A246 CLINICAL CHEMISTRY, Vol. 56, No.

6, Supplement, 2010
Thursday, July 29, 9:00 am - 12:00 pm TDM/Toxicology/DAU
E-124
Serum Uric Acid Levels as an adjunct in the assessment of certain
Psychiatric Disorders
J. R. Peela
1
, A. M. Jarari
1
, S. S. Dastagir
1
, E. A. Salh
2
, A. Hai
1
, A. K. Rawal
1
.
1
Department of Biochemistry, Faculty of Meaicine,Al-Arab Meaical University,,
Bengha:i, Libyan Arab Jamahiriya,
2
Department of Biochemistry, Faculty of
Pharmacy,Al-Arab Meaical University,, Bengha:i, Libyan Arab Jamahiriya,
The increasing number oI latent and maniIest hyperuricemia is important concerning
diIIerential diagnosis in neurological and psychiatric diseases. The pathological
importance oI hyperuricemia in these diseases is particularly unknown. Previous studies
have shown that uric acid estimation in cerebrospinal fuid was made with neurological
and psychiatric diseases. On an average the uric acid level in cerebrospinal fuid is 1:10
oI the blood uric acid level. High levels Ior example were Iound aIter L-Dopa treatment
and epileptic seizures and parkinsonism. Monitoring serum uric acid levels, a relatively
inexpensive and easily available test, may prove to be a useIul adjunct in the assessment
oI certain indices oI certain psychiatric illness. There is evidence oI dysregulation oI the
antioxidant deIense system in schizophrenia. Some studies have shown uric acid, a potent
antioxidant, is reduced in the plasma oI patients with schizophrenia. Tranquilizers like
1,4 -benzodiazepine (purinergic) are shown to decrease xanthine oxidase activity initially
and may cause fuctuations in serum uric acid levels. In the present study an attempt
was made to understand the eIIect oI tranquilizers on serum uric acid levels in diIIerent
psychiatric conditions.40 cases ( 22 males and 18 Iemales) oI MDP ( Maniac depressive
psychosis) and Schizophrenia who were undergoing treatment with diIIerent tranquilizers
Irom the psychiatric hospital, Benghazi, Libya and 20 healthy controls ( 12 males and 8
Iemales ) not using any tranquilizers were taken Ior study. The serum uric acid levels were
signifcantly elevated ( p 0.05 ) in these patients receiving treatment with tranquilizers.
The levels oI uric acid in male patients were signifcantly higher when compared with
Iemales and also over male controls ( p 0.01 ).In Iemale patients uric acids levels are
raised but not signifcant ( p ~ 0.05 ) over control Iemales. This preliminary study does
indicate that serum uric acid levels do change with the administration oI these drugs. The
nature, gender, duration and the type oI drugs used and their individual eIIects on diIIerent
psychiatric disorders will be discussed.
Irom human plasma was achieved using Waters Sep-pak C-18 cartridges, washing with
methanol and 1M HCL, and eluted in 75 methanol in water.
Results: Chromatography was initially evaluated Ior both methods using a combination oI
unextracted racemic warIarin and IS standards. Under published mobile phase conditions
(40 ACN : 60 HOAc), warIarin and IS were retained on the column. Increasing ACN
between 80-90 caused co-elution oI R-warIarin and IS. Optimum chromatographic
separation oI warIarin enantiomers was achieved with conditions set at 30 HOAc/70
ACN. Under these conditions S-warIarin, R-warIarin and IS eluted at 6.3, 7.1 and 8.0
minutes, respectively. Peak width at the highest concentration tested (5 mg/L) was less
than one minute. Chromatography results based on the published solid phase extraction
technique yielded artiIacts which co-eluted or interIered with IS detection. The optimized
extraction method resulted in an extraction eIfciency oI 65 Ior S-warIarin with no
baseline derangements in blank plasma over the time Irame where peaks oI interest elute.
A standard curve was generated Ior extracted plasma standards Irom 0.1 to 5 mg/L giving
a correlation coeIfcient oI 0.9998 (conc 2.2934(peak area) - 0.1065).
Conclusions: We have improved upon the literature methods in terms oI reproducibility
oI chiral chromatography results, ease and pureness oI extraction Irom human plasma,
and employed a commercially available internal standard used previously by Osman et
al. now optimized with a solid phase extraction method. Together, we have developed
an assay Ior chiral separation and quantitation oI S-warIarin that suits the needs oI our
pharmacogenomics research.
E-123
Drug Monitoring and Toxicology: A Rapid Method for the Monitoring of
Runamide by HPLC-UV
P. H. Tang. Cincinnati Chilarens Hospital Meaical Center, Cincinnati, OH,
Background: Rufnamide is a new antiepileptic drug with a triazole derivative structure. It
has shown promise as adjunctive treatment Ior Lennox-Gastaut Syndrome and may have
some role in localization related epilepsies as well. US FDA has recently approved the use
oI drug Ior the adjunctive treatment oI seizures associated with Lennox-Gastaut Syndrome
and as adjunctive treatment Ior partial-onset seizures. Therapeutic drug monitoring oI
antiepileptic drug concentration in plasma is helpIul to physicians in evaluating patient
compliance with treatment, in providing guidance to achieve well-tolerated and eIIective
dosing, and in identiIying drug-drug interactions when drugs are given as polytherapy.
Plasma rufnamide is generally measured by high-perIormance liquid chromatographic
(HPLC) method. In some HPLC methods, multi-step extraction techniques and extensive
sample pretreatment are used. Simple and reliable HPLC procedures based on direct
HPLC injection aIter sample deproteinization or even without sample pretreatment
have not been reported. Previously, measurement oI plasma rufnamide ordered by the
physicians here at the Cincinnati Children`s Hospital Medical Center was perIormed at the
reIerence laboratory. The results turnaround time were not always satisfed and service oI
therapeutic drug monitoring was, thereIore, lagging.
Objective: The need Ior a quick measurement oI rufnamide in plasma samples in a
simplifed manner and the need Ior a cost-eIIective procedure prompted the development
oI a rapid HPLC method. Here, a simple and reliable HPLC method is described Ior
determination oI rufnamide concentrations in a small volume (100L) oI plasma that is
suitable in pediatric practice.
Methods: Sample was vortex-mixed with the internal standard (trimethadione) and
methanol Ior 1 minute and centriIuged at 10,350 g Ior 10 minutes at room temperature.
The supernatant was transIerred to an autosampler vial, a portion oI supernatant (20L)
was injected directly onto the HPLC system. Using a mixture oI phosphate buIIer,
methanol, and acetonitrile as mobile phase, the derivatization products were analyzed on
a 5-m Microsorb-MV reversed-phase C18 column (250 x 4.6 mm) column using UV
detection at 199 nm. Quantifcation was based on a 5-point calibration using peak height
measurements oI rufnamide and internal standard.
Results and Discussion: The method achieved a linear concentration range oI 1-50 mg/L,
which covered the proposed therapeutic range oI 5-48 mg/L. The limit oI detection was
0.2 mg/L. Both within-run and between-run precision Ior three Iortifed controls (10, 20,
and 40 mg/L) in plasma were lower than 5. Analytical recoveries were greater than
95. No interIerence was observed Irom the most commonly administered antiepileptic
drugs (primidone, carbamazepine, ethosuximide, Ielbamate, lamotrigine, levetiracetam,
oxcarbazepine, phenobarbital, phenytoin, topiramate, valproic acid, and zonisamide). The
method was compared to a reIerence laboratory HPLC assay using 30 samples ranging
Irom 4 to 45 mg/L. The correlation showed a slope oI 1.04, an intercept oI 0.2 mg/L and
an r oI 0.93.
Conclusion: This method is simple and easy to perIorm with excellent reproducibility,
requires no solid-phase extraction and one step deproteinization prior to chromatography.
It is suitable Ior routine analysis oI rufnamide in human plasma samples.