72 adrenal carcinomas, 148 11β-hydroxylase deficiency, 60–62 adrenal incidentaloma, 152 21-hydroxylase deficiency, 48–52 biochemical assessment, 149 11β-hydroxysteroid dehydrogenase Cushing’s disease, 147 (11β-HSD) deficiency, 126–128 diagnosis, 151–152 17β-hydroxysteroid dehydrogenase ectopic ACTH secretion, 147 deficiency, 108 genetics, 150 adrenal insufficiency, 173–175 histology, 149 Index 197

hypercortisolism, 148 Tetrahydrocortisone (THE), 32, 35 imaging, 148 Tetrahydrodeoxycorticosterone (THDOC), 59, 83 incidentalomas, 147 Trimethylsilyl ethers (TMS), 30 modified Weiss system, 149 Triple A deficiency, 170 presentation, 150 rapid-onset virilisation, 148 steroid profile, 153–155 U testicular adrenal rest tumours, 148 Unconjugated oestrogens, 132 testicular tumours, 148 Upper most layer (ZG), 3 treatment, 155, 156 Synacthen stimulation test, 47, 82, 172, 173 V Virilisation, 56 T T box 19 transcription factor (TBX19), 170 Tertiary adrenal insufficiency, 167, 170 W Testicular adrenal rest tumours (TARTs), 45, 148 Wnt/β catechin pathway, 150 Testicular tumours, 148 Wolffian duct development, 105 5α-Tetrahydrocortisol (5αTHF), 30 Wolman’s disease, 139