index [link.springer.com]978-94-017-3078...index absolute configuration 3, 18, 20 accreditation 663...
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Index
absolute configuration 3, 18, 20 accreditation 663 accuracy 438, 649 see also inaccuracy; bias
accurate mass 94 acetonides, derivatives for mass spectrometry 106
17-acetoxy-6a-methylpregn-4-ene-2,30-dione 14, 83, 88
17 fJ-acetoxyandrost-4-en-3-one 15 acid hydrolysis 429,447, 449
of steroid sulphates 122-124 ACTH test 232, 244, 249 adrenal function test 232 adrenocorticotrophin hormone see ACTH
adrenosterone 51 affini ty 444 affinity chromatography in C19 steroid isolation 321, 335
affinity column, use in extraction of vitamin D metabolites 574
agarose 448 agglutination inhibition 434 albumin 451,452 aldosterone 7, 11, 660, 688, 689, 691
assays for 250-252 excretion rate, measurement of urinary
metabolites 252 18-glucuronide assay 252 synthesis from cholesterol 230
alkaline phosphatase 439 alkane boronates, derivatives for mass spectrometry 106
alkenes 18, 23, 46, 47, 61, 84 alkynes 46, 47, 52, 61, 84 All-Laboratory Trimmed Mean 667, 671,
672,673 allocholane 4 allopregnane 4 allotetrahydrocortisol 690 ALTM see All-Laboratory Trimmed
Mean aluminium oxide, in column chromatography 131-132
amberlite XAD resins 528, 529, 539 amniotic fluid, cortisol in 242 anabolic steroids
biological fluids for analysis of 493
conjugates, hydrolysis of 489--491 derivatives of, and GC-MS 498-509,
511 extraction and purification of 485--492
HPLC of 144 immunoassays, anabolic steroids for
493--497 immunochromatography of 489 in food, extraction of 120 liquid-phase extraction of 487--488 measurement by GC-MS 165 metabolism of 475--485 mixed-phase extraction 488--489 natural hormone administration,
detection of 509-518 preparation of 475 screening analysis of 492-503 solid-phase extraction of 486--487 structural requirements for 474--475 synthetic, confirmatory analysis of
503-509 TLC of 489 trivial, systematic and trade names of 469
undesirable effects of 473--474 use and abuse 468--472 see also individual steroids
analysis, steps in 115 et seq. analytical goals 656, 657 androgen, biosynthetic pathway 230 androgen glucuronides, HPLC-MS of
176 androgen sulphate 446 androgens
biosynthesis of 268-269 characterisation of 280-291, 294-317,
324-332 by GC profiling 301-303 by HPLC 295-301 by MS 305-318 by specific radioactivity measurements 294-295
detection and quantification by UV, NMR 273-274
extraction of 276-279, 280-291, 292, 324-332 by liquid-phase 277-279, 292 by solid-phase 276-277
698
androgens (contd) immunoassays (lAs) for 323-353
chemiluminescent assays 343-346 ELISAs and Els 340-343 f1uorometric assays 346-349 problems with direct methods 336-337
radioimmunoassays 349-353 sample preparation in 334-335
measurement in plasma by GC-MS 164
metabolism of 269-272 nature and sources of 268 physiological actions of 273 purification of 280-291, 292-294,
324-332 by C18 Sep-Pak cartridges 292-293 by Lipidex 293 triethylaminohydroxypropyl (TEAP)-LH20 292-294
separation by TLC 275-276,280-291, 324-332
androst-4-en-19-al-3,17-dione 53 androst-5-ene-3a, 17 fi-diol 17 -acetate 15 androst-5-ene-3fi, 17 fi-diol 17-benzoate
51 androst-5-ene-3a, 17,B-diyl acetate 15 androstane 2, 4, 12 5a-androstane-3a 17 fi-diol 13 5a-androstane-3a,16a-17 fi-triol, characterisation of by GC-MS in Sertoli cells 313
androstanediols derivatives for, in GC-MS 500, 507 formation from testosterone 477 fragmentation of, in GC-MS 505 mass spectra of 503, 506, 508 selected ions for, in GC-MS 501 separation of, from boldenone 508
5a-androstanediols formation of 269-271, 313 glucuronidation of 271 measurement of by GC-MS 280-291,
307, 311-312 RIAs for glucuronides of 324-332,
352-353 5-androstene-5fi,17 fi-diol
assays for 280-291, 324-332 biosynthesis of 269 3-sulphate of RIA for 353 sulpho-conjugation of 352
androstenedione 6, 86, 437, 660, 688, 691 assays for 246-247
4-androstenedione androgenicity of 273 as internal standard 310 assay of by GC-MS 280-291, 314 assay of by lAs 324-332, 342-343
INDEX
formation of 268-269 hydroxy derivatives HPLC of 297 metabolism of 269-270
16-androstenes, measurement by GC-MS 164
androsterone 11, 660, 690 anisotropic effects in NMR spectroscopy 62
anti-estrogens 441 antibodies 433 antibody 652, 658 antigen 439 antigenic protein 436 aromatic steroids 5, 11, 12, 14, 35, 47, 53,
61, 62, 79, 80, 84 ascorbic acid 449 assisted conception 649 AS TED, automated system for cortisol
analysis 239 audit 662 automated immunoanalyser 439, 456 avidity 441 axial bonds 9, 63
Baljet reaction 625 BCR see Community Bureau of Reference
beads, coated 439 benzo-15-crown-5 ether, in LC-MS of vitamin D metabolites 584
Bergmeyer Conferences 650 bias 649, 659, 670 see also inaccuracy bile acid
antiserum 550 breath test 552 glucuronides 527, 528, 545, 547-549 nomenclature 528 sulphates 539, 545-547
bile acid derivatives l-anthroyl nitrite 536 bromoacetylpyrene 536, 540 bromoethyl-7-methoxy coumarin 536 dansylhydrazones 536 I -125 histamine 549 1-125 tyramine 549 methyl esters 545, 548 oximes 537 pre-column 536 trifluoroacetate 548 trimethylsilyl ethers 546-548
bile acid esters, use in HPLC 157 bile acid HPLC, detection by use of 3c£hydroxysteroid dehydrogenase 154-155, 157
bile acid metabolism biosynthesis 527 deconjugation 527 enterohepatic circulation 527, 528
inborn errors 551 primary 527 secondary 527
bile acid methods affinity chromatography 550 breath test 552 enzymatic assay 531-535 enzyme electrode 530 extraction 529 gas-liquid chromatography 545-549 high performance liquid
chromatography 535-545 hydrolysis 546, 548 immunoassay 549-551 mass spectrometry 551-552 SeHCAT 552 separation 530, 535 solvolysis 546, 548
bile acids 8, 11, 68 chenodeoxycholic 527, 528, 549 cholic 527, 528, 549 deoxycholic 527, 528 detection after HPLC 154-155 7-,12-dihydroxy-5-cholanic 538 glycine-conjugated 527, 528, 536, 538 HPLC of 144, 145, 151 hydrolysis of conjugates 124 3,B-hydroxy-5-cholenic acid 530 1-P-hydroxycholic 549 in bile 532-533, 540--545 in faeces 529, 533-534, 552 in serum 529, 532-533, 552 in urine 528, 552 lithocholic 527, 530, 538, 539, 549 ,B-muricholic 549 23-selenium-25-homotaurocholic
[SeHCA T] 552 taurine-conjugated 527, 528, 538 unconjugated 536, 538, 541 ursodeoxycholic 538, 539, 549
binding proteins 650 bioassay 432, 457 biological variation 657, 662 biologically active estrogen 443, 452 bioluminescent assay 439 blank values 440, 445 blood spot, assay for 17-hydroxyprogesterone 244
Blood Transfusion Service 658, 665, 684 blunders 684 boldenone
conjugation of 485 mass spectrum (MO-TMS) of 507 metabolism of, in equine and human
478-479,485 separation from androstanediol 508 solid-phase extraction of 486 structure of 471
INDEX 699
boldenone sulphate, HPLC-MS of 181 Bond-Elut 572-573
C1s-OH cartridges, for extraction and separation 575
C1s-OH, in vitamin D metabolite extraction 572-573
C18, use in extraction of vitamin D metabolites 598
bone gla protein see osteocalcin 566 bovine serum albumin see BSA box and whisker plots 671, 676 brassinolide 635 brassinosteroids 635-637 British Laboratory Ware Association
656 broad band decoupling in 13C NMR spectra 85
BSA antigenic protein 448 methylated 446
bufodienolide, definition 621 bufotoxin 621, 622 butylboronate, formation of with vitamin D metabolites prior to GC-MS 596
t-butyldimethylsilyl ethers, derivatives for mass spectrometry 106
t-butyldimethylsilyl oxime, derivatives for mass spectrometry 107
13C NMR spectroscopy 84 C19 steroid conjugates
chemiluminescent assay for 345 formation of 271-272, 319 hydrolysis of 318-319 immunoassays for 341 RIAs for 352-353 separation of by FAB 320--323 separation of by HPLC 280--291, 319-320, 324-332
CAH need for extraction in assay for 17-hydroxyprogesterone 121-122
use of column chromatography 131 calbindin-9K gene 567 calcidiol 20 see also 25-hydroxyvitamin
D3 calciferol BP 562 calciol 20 calcipotriol (MC903), structure 568 calcipotriol
assay of in human plasma 593 NMR of metabolite of 608 structure 608 therapeutic effects 570
calcitetrol 20 calcitriol 20 see also 1,25-dihydroxyvitamin D3
calcitroic acid, structure 568
700
calf thymus, use in vitamin D metabolite assay 589
calibrated standards, availability of 234 calibration 441, 650 campestane 22 CAP see College of American
Pathologists carbonyl compounds 33, 49, 50, 54 carboxylic acids 16, 55 cardenolide, definition 621 cardiotonic steroids 621-626 catechol estrogens 448, 454
pregnancy urine 449 RIA 449
catechol-O-methyl transferase 449 CBG see corticosteroid binding globulin celite 430, 431
chromatography 442 columns 442,451
centrifugal ultrafiltration 338 dialysis 394-395 testosterone assay by 339
cerebroteninous xanthomotosis, use of GC-MS 166
Certified Reference Materials 679-680, 681 see also Community Bureau of Reference
chair conformation 9 charcoal 440, 442 chemical analogues of vitamin D 569 chemical ionisation (CI) 94, 95, 158-159
in mass spectrometry 94-95, 97 chemical shift in NMR spectroscopy 59 chemiluminescence 439
as enzymeimmunoassay endpoint 389-390
assay of pregnanediol 3o:-glucuronide in urine 401
assay of progesterone 390 chemiluminescent immunoassays 343-346
commercially available kits for 324-332
isoluminol derivatives in 344-345 testosterone glucuronide measurement by 345-346
testosterone measurement by 346 chenodeoxycholic acid 8, 11 chiroptical properties 36 chloramine T 443 cholane 2, 3, 4 cholecalciferol 8, 20 see vitamin D3 cholestane 2, 4 cholest-4-en-3-one 13 cholest-5-en-3fJ-ol 8, 11 cholest-5-ene 12 cholesterol 8, 11, 86, 96, 371
conversion into steroid hormones 230 EI mass spectrum of 96
INDEX
GC-MS for validating assay of 166 in food, extraction of 120 sulphate, HPLC-MS of 179
cholic acid 8, 11 cholylglycine hydrolase 546, 548 chromatography 442, 446
partition 443 prior to immunoassay 234-237
CIA see chemiluminescent immunoassay circular dichroism 36 CLlA see Clinical Laboratory
Improvement Act Clin-Elut 573 Clinical Laboratory Improvement Act 691
Clinical Pathology Accreditation (UK) Ltd 663
closed systems see automated immunoanalysers
CN cartridges, for separation of oxosteroids 577
collagen type-I gene 567 College of American Pathologists 664, 691
collisionally activated decomposition, in mass spectrometry 94
colorimetry 432, 433 column chromatography 129-135,444, 457
adsorption 131 effect of packing on resolution 130 review of 131 separation of androstanedione and androstenedione 130
column packing material for HPLC 142-152
column switching, during HPLC 157-158
columns used for GLC 138 Community Bureau of Reference 663,
679 see also Certified Reference Materials
commutability 666 competitive assays 433 competitive binding assay 442 competitive protein binding 374-375,408, 409 see also CPB
configuration 3, 18, 20 confirmatory analysis
by MS-MS 509 criteria for 503-505 of methenolone 504-505 of natural anabolic steroids 492,
509-518 of synthetic anabolic steroids 503-509
conformation 8, 10, 33 congenital adrenal hyperplasia (CAH) 231-232, 246, 251, 257 see also CAH
conjugated dienes 26 estrogens 428 trienes 26 lX,fJ-unsaturated ketones 26
conjugates 5, 15 Conn's syndrome, steroids assays in 254 Continuous Quality Improvement 648 contour plots in 1 H NMR spectroscopy
88 control rules 661 coprostane 4 cortical hormones 7, 11 corticosteroid binding globulin 375, 379,
391 corticosteroid glucuronides, HPLC~MS
of 177~179
corticosteroids definition 229 HPLC of 145 measurement in plasma by GC~MS 163~165
separation by aluminium oxide chromatography 132
synthetic 243 corticosterone 7, 11, 86
assays for 257~258
HPLC of 144 metabolites in urine 257
cortisol 7, 11, 86 analytical goals 657 Certified Reference Materials 679~680,
681 EQA serum concentration 666, 668 estimation of 237~242
external quality assurance of assays 238 GC~MS for validating assay of 166 HPLC of 145, 146 HPLC~MS of 170-172 in amniotic fluid 242 in saliva 237 in urine 241 IQC materials 660 measurement
by fluorimetry or colorimetry 232 by GC~MS 163 by polarisation immunoassay 240 by scintillation proximity assay 240
methods 651 specificity of immunoassay of 240 standardisation 650 synthesis from cholesterol 230 UK NEQAS GC~MS targeting exercise 682, 683, 684-685
UK NEQAS participation 654 UK NEQAS performance 671, 674,
675, 676
INDEX
urinary free 685, 660 WHO EQAS 692
cortisone 7, 11 21-acetate 71, 90 HPLC of 146 measurement by GC~MS 163
cortoic acids HPLC of 146 HPLC~MS of 175~176
fJ-cortolic acid, HPLC~MS of 175
701
fJ-cortolonic acid, HPLC~MS of 175 cortolonoic acids, HPLC of 146 COSY NMR spectra 87 counter-current distribution 430, 431 CPA (UK) Ltd see Clinical Pathology Accreditation (UK) Ltd
CPB 433, 446 see also competitive protein binding
CQI see Continuous Quality Improvement
CRM see Certified Reference Materials cross-reaction 436, 450, 451
synergism 438 cross-reactivity
of testosterone and 51X-DHT, in immunoassays 340-341
of testosterone and 51X-DHT, in RIAs 351
with commercial immunoassay kits 233
CUSUM 661 cutaneous androgen metabolism, markers for 269~270, 272
cyanogen bromide, coupling of antisera 387
cyc10dextrin derivatives with vitamin D metabolites 609~61O
in steroid extraction 128 in steroid HPLC 143
cyc10dextrins in HPLC 152~154
in HPLC mobile phase 579 cytochrome P450
in 11X-hydroxylation 566 in 25-hydroxylation 565
cytoreceptor assay, in measurement of 1,25-dihydroxyvitamin D 590
cytosol 442 cytosol sheep uterus 444 cytosol-target tissue 441
Daivonex see calcipotriol Danazol 441 data 661, 667 daturametelin-F 628, 629 DBP (vitamin D binding protein) 564
antibody to 574
702
DBP (vitamin D binding protein) (contd) as binding protein in saturation analysis
of vitamin D metabolites 588 binding affinities of vitamin D3 564 precipitation with ammonium sulphate 564
use in extraction of 1,25-dihydroxyvitamin D 574
use in extraction of metabolites from plasma 564
dehydrated vitamin D metabolites, use in GC~MS 601
7-dehydrocholesterol 562 measurement in skin 586 UV spectrum 564
dehydroepiandrosterone 6, 11, 660, 690 see also DHA
dehydroepiandrosterone (DHA) biosynthesis of 268~269
GC profiling of 301~303 immunoassays for 324~332
metabolism of in equine testis 314-315 separation of by HPLC 295~300 sulphoconjugation of 273~274
dehydroepiandrosterone (DHEA), effect of enzyme hydrolysis 124
dehydroepiandrosterone sulphate 660, 688, 689, 691 assays for 248~249
HPLC~MS of 180 delayed fluorescence assay 433 21-deoxyaldosterone, assays for 249~250 deoxycholic acid 8, 11 deoxycorticosterone 7, 11 ll-deoxycorticosterone, assays for 258 21-deoxycorticosterone, assays for 249~250
ll-deoxycortisol 660, 691 assays for 245~246
HPLC of 145 21-deoxycortisol, assays for 249~250
I1-deoxytetrahydrocortisol 690 Department of Health 655 DEPT NMS spectra 85 derivatisation
for mass spectrometry 94, 95, 105 in mass spectrometry 94, 105
deuteriated steroids, as internal standards 116
deuterium labelled internal standards 575 deuterium NMR spectroscopy 91 deuterium-proton exchange 80 Deutsche Gesellschaft fUr Klinische Chemie 690
dexamethasone assay of 243 determination by HPLC 157 suppression test 232
INDEX
dextran coated charcoal 433, 437, 443 see also charcoal
DGKC see Deutsche Gesellschaft fUr Klinische Chemie
DHA sulphate 446 assay of by GC~FAB 280~291,
320-323 formation of 271 hydrolysis of 318~319 immunoassays for 335~336, 343 occurrence in urine 272 RIAs for 324-332 separation of by HPLC 319~320
separation of by TLC 319 DHEA see dehydroepiandrosterone DHEAS see dehydroepiandrosterone sulphate
50:-DHT 441 dialysis see centrifugal ultrafiltrationdialysis; equilibrium dialysis; rate dialysis
dianabol, determination by HPLC 157 diethyl ether 428, 443 diethyl stilbestrol 5, 441 differential ammonium sulphate technique, testosterone assay by 339
digitalis steroids 621 ~626 digoxin 621, 622, 625, 626
HPLC of 144 dihydroequilenin-17 0: 451 dihydroequilenin-17 {3 451 170:-dihydroequilenin sulphate 454 dihydroprogesterone 408-409
in urine 398~399
dihydrotachysterol GC~MS in study of metabolism 602~604
HPLC of metabolites 582 HPLC separation of metabolites 582 UV spectrum 582
dihydrotachysterol3 , structure of 569 dihydrotesterone 6
detection of 515~516
effects on TIE ratio of 516 50:-dihydrotestosterone see 50:-DHT 50:-dihydrotestosterone (50:-DHT)
19-norderivative as internal standard 310
cross-reaction of with testosterone in ELISA 340-341, 351
formation from testosterone 299 glucuronidation of 270 measurement of by GC~MS 280-291 metabolism of 271
22,23-dihydroxyergosterol 17 10:, 25-dihydroxycholeca1ciferol 20 30:, 70:-dihydroxy-5{3-cholan-24-oic acid
11
3tX, 7 P-dihydroxy-5p-cholan-24-oic acid 11
(24R)-24, 25-dihydroxycholesterol 17 1,25-dihydroxy-16-ene-vitamin D 3, structure of 569
1 tX,25-dih ydroxydih ydrotachysterol mass spectrum of per-trimethylsilyl
ether 603 structure 603
lIP, 21-dihydroxypregn-4-ene-3,20-dione 7,11,86
17,21-dihydroxypregn-4-ene-3,11,20-trione 7, 11
15P,17tX-dihydroxypregnanolone, in urine as marker for 21-hydroxylase deficiency 245
1,25-dihydroxyvitamin D measurement by cytoreceptor assay
590 measurement by GC-MS 596-597
24,25-dihydroxyvitamin D, measurement by GC-MS 596-597
25,26-dihydroxyvitamin D, measurement by GC-MS 596-597
1,25-dihydroxyvitamin D3 24-hydroxylation of 567 26-hydroxylation of 567 binding to the VDR 566 catabolism of 567-569 control of production 566 in cell differentiation 567 in tumour regression 567 measurement by scintillation proximity
assay 591 metabolites of 568-569 molecular events in target cells
566-567 physiological effects 566 receptors in skin 567 stimulation of production of
osteocalcin 566 structure 568 26,23-lactone, structure 568
24,25-dihydroxyvitamin D3 567-568 structure 568
25,26-dihydroxyvitamin D 3, structure 568 dimethyldichlorosilane, used to treat glass
surfaces 118 23,24-dinor-5p-cholan-22-oic acid 19 direct assay 438, 447 direct systems 441 dissociation-enhanced lanthanides, f1uoroimmunoassay (DELFIA) 348
double-antibody separation 437 double-isotope derivatisation 433 drift 659
EC see European Union
INDEX
ecdysone 631 ecdysteroids 631-634 ecdysterone 631, 632
703
EIA see enzyme immunoassay electrochemical detectors for HPLC 154 electron 94, 95
impact, in mass spectrometry 94, 96 ionisation (EI), in mass spectrometry
94, 95, 158 et seq. electron capture 94, 95
in mass spectrometry 94--95, 97 electrospray 94, 95 electrospray MS 166 et seq. electrospray-mass spectrometry 94-95, 97 ELISA 439 see also enzyme-linked immunosorbent assays
end-capping of microparticulate silica 572,575
endogenous binding proteins interference 441
enol ether, derivative for mass spectrometry 107
enzyme hydrolysis 446,447,451 of steroid glucuronides 122-124
enzyme immunoassay 385-389,401-402, 415
for 25-hydroxyvitamin D3 574 enzyme immunoassays 324--332, 340-343
4-androstenedione 342-343 for DHA sulphate 343 for testosterone 340-342
enzyme labelling 433 enzyme-linked immunosorbent assays
324--332, 340-343 C19 steroid cross-reactivities in
341-342 effect of hapten heterology on 341-342 for 4-androstenedione 342-343 for testosterone 340-342 principles of 340 structure of haptens in 340-341 see also ELISA
enzymes in steroid synthesis 230 16-epi-estriol 431 epimerisation, mechanism of 482-483 epitestosterone
excretion of 511 testicular production of 510 see also hormone ratios
epitopes 433 11 p,18-epoxy-18-hydroxypregn-4-ene-3,20-dione 11
EQA see External Quality Assessment EQASs see External Quality Assessment, Schemes
equatorial bonds 9, 63 equilenin 5, 450, 451, 454 equilenin sulphate 450
704
equilibrium dialysis (EQD) 392-393, 452 principles of 338 testosterone measurement by 338
equilin 450 3H-equilin 451 equilin 3-hemisuccinate-BSA 451 equilin 3-sulphate 451 equilin sulphate 450 equilin-17 -( O-carboxy-methyl)-oxime-BSA 450
equilin-17 -( O-carboxymethyl)-oxime-[125IJiodohistamine 450
equine estrogens 441 ergocalciferol 8, 20 see vitamin D2 ergosta-5,7-dien-3tJ-ol 17 (22E)-ergosta-5, 7, 22-trien-3tJ-ol 17 ergostane 22 ergosterol 17 error 656, 657 erythrocyte 449 esters 14, 50, 56 estetrol 453, 454 estradiol 5, II, 428, 431, 435, 436, 437, 439,440,441,442,443,446,447,452,453, 454
-17a 435,441,454 -17 tJ see estradiol analytical goals 657 CAP PT scheme 691 -6-( O-carboxymethyl)-oxime-BSA 443, 444, 445
-6-( O-carboxymethyl)-oxime-(2-[125IJ)iodohistamine 443
conjugates 439 EQA serum concentration 666, 668 extraction using affinity column 574 fatty acid esters, measurement by GC-MS 165
-17 fatty esters 456 glucuronide, urine 429 17tJ-glucuronoside 447,448 3-glucuronoside 447, 448 3-hemisuccinate-BSA 436 -lla-hemisuccinate-BSA 436 -17 tJ hemisuccinate-BSA 436, 444, 445 HPLC-MS of fatty esters 172-173 2-hydroxylase 432 4-hydroxylase 432 IQC materials 660 Murex EQA scheme 689 oral HRT effects on results 685, 686, 687
reduction 445 RIA 454 saliva 443 -17a serum 444 serum concentration 444, 649
men 442
INDEX
postmenopausal women 442, 443 sulphate 452 -17 tJ sulphate 455 UK NEQAS participation 654 UK NEQAS performance 671, 672, 674, 675, 676
UK NEQAS recovery experiment 678, 679
urine 442 WHO NEQAS 692
17 tJ-estradiol see estradiol 17 tJ-estradiol-6-(C-carboxymethyl)-oxime-BSA 435
3H-estradiol 442, 444, 445 estrane 2,4 estranediols
formation of, from nandrolone 477, 516-517
GC-MS fragmentation of 505 selected ions for, in GC-MS 501 structures of 477 urinary excretion of 517
estra-5(1O)-ene 12 estra-l,3,5(1O)-triene 12 estra-l,3,5(1O)-triene-3,17a-diol 11 estra-l,3,5(1O)-triene-3,17 tJ-diol 5, 11 estra-l,3,5(1O)-triene-3,16a, 17tJ-triol 5, 11 5( 10)-estrenediol
in eq uine testis 517 selected ions for in GC-MS 501 structure of 516 use of, in nandrolone administration 517-518
estriol 5,86,428,431,441,446,447,448, 660,691 -3-(O-carboxymethyl)-oxime-BSA 445 -6-( O-carboxymethyl)-oxime-BSA 445, 446
glucuronide, urine 429 3-glucuronide 445, 455 16-glucuronide 455 17 -glucuronide 455 16a-glucuronide, urine 429 3-glucuronoside 447, 448 16a-glucuronoside 447,448 16tJ-glucuronoside 447 16-glucuronoside urine, non-pregnant women 448
saliva 453, 454 serum 445 3-sulphate 445 3,16,17-trihemisuccinate-BSA 451
3H-estriol 446, 447 3H-estriol 16-glucuronoside 448 estrogen glucuronide
plasma 447 urine 447
estrogen glucuronoside 452
3H-estrogen 447 estrogen-protein conjugates 434 estrogens 432, 660
extraction of esters 120 GLC amniotic fluid 432 GLC blood 432 GLC urine 432 HPLC of 148-150 separation by solvent extraction 121
estrone 5, 86, 428,431, 435, 436, 437, 439, 443,444,445,446,447,448,450,453,454
acetate 53 -3-( O-carboxymethyl)-oxime-BSA 446 -6-( O-carboxymethyl)-oxime-BSA 445 conjugates 439 3-glucosiduronate-bovine thyroglobulin 446
3-glucuronide 692 glucuronide amniotic fluid 429 glucuronide urine 429 3-glucuronoside 447, 448 3-hemisuccinate-BSA 437 3-methylphosphothionate 446 3-sulphate 451 3-sulphate 60(-hemisuccinate-BSA 446
3H-estrone 445 estrone sulphate 446, 452, 454
amniotic fluid 429 cow's milk 446 extraction ether-ethanol 430 plasma 446 serum 446 sow serum 446 urine 446
[3HJ-estrone 446 sulphate 446
ethers 14 ethinyl estradiol 441, 451, 453, 454 ethinylestradiol 3-methyl ether 451 ethisterone 52 ethynylestradiol 19 170(-ethynyl-17 -hydroxyestr-4-en-3-one 19 170(-ethynylestra-1, 3, 5(1O)-triene-3, 17 p-diol 19
etiocholanone 690 EU see European Union European Commission see European Union
European Union 656,663,693 exocyclic alkenes 32 External Quality Assessment 662-693
data processing 667, 670 definition 662 performance evaluation 667, 671-685 principles and practice 664--670 samples 665-666 Schemes 662, 689-692 survey method 664
INDEX
target values 666-667 see also UK NEQAS
extraction
705
selectivity by choice of solvents 121 use of ion exchange columns 134-135
extraction by solid phase material 124 et seq.
extraction of anabolic steroids by immunoaffinity chromatography 489
by liquid-phase 487-489 by solid-phase 486-489 by thin-layer chromatography 489
extraction of estrogens 428 extraction of steroids 117 et seq.
use of ammonium sulphate 118 use of solvents 118 et seq.
extraction of vitamin D 571-575 extraction of vitamin D metabolites
from plasma, use of DBP 564 prior to GC-MS 596, 598
Extrelut 573 E/Z-convention for alkenes 23
F AB/LSIMS 95 fast atom bombardment (F AB) 94 fast atom bombardment (F AB), mass spectrometry 94--95, 97
fatty acid esters 440 of steroids, measurement by GC-MS
165 fatty acids, removal from vitamin D extracts 571
FDA see Food & Drug Administration Federal Medical Society of Germany 689-690
feto-placental function 447 FIA see fluoroimmunoassay Florisil, in column chromatography 131 fluorescence 35
detection during HPLC 154 fluorimetric detection 453 fluorimetry 432, 433 fluoro-steroids 92 fluoroimmunoassays (FlAs) 346-349
commercially available kits for 324-332, 349
DELFIA enhancement in 348 europium 3H-liposome encapsulation in 349
17 P-hydroxysteroid dehydrogenase measurement by 348-349
lanthanide chelates in 348-349 linkage of steroids to fluorescent labels in 348-349
principles of 346-347 testosterone measurement by 347-348
follicular function 448
706
Food & Drug Administration 656 food steroids, extraction of 120 Fourier transform
IR spectroscopy 43 NMR spectroscopy 58
free progesterone determination by centrifugal ultrafiltration-dialysis 394-395
by equilibrium dialysis 392-393 by rate dialysis 394 by steady state gel filtration 393-394 by ultracentrifugation 394
free progesterone equivalence to saliva concentration 396
free unbound plasma steroid 566 functional group nomenclature 5, 13
gas chromatography coupling of with mass spectrometry
305-316 derivatives used in 303-304 separation of C19 steroids by 301-303 see also GLC
gas chromatography-mass spectrometry 94-95, 100, 103, 108,234 derivation of target values 666-667 derivatives of anabolic steroids for 498-503
full scan spectra of anabolic steroids 505-509
selected ions of anabolic steroids 499, 501
UK NEQAS studies 678-685, 682, 687 use in German EQA schemes 690 use in standardisation 650 see also GC-MS
gas liquid chromatography see GLC GC-MS 94,161-166,433,438,455,457
for hexafluoro-l,25-dih ydroxyvitamin D3 592
measurement of dehydrated vitamin D metabolites 601
of cortisol and 18-hydroxycortisol in urine 255
of medrogestone 409--410 of medroxyprogesterone acetate 411--412
of norethisterone 413--414 of pregnanediol 417 of progesterone 404--405 use for obtaining target values of vitamin D metabolites 600
use in determination of vitamin D metabolite structure 602 et seq.
use in studies of vitamin D metabolising enzymes 599
use of double focussing instrument in vitamin D measurement 601
INDEX
see also gas chromatography-mass spectrometry
gel filtration 428, 452 glass surfaces
binding of steroids to 118 destruction of steroids on 116
glassware, silanisation to prevent adsorption 573
GLC 137-141, 373-374, 401, 405, 432--433
capillary 433 derivatisation 433 derivatives used in 140-141 electron capture 433 flame ionisation 433 for aldosterone 252 methods of detection 141 of vitamin D metabolites 595 et seq. urinary steroid profile in normal man
142 use in assay of 18-hydroxycorticosterone 253
GLC-MS of vitamin D metabolites 594 et seq.
y-globulin sheep anti-rabbit 443 GLP see Good Laboratory Practice glucocorticoid, biosynthetic pathway 230 glucuronic acid 448
conjugation 449 fJ-glucuronidases 547 glucuronide 437, 439 17 fJ-glucuronides hydrolysis of 489--492 glycyrrhetinic acid, HPLC-MS of 174 glycyrrhizic acid, HPLC-MS of 174 GMP see Good Manufacturing Practice goat 443 gonadotrophins 434, 448 Good Laboratory Practice 648 Good Manufacturing Practice 648 graphitised carbon in steroid extraction
127-128
haemocyanin 436 hapten 435, 436, 442, 447 Helix pomatia, in steroid conjugate hydrolysis 124
hemiacetals 55 hepatitis Band C 658, 665 heteroannular dienes 30 heterodimers in activation of vitamin-D responsive genes 567
heteronuclear spin-spin coupling in NMR spectroscopy 86, 91, 92
hexafluoro-1,25-dihydroxyvitamin D 3,
measurement by GC-MS 165,592,596 high performance liquid chromatography
coupling with MS (thermospray) 316-317
separation of C19 steroid conjugates by 319-320
separation of C19 steroids by 295-301 high performance thin layer chromatography 577-578
high pressure liquid chromatography see HPLC
histamine 438 [125I]-histamine 445 HIV 658,665 homoannular dienes 30, 32 homodimers in activation of vitamin-D responsive genes 567
homonuclear spin-spin coupling in NMR spectroscopy 81
homosteroids 20 Hooker-Forbes test 372 hormone ratios
detection of testosterone administration and 510-515
dihydrotestosterone administration and 515-516
in biological outliers 514-515 isotope mass spectrometry and
513-514 testosterone/dehydroepiandrosterone and equine 518
testosterone/epitestosterone (T /E) 510-516
testosteronejluteinising hormone 510-513
hormone replacement therapy 655, 658, 685, 686, 687
24 hour urine collection, necessity for 232
HPLC 141-158,432,433,453,454,457 assessment of homogeneity of emerging peaks 579-581
automated 454 autosampling 454 box-car 454 catechol estrogens 453 choice of mobile phase 152-153 columns available 142-152 detection limit 454 detection of steroids 154-155 estrogens 454 identification of steroids 155-156 improved resolution of vitamin D2 and
tachysterolz 563-564 incorporation of cyclodextrins in mobile phase 579
ion-pair chromatography 153 linked to mass spectrometer 581 et seq. micellar chromatography 153 modern methodology 144-150 precolumn derivatisation 156--157 prior to immunoassay 235
INDEX 707
resolution of skin irradiation products 563
review of methods up to 1987 142 sample injection 153-154 separation of irradiation products of ergosterol 565
systems for vitamin D separation 578-579
use of cyano columns for isolation of vitamin D lactones 578
use of electrochemical methods for vitamin D metabolite analysis 586
use of photodiode array detection 579-583
see also high performance liquid chromatography
HPLC-MS 156 electrospray of vitamin D metabolites
585 of androgen glucuronides 176 of boldenone sulphate 181 of cholesterol sulphate 179 of corticosteroid glucuronides 177-179 of cortisol 170-172 of ,B-cortolic acid 175 of ,B-cortolonic acid 175 of dehydroepiandrosterone sulphate
180 of estradiol of fatty esters 172-173 of etrahydrocortisone-3-glucuronide
178 of glycyrrhetinic acid 174 of glycyrrhizic acid 174 of 4-hydroxyandrostene-3,17-dione 172 of 17-hydroxypregnenolone sulphate
182 of 5-hydroxyvitamin D3 172 of ortoic acids of 175-176 of 3-oxo-4-ene-steroids 169-172 of placental sulphatase deficiency in investigation 183
of steroid acids 174-176 of steroid conjugates 176--184 of steroid fatty acid esters 172-174 of steroid sulphates 179-184 of vitamin D metabolites 601 thermospray of triazoline adducts of vitamin D metabolites 585
use in studies of vitamin D metabolising enzymes 599
HRT see hormone replacement therapy human chorionic gonadotropin see testosterone and hCG
human pre-pro gene 567 hybrid mass spectrometer 105 hydrolysis
conjugates 428 enzymes 430
708
hydrolysis (contd) glucuronidase 430 fJ-glucuronidase 432 of estrogens 429, 430 steroid sulphatase 430
phosphate 430 saccharolactone inhibition 430
sulphatase 432 (IS)-I-hydroxy calciol 20 17-hydroxy corticosteroids 660 18-hydroxy-ll-deoxycorticosterone, assays for 256-257
IlfJ-hydroxyandrostenedione, assays for 247-248
17 fJ-hydroxy-5a-androstan-3-one 6 17 fJ-hydroxyandrost-4-en-3-one 6, 11 4-hydroxyandrostene-3, 17 -dione, HPLC-MS of 172
4-hydroxyandrostenedione, metabolites of in urine 122
3fJ-hydroxyandrost-5-en-17-one 6, 11 I1-hydroxyandrosterone 660, 690 25-hydroxy cholecalciferol 20 18-hydroxycorticosterone metabolites in urine 254 assays for 253-254 interaction with alcoholic solvents 253
18-hydroxycortisol, assays for 254-255 17 fJ-hydroxyestr-4-en-3-one 19 3-hydroxyestra-I,3,5(1O)-trien-17-one 5 3-hydroxyestra-l,3,5,7,9-pentaen-17-one 5 4-hydroxyestradiol 453, 456 2-hydroxyestradiol 453, 456 2-hydroxyestradiol plasma 450 2-hydroxyestradiol-17 fJ 449 15a-hydroxyestriol plasma 450 15a-hydroxyestriol serum 450 15a-hydroxyestriol-BSA 450 2-hydroxy estrogen 431 2-hydroxyestrogens 449 2-hydroxyestrone 14, 428, 449 2-hydroxyestrone plasma 450 2-hydroxyestrone-17 -( O-carboxymethyl)-[125IJiodohistamine 449
2-hydroxyestrone-17-( O-carboxymethyl)oxime-BSA 449
ll-hydroxyetiocholanolone 660, 690 23-hydroxy-24-homo-la,25-dihydroxyvitamin D3 mass spectrum of per-trimethylsilyl
ether 605 structure of per-trimethylsilyl ether 605
I1fJ-hydroxylase deficiency 246 21-hydroxylase deficiency, 17-hydroxyprogesterone as marker for 246
17-hydroxylase deficiency 251, 257 hydroxylase enzymes, in steroid synthesis 230
INDEX
18-hydroxylated corticosteroids, measurement by GC-MS 163
18-hydroxylated steroids, problems with HPLC of 151
hydroxylated vitamin D metabolites, assignment of structure by NMR 609
17{i-hydroxy-17a-methyl androgens conjugation of 483 demethylation of 481--482 epimerisation of 480--481 metabolism of 475,478--483 see also methandrostenolone, methandienone, stanozolol
17 {i-hydroxy-19-nor-17a-pregn-4-en-20-yn-3-one 19
21-hydroxypregn-4-ene-3,20-dione 7, 11 3fJ-hydroxypregn-5-en-2-one 11, 109 17-hydroxypregnenolone sulphate, HPLC-MS of 182
17a-hydroxy progesterone CAP PT scheme 666, 669 EQA serum concentration 666, 669 IQC materials 660 Murex EQA scheme 689 UK NEQAS participation 654 UK NEQAS performance 671, 674
17-hydroxypregnenolone assays for 249 17-hydroxyprogesterone
assays for 243-245 concentration in plasma 243 immunoassay using extraction 122 interference by sulphate conjugates in
assay of 121-122 measurement in saliva 245 use of solvent extraction to prevent interference 122
hydroxysteroid dehydrogenase 3-a 530, 533, 539 7-a 531 12-a 531
hydroxysteroids 5, 6, 7, 8, 11, 13, 15, 17, 19, 22, 48, 79, 84
3a-hydroxysteroid dehydrogenase, use in detection of bile acids during HPLC 155, 157
17 fJ-hydroxysteroid dehydrogenase 444 3fJ-hydroxysteroid oxidases 108 25 hydroxy vitamin D 660, 688 25-hydroxyvitamin D
immunoassays for 588 measurement by GC-MS 596 NMR of 608
25-hydroxyvitamin D-la-hydroxylase 566 25-hydroxyvitamin D3
in human plasma 565 enzyme immunoassay for 574 enzyme labelled immunoassay for 587 HPLC-MS of 172
measurement by HPLC 579-581 structure 568
25-hydroxyvitamin D3-1-hydroxylase, in vitro studies using GC-MS 599
25-hydroxyvitamin D3-24-hydroxylase, in vitro studies using GC-MS 599
25-hydroxyvitamin D 3 - 26,23-lactone, structure 568
la-hydroxyvitamin D 3, structure of 569
1251 433 125I-labelled steroid 438 1251 NaI see 1251 sodium iodide 1251 sodium iodide 443 125I-histamine 448 125I-label 441 ID-GC-MS see gas
chromatography-mass spectrometry idiometric assay 434 immobilised antibodies in steroid extraction 129
immobilised antigen 439 immunisation 435, 436 immunoaffinity see extraction of anabolic
steroids immunoassay 430
accuracy 205,206,213 acridinium derivatives 197 affinity 189, 190 alphatype 210 anabolic steroids for 493-497 anti-allotype 210 antibody 186
characterisation 189 occupancy 209 titre 190
antigen binding site 209 antigenic response 187 automation 653 avidity 190, 191, 192, 204 background 199 betatype 210 biotin/streptavidin 205 bridge recognition 192, 193 calibration curve 205 captured analyte 209 carrier molecule 187 chelates 200 chemiluminescence 195, 212
enzyme assay 198
INDEX
competitive binding assay 185, 189, 198 correlation 207 cross-reactivIty 192, 202
antisera in 496-497 data 661 design of immunogens for 495-497 direct assay 201 directory 652
disassociation 199 displacing agents 202 enhanced 198, 201 enzymes 194 epitope 186, 187, 209, 210 excess reagent 205 external quality assurance 207 extraction 201 fertile period 203 FITC 213 fluorescence 198 fluorescence polarisation 213 fluoroimmunoassay 198, 213 for cortisol 238 et seq. free steroids 207 hapten 187, 188, 209
709
homogeneous immunoassay 212 hormone replacement therapy 202, 204 idiometric assay 210 idiotype 210 immunisation protocol 188 immunogen 187, 192, 193 immunogenic 187 immunoglobulins 186 immunometric 208 incubation timing 233 interference 206 isoluminol 195, 212 labels 192, 199 lanthanides 199, 203 matrix effect 201, 202 miscellaneous controls 206 monoclonal antibody production 188 non-competitive assay 208, 210 non-radioisotopic labels 194, 212 non-separation assay 210, 213 non-specific binding 204, 207, 208 of vitamin D metabolites 588-591 ovarian stimulation 204 photodetectors 195 polyclonal 187 precision 205, 206, 208, 210, 213 premenstrual syndrome 204 protein binding agents 202 quality control 205 quenching 193, 200 radioimmunoassay 185 radioisotopes 193 receptors 185, 189 recovery 206 reproducibility 206 saliva 203 sample preparation 201 saturation analysis 185, 189 scintillation 193, 211 scintillation proximity assay 210 screening assays 190, 210 second antibody assay 205
710
immunoassay (contd) sensitivity 190, 205, 208, 213 separation methods 204 separation of bound and free hormone 233
separation procedures 233 serum binding proteins 185, 202, 208 SHBG 202 simultaneous assay 203 solid phase 205, 208 specific activity 193 specific binding 202 specificity 189, 190, 191, 201, 202,
213 standardisation 201, 650 standards 206 steric hindrance 209 steroid analogues 208 steroid conjugates 195, 202 steroid derivatives 192 two-site assay 209 types 652, 653 urinary metabolites 203 urine 203 validation 205
immunoradiometric assay 453 imprecision 649, 657 see also precision inaccuracy 649, 657, 662 see also
accuracy; bias INADEQUATE NMR spectra 91 incubation time 437 indirect assay 448 infertile women 448 infertility 427 infrared spectra 38 injection onto GLC, methods available
139-140 injection onto HPLC columns
153-154 INSTAND 690 interassay variations 231 interfaces for LC-MS 166 et seq. interference 440
lipid 440 paper fibres 442 steroids 441 urate 432
Internal Quality Control 656-662 analyte concentration 658 data presentation 661 materials 658-659, 660 principles 657 procedures 659
internal standard use in mass spectrometric measurements
of vitamin D metabolites 575 use of deuterium labelled metabolites
575
INDEX
internal standards 115 et seq. use of in C19 steroid quantification
310-315 ion detection in mass spectrometry 103 ion-exchange columns, use in steroid
extraction and separation 134-135 ion-pair chromatography, in HPLC 153 ion-pairing agents 453 ion separation 99 ion trap mass spectrometry 94, 100, 105,
159-160 ionisation, in mass spectrometry 93 IQC see Internal Quality Control isofiavenoid phytoestrogens 456 isotachysterol isomers
formation 581, 595 use in HPLC assay for 25-hydroxyvitamin D3 581
isotope dilution in mass spectrometry 94 mass spectrometry see GC-MS see also gas chromatography-mass spectrometry
Karplus equation 75 Keller-Liliani reaction 624, 625 ketoconazole
effects on TIE ratios, of 514-515 use of, in differentiation of biological outliers to athletes using testosterone 515
ll-ketoandrosterone 660, 690 ll-ketoetiocholanolone 660, 690 17-ketogenic steroids 660 17-ketosteroids 660 keyhole limpet haemocyanin antigenic protein 448
Kieselguhr, in steroid extraction 125 et seq.
14C-labelled steroid 438 3H-Iabels 449 Laboratoire National de Sante 690 lactones 49, 56 Landelijke Werkgroep voor Bindingsanalysen 690
lanthanide ions and chelates use in fiuoroimmunoassays 346-349 use of in detection of anabolic steroids in HPLC 299
lanthanide shift reagents in NMR spectroscopy 67
latex particles 434 LC-MS 94, 166-184 see also HPLC-MS
levonorgestrel 692 LH20 use in steroid extraction and separation 134-135
Lichrosorb DIOL 454 LIDIA see ligand differentiation immunoassay
ligand binding assay 433
INDEX
ligand differentiation immunoassay, use in pseudohomogenous RIA 351
linearity 663, 675 linked scanning in mass spectrometry 104 Lipidex 1000 529 Lipidex columns 432 Lipidex-DEAP 530, 535 Lipidex use in steroid extraction and separation 134--135
lipoprotein disruption before efficient extraction
572-573 vitamin D incorporation into 573
liquid chromatography-mass spectrometry 94, 99
liquid secondary ion mass spectrometry (LSIMS) 97
lithocholic acid 11 LNS see Laboratoire National de Sante long-range spin-spin coupling in NMR spectroscopy 77, 92
luminescence, enhanced 439 lumistero12, HPLC of 565 lumisterol3
formation in skin 563 UV spectrum 564
luteal function 448 LWBA see Landelijke Werkgroep voor Bindingsanalysen
lyophilisation 658, 666
magnesium oxide, minicolumns in extraction 120
magnetic particles 439 magnetic sector instruments 100 magnetic sector mass spectrometers
99-100 mass fragmentography, of vitamin D metabolites 596 et seq.
mass spectrometric fragmentation 108 mass spectrometric resolution 102 mass spectrometry 93, 97-100
characteristic fragmentations by EI 108 combined GC-MS (SIM) 305-309 comparison of with RIA results 353 data handling 103 definition of common terms used in
94--95 derivatisation
of hydroxyl groups 106-107 of oxo groups 107-108
derivatisation techniques in 105-109 electron impact ionisation 94, 96 fragmentation 108
HPLC (thermospray) 316-317 ion detection and data handling
103-104 ion separation 99-100 ionisation 95
711
magnetic sector instruments 100--101 metastable peak monitoring in 309 negative ions CI of vitamin D3 in skin
594 of vitamin D metabolites and structural
analogues 594 et seq. principle ionisation and sample
introduction techniques 95 quadrupole ion trap 100 quadrupole mass filter 100 resolution of ions 94, 100, 102-103 sample introduction and ionisation 93 stable isotopes use in 309-310,311-315 static F AB-MS of vitamin D
conjugates 594 tandem (or MS-MS) 315-316 terminology 94 thermospray 595 time-of-flight (TOF) analysers 100--101 types of 305-318 use in measurement of DNA sulphate
249 useful ions for monitoring in 305-308 using chemical ionisation 94--95, 97
materials certified reference 680, 681 External Quality Assessment 665-666 Internal Quality Control 658, 659, 660
matrix 650, 658 matrix-assisted laser desorption/ionisation (MALDI), in mass spectrometry 99
MC903 see calcipotriol Medical Devices Directorate 655 Medipolar ODS 454 medroxyprogesterone acetate 692
determination by GC-MS 411--412 determination by RIA 410--411 structure 370
melengesterol acetate, determination by HPLC 157
menopause 427 mesteralone 441 metabolites 442 metastable ions, in mass spectrometry 94 methandienone
ions of metabolite 499 phase I metabolism 480--482 phase II metabolism 483 solid-phase extraction 486 structure 471
methandrostenolone see methandienone methanolysis comparison with enzymic
hydrolysis/solvolysis 491
712
methenolone confirmatory analysis of 504-505
method comparison 655 definitive 657, 666 evaluation 650-656 reference 650, 657, 662, 666, 678-685 selection 650-656 trouble-shooting 655 validation 655
methodology for vitamin D analysis 598-599
2-methoxy estrogen 431 2-methoxy[3HJestrone 449 2-methoxyestrone 449 methyl ether, derivative for mass
spectrometry 106 2-methyl ether 449 methyloxime, derivative for mass spectrometry 107
metyrapone 245 micellar chromatography, in HPLC 153 micro particle immunoassay 439 micro particulate silica
in steroid extraction 125 et seq. in vitamin D metabolite extraction 572
micro titre plate assay 439 mifepristone
anti glucocorticoid 412 antiprogestin 412 determination by HPLC 413 metabolism 412-413 pharmacokinetics 412 structure 370
milk, vitamin D in 562 mineralocorticoid, biosynthetic pathway 230
mobile phases in HPLC 152-153 statistical optimisation 152
molecular events in target cells of 1,25-dihydroxyvitamin D3 566-567
molecular ion 94 in mass spectrometry 96
monoclonal antibodies 391, 397, 435, 437 monoclonal antibody, use in immunoassay
of 1,25-dihydroxyvitamin D3 589 MO-TMS derivatives see gas chromatography-mass spectrometry
mouse osteopontin gene 567 MP A see medroxyprogesterone acetate MS 158-184 MS-MS 160-161 see also mass spectrometry, tandem
Murex Quality Assessment Programmes 689
NaBH4 see sodium borohydride
INDEX
nandrolone biosynthesis of 510, 517 ester formulations of 475 immunoassay of 494, 496-497 liquid-phase extraction of 488 metabolism of 476-477,484,494-495,
516-517 methanolysis of conjugates of 491 misuse of 468, 470 structure of 471
National Institute for Biological Standards & Control 663
natural binding proteins 433 network of steroid reference laboratories
692-693 neutral steroids, HPLC of 169-172 NH2 cartridges, for separation of vitamin
D metabolites 577 NHz columns, use in purification of estrogens 127
NIBSC see National Institute for Biological Standards & Control
NMR high field spectra of steroid hormones
610 of calcipotriol 608 of 25-hydroxyvitamin D 607 of vitamin D metabolites 606-610
NOESY NMR spectra 91 nomenclature 10 non-competitive assay see idiometric
assay non-extraction methods 441 non-invasive methods 445 non-invasive sampling 456 non-protein bound cortisol 452 non-protein bound estradiol
centrifugal analysis 453 flowing dialysis measurement 453 men 452 rats 452 women 452
non-protein bound estrogens 451 non-protein bound steroid 444 A-nor-5il(-androstane 19 4-nor-5il(-androstane 19 19-nor-5il(-pregnane 19 24-nor-5fJ-cholan-23-oic acid 19 nor-steroids 18 norethisterone 692
determination by GC-MS 413 determination by RIA 413-414 metabolism 415 structure 370
19-norethisterone 19 norgestrel
determination by RIA 415-416 determination in milk 416
structure 370 19-nortesterone 19 19-nortestosterone see nandrolone novel application 439 nuclear accessory factor (NAF) 566 nuclear magnetic resonance see NMR nuclear magnetic resonance spectra 56 nuclear Overhauser effect 82
o-catechol 449 OCT
antisera to 593 see also 22-oxacalcitriol
octadecylsilane-bonded silica 529, 530 oestradiol see estradiol off-resonance decoupling in 13C NMR spectra 85
16o:-0H estrone 431 250HD see 25 hydroxy vitamin D 170HP see 17o:-hydroxy progesterone on-line mass spectrometry 432 optical rotation 36 optical rotatory dispersion 36 oral contraceptives 451 ouabain 623, 624, 625 outliers 667 ovulation 439 22-oxa-l,25-dihydroxyvitamin D3 see 22-oxacalcitriol
22-oxacalcitriol (OCT), structure of 569 oxidation 443 oximes
derivatives for mass spectrometry 107 in mass spectrometry 107
18-oxocortisol, assays for 254--256 24-oxo-23,25-dihydroxydihydrotachysterol, structure 604
3-oxo-4-ene-steroids, HPLC-MS of 169-172
6-oxo-estradiol-17 fJ 428 6-oxoestradiol 431 16 oxo-estradiol 431 oxo-steroids 13, 54, 84 17 -oxosteroids
assays for GC-MS 280-291 estimation of by Zimmerman reaction
274 formation and conjugation of 271-272 separation of by GC profiling 301-303
paper chromatography 136-137,430,431 in assay of 1,25-dihydroxyvitamin D3
578 partition chromatography 442, 445, 446 Patella vulgata, in steroid conjugate hydrolysis 124
perfluoroacyl esters, derivatives for mass spectrometry 106
INDEX 713
peroxidase 439 phase I metabolism of anabolic steroids 475--483
phase II metabolism of anabolic steroids 483--485
'phase switching' 575 in vitamin D metabolite extraction 573
photodiode array detection, use in identification of steroids after GLC 155
phthalates interference in GC-MS of vitamin D metabolites 602
interference in mass spectrometry 587 phytoestrogens 441 placental insufficiency 445 placental sulphatase deficiency, HPLC-MS in investigation of 183
plasma concentrations of steroid hormones 231 of vitamin D metabolites by GC-MS
595 et seq. plasma estrogens 428 plasma steroids, measurement by GC-MS 164--165
polarisation immunoassay, for cortisol 240
polycystic ovary syndrome, androstenedione production in 247
ponasterone-A 632 postpartum blues 371,416 potato breeding, steroid analysis for 638-640
precision 438, 649, 659 see also imprecision
precision profile 659 prednisolone, HPLC of 145, 146 prednisone, HPLC of 145, 146 pregnancy 434
urine 447, 449 pregnane 2, 4 (20R)-5f:1-pregnane-3o:,20,21-triol 17 (20S)-5fJ-pregnane-3,20-diol 17 pregnanediol 17,660,690
determination by GC-MS 417 determination in milk 417 determination in serum 416--417 detennination in urine 372 glucuronide 692
5o:-pregnanedione see dihydroprogesterone
pregnanetriol 660, 690 metabolite of 17-hydroxyprogesterone
245 pregnanolone 399--400, 416 pregnant mare 450 pregnenolone 11, 14 5-pregnen-3fJ-ol-20-one-trimethylsilyl ether-methyloxime, EI mass spectrum of 109
714
Premarin 451 preservatives 658, 666 see also lyophilisation; sodium azide
previtamin D2, HPLC of 565 previtamin D3 562 PROBIOQUAL 690 Process Control see Internal Quality Control
Product Cycle 652 Proficiency Testing 662, 663, 691 see
also External Quality Assessment progesterone 7, 11, 86
antisera to 376-377 biosynthesis 291 CAP PT scheme 691 l1o:-carboxymethyl ether 398 determination of
by bioassay 372 by competitive protein binding
374-375 by double isotope derivative 374 byenzymeimmunoassay 385-389 by fluorimetry 373 by fluoroimmunoassay 390 by gas-liquid chromatography
373-374 by radioimmunoassay 375-385 by spectrophotometry 372-373
EQA serum concentration 666, 668 GC-MS for validating assay of 166 11o:-glucuronide 384, 397 l1o:-hemiphthalate 383-384 l1o:-hernisuccinate 376, 382-384, 397 12-hemisuccinate 382-383 IQC materials 660 metabolites, use of solid phase cartridges
for separation 133 Murex EQA scheme 689 solvent extraction 378-379 UK NEQAS participation 654 WHO EQAS 692
progestogens 369, 426 see also progesterone; medroxyprogesterone acetate; norethisterone; norgestrel
protein binding agents for disruption of 121
proton NMR spectroscopy 59 provitamin D3 see 7-dehydrocholesterol pseudohomogenous RIA for testosterone
350-351 PT see Proficiency Testing
QA see quality assurance quadrupole ion trap 100 quadrupole mass filter 100 quadrupole mass spectrometer 94, 99-100, 102
quality 647, 648
INDEX
quality assessment see External Quality Assessment
quality assurance 403-404, 438, 648 for 25-hydroxyvitamin D3 575, 587
quality control 403 see also Internal Quality Control
quality goals 648, 656, 662 quality manual 656 quality system 656
RAC see repeat analytical controls radioimmunoassay 324-332, 349-353
alternative methodologies for 332-334
commercially available kits for 324-332, 337
comparison of with GC-MS 353 of cyproterone acetate 407-408 of norethisterone 413-415 of norgestrel 415-416 of plasma progesterone 375-385 of salivary progesterone 396--398 pseudo homogeneous for testosterone
350-351 separation of bound/free steroid in
350-351 use in measurement of aldosterone 250-252
see also RIA radiolabelled steroids
as internal standards 117 use III Immunoassay 232, 233
Randox International Quality Assessment Scheme 689
rare earth chelates 439 rate dialysis 394 reagent gas 97 receptor auxiliary factor (RAF) 566 receptors for 1,25-(OH)2D3
in skin 567 widespread distribution 567
recovery 438, 663, 675, 678, 679, 683 5o:-reductase deficiency 242 reference interval 657 reference method see method refractive index, as method of detection during HPLC 154
repeat analytical controls 659 Repetends 659 see also repeat analytical controls
reproducibility 659 9-cis-retinoic acid 567 retinoid-X receptor (RXR) 566
involvement in tight VDR-VDRE binding and transcriptional activation 567
RIA 430, 433, 446, 456 see radioimmunoassay
INDEX
ring closure, in vitamin D metabolites during GC 594
ring residues in UV spectroscopy 31 R/S-nomenc1ature for chiral centres 20-23 RU486 see mifepristone
saliva analysis, as alternative to plasma 237 collection of 237 determination of progesterone in
396--398 equivalence to plasma free levels 396 17-hydroxyprogesterone in 245
saponification 120 scintillation counting 452 scintillation proximity assays 351, 433, 437 for cortisol 240 use in measurement of 1,25-dihydroxyvitamin D3 591
use of in eliminating separation of free/ bound steroid 351
screening analysis for anabolic steroids by GC-MS 498-503 by immunoassay 493-498 of natural hormones 509-518 of synthetic hormones 492-503
seco steroids 3, 20 (5Z, 7 E)-(lS,3R, 24R)-9,10-secocholesta-
5,7,10(19)-triene-1,3,24,25-tetrol 20 (5Z,7 E)-(3S)-9,1 0-secocholesta-5, 7,10(19)
triene-3,25-diol 20 (5Z, 7 E)-(3S)-9, 1 0-secocholesta-5, 7,1 O( 19)
triene-3-ol 20 (5Z, 7 E)-( 1 S,3R )-9,1 O-secocholesta-
5,7,10(19)-triene-1,3,25-triol 20 secocholestane 2 second antibody separation 433, 443, 448
in immunoassay 387 see also separation of free and bound
analyte SECSY NMR spectra 91 selected ion monitoring 94, 104 selected reaction monitoring 94, 104 Sep-Pak 572 Sep-Pak cartridges 432, 453 Sep-Pak SIL cartridges
for steroid purification 575-576 use in extraction of vitamin D metabolites 598
separation of free and bound analyte 443
dextran-coated charcoal 433 second antibody 433 solid phase 433, 437
of ions in mass spectrometry 99 separators for mass spectrometry 94 Sephacryl, use in affinity columns 574
Sephadex 431 Sephadex diethylaminoethyl 428 Sephadex G-25 429
715
Sephadex LH-20 430, 431, 441, 442, 443, 445,449,450,451,577
Sephadex, use in fractionation of neutral steroids 134
Sepharose-4B 445 sequence rules 21 serum see materials sex hormone binding globulin see SHBG SHBG 428, 430, 441, 451, 452 SHBG-bound testosterone, assays of 337-339
SHBG-effect in 'direct' assays for testosterone 336-337
silanisation of glassware 573 silica cartridges for removal of excess derivatising agents 577
silica-gel 442 silyl ethers in mass spectrometry 106 skin
effect of UV radiation 563 formation of lumisterol3 563 formation of tachysterol3 563 formation of vitamin D3 562 vitamin D3 in 594
SKZL see Stichting Kwaliteitsbewaking Klinische Chemische Ziekenhuislaboratoria
sodium azide 658, 665-666 sodium borohydride 445 soft ionisation 98
techniques in MS 159 fj-solamarine 637 solasodine 638 solid-phase antibody 442 solid-phase columns, use for separation of corticosteroids 13 3
solid-phase extraction 124 et seq., 572 details of available sorbents 126
solid phases Amberlite XAD2 use of 277-278 CIS by Sep-Pak cartridges use of 279 Extrelut use of 292 use in C 19 steroid extraction 277-279,
280-291, 324-332 solvent extraction 428, 443
of vitamin D and metabolites 571-572 selectivity of 121
solvent residues 440 solvents 440
available for steroid extraction 119 for IR spectroscopy 42 for NMR spectroscopy 66 for UV spectroscopy 29
solvolysis 429, 446 sorbents, for solid-phase extraction 126
716
specific rotation 36 specificity 435, 438, 441, 443 spin decoupling in NMR spectroscopy 81 spin-spin coupling in NMR spectroscopy 69,75
spironolactone 52 stable isotope labelled steroid, use of as internal standard in GC-MS 311-315
stable isotope labelling 108 stacked plot in 1 H NMR spectroscopy
88 standardisation 650 stanozolol
phase I metabolism of 478--480 phase II metabolism of 483 structure of 471 see also 17 fJ-hydroxy-171X-methyl androgens
stationary phases for GLC 137-138 steady state gel filtration 393-394 steric hindrance 433 steroid
acids, HPLC-MS of 174-176 analogues, as internal standards 116 binding proteins, interaction and extraction 114-115
conjugates, HPLC-MS of 176--184 hydrolysis of 122-124
derivatisation prior to GLC 140 fatty acid esters, HPLC-MS of
172-174 glucuronide profiles, in urine by negative ion LCIMS 98
glucuronides, enzyme hydrolysis 122-124
hydrazones, use in HPLC 156-157 sulphates
acid hydrolysis 122-124 HPLC-MS of 179-184 in plasma of newborn infants 244
steroidal alkaloids 637-640 Stichting Kwaliteitsbewaking Klinische Chemische Ziekenhuislaboratoria 690
stimulation and suppression tests 232 streptavidin 439 sulphatases 547 sulphate 15, 437, 439 17 fJ-sulphates
chemical synthesis of 482 epimerisation of 482 hydrolysis of 489--491 preferential formation of 483--484 quantification of intact conjugates of 492
urinary excretion of 476, 485 sulpho-conjugates see 17 fJ-sulphates sulphuric acid conjugation 449 Supe1co C18 454
INDEX
symmetric dialysis (SYD), saliva testosterone assay by 338-339
synergism 438 synthetic corticosteroids, assays for 243
tachysterol2, HPLC of 565 tachysterol3
formation in skin 563 structure 563 UV spectrum 564
tandem mass spectrometry 95, 104, 160-161
TIE ratio see hormone ratios testosterone 6, 11, 60, 86, 87, 437, 452
and hCG, effect on TIE test 515 and ketoconazole administration of
514-515 assays for conjugates 345 biosynthesis of 268 CAP PT scheme 691 CIAs for 346 cross-reactions of with 51X-DHT 340,
351 detection of administration of
in equine 518 in human 510-515
ELISAs for 340-341 EQA serum concentration 666,669 FIAs for 347-348 forms of in plasma, assay of 337-339 formulations of 475 hydrolysis of conjugates of 489--490,
511 hydroxy derivatives of, separation by HPLC 297-298
hydroxylation of 272 IQC materials 660 mass spectrum of
as MO-HFB 509 as oxime-bis-TBDMS 502
measurement of 337-339 by GC-MS 280-291,309
metabolism of 269-272, 476--477, 484--485
misuse of 470, 510 Murex EQA scheme 689 pseudohomogeneous RIA for 350-351 RIAs for 350-351 separation of, from epitestosterone and
llfJ-hydroxyandrosterone 511-512 specificity of tests for doping with
512-515 structural modifications of 474 total ion chromatogram for 512 UK NEQAS participation 654 urinary metabolites of, in equine 518 WHO EQAS 692 see also hormone ratios
tetrahydroaldosterone, measurement in urine 252
tetrahydrocortisol 690 tetrahydrocortisone 690 tetrahydrocortisone-3-glucuronide, HPLC-MS of 178
tetrahydro-ll-deoxycortisol 246 tetrahydro-18-hydroxy DOC 256 thermospray 95
HPLC-MS, use in analysis of steroid sulphates 248-249
mass spectrometry 95, 97, 99 MS 166--167
thin-layer chromatography 135-136 in separation of androgens 274--276,
280-291, 324--332, 336 in separation of C 19 steroid conjugates
319 of anabolic steroids 489 review of use 136 see also TLC
thyroglobulin-antigenic protein 447 time-of-flight analysers 101 time-resolved fluorescence 439 tissue extraction 121 TLC 430,431,442,443 TMS derivatives and anabolic steroids 498-500
total ion current 103 in mass spectrometry 103
Total Quality Management 648 TQM see Total Quality Management transfec1ed COS cell, use in assessment of biological activity 593
trenbolone structure of 471 triazoline adducts, use in measurement of
vitamin D metabolites 583-585 1,24,25-trihydroxyvitamin D3, structure 568
11{3,17,21-trihydroxypregn-4-ene-3,20-dione 7, 11, 86
trimethylsilyl ether, derivatives for mass spectrometry 106
trimethylsilyl ethers, formation from vitamin D metabolites prior to GC-MS 597
trimethylsilyl oximes, derivatives for mass spectrometry 107
tritium NMR spectroscopy 91 two-dimensional NMR spectroscopy
86 tyrosine 438 tyrosine methyl ester 443
UK NEQAS advisory panels 663 cortisol methods 651 data presentation 670
INDEX
GC-MS studies 678-685 linearity studies 675 oestradiol method types 653 of cortisol 238 participation 653, 654, 664 performance criteria 670
717
performance evaluation 667, 671-675, 676
recovery studies 677-678, 679 reports 670 samples 665 steering committee 663 surveys 664 target values 667, 671, 672, 673 urinary free cortisol 685, 687-688
ultrafiltration 394 ultraviolet
absorption spectra 26 detection of vitamin D metabolites after
HPLC 579 unconjugated estrogens 428 1X,{3-unsaturated ketones, detection after
HPLC 154-155 urinary steroids
analysis by LSIMS 98-99 assay in investigation of steroid abnormalities 236-237
in normal man by GLC 142 profile by GC-MS 162
urine estrogens 428 human pregnancy 431,432 profile analysis 242
uterus cytosol binding rabbit 430, 441, 442 rat 445
UV spectrum of 7-dehydrocholesterol 504 of lumisterol3 564 of tachysterol3 564 of vitamin D3 564
vaginal cornification 433 V AR see variability of the bias variability of the bias 670 see also UK
NEQAS, performance criteria vibrational spectra 38 vitamin D 8, 20, 33, 562 et seq.
assay of metabolites by saturation analysis 588-591
assays, use of internal standard 574
binding protein see DBP chemical analogues of 569 chemistry 562 et seq. esters 573 extraction 571-575 in food, extraction of 120
718
vitamin D (contd) in milk 562 measurement 581
by GC-MS 596 metabolites, assay by mass spectrometry 594 et seq.
metabolites, formation of N-Omethyloximes for GLC 595
metabolites, in plasma by GC-MS 596 et seq.
metabolites, measurement by HPLC with electrochemical detection 586
metabolites, separation on SIL cartridges 576, 577
metabolites, solid-phase extractions 572-573
metabolites, storage under nitrogen 573-574
metabolites, structural investigations by NMR 606-610
metabolites, structures 568-569 metabolites, use of affinity column for
extraction 574 metabolites, use of amber glass for
storage 574 metabolites, use of triazoline adducts in
measurement 583-585 nomenclature 562 receptor (VDR) 566
INDEX
reviews of analytical methodology for 571
solvents used for extraction 572 stages in analysis of 570 structure 562 et seq.
vitamin D-responsive element 567 vitamin D 2, in plants 562 vitamin D3
conjugates, use of static F AB 583 formation in skin 562 UV spectrum 564
vitamin D3-25-hydroxylase 565 vitamin D3-25-hydroxylase, in vitro studies
using GC-MS 599
Wakosil 5C18 454 walk-away automation see automated immunoanalysers
Wellcome Quality Assessment Scheme see Murex Quality Assessment Programmes
WHO see World Health Organisation withaferin-A 626, 627 withametelin 628 withanolides 626-631 World Health Organisation 663, 691-692
Zorbax SIL, use in HPLC separation of vitamin D2 isomers 565