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is given above.

For systematic toxicological screening of blood (serum,


plasma) samples by HPIC-DAD, the measurement of two extracts
obtained at pH 2 and pH 9 with dichloromethane and of the supernatant
of a protein precipitation by acetonitrile has proved to be very useful
(Pragst et u|. 2002).
Preparaticn cf a basic and an acidic methyIene chIcride extract
1. Dispense 500 I of whole blood, serum or plasma into two 1.5-mI
vials.
2. To vial 1 add 100 I of a 0.2 mol/I solution of tri(hydroxymethyl)
amine (basic extract).
3. To vial 2 add 100 I of 0.1 mol/I hydrochloric acid (acidic extract).
4. To both vials add 400 I of dichloromethane.
5. Vortex mix the vials for 1 min and centrifuge.
6. Withdraw 200 I of the dichloromethane extract and evaporate the
solvent at room temperature under a stream of nitrogen.
7. Dissolve the residue in 100 I of mobile phase.
8. Analyse 50 I of each extract (basic extract in mobile phase A and
acidic extract in mobile phase B).
Prctein precipitaticn by acetcnitriIe
1. To 500 I of whole blood, serum or plasma add 500 I of aceto-
nitrile.
2. Vortex the mixture for 2 min and centrifuge.
3. Separate off the supernatant.
4. Analyse 50 I in mobile phase A.
Protein precipitation is particularly useful for hydrophilic drugs, which
are extracted poorly by the procedure mentioned above. These include
paracetamol, salicylic acid and lamotrigine. The limits of detection are
between 0.01 and 0.1 g/mI for dichloromethane extraction (depend-
ing on the extinction coefficient and on the extraction yield) and
between 0.1 and 1 g/mI for protein precipitation.
Applicution exumple
In STA, the library search must be applied to all peaks of the HPIC-DAD
chromatogram. As an example, the chromatogram at 225 nm of the
basic extract from the blood sample of a lethal drug poisoning case
and the UV spectra of the highest peaks are shown in Figure 41.14. To
determine RRT, the standard compound (MPPH, peak no. 10,
RRT 1.000) was added. From the remaining 11 peaks of the chrom-
atogram, 7 could be identified by both UV spectrum and RRT. As the
result, a high overdose of trimipramine and promethazine was found to
be the cause of death. The extensive metabolism indicated that there had
been a long survival time after drug ingestion. The similarities between
the UV spectra of the parent drugs (peaks 9 and 12) and some of their
metabolites (peak 8, and peaks 6 and 11, respectively) are also demon-
strated in this case. On the other hand, the sulfoxides of promethazine
(peak 3) and desmethylpromethazine (peak 2) show completely chan-
ged spectra because of the transformation that takes place directly at the
UV-absorbing phenothiazine ring. Caffeine (peak 1) is found in almost
all samples. The poor separation of peaks 4, 5 and 7 meant that the UV
spectra were not suitable for a library search.
Recommended HPLC systems
There are general screening methods based on gradient elution and
retention indices that have proven value by many laboratories, and
data from these are listed below (systems HA, HX, HZ, HYand HAA).
Another (system HBK) is based on a combination of isocratic systems.
The tabulated data are derived from systems in which groups of
compounds have been chromatographed either as part of a general
screening procedure or from systems that have been used specifically
for that group of compounds. Other systems for the chromatography
of individual compounds, especially those used for quantification, are
given in the monographs.
Chromatographic retention data are presented as k values as well as
retention times (RTs), retention indices (RIs) and relative retention
times (RRTs).
Ncte In the tables, a dash indicates that no value is available for the
compound, not that it does not elute.
CeneraI screens
5ystem HA
Jane I et u|. (1985). j Chrcmutcgr 323: 191-225.
 Cc|umn. Silica Spherisorb S5W (125 4.9 mm i.d., 5 m).
 Mcbi|e phuse. Solution containing 1.175 g (0.01 mol/I) ammonium
perchlorate in 1 I methanol; adjust to pH 6.7 by the addition of 1 mI
0.1 mol/I sodium hydroxide in methanol.
 k vu|ues. Values for drugs in this system will be found in drug
monographs and in the Indexes to Analytical Data; they are also
included in the systems for specific groups of drugs that follow.
5ystem HX
J Hartstra, JP Franke, RA de Zeeuw, personal communication.
 Cc|umn. Iichrospher 60 RP-Select B (125 4.0 mm i.d., 5 m) with
precolumn Iichrospher 60 RP-Select B (4 4.0 mm i.d., 5 m).
 Mcbi|e phuse. (A:B) triethylammonium phosphate buffer (25 mmol/
I, pH 3.0)-acetonitrile.
 L|uticn prcgrumme. (A:B) (100 : 0) to (30 : 70) in 30 min, hold
10 min, back to initial conditions in 3 min with equilibration for
10 min before next injection.
 I|cw rute. 1 mI/min.
 Detecticn. DAD.
 Stundurds. Nitro-n-alkanes (C to C ) 10 I in 10 mI acetonitrile.
 RI vu|ues. Values for drugs in this system will be found in the mono-
graphs and in the Indexes to Analytical Data; they are also included in
the systems for specific groups of drugs that follow.
5ystem HY
RK Watt, RA Waters, AC Moffat, unpublished information.
 Cc|umn. C symmetry (250 4.6 mm i.d., 5 m).
 Cc|umn temperuture. 40 C.
 Mcbi|e phuse. (A:B) sulfuric acid (0.5 mI 2.5 mol/I) in water
(500 mI)-sulfuric acid (0.5 mI 2.5 mol/I) in acetonitrile (500 mI).
 L|uticn prcgrumme. (98 : 2) for 3 min to (2 : 98) over 23 min, hold for
10 min, back to initial conditions over 2 min with equilibration of
8 min before next injection.
 Detecticn. DAD.
 Stundurds. Nitro-n-alkanes (C to C ) 10 I in 10 mI acetonitrile.
 RI vu|ues. Values for drugs in this system will be found in the
monographs and in the Indexes to Analytical Data; they are
also included in the systems for specific groups of drugs that
follow.
5ystem HZ
Conemans JMH et u|. http://www.zanob.nl/pages/ISShowElements
Page_v2.asp?IistID1650&elemid29275&articleid133751&token
(accessed 14 December 2010).
 Cc|umn. C end-capped IiChrospher 100 RP-18e (125 4.0 mm
i.d., 5 m), with precolumn IiChrocart 124-4.
 Mcbi|e phuse. Add 146 I triethylamine and about 750 I phos-
phoric acid to 530 mI water. Adjust pH to 3.3 using a 10% potassium
hydroxide solution and finally add 470 mI acetonitrile.
 I|cw rute. 0.6 mI/min.
 Detecticn. DAD.
 Retenticn times. Values for drugs in this system will be found in
the monographs and in the Indexes to Analytical Data; they are
also included in the systems for specific groups of drugs that
follow.
5ystem HAA
Gaillard Y, Pepin G. (1997). j Chrcmutcgr A 763: 149-163.
 Cc|umn. C Symmetry (250 4.6 mm i.d., 5 m) with Symmetry C
precolumn (20 mm).
 Cc|umn temperuture. 30 C.
 Mcbi|e phuse. (A:B) phosphate buffer (pH 3.8)-acetonitrile.
 L|uticn prcgrumme. (85 : 15) for 6.5 min to (65 : 35) until 25 min to
(20 : 80) for 3 min, and back to initial conditions for equilibration for
7 min.
Recommended HPLC sysLems 7J7
 I|cw rute. 1 mI/min for 6.5 min, then linear increase to 1.5 mI/min
for 6.5-25 min and hold for 3 min (re-equilibration is made at
1.5 mI/min).
 Detecticn. DAD.
 Retenticn times. Values for drugs in this system will be found in
the monographs and in the Indexes to Analytical Data; they are
also included in the systems for specific groups of drugs that
follow.
Figure 41.14 HPLCD^D invesLigaLion of a combined LrimiþramineþromeLhazine þoisoning. ChromaLogram of a basic exLracL of a venous blood samþle,
Uv sþecLra of Lhe highesL þeaks, resulLs of Lhe library search and semiquanLiLaLively deLermined concenLraLions.
7J8 High Performance Liquid ChromaLograþhy
5ystem H8K
Pragst F et u|. (2001) UV Spectru cf Tcxic Ccmpcunds. Heppenheim:
Verlag Dr Dieter Helm.
 Cc|umn. Iichrospher RP-8ec (250 4.0 mm i.d., 5 m).
 Mcbi|e phuse. Three different composition are used: A: acetoni-
trile-phosphate buffer pH 2.3 (33 : 67). Internal standard: 5-(4-
methylphenyl)-5-phenylhydantoin (for compounds eluting within
30 min). B: acetonitrile-phosphate buffer pH 2.3 (67 : 33). Internal
standard: 4-phenylbenzophenone (for compounds eluting after
30 min). C: acetonitrile-phosphate buffer pH 2.3 (20 : 80).
Internal standard: salicylamide (for compounds with RRTs below
0.2).
 I|cw rute. 1 mI/min.
 Detecticn. DAD.
 ^cte. The phosphate buffer is prepared by dissolving 4.8 g phospho-
ric acid (85%) and 6.66 g potassium dihydrogenphosphate in 1 I
water, adjust pH to 2.3. Values for drugs in this system will only be
found in the Indexes to Analytical Data.
Amfetamines, cther stimuIants and ancrectics
Systems HA, HX or HY previously described may be used, or Systems
HB or HC below.
5ystem H8
Gill R et u|. (1981). j Chrcmutcgr 218: 639-646.
 Cc|umn. ODS Hypersil (250 5 mm i.d., 5 m).
 Mcbi|e phuse. Solution containing 19.60 g (0.2 mol/I) phosphoric
acid and 7.314 g (0.1 mol/I) diethylamine in 1 I of a 10% v/v solu-
tion of methanol; adjust the pH to 3.15 by the addition of sodium
hydroxide solution.
5ystem HC
Iaw B et u|. (1984). j Chrcmutcgr 301: 165-172.
 Cc|umn. Silica Spherisorb (250 5 mm i.d., 5 m).
 Mcbi|e phuse. Methanol-ammonium nitrate buffer solution (90 : 10).
To prepare the buffer solution add 94 mI strong ammonia solution
and 21.5 mI nitric acid to 884 mI water and adjust to pH 10 by the
addition of strong ammonia solution.
Amfetamines, cther stimuIants and ancrectics
HA H8 HC HX HY
k k k Rl Rl
^drenaline   0.53  
^mfeLamine 0.9 8.48 0.98 244 
BenzfeLamine 1.2  0.15  
Brucine 11.1   312 257
Caffeine 0.2  0.25  
CaLhine 1 4.39 0.83  
ChlorþhenLermine 0.9  0.82  
DieLhylþroþion 1.7  0.15  230
DimeLhylamfeLamine  11.08 1.89  
D0M   1.13  
Eþhedrine 1.0 5.58 1.79  
Fencamfamin 1.3  0.72 354 309
FeneLhylline   0.27  
Fenfluramine 1.3  0.88 371 315
Norfenfluramine 1    
Fenþroþorex     225
Hordenine  2.00   
HydroxyamfeLamine  2.24 1.11  
Hydroxyeþhedrine  0.73   
Mazindol 1.8  0.2 357 285
MeþhenLermine 1.5  2.48  
Mescaline 1.3 15.82 2.17  
MeLamfeLamine 2 10.52 2.07 252 215
MeLhoxyamfeLamine  14.95   
MeLhoxyþhenamine 1.7 32.17   
MeLhylamfeLamine 2.0 10.52 2.07  
MeLhylenedioxymeLhamfeLamine    278 252
MeLhyleþhedrine 2.3  1.83  
MeLhylþhenidaLe 1.7  0.35  277
Noradrenaline  0.10   
NormeLaneþhrine   1.08  
0xedrine  0.27   
Pemoline 0.2  0.1 307 271
PhendimeLra zine 0.9  0.3 253 218
Phenelzine 1.0 5.91 0.37  
tab|e ccntinued
Recommended HPLC sysLems 7J9
AnaIgesics, ncn-stercidaI anti-infIammatcry drugs
5ystem HD
HM Stevens, R Gill, unpublished data.
 Cc|umn. ODS Hypersil (160 5 mm i.d., 5 m).
 Mcbi|e phuse. Isopropyl alcohol-formic acid-0.1 mol/I potassium
dihydrogenphosphate (13.61 g/I; 540 : 1 : 1000).
5ystem Hv
 Cc|umn. ODS Spherisorb (200 4.6 mm i.d., 5 m).
 Mcbi|e phuse. Acetronitrile-acetic acid (45 : 55) for 2 min, to (75 : 25)
at 3%/min, for 6 min.
 I|cw rute. 1.7 mI/min.
5ystem Hw
HM Stevens, R Gill, unpublished data.
 Cc|umn. As for System HD, above.
 Mcbi|e phuse. Isopropyl alcohol-formic acid-0.1 mol/I potassium
dihydrogenphosphate (13.61 g/I; 176 : 1 : 1000).
AntifungaIs
The general screening systems, previously described, may be used.
Amfetamines, cther stimuIants and ancrectics, ccntinued
HA H8 HC HX HY
k k k Rl Rl
PheneLhylamine 1.2 3.54 1.31  
PhenmeLrazine 1.7   258 241
PhenLermine 0.5 19.45 0.85  245
Phenyleþhrine 1.3  1.54  
Phenylþroþanolamine 0.9 3.87 0.70  
Piþradrol 1.2  0.59 355 
ProlinLane 2  1.3 370 
Pseudoeþhedrine 1.2 5.90 1.77  
Tranylcyþromine 1.0  0.25  
TrimeLhoxyamfeLamine   1.48  
Tyramine 1.2 0.81 1.47  
AnaIgesics, N5AlDs
HD Hv Hw HX HY HZ HAA
k RR7 k Rl Rl R7 R7
^ceLanilide 0.5  2.3  281  
ParaceLamol 0.1  0.32    
^lclofenac 2.5 0.51     
^minoþhenazone 0.2  0.32 252 204 2.1 
^sþirin 0.5  2.7 350 318 2.7 
Salicylic acid 0.7  4.5    
BenorilaLe 0.7  22.4    
^sþirin 0.5  2.7    
ParaceLamol 0.1  0.32    
Benoxaþrofen 11.3 0.98     
Clonixin  0.87   345  
Diclofenac 11.5 0.85  515 592 14.8 22.1
Diflunisal 4.1 0.77  508 583 5.4 
Diþyrone 0.1  0.45 315 194 1.4 
ELenzamide 0.55  4.5  303  
Fenbufen 4 0.81  520 451  19.3
Fenoþrofen 7.9   574 524 10.9 21.2
FlocLafenine      4.4 17.2
Flufenamic acid 19.7 1  571 557  
Flunixin  0.99   414  
AntifungaIs
HX HY HZ HAA
Rl Rl R7 R7
Econazole 525 385  20.1
Fluconazole 340 289  11.4
FlucyLosine 72  1.5 3.1
Criseofulvin  488  18.4
KeLoconazole 439 454 5.2 15.7
740 High Performance Liquid ChromaLograþhy
AntibacteriaIs
The general screening systems, previously described, may be used.
AnaIgesics, N5AlDs, ccntinued
HD Hv Hw HX HY HZ HAA
k RR7 k Rl Rl R7 R7
Flurbiþrofen  0.89  585  11.8 21.3
Clafenine    372 275 2.3 
lbuþrofen 15.1   515 598 15.5 23.8
lndomeLacin 5.95 0.87  507 590 14.4 21.7
lndoþrofen 1.2 0.52   405  
KeLoþrofen 2.4 0.55  495  5.4 19.5
KeLorolac      4.1 
Meclofenamic acid    553 590  
Mefenamic acid 21.1 0.95  551 585  
MeLhyl salicylaLe 3.9   480 449  
Salicylic acid 0.7      
Morazone 0.4  2.05  294  
Naþroxen 3.3   501 458 5.8 
Nefoþam     313  12.7
Nifenazone 0.1  0.45 310   
Niflumic acid  0.93  595 530  22
0xyþhenbuLazone 1.95 0.59  501 459 5.7 
ParaceLamol 0.1  0.32 254 241 1.9 5.5
PhenaceLin 0.5  4.4 377 335 3.0 
ParaceLamol 0.1  0.3 254 241 1.9 
Phenazone 0.1  0.95 333 299 2.1 
PhenylbuLazone 5.5 0.95  572 543 19.5 24.1
0xyþhenbuLazone 1.95 0.7  501 459 5.7 
Piroxicam 0.5  7.7 431 382 4.9 15.5
M (5-hydroxy)     445  
Proþyþhenazone 1.3  11 441 370 4.7 
Salicylamide 0.4  2.5 327 289  
SalsalaLe 3.5 0.59     
Sulindac 1.25 0.78  488 452 3.9 15.5
Sulindac sulfoxide      7.2 
Tenoxicam    355   12.7
Tiaþrofenic acid    484 452 5.8 17.5
Tolfenamic acid    590  37.9 
TolmeLin 2.05 0.50 and 0.99  470 434 5.4 
Zomeþirac 3.7    495  
AntibacteriaIs
HX HY HAA
Rl Rl R7
^moxicillin  225 3.1
^mþicillin  250 3.8
^ziLhromycin   
CefLriaxone 239  5.3
Chloramþhenicol 390 335 14.1
Ciþrofloxacin 318 250 9.1
ClariLhromycin   
Clindamycin 354 291 12
Furazolidone 335  12.2
tab|e ccntinued
AntibacteriaIs, ccntinued
HX HY HAA
Rl Rl R7
lsoniazid  245 
MeLronidazole 257 225 5.8
Minocycline  240 22.5
Nalidixic acid  380 15
NiLrofuranLoin 319 288 
0floxacin 314 250 8.5
0xyLeLracycline dehydraLe 299 250 
Rifamþicin  417 15.2
RoxiLhromycin   15.8
TeLracycline 314 255 9.9
TrimeLhoþrim 299 254 8.3
Recommended HPLC sysLems 741
AntichcIinergics
The general screening systems, previously described, may be used or
systems HAX and HAY below.
5ystem HAX
Koves EM (1995). j Chrcmutcgr A 692: 103-119.
 Cc|umn. Supelcosil IC-DP (250 4.6 mm i.d., 5 m).
 L|uent. (A : B : C) Acetonitrile-phosphoric acid (0.025% v/v)-
triethylamine buffer.
 Isccrutic e|uticn. (25 : 10 : 5).
 I|cw rute. 0.6 mI/min.
 Detecticn. DAD ( 229 nm).
 ^cte. The triethylamine (TEA) buffer is prepared by adding 9 mI
concentrated phosphoric acid and 10 mITEAto 900 mIwater, adjusted
to pH 3.4 with diluted phosphoric acid and made up to 1 I with water.
5ystem HAY
Koves EM (1995). j Chrcmutcgr A 692: 103-119.
 Cc|umn. IiChrospher 100 RP-8 (250 4.0 mm i.d., 5 m).
 L|uent. (A : B : C) as for System HAX.
 Isccrutic e|uticn. (60 : 25 : 15).
 I|cw rute. 0.6 mI/min.
 Detecticn. DAD ( 229 nm).
AnticcnvuIsants and barbiturates
5ystem HE
Christofides JA, Fry DE (1980). C|in Chem 26: 499-501.
 Cc|umn. Alkyl-silica SAS-Hypersil (125 4.5 mm i.d., 5 m).
 Mcbi|e phuse. Acetonitrile-tetrabutylammonium phosphate,
0.005 mol/I, pH 7.5 (20 : 80).
5ystem HC
Gill R et u|. (1981). j Chrcmutcgr 204: 275-284.
 Cc|umn. ODS Hypersil (150 4.6 mm i.d., 5 m).
 Mcbi|e phuse. Methanol-0.1 mol/I sodium dihydrogenphosphate
(11.998 g/I) (40 : 60); adjust to pH 3.5 by the addition of phosphoric
acid.
5ystem HH
Gill R et u|. (1981). j Chrcmutcgr 226; Bicmed App| 15: 117-123.
 Cc|umn. As for System HG, above.
 Mcbi|e phuse. As for System HG except that the mixture is
adjusted to pH 8.5 by the addition of sodium hydroxide
solution.
AntichcIinergics
HA HX HY HZ HAA HAX HAY
k Rl Rl R7 R7 R7 R7
^diþhenine 1.8 422     
^Lroþine 3.9 305 251 2.2 10.4 7 3.8
Biþeriden    5.4 14.8  
Chlorþhenoxamine 2.9  345    
Clidinium  379     
Clidinium bromide     13.3  
CycloþenLolaLe 1.5 353 287 3.2   
Dicycloverine 1.1  575    
DieLhazine 3.4     15.1 7.4
Emeþronium bromide 5.2 420     
HomaLroþine 4.2 272 223   5.8 3.5
Hyoscine 1.1 270 253  7.4 7 3.7
Hyoscyamine 3.7    9.7  
lsoþroþamide lodide 2.4 379     
MeLixene 3.5 451     
0rþhenadrine 3 418 323 5   
N-MonodesmeLhylorþhenadrine 1.7      
N-0xide 1.1      
0xyþhencyclimine 2.8 424     
0xyþhenonium bromide 2.5 424     
PiþeridolaLe 1.7 429     
Procyclidine 2 405  5.2  20 4.7
Profenamine 2.4 444 338   15.5 8.3
ProþanLheline bromide 4.4 454     
XanLhanoic acid  499     
Trihexyþhenidyl 1.8 429 381 7.5 15.3  
742 High Performance Liquid ChromaLograþhy
Antidepressants
The general screening systems, previously described, may be used or
systems HF and HAZ below.
5ystem HF
R Gill, unpublished data, after Kabra PM et u|. (1981). C|in Chim Actu
111: 123-132.
 Cc|umn. ODS Hypersil (160 5 mm i.d., 5 m).
 Mcbi|e phuse. Acetonitrile-phosphate buffer (pH 3.0; 30 : 70). To
prepare the phosphate buffer, add 0.6 mI nonylamine to 1 I
0.01 mol/I sodium dihydrogenphosphate (1.1998 g/I) and adjust
the pH to 3.0 by the addition of phosphoric acid.
5ystem HAZ
Chiba K et u|. (1995). j Chrcmutcgr B 668: 77-84.
AnticcnvuIsants, barbiturates and antiepiIeptics
HC HH HX HY HZ
k k Rl Rl R7
^llobarbiLal 2.45 1.33 345  2.7
^mobarbiLal 10.91 7.05 424 374 4
^þrobarbiLal 3.42 2.22 357 319 2.8
BarbiLal 1.11 0.53 308 258 2.2
BenacLyzine   382  
BrallobarbiLal 3.09 1.72 371 335 3
BuLalbiLal 5.17 3.48 394 342 3.4
BuLeLamaLe   390  
BuLobarbiLal 5.43 3.42 384 355 3.2
Carbamazeþine   418 358 
Clonazeþam   455 403 4.5
CyclobarbiLal 5.25 2.51 384 352 3.2
CycloþenLobarbiLal 5 3.84 391 352 
Enallylþroþymal 8.55 5.95  394 
ELhosuximide   301 275 2.3
FlavoxaLe     
HeþLabarb 9.9 4.93 415 377 3.9
HexeLhal 34.28 20.39  451 
HexobarbiLal 7.37 5.57 419 242 4.3
lbomal 4.01 2.58 379 352 
ldobuLal 8.12 4.77  357 
Mebeverine   448  7.1
MeþhenyLoin    355 3.7
Mesuximide    387 4.8
MeLharbiLal 2.59 1.99 435 324 
BarbiLal 1.11 0.53   
MeLhylþhenobarbiLal 7.27 3.84 435 395 4.5
NealbarbiLal 10.22 5.19 417 382 
Paþaverine   353 295 
PenLobarbiLal 10.95 8.07 424 383 4.1
Phenacemide   339 255 
PhenobarbiLal 3.09 1.23 379 335 3
PhenyLoin   431 381 3.7
Primidone   322 288 2.1
SecbuLabarbiLal 4.9 3.3 377 331 
SecobarbiLal 15.28 11.47 437 407 4.7
SulLiame   344 275 
TalbuLal 7.2 4.7 403 370 
Thiamylal   515 475 
ThioþenLal   485 433 5.9
vinbarbiLal 4.83 2.32 379 353 
vinylbiLal   424  4.1
Recommended HPLC sysLems 74J
 Cc|umn. C (250 4.0 mm i.d., 5 m).
 Mcbi|e phuse. (A : B : C) Water-methanol-triethylamine adjusted to
pH 5.5 with phosphoric acid.
 Isccrutic e|uticn. (70 : 30 : 0.1).
 I|cw rute. 0.7 mI/min.
 Detecticn. UV ( 240 nm).
Antidepressants and antipsychctics
HA HF HX HY HZ HAA HAX HAZ
k k Rl Rl R7 R7 R7 k
^miLriþLyline 3.3 5.42 440 375 7.5 15.9 15.8 1.75
10-HydroxyamiLriþLyline 2.9       
10-HydroxynorLriþLyline 1.8       
NorLriþLyline 2 4.58      1.71
^moxaþine   398   14.2  
Benþeridol 1.1  393 324 3.5   
BuLriþLyline 2.7 7.33  359    
NorbuLriþLyline 1.7       
CiLaloþram   403  4.5   
DesmeLhylciLaloþram     3.7   
Clomiþramine 3.4 9.92 452 405 10.2 15.4  
MonodesmeLhylclomiþramine 2       
Desiþramine 2.1 3.5 424 351 5.9 14.9 13 1.52
DidesmeLhylimiþramine 1.3       
2-Hydroxydesiþramine 1.2       
M (2-0H-)        0.39
Dibenzeþin 2.8 0.5 351 300    
Dosuleþin 3.2 3.5 428 357 5.7   
M (sulfoxide) 4.5       
M (nor-) 2.2       
Doxeþin 3.7 2.27 404 315 5 14.1 12.9 
M (nor-) 2.2    4.5   
FluoxeLine    400 7.5 15.2 12.2 
DesmeLhylfluoxeLine     5.7   
Fluvoxamine   430 353 5.5 15.3 10 
lmiþramine 4.2 4.17 437 335 5.7 15.1 14.7 1.52
Desiþramine 2.1 3.5      
2-Hydroxydesiþramine 1.2       
2-Hydroxyimiþramine 3.1       
M (10-0H-)        0.39
M (2-0H-)        0.39
M (N-oxide)        1.85
lþrindole 4.1 10.83      
lsocarboxazid   392 353    
MaþroLiline 2.2 4.92 438 389 5.5 15.5  1.44
DesmeLhylmaþroLiline 1.1       
Mianserin 1.8  391 342 4.5 13.8  1.18
M(nor-) 2.4       
M (nor-)        0.88
M (N-oxide)        0.53
M (8-0H-)        0.19
Moclobemide   295  2.4 10.2 5.9 
Nialamide 1.2  334     
Nomifensine 0.9 0.42 349 295    
NorLriþLyline 2 4.58  338 5.5 15.5 13.7 1.71
10-HydroxynorLriþLyline 1.8       
NoxiþLiline  1.53  330    
0þiþramol 2.2 1.53 377 340 3.9 14.2  
744 High Performance Liquid ChromaLograþhy
Antihistamines
The general screening systems, previously described, may be used.
AntimaIariaIs
The general screening systems, previously described, may be used.
Antidepressants and antipsychctics, ccntinued
HA HF HX HY HZ HAA HAX HAZ
k k Rl Rl R7 R7 R7 k
ParoxeLine   425 337 5.5 15.3 11.1 
Phenelzine 1  184     
ProLriþLyline 2.1 3.5 418 352    
Remoxiþride   334  3  8.8 
M (FL^-838)   315     
M (NCM-001)   354     
M (NCM-009)   341     
SerLraline   450  8.2  14.5 
(desmeLhylserLraline)     7.0   
Tofenacin 1.7    5.3   
Trazodone 0.5  378 305 3.3 12.7  
Trimiþramine 2.7 5.17 454 345 8.3 15.9 15.5 
M (nor-) 1.8       
viloxazine  2.7 325 273  11  
Zimeldine 3.2 0.57  270    
M (nor-) 2.9       
Antihistamines
HA HX HY HZ HAA HAX HAY
k Rl Rl R7 R7 R7 R7
^limemazine 3.1 420    14.9 7.1
^nLazoline 1.8 383 294    
^sLemizole   285 3.9 13.2  
(asLemizole)  383     
(M-nor)  351     
Bromazine 2.7 444     
Bromþheniramine 4.1  257  13.9  
Buclizine 0.7  454    
Carbinoxamine 4.7 359   12.8  
CeLirizine    3.5 15.7 8.89 5.29
Chlorcyclizine 2.3  340    
Chlorþhenamine 3.9 355 254 3.5 12.9 10.8 5.3
Cinnarizine 0.8 550  22 19.3  
ClemasLine 3.7 501  14   
Clemizole 4.8 420     
Cyclizine 2.9 405  4.8  12.4 5.8
Norcyclizine 2.2      
CyþroheþLadine 3.2  354 5.5 15  
DeþLroþine 5 471  10.3   
DimeLindene 5.1 338 288    
Diþhenhydramine 3.3 393 335   12.2 5
Diþhenylþyraline 3.3 401     
Doxylamine 4.4  259  11.1  
Hydroxyzine 1.4 437 325 5.7 15.3 11.4 5.3
lsoLhiþendyl 3.8 390   13.5  
LoraLadine  523 352 14.5 22.9 10.9 13.3
tab|e ccntinued
Recommended HPLC sysLems 745
AntinecpIastics
The general screening systems, previously described, may be used.
Antitussives
The general screening systems, previously described, may be used.
AntiviraIs
The general screening systems, previously described, may be used or
systems HAB and HAC below.
Antihistamines, ccntinued
HA HX HY HZ HAA HAX HAY
k Rl Rl R7 R7 R7 R7
Mebhydrolin 3 411  5.3   
Meclozine 0.7 587 398  20  
Meþyramine 3.9 448 257    
MeLhaþyrilene 4.1 342 197    
MeLhdilazine 5     15.2 5.7
Phenindamine 2.5 397     
Pheniramine 4.1 283 205   9.5 4.5
PhenylLoloxamine 3.1 415     
PizoLifen 3.4 435  5.5 15.2  
PromeLhazine 5 409 324 5.7 14.5 13.2 5.4
Proþiomazine 2.1 440 359   14.1 7.1
PyrrobuLamine 2.8 477     
Thenyldiamine 4 317     
Thiazinamium
meLilsulfaLe
   5.4   
TrimeLhobenzamide 4.7 347     
Triþelennamine 3.5 335 255    
Triþrolidine 3.2 388 270  13.1  
AntimaIariaIs
HA HX HY HZ HAA HAX HAY
k Rl Rl R7 R7 R7 R7
Chloroquine 15.2 282 245 2.1 5.4 12.7 3.5
Cinchonidine 3.1 305 214    
Cinchonine  304 209  10.2  
HalofanLrine  800   23  
Hydroxychloroquine  280  1.9  9.5 3.2
Primaquine 1.4  275    
Proguanil  379  3.8 13.5  
PyrimeLhamine 1  289  12.5  
Çuinine 2.4 327 245 2.5 11.3 8.3 4.5
AntinecpIastics
HX HAA
Rl R7
DieLhylsLilbesLrol 592 20.9
Doxorubicin 370 12.1
Fluorouracil 70 3.4
MeLhoLrexaLe 292 
vinblasLine  8.4
Antitussives
HA HX HY HAA
k Rl Rl Rl
Bromhexine 0.4 417 334 
DexLromeLhorþhan 5.5 377 298 13.3
DexLrorþhan 4.7   
DexLrorþhan  325  
Droþroþizine  240  7.2
Cuaifenesin  328 252 11.4
Noscaþine 0.3 358 289 12.8
Pholcodine 5 55 92 2.7
PiþazeLaLe 5.4 385  
746 High Performance Liquid ChromaLograþhy
5ystem HA8
Sparidans RW et u|. (2000). j Chrcmutcgr B Bicmed Sci App| 742:
185-192.
 Cc|umn. C Symmetry (100 4.6 mm i.d., 3.5 m) with Symmetry
C precolumn (20 3.8 mm, 5 m).
 Mcbi|e phuse. Acetonitrile-sodium phosphate buffer (25 mmol/I,
pH 6.8) (40 : 60).
 I|cw rute. 1.5 mI/min.
 Detecticn. Fluorescence ( 270 nm,  340 nm).
 ^cte. 8 min after each injection, flush column for 5 min at 1.5 mI/
min with acetonitrile-water (30 : 70). Equilibrate for about 8 min
with the original eluent before injecting the next sample.
5ystem HAC
Aymard G et u|. (2000). j Chrcmutcgr B Bicmed Sci App| 744:
227-240.
 Cc|umn. C Symmetry (250 4.6 mm i.d., 5 m) with C precol-
umn (Guard-Pak, Bondapak).
 Cc|umn temperuture. 37 C.
 Mcbi|e phuse. (A : B) Disodium hydrogenphosphate (0.04 mol/I)
with 4% (v/v) octane sulfonic acid (0.25 mol/I)-acetonitrile.
 Isccrutic e|uticn. (50 : 50).
 I|cw rute. 1.3 mI/min.
 Detecticn. DAD ( 261 nm between time 0 and 9 min;  241 nm
between time 9 and 20 min;  254 nm between time 20 and end of
the run (32 min).
Benzcdiazepines
5ystem Hl
R Gill, unpublished data.
 Cc|umn. ODS Hypersil (200 5 mm i.d., 5 m).
 Mcbi|e phuse. Methanol-water-phosphate buffer (55 : 25 : 20). To
prepare the phosphate buffer, dissolve 11.038 g (0.092 mol/I)
sodium dihydrogenphosphate and 1.136 g (0.008 mol/I) disodium
hydrogenphosphate in sufficient water to produce 1 I.
5ystem Hl
R Gill, unpublished data.
 Cc|umn. As for System HI, above.
 Mcbi|e phuse. Methanol-water-phosphate buffer (as in System HI)
(70 : 10 : 20).
5ystem HK
R Gill, unpublished data, after RJ Flanagan et u|. (1980). j Chrcmutcgr
187: 391-398.
 Cc|umn. Silica Spherisorb (250 5 mm i.d., 5 m).
 Mcbi|e phuse. Methanol to which has been added 100 I perchloric
acid per litre.
Cannabincids
5ystem HL
Baker PB et u|. (1980). j Anu| Tcxicc| 4: 145-152.
 Cc|umn. ODS Spherisorb (250 4.6 mm i.d., 5 m).
 Mcbi|e phuse. 0.01 mol/I sulfuric acid-methanol-acetonitrile
(7 : 8 : 9).
Cardiac gIyccsides
5ystem HM
Cobb PH (1976). Anu|yst (Icnd) 101: 768-776.
 Cc|umn. Silica IiChrosorb SI60 (250 4 mm i.d., 10 m).
 Mcbi|e phuse. Cyclohexane-ethanol-acetic acid (60 : 9 : 1).
AntiviraIs
HA8 HAC
R7 k
^bacavir 1 
^mþrenavir 4 2.5
Efavirenz  8.5
lndinavir 4.2 2
Benzcdiazepines
Hl Hl HK HX HY HZ HAA HAX HAY
k k k Rl Rl R7 R7 R7 R7
^cecarbromal    429 374    
^lþrazolam   2.79      
Bromazeþam   2.99      
tab|e ccntinued
Cannabincids
5ystem HL
k
Cannabichromene 19.09
Cannabicyclol 14.78
Cannabidiol 7.47
Cannabidiolic acid 8.75
Cannabigerol 8.18
Cannabinol 11.77
Cannabivarin 7.47
 -TeLrahydrocannabinol 14.07
 -TeLrahydrocannabinol 13.35
TeLrahydrocannabinolic acid 25.83
TeLrahydrocannabivaric acid 14.54
TeLrahydrocannabivarin 8.18
Cardiac gIyccsides
5ystem HM
k
DigiLoxigenin 2.0
DigiLoxigenin bisdigiLoxoside 3.9
DigiLoxigenin monodigiLoxoside 2.8
DigiLoxin 5.4
Digoxigenin 4.5
Digoxigenin bisdigiLoxoside 8.2
Digoxigenin monodigiLoxoside 5.5
Digoxin 11.3
CiLaloxin 5.8
CiLoxigenin 3.7
CiLoxigenin bisdigiLoxoside 5.5
CiLoxigenin monodigiLoxoside 4.5
CiLoxin 8.5
LanaLoside ^ 17.9
LanaLoside B 31.8
LanaLoside C 39.5
Recommended HPLC sysLems 747
Cardicactive drugs
The general screening systems, previously described, may be used.
Benzcdiazepines, ccntinued
Hl Hl HK HX HY HZ HAA HAX HAY
k k k Rl Rl R7 R7 R7 R7
Bromisoval    355 307 2.9   
BroLizolam    484  4.5  7.4 7.9
Carbromal    410 377 3.9   
Chlordiazeþoxide   2.87      
Clobazam   0.03      
ClomeLhiazole    395 292  15  
Clonazeþam   0.35      
Clorazeþic acid   2.00      
Demoxeþam   0.03      
Diazeþam   2.49      
Flumazenil    387 327 2.5   
FluniLrazeþam 3.15  0.47 483 305 5.5 18.5  
Flurazeþam  3.19 5.5 397 305 4.2  10.5 5.5
CluLeLhimide    435 401 4.8  5.5 5.2
KeLazolam   0.04      
Loþrazolam    388   13.4  
Lorazeþam   0.14      
LormeLazeþam 5.32  0.08 487 453 5.2   
Medazeþam   4.44      
MeLhaqualone    459 400 5.4  5.8 7.4
MeLhyþrylon    347 302    
Midazolam 9.75 2.1 5.9 399 305 4.2 14.9 10.2 5.3
NiLrazeþam 2.95  1.49 448 370 4.2 15.9 5.3 5
Nordazeþam   1.99      
0xazeþam 4.52  0.73      
Prazeþam   2.19      
Çuazeþam     755 37.5  11.9 17.7
Temazeþam 5.58  0.5 472 438 5.5 18.5 8.9 5.7
0xazeþam   0.73      
Triazolam 4.38  1.83 475 390 4.2 17.4 5.4 5.7
NoL deLecLed         
Zolþidem     291 3.2 11.9  
Zoþiclone    331 259 2.3  7.5 3.8
Cardicactive drugs
HA HX HY HZ HAA
k Rl Rl R7 R7
^|maline 2.8  277  
^lfuzosin    2.4 10.4
^miodarone 2.4 583 475 90.4 
MonodeseLhyl-
amiodarone
1.8    
^þrindine  433   17
BameLhan 0.9 250   5.9
BenzLhiazide   415  
BeLahisLine 3.1    3.2
BreLylium
LosilaLe
4.3  275  
Cardicactive drugs, ccntinued
HA HX HY HZ HAA
k Rl Rl R7 R7
Buþhenine 0.9 370   
CaþLoþril  315 283 2.1 9.7
Cilazaþril  420  4.5 14.4
CilazaþrilaLe    1.7 
Clonidine 1.2 258 194 2.5 5.1
Cloþamide  377 310  
Debrisoquine 1.2  245  
DilLiazem   351 4.5 14
DeaceLyl-
dilLiazem
    
DesmeLhyl-
dilLiazem
    
748 High Performance Liquid ChromaLograþhy
Diuretics
5ystem HN
R Gill et u|., unpublished data, after Tisdall PA et u|. (1980). C|in Chem
26: 702-706.
 Cc|umn. ODS Hypersil (160 5 mm i.d., 5 m).
 Mcbi|e phuse. Acetonitrile-water containing 10 mI/I acetic acid
(30 : 70).
Drugs cf abuse
A comprehensive HPIC method for the screening of common drugs of
abuse is described in Chapter 1, Table 1.24. Furthermore, an additional
eight systems (HBC, HBD, HBE, HBF, HBG, HBI and HBJ) are provided
in Chapter 11, Table 11.4.
Cardicactive drugs, ccntinued
HA HX HY HZ HAA
k Rl Rl R7 R7
DesaceLyldil-
Liazem
    
Disoþyramide 2.4 345 281 3 11.4
N-Monodesiso-
þroþyldisoþyramide
1.8    
Enalaþril  201  1.5 3.4
Encainide  353   
Felodiþine  590  25.8 24.4
Flecainide  419 355 5.2 
Hydralazine  193 132 1.9 
lsoxsuþrine 0.8 353 301  
LabeLalol 1.7 355 290 3 
Lidoflazine 0.5 530   
Lisinoþril  271 250 1.5 
Lorcainide 1.8 425  5.5 
MeLhyldoþa  59  1.4 3
MexileLine 1.2 329 278  11.5
Minoxidil  297  2.4 9.8
NafLidrofuryl
oxalaLe
  409  15.8
Nifediþine 0.2 527 454 7.2 19.5
Pargyline 0.2  203  
PenLaeriLhriLyl
LeLraniLraLe
 553   
(þenLaeriLhriLyl)     23.1
PenLoxifylline  355 274 2.1 11.5
Perindoþril    1.5 13.7
(þerindoþrilaL)  314   
Phenoxybenzamine 0.1 395   
PhenLolamine 1.7 358  3 
Pra|malium
biLarLraLe
2.2  340  
Prazosin 0.8 352  2.5 10.5
Procainamide 1.3 208 150 1.9 
N-^ceLylþro-
cainamide
3   1.8 
Çuinaþril    5.4 15.8
Çuinidine 2.1 322 245 2.5 11
Ramiþril    4.2 15.7
Rescinnamine 0.5 495 407  
Reserþine  457 351  15.4
SoLalol 1.2 225  2 3.8
Tocainide 1.2 247 208 2.1 
Tolazoline 2.1 225 179  
Trandolaþril    5.1 17
TrandolaþrilaL    2.1 
TrimeLazidine 3    5.1
veraþamil 2.5 447 385 7 15.4
M (nor-) 1.7   5.5 
Diuretics
HN HX HY HAA
k Rl Rl R7
^ceLazolamide  258 225 5.9
^miloride  257 190 3.5
BendroflumeLhiazide 15.35 508  18.5
BenzLhiazide 9.32  415 
ChloroLhiazide 0.54  239 
ChlorLalidone 1.28 357 308 
Cloþamide 4.01 377 310 
Clorexolone 7.25  391 
CycloþenLhiazide 15.45  453 
CycloLhiazide 10.78, 11.91,
and 12.81
 433 
ELacrynic acid  521 497 
Furosemide  435 380 15.2
HydrochloroLhiazide 0.7 294 255 
Mefruside 8.57  417 
MeLhycloLhiazide 3.82  354 15.4
MeLolazone 4.89  371 
SþironolacLone  592 539 20.7
TriamLerene  298 253 8.7
TrichlormeLhiazide 3.1  341 14.9
Xiþamide  488  18.8
Drugs cf abuse
HA HC HX HY HZ HAA
k k Rl Rl Rl k
5-MeLhylLryþL-
amine
     
^mfeLamine 0.9 0.98 244   3.7
BenzfeLamine 1.2 0.15    
Benzoylecgonine 0.9   235 1.7 9.7
BufoLenine 3.1   181  
Cannabidiol   990 902  
Cannabinol   1080 1028  
Cocaine 2.8  348 289 3.3 11.9
 -THC      
Diamorþhine 3 0.55 340 282  
DieLhylLryþL-
amine
     
DimeLhylLryþL-
amine
   228  
D0M  1.13 340   
KeLamine   311 252 2.4 9.5
Lysergic acid 0.8   235  
Lysergide 0.7  352   12
tab|e ccntinued
Recommended HPLC sysLems 749
Ergct aIkaIcids
System HA, previously described, may be used or System HP, below.
5ystem HP
R Gill et u|., unpublished data, after Twitchett PJ et u|. (1978). j
Chrcmutcgr 150: 73-84.
 Cc|umn. ODS Hypersil (100 5 mm i.d., 5 m).
 Mcbi|e phuse. Methanol-phosphate buffer (60 : 40). To prepare the
phosphate buffer, dissolve 3.43 g (0.022 mol/I) sodium dihydrogen-
phosphate and 10.03 g (0.028 mol/I) disodium hydrogenphosphate
in sufficient water to produce 1 I.
LccaI anaesthetics
The general screening systems, previously described may be used, as well
as system HQ or HR, below.
5ystem HÇ
Gill R et u|. (1984). j Chrcmutcgr 301: 155-163.
 Cc|umn. ODS Hypersil (160 5 mm i.d., 5 m).
 Mcbi|e phuse. Methanol-water-1% v/v solution of phosphoric acid-
hexylamine (30 : 70 : 100 : 1.4).
5ystem HR
Gill R et u|. (1984). j Chrcmutcgr 301: 155-163.
 Cc|umn. As for System HQ above.
 Mcbi|e phuse. Methanol-1% v/v solution of phosphoric acid-hexyl-
amine (100 : 100 : 1.4).
Narcctic anaIgesics
Systems HA or HC, previously described, may be used or System HS,
below.
5ystem H5
Baker PB, Gough TA (1981). j Chrcmutcgr Sci 19: 483-489.
 Cc|umn. Aminopropyl-bonded silica Spherisorb S5NH (250 4 mm
i.d., 5 m).
 Mcbi|e phuse. Acetonitrile-tetrabutylammonium phosphate,
0.005 mol/I, pH 7.5 (85 : 15).
Drugs cf abuse, ccntinued
HA HC HX HY HZ HAA
k k Rl Rl Rl k
Mescaline 1.3 2.17 272 243  
MeLamfeLamine 2 2.07 252 215 2.4 8.4
MeLhadone 2.2 1.03 440 343 8.5 15.8
MeLhylenedioxy-
amfeLamine
 0.98 255 248 2.1 8.1
MeLhylenedioxy-
meLhamfeLamine
  278 252 2.2 9.1
MonoaceLyl-
morþhine
3.5 0.8    7.3
Morþhine 3.8 1.3 200 182 1.8 3.3
N-MeLhylLryþL-
amine
     
p-MeLhoxyam-
feLamine
     
Psilocin 3.1  240 225  
Psilocybine    185  
Ergct aIkaIcids
HA HP
k k
BromocriþLine  44.3
DihydroergocrisLine  18.3
DihydroergocryþLine  15.9
DihydroergoLamine 0.5 11.4
Ergocornine 0.4 10.2
ErgocrisLine 0.3 17.3
ErgocryþLine 0.4 15.2
ErgomeLrine 0.4 0.50
Ergosine 0.3 7.08
Ergosinine 0.3 17.7
ErgoLamine 0.4 9.58
lso-lysergic acid  0.83
lso-lysergide 2.5 0.0
Lysergamide 0.5 0.33
Lysergic acid 0.8 0.0
Lysergic acid meLhylþroþylamide  1.98
Lysergide 0.7 1.83
Lysergol 1.1 0.83
MeLhylergomeLrine 0.4 0.83
MeLhysergide 0.4 2.33
2-0xylysergide  0.92
LccaI anaesthetics
HA HÇ HR HX HY HZ
k k k Rl Rl R7
Benzocaine 0.1 20.05 1.51 404 358 4.3
Buþivacaine 0.9 7.19 0.85 355 310 4.1
BuLacaine 1.2 8.97  392 331 
BuLanilicaine  4.42   280 
Chloroþrocaine  0.24   250 
Cinchocaine 1.9  5.51  371 
Cocaine 2.8 2.58  348 289 3.3
Benzoylecgonine 0.9 5.58    
Ecgonine 1.1     
CyclomeLhycaine   10.31  413 
Dyclonine   2.78  347 
ELomidaLe   475 417 
KeLamine    311 252 2.4
Lidocaine 0.5 0.79  288 258 2.5
M (monoeLhyl-
glycinexylidide)
1.2     
Meþivacaine 0.9 1.09  295 250 2.5
MeLhohexiLal    503 484 
0xybuþrocaine  15.25 0.85 405  
Piþerocaine  4.59  357 312 
Pramocaine 0.5  2.48 415  5.5
Prilocaine 1 1.38    2.7
Procaine 1.9   254 225 
Proþofol      35
ProxymeLacaine 2.1 1.38   259 
Çuinisocaine 2.2  11.24   
TeLracaine 2 15.25 1.33 389 321 4.4
750 High Performance Liquid ChromaLograþhy
Narcctic anaIgesics and narcctic antagcnists
HA HC H5 HX HY HZ HAA HAX HAY
k k k Rl Rl R7 R7 R7 R7
^lþhaþrodine 2.8   353 317    
BeziLramide 0.2   554  22.5   
Buþrenorþhine 0.4 0.05  397 339 5 14  
Codeine 4.8 1.21 1.9 255 237 1.9 5 5.1 3.4
Morþhine 3.8 1.3 5.15      
M (nor-) 3.1 3.51       
Cyclazocine 2.1    289    
DexLromoramide 0.7 0.09  440 390  15.8  
DexLroþroþoxyþhene 1.9 0.19   374 7.5 15.8  
Norþroþoxyþhene 1.3        
Diamorþhine 3 0.55 0.35 340 282   7.9 4.1
5-MonoaceLylmorþhine 3.5 0.8 1      
Morþhine 3.8 1.3 5.15      
Dihydrocodeine 7.2 2.5  251 208 2 4.7  
Dihydromorþhine 5.7 2.75  237 155    
Diþiþanone 2.2 1.51  500 353    
ELhoheþLazine 3.3 1.55  359     
ELhylmorþhine 3.7 1.05 1.45 291 244   5.7 3.5
FenLanyl 0.8 1.11  373 299  14.2 11.4 5
Hydromorþhone 7.9   240 187   5.8 3.4
KeLobemidone 2.8   294 245    
Levallorþhan 1.9 1.45  355 291    
Levorþhanol 4.4 3.2   255    
MeþLazinol 3.1    259    
MeLhadone 2.2 1.03  440 343 8.5 15.8 15.5 8.4
M (EDDP) 2.8        
M (EMDP) 0.2        
Morþhine 3.8 1.30 5.15 200 182 1.8 3.3 5.5 3.2
Morþhine-
3-glucuronide
 1.55       
N-oxide 3.2        
Nalorþhine 1 0.29  250 237  4.8  
Naloxone 1.4 0.17   238 2 14  
Norcodeine 3.1 3.51   235    
NormeLhadone  0.53   355    
Normorþhine 2.9 3.92   133    
Norþiþanone  0.35  455     
0xycodone 5.9 0.85  277 245   5.5 5.8
0xymorþhone 5.7        
0xymorþhone 5.7   217 184    
PenLazocine 1.8 0.57  372 288 3.8 12.5 9.9 5.5
PeLhidine 2.8 0.55  345 281 3.2 11.8 9.2 4.8
M (nor-) 1.7 2.04       
PeLhidinic acid 2.8        
Phenazocine 1.3 0.3  409 299    
Phenoþeridine 0.8 0.1  434     
NorþeLhidine 1.7 2.04       
PeLhidine 2.8 0.55       
PiriLramide 0.5 0.1  377 343    
Thebacon 3.7 0.85  333     
Tramadol    328 257 2.9   
Recommended HPLC sysLems 751
OraI hypcgIyacemics and antidiabetics
The general screening systems, previously described, may be used.
Pesticides
5ystem HA0
Osselton MD, Snelling RD (1986). j Chrcmutcgr 368: 265-271.
 Cc|umn. ODS Hypersil (160 5 mm i.d., 5 m), stainless steel.
 Mcbi|e phuse. Acetonitrile-water (60 : 40).
 I|cw rute. 2 mI/min.
 Detecticn. DAD (200-450 nm).
5ystem HAP
Osselton MD, Snelling RD (1986). j Chrcmutcgr 368: 265-271.
 Cc|umn. Silica Spherisorb S5W (250 5 mm i.d.).
 Mcbi|e phuse. Dichloromethane-isoctane (60 : 40).
 I|cw rute. 2 mI/min.
 Detecticn. DAD (200-450 nm).
For more information on screening pesticides, see Chapter 16,
Table 16.1.
Phencthiazines and cther tranquiIIisers
The general screening systems, previously described, may be used.
OraI hypcgIyacemics and antidiabetics
HX HY HZ HAA
Rl Rl R7 R7
CarbuLamide  321  14.5
Chlorþroþamide 450 411 and
413
5 17.7
Clibenclamide 537 571 14.4 22
Cliclazide 535 483 8.8 20.5
Cliþizide 478 423 4.5 17.5
MeLformin 50  1.7 2.8
Tolazamide 452 445 5.8 
TolbuLamide 477 424 5.9 
Phencthiazines and cther tranquiIIisers
HA HX HY HZ HAA HAX HAY HAZ
k Rl Rl R7 R7 R7 R7 k
^ceþromazine 4.1  350  10.8   
^zacyclonol 1.2     8.7 4.5 
BenzocLamine 1.7 380 322     
BuLaþerazine 3.4 454 405     
CaþLodiame  551   20.2   
Chlordiazeþoxide  353 285 3.2 15.2 5.9 5.3 1.58
Chlormezanone   334  15.5 5 5.3 
Chlorþromazine 4.1 455 350 9.1 15 17 B^SE 2.54
M (nor-) 2.2       
M (sulfoxide)      8.4 4.3 0.52
ChlorþroLhixene 3 459 353 10.1  17.5 8.3 
CloþenLhixol  448 411     
Clorazeþic acid  475 388 5.5    
ClorazeþaLe     18.4   
Fluanisone  423 349     
FluþenLixol 1.2 475 435 10.7 17.4 13.7 7.5 
Sulfoxide 1.3       
Fluþhenazine 1.2 452 471 10.1 17.4 13.5 7.2 
Flusþirilene  538    18.3 9.8 
Haloþeridol 1.2 421 315 5.8 14.4 11.1 5.2 0.72
Levomeþromazine 3.2 435 381 7.5  15.2 7.2 1.82
Loxaþine 1.1 407 335  14.5   
Mesoridazine 5  337 3.4  10.1 5 
0xyþerLine 0.7 402      
Pecazine 3.9 443 382   15.3 7 
Penfluridol  559 555 43.4 20.2   
Perazine  403 371 5.3    
Pericyazine 1.3 410 355 4.4  10.2 5.1 
Perþhenazine 1.9 428 395 7.2 15 13.1 5.3 3.28
Pimozide 0.7 504  11.9 17.2   
Piþamþerone  299 241 2.7 10.9   
PiþoLiazine  431   14.7   
Prochlorþerazine 3.9 450 323 10.4    
Promazine 5.9 407 325 5.9    
752 High Performance Liquid ChromaLograþhy
5tercids
5ystem HA7u
Walters MJ et u|. (1990). j Asscc Off Anu| Chem 73: 904-926.
 Cc|umn. ODS Zorbax (250 4.6 mm i.d., 5 m), stainless steel.
 L|uent. (A) Methanol.
 Isccrutic e|uticn. (100).
 I|cw rute. 1.5 mI/min.
 Detecticn. UV ( 240, 210 and 280 nm).
5ystem HA7b
Walters MJ et u|. (1990). j Asscc Off Anu| Chem 73: 904-926.
 Cc|umn. ODS Zorbax (250 4.6 mm i.d., 5 m), stainless steel.
 L|uent. (A : B) Methanol-water.
 Isccrutic e|uticn. (75 : 25).
 I|cw rute. 1.5 mI/min.
 Detecticn. UV ( 240, 210 and 280 nm).
5ystem HAR
Iurie I et u|. (1994). j Icrensic Sci 39: 74-85.
 Cc|umn. ODS Zorbax (250 4.6 mm i.d., 5 m).
 Mcbi|e phuse. (A : B) Water-methanol.
 Grudient e|uticn. (30 : 70) to (0 : 100) over 15 min with 15 min hold.
 I|cw rute. 1.0 mI/min.
 Detecticn. DAD.
5ystem H7
Rose JQ, Jusko WJ (1979). j Chrcmutcgr Bicmed App| 162: 273-280.
 Cc|umn. Silica Zorbax SII (250 4.6 mm i.d., 5 m).
 Mcbi|e phuse. Methylene chloride-methanol (97 : 3).
Phencthiazines and cther tranquiIIisers, ccntinued
HA HX HY HZ HAA HAX HAY HAZ
k Rl Rl R7 R7 R7 R7 k
ProLhiþendyl 4.4 388      
Sulforidazine  421  4.8    
Sulþiride  259 235 2 3.9   0.02
ThioþroþazaLe 1 483      
Thioþroþerazine 4.1 427 305 15.4 15.2   
Thioridazine 5.2 490 427 13.5 17.2  9.8 3.88
Mesoridazine 5       
TioLixene 3.8 442 374 5.8    
Trifluþromazine 2.7 484 454 12.3  17.3 8.9 
5tercids
H7 HX HY HZ HAA HAR HA7u HA7b
k Rl Rl R7 R7 RR7 RR7 RR7
BeclomeLasone 4.2 444      
DiþroþionaLe   711     
BeLameLhasone    14.2 13.3   
BeLameLhasone valeraLe   584     
Boldenone      0.74  0.75
UndecylenaLe       1.94 
CorLisone 2.4  372     
DexameLhasone 4.8  381 3.4 13.1   
FluoxymesLerone   427   0.78  0.7
HydrocorLisone 5.8 403 349  17.7   
HydroxyþrogesLerone  1054      
MeLenolone        
^ceLaLe       1.25 3.54
EnanLaLe       1.87 
MeLhandienone      0.85  0.87
MeLhandriol      1.25  1.29
DiþroþionaLe        2.75
MeLhylþrednisolone 7.5 425 390  18.9   
MeLhylLesLosLerone   587   1.17  1.27
Nandrolone      0.84  0.92
NoreLhisLerone  535 575  24   
Prednisolone 8.4 401 351 2.5 14.1   
Prednisone 3.4 250 340 2.5 14.2   
ProgesLerone  572 598  23.8   
tab|e ccntinued
Recommended HPLC sysLems 75J
5uIfcnamides
5ystem HU
Cobb PH, Hill GT (1976). j Chrcmutcgr 123: 444-447.
 Cc|umn. Silica Spherisorb (250 4 mm i.d., 5 m).
 Mcbi|e phuse. Cyclohexane-ethanol-acetic acid (85.7 : 11.4 : 2.9).
Xanthine stimuIants
The general screening systems, previously described, may be used.
AdditicnaI systems
5ystem HAD
Aymard et u|. (2000). j Chrcmutcgr Bicmed Sci App| 744: 227-240.
 Cc|umn. C Symmetry Shield (250 4.6 mm i.d., 5 m) protected
by 2 m Upchurch filter.
 Cc|umn temperuture. 30 C.
 Mcbi|e phuse. (A : B) M/15 potassium dihydrogenphosphate with 1%
(v/v) octane sulfonic acid : acetonitrile. Mobile phase (MP) 1: (95 : 5)
at flow rate 1 mI/min; MP 2: (80 : 20) at flow rate 1 mI/min; MP 3:
(30 : 70) at flow rate 1.2 mI/min.
 L|uent switching prcgrumme. At injection, MP1 to the column. From
time 12 min to 30 min, MP2 to the column. From time 30 min, MP3
to the column to rinse it. From time 35 min to 40 min, equilibration
with MP1.
 Detecticn. DAD ( 260 nm).
5tercids, ccntinued
H7 HX HY HZ HAA HAR HA7u HA7b
k Rl Rl R7 R7 RR7 RR7 RR7
TesLosLerone  534 508     
^ceLaLe   894   1.75  2.59
ProþionaLe   1003   2.01 1.31 4.05
MeLhylLesLosLerone      1.17  1.27
lsobuLyraLe      2.17  
CiþionaLe      2.53  
EnanLaLe      2.5 1.8 
UndecanoaLe      3.18  
PhenylþroþionaLe       1.48 
lsocaþroaLe       1.52 
CiþionaLe       2.05 
UndecenoaLe       2.53 
DecanoaLe       2.78 
UndecylaLe       3.27 
Triamcinolone  438 312     
^ceLonide 2.5       
Trenbelone        
HexahydrobenzylcarbonaLe       1.55 
^ceLaLe        1.71
5uIfcnamides
HU
k
PhLhalylsulfaLhiazole 14.0
SuccinylsulfaLhiazole 15.8
Sulfadoxine 4.4
Sulfamerazine 8.1
Sulfaquinoxaline 4.8
SulfaceLamide 7.7
Sulfachlorþyridazine 3.3
Sulfadiazine 8.7
Sulfadimidine 7.1
Sulfafurazole 5.0
SulfameLhoxazole 4.8
SulfameLhoxydiazine 8.2
SulfameLhoxyþyridazine 7.5
Sulfamoxole 12.5
Sulfanilamide 8.9
Sulfaþyridine 3.8
SulfaLhiazole 13.4
Xanthine stimuIants
HA HX HY HZ HAA
k Rl Rl R7 R7
Caffeine 0.2 305 259 1.9 5.7
Diþroþhylline  275 227  3.5
FeneLylline  335 277  
Proxyþhylline 0.1 293   
Theobromine 0.1 252 201 1.5 3.8
Theoþhylline 0.1 275 249 1.7 4.9
k CompounJ
2.7 Lamivudine
3.2 Didanosine
3.8 SLavudine
5.5 Zidovudine
8.1 ^bacavir
11.1 Neviraþine
754 High Performance Liquid ChromaLograþhy
5ystem HAF
Tanaka E et u|. (1996). j Chrcmutcgr B Bicmed Sci App| 682: 173-178.
 Cc|umn. ODS TSK-gel Super (100 4.6 mm i.d., 2 m).
 Mcbi|e phuse. (A : B) Acetonitrile-5 mmol/I sodium dihydrogen-
phosphate (pH 6).
 Isccrutic e|uticn. (45 : 55).
 I|cw rute. 0.65 mI/min.
 Detecticn. UV ( 254 nm).
5ystem HAv
Rutledge DR et u|. (1994). j Ihurm Bicmed Anu| 12: 135-140.
 Cc|umn. RP-short alkyl chain, silanol-deactivated (SCD 100;
250 4.6 mm i.d.), stainless steel.
 Mcbi|e phuse. (A : B) Methanol-0.04 mol/I dibasic potassium phos-
phate (pH 5.5).
 Isccrutic e|uticn. (50 : 50).
 I|cw rute. 1 mI/min.
 Detecticn. UV ( 237 nm).
5ystem H8A
Sastre-Toran¯o J, Guchelaar H-J (1998). j Chrcmutcgr B Bicmed Sci App|
720: 89-97.
 Cc|umn. C base-deactivated silica (125 4.6 mm i.d., 5 m) with
base-deactivated C precolumn (20 4.6 mm i.d., 5 m).
 L|uent. (A : B) Acetonitrile-50 mmol/I potassium dihydrogen-
phosphate (pH 7.5, containing 500 I triethylamine).
 Isccrutic e|uticn. (60 : 40).
 I|cw rute. 2 mI/min.
 Detecticn. Fluorescence ( 255 nm,  315 nm).
5ystem H88
Taninaka C et u|. (2000). j Chrcmutcgr B Bicmed Sci App| 738: 405-411.
 Cc|umn. C (250 6.0 mm i.d., 5 m).
 L|uent. (A : B) Acetonitrile-50 mmol/I phosphate buffer (pH 7.2).
 Isccrutic e|uticn. (43 : 57).
 I|cw rute. 1.7 mI/min.
 Detecticn. Electrochemical (working electrode: glassy carbon; refer-
ence electrode: Ag/AgCl).
5ystem HAE
Proust V et u|. (2000). j Chrcmutcgr B Bicmed Sci App| 742: 453-458.
 Cc|umn. C (Iichrospher, 100 RP-18, 5 m) with C precolumn
(Iichrospher RP-18, 5 m).
 Mcbi|e phuse. (A : B) Acetonitrile-25 mmol/I sodium phosphate
modified with diethylamine (0.9%) and tetrahydrofuran (1%), pH
3.0.
 Isccrutic e|uticn. (44.8 : 55.2).
 I|cw rute. 0.5 mI/min.
 Detecticn. UV ( 260 nm).
5ystem HAK
Ie Guellec C et u|. (1988). j Chrcmutcgr Sci App| 719: 227-233.
 Cc|umn. C Symmetry (250 4.6 mm i.d., 5 m) with C precol-
umn Symmetry sentry.
 Mcbi|e phuse. (A : B) Acetonitrile-20 mmol/I potassium di-
hydrogenphosphate.
 L|uticn prcgrumme. (50 : 50) to (70 : 30) in 15 min.
 I|cw rute. 1 mI/min.
 Detecticn. UV ( 313 nm).
5ystem HAL
Boukhabza A et u|. (1990). j Chrcmutcgr 529: 210-216.
 Cc|umn. C Novapak (150 4.6 mm i.d., 5 m).
Retention time (min) CompounJ
5.3 Clonazeþam
5.5 Bromazeþam
9.1 NiLrazeþam
13.7 Triazolam
15.0 Lorazeþam
18.4 ELizolam
21.0 Chlordiazeþoxide
29.8 Diazeþam
32.2 FluLazolam
k CompounJ
2.2 Celiþrolol
2.3 Proþranolol
3.5 DilLiazem deaceLyldilLiazem
5.1 DilLiazem desmeLhyldilLiazem
5.1 DilLiazem
5.4 lmiþramine
8.2 veraþamil
Retention time (min) CompounJ
8.8 EryLhromycin
15.7 ClariLhromycin
17.1 RoxiLhromycin
20.7 ^ziLhromycin
Retention time (min) CompounJ
5.8 ClariLhromycin
5.8 EryLhromycin
9.5 ^ziLhromycin
15.3 RoxiLhromycin
Retention time (min) CompounJ
5.3 Delavirdine
7.0 Saquinavir
8.0 Nelfinavir
9.4 ^mþrenavir
22.2 RiLonavir
28.5 Efavirenz
Retention time (min) CompounJ
4.7 Carbamazeþine
5.2 Clonazeþam
7.5 Nordazeþam
9.3 Clobazamm
NoL deLecLed PhenobarbiLal
NoL deLecLed PhenyLoin
Recommended HPLC sysLems 755