tuesday am, july 29 - mendeluweb2.mendelu.cz/af_239_nanotech/data/08/publikace/2008/...aqu&rus...

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Tuesday, Ju,Iy 29,10:00 am - 12:30 pm Tuesday AM, July 29 PosterSession: 10:00 am - 12:30 pm TDM/Toxicology/DAU fa-l I Evaluation of Bio-Rad To)íSeeTM oxycodone Rapid Urine Drug Screen C. B. Taool]'r, B. C. Yu2, H. K. Lee3. tDartmouth-Hitchcock Medical Centet Lebanon, NH, 2Dartmouth College, Hanove1 NH, sDartmouth Medical School & Dartmouth-Hitchcock Medical Center Lebanon, NH, Background: Urine drugs-of-abuse (DAU) testing devices re gaining popularity in hospital laboratories, emergency rooms and clinics. Cunently, our institute's DAU screen does not detect oxycodone at therapeutic doses and there was a request b.v the emergency department to souce for an oxycodone-specific urine drug screen due to the large number of people who were abusing oxycodone. Objectives: We eyaluated the Bio-Rad TOX/See Oxycodone Rapid Urine Drug Screen to investigate the accuacy, sensitivity, specihcity and hydrocodone cross-reactivity of this test. Methods: The TOX/See drug screen is a single-step, lateral flou chromatographic immunoassay for the qualitative detection of drugs and metabolites in urine. It provides the test result in 5 milutes with a detection cutoff of oxycodone at I 00 ng/ mL. The first part oíthe study was to evaluate the accuacy' sensitii'ity and specificity of oxycodone/oxymorphone detection using the TOXSee drug screen by testing 50 uine specimens with known GC/MS results. The second paň of the sfudy was to check the cross-reactivity ofhydrocodone and the sensitivity of TOX,/See drug screen ner cutoff level by testing 38 urine specimens with knorvn GC/MS confimed positive oxycodone/oxymorphone or hydrocodone drug levels. Results: In part one of the study, the sensitivity ofthe TOX/See drug screen u'as found to be 91.7 % with a specificity of l00 %. Twelve out ofthe 50 samples were positive for oxycodone mď or oxymorphone and only I false negative was obsened when the oxycodone level was only 175 ng/ml with no detectable oxymorphone. The overall accuacy ofTOX/ See drug screen was 98 %. In part two ofthe study, 516(83 %) hydrocodone positive uine samples, ranging from 559 to 7,045 ng/ml-, were detected as negative in the Bio-Rad TOX/See drug screen. The sample that was detected as positive oxycodone had a hydrocodone level of 23,755 nýmL, which was much higher than the hydrocodone cross.Ieacting level of 1562 nýml. Three samples, near oxycodone cutofflevels of 102 to l24nglmL, were detected as positive in the screen. Two samples with oxymorphone levels of 352 and 86l nýml were a|so detected as positive in the screen although the detection cutoff for oxymorphone was 1562 ng/ mL. Conclusions: The Bio-Rad TOX/See Oxycodone Rapid Urine Drug Screen is accuate, sensitive md specific for detecting oxycodone and its metabolite, oxymorphone. Hydrocodone cross-reactivity was obserued at a concentration much higher thm that claimed by the mmufacturer. t At-l Blood Lead Levels in Southeastern Wisconsin Children: A Response to the Lead Toy Controversy A. A. Tobinr, J. A. CarP, D. M. Breitenfeld2, P. J. Jannettor, S. Koether. tMedical College of Wisconsin, Milwaukee, ll'1, 2Dynacare Laboratories, Milwaukee. ývI. Lead is a heavy metal and a common environmental contaminant. Human lead exposue may occur thÍough contaminated dust' soil, air or water. An elevated body lead burden can cause toxicity in humans, especially in children. In the United States, a lead paint hazard wming was issued regrding certain children's toys sold between May-August of 2007. We hypothesized that lead contamination in toys newly imported to Southeastem Wisconsin would be reflected in increased blood lead levels (BLL) in resident children. A retrospective study was conducted using BLLs (venous md capillary) from Southeastem Wisconsin children mder six years of age. Lead levels were measured using a graphite fumace atomic absorption spectrometeÍ. The quality assuance and proficiency testing oťthis method in 2006 was comparable to testing performed in 2007 ' Test Íesults collected by Dynacare Laboratories, Milwaukee, WI, between July and October 2007 (n:9,139) were compared to the previous year's values (n : 7,528). The volume of lead tests perfomed was significmtly greateÍ(p < 0.00l) in l-year.olds (26.30/o),Z.year-o|ds (22.2%,) anď in children under 6 years ofage (21.4%) in 2007, compred to the same period in 2006. A significant decline ín the incidence ofelevated BLLs ( > 10 pýdl) was obserued in TDI\{/Toxicology/DAU all children less than 6 years ofage (6.3%; p < 0.001), especially in 2-year-olds (9.9%, p : 0.04) in 2007, compared to the prior year (8.2%o and 13.5%o, respectively). The results show m increase in the volume of lead testing in Southeastem Wisconsin, possibly in response to the lead toy hazard waming. A concomitant reduction in the frequency ofelevated BLLs occmed oveÍ the same period, particularly in the 2-year- old children tested. The increase in lead testing between 2006 and 2007 may have been in response to the concem that children were exposed to an increased lead burden due to the influx of lead-contaminated toys. The decreased incidence of elevated BLLs in 2007 compared to 2006 suggests that while the paint on some toys may have higher than desirable levels of lead, this lead is not being ingested by resident children. tA{ Plasma and urine strontium and serum free thiols levels at patients with osteoporosis J. Kukackar, I. Sotomik2, V Adam3, R. Prusar, D. Vajtrr, K. Kotaskal, M. Bunesoval, R. Kizek3. ]Charles Universiý 2nd Medical School and Un ivers ih, Hospital Motol, Prague, Czech Republic, 2 Institute for Clinical and Experimental Medicine, Prague, Czech Republic, 3Mendel Universiý ofAgriculture and Forestry, Brno, Czech Republic, InÚoduction. one ofthe substancesused for the treatment ofosteoporosis is strontium ranelate (SrR). It was shown that it lowers risk of fractues and it increases bone mineral density. Toxic symptoms due to overdosing have not been reported in man. However, data on Sr concenlrations at tÍeated patients or effects ofstrontium on other metabolic pathways remain mexplored properly. Objective. Our goal was to investigate changes of strontium concentration and free thiols (metallothionein, reduced glutathion, cysteine, and homocysteine)level in plasma, serum, and urine samples ofpatients tÍeated with strontium ranelate. Materials and methods. Blood md 24 hours urine samples were collected during routine biochemical examination from 28 patients with osteoporosis before SrR administration (Z e/day) and at 30th day of the therapy. Differential pulse voltammetry (Brdicka reaction) coupled with an adsorptive transfer stripping technique was used for metallothionein (MT) detection in sem using Autolab instment (EcoChemie, Netherlmd). For detection of reduced glutathione (GSH), cystein (C), and homocystein (HC) was used high perfomance liquid chromato$aphy with electrochemical detection (ESA, USA). Strontium concentration in plasma was measured by atomic absorption spectrometry (Varian, Australia). Results. Strontium concentration in plasma was 55+5 pgll- (mean+standard enor) before and 1046311376 yglL at 30th day of SrR administration, respectively. Moreover, we found signiírcant conelation between stlontium levels in plasma and serum (R=0.98). Mean uine concenhation of strontium was22,2+3.3 mýL and mean daily uine strontium output was 46.9+6.7 mg/24 hours. Levels of metallothionein in serum ranged from 0.l4 to the híghest 6.43 pmol/L (nomal values < 1 pmol/L). The mean serum concentration of cystein was 2 times higher (542+77 pmol/L) thm physiological one. Serum level of HC wm increased (22.5Í6.2 pmol/L; nomal values 5-12 pmoVl). We found GSH concentration 5.7+0.8 pmol/L in patients after treatment with SrR. Conclusions. Plasma strontium is extremely elevated (2,000 times) at patients treated u'ith stontium ranelate' We proved very good coÍrelation between plasma and serum le\ els of strontium. Metallothionein and other thiols are also highly elevated as direct chelating molecules to eliminate potential toxicity at patients treated with strontium ranelate. Analt'tes Recovery from deproteinized serum for HPLC Z. K. Shihabi. Wake Foresí UniversiÍt, Baptist Medical Center,, I|inslon- Sa/err. .\-C Serum deproteinization is often med as a rapid method for malysis of drugs and small molecules in serum in HPLC. Acetonitrile is a common method for deproteinization not just tor HPLC but also in capillary electrophoresis. The objective of this study is to demonsrrate that acetonitrile present in the sample limits the amount of small molecules injected on the HPLC column and also to offer other altematives. When the rr'corcn ofcompounds in serum is calculated based on peak height ofaqueous standards rreated also by the acetonitrile similar to s.'rum. a high recovery is obtained. Houc'rcr. \\hen these drugs treated with the acrrtrnitrile are calculated based on aqu&rus srandardsdiluted in the mobile phase rpump soh'ent) a low recovery is obl3in.'d. Diilťrenl tests (caffeine, voriconazole. lamtrtrigine. and pentobarbital) with L2 CLINICAL CHEMISTRY, Vol. 54 No. 6, Supplement, 2008

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Page 1: Tuesday AM, July 29 - MENDELUweb2.mendelu.cz/af_239_nanotech/data/08/publikace/2008/...aqu&rus srandards diluted in the mobile phase rpump soh'ent) a low recovery is obl3in.'d. Diilťrenl

Tuesday, Ju,Iy 29,10:00 am - 12:30 pm

Tuesday AM, July 29

Poster Session: 10:00 am - 12:30 pmTDM/Toxicology/DAU

fa-l IEvaluation of Bio-Rad To)íSeeTM oxycodone Rapid Urine Drug

Screen

C. B. Taool]'r, B. C. Yu2, H. K. Lee3. tDartmouth-Hitchcock Medical

Centet Lebanon, NH, 2Dartmouth College, Hanove1 NH, sDartmouth

Medical School & Dartmouth-Hitchcock Medical Center Lebanon, NH,

Background: Urine drugs-of-abuse (DAU) testing devices re gaining popularity inhospital laboratories, emergency rooms and clinics. Cunently, our institute's DAU

screen does not detect oxycodone at therapeutic doses and there was a request b.v the

emergency department to souce for an oxycodone-specific urine drug screen due tothe large number of people who were abusing oxycodone. Objectives: We eyaluated

the Bio-Rad TOX/See Oxycodone Rapid Urine Drug Screen to investigate theaccuacy, sensitivity, specihcity and hydrocodone cross-reactivity of this test.Methods: The TOX/See drug screen is a single-step, lateral flou chromatographicimmunoassay for the qualitative detection of drugs and metabolites in urine. Itprovides the test result in 5 milutes with a detection cutoff of oxycodone at I 00 ng/

mL. The first part oíthe study was to evaluate the accuacy' sensitii'ity and specificity

of oxycodone/oxymorphone detection using the TOXSee drug screen by testing 50uine specimens with known GC/MS results. The second paň of the sfudy was tocheck the cross-reactivity ofhydrocodone and the sensitivity of TOX,/See drug screenner cutoff level by testing 38 urine specimens with knorvn GC/MS confimedpositive oxycodone/oxymorphone or hydrocodone drug levels. Results: In part one ofthe study, the sensitivity ofthe TOX/See drug screen u'as found to be 91.7 % with aspecificity of l00 %. Twelve out ofthe 50 samples were positive for oxycodone mď

or oxymorphone and only I false negative was obsened when the oxycodone levelwas only 175 ng/ml with no detectable oxymorphone. The overall accuacy ofTOX/See drug screen was 98 %. In part two ofthe study, 516 (83 %) hydrocodone positive

uine samples, ranging from 559 to 7,045 ng/ml-, were detected as negative in theBio-Rad TOX/See drug screen. The sample that was detected as positive oxycodonehad a hydrocodone level of 23,755 nýmL, which was much higher than thehydrocodone cross.Ieacting level of 1562 nýml. Three samples, near oxycodonecutofflevels of 102 to l24nglmL, were detected as positive in the screen. Twosamples with oxymorphone levels of 352 and 86l nýml were a|so detected aspositive in the screen although the detection cutoff for oxymorphone was 1562 ng/mL. Conclusions: The Bio-Rad TOX/See Oxycodone Rapid Urine Drug Screen isaccuate, sensitive md specific for detecting oxycodone and its metabolite,oxymorphone. Hydrocodone cross-reactivity was obserued at a concentration much

higher thm that claimed by the mmufacturer.

t At- lBlood Lead Levels in Southeastern Wisconsin Children: A Response to

the Lead Toy Controversy

A. A. Tobinr, J. A. CarP, D. M. Breitenfeld2, P. J. Jannettor, S. Koether.tMedical College of Wisconsin, Milwaukee, ll'1, 2Dynacare Laboratories,

Milwaukee. ývI.

Lead is a heavy metal and a common environmental contaminant. Human leadexposue may occur thÍough contaminated dust' soil, air or water. An elevated bodylead burden can cause toxicity in humans, especially in children. In the United States,a lead paint hazard wming was issued regrding certain children's toys sold betweenMay-August of 2007. We hypothesized that lead contamination in toys newlyimported to Southeastem Wisconsin would be reflected in increased blood lead levels(BLL) in resident children. A retrospective study was conducted using BLLs (venous

md capillary) from Southeastem Wisconsin children mder six years of age. Leadlevels were measured using a graphite fumace atomic absorption spectrometeÍ. Thequality assuance and proficiency testing oťthis method in 2006 was comparable totesting performed in 2007 ' Test Íesults collected by Dynacare Laboratories,Milwaukee, WI, between July and October 2007 (n:9,139) were compared to theprevious year's values (n : 7,528). The volume of lead tests perfomed wassignificmtly greateÍ (p < 0.00l) in l-year.olds (26.30/o),Z.year-o|ds (22.2%,) anď inchildren under 6 years ofage (21.4%) in 2007, compred to the same period in 2006.A significant decline ín the incidence ofelevated BLLs ( > 10 pýdl) was obserued in

TDI\{/Toxicology/DAUall children less than 6 years ofage (6.3%; p < 0.001), especially in 2-year-olds (9.9%,

p : 0.04) in 2007, compared to the prior year (8.2%o and 13.5%o, respectively). The

results show m increase in the volume of lead testing in Southeastem Wisconsin,possibly in response to the lead toy hazard waming. A concomitant reduction in the

frequency ofelevated BLLs occmed oveÍ the same period, particularly in the 2-year-

old children tested. The increase in lead testing between 2006 and 2007 may have

been in response to the concem that children were exposed to an increased lead

burden due to the influx of lead-contaminated toys. The decreased incidence of

elevated BLLs in 2007 compared to 2006 suggests that while the paint on some toys

may have higher than desirable levels of lead, this lead is not being ingested by

resident children.

tA{Plasma and urine strontium and serum free thiols levels at patients

with osteoporosis

J. Kukackar, I. Sotomik2, V Adam3, R. Prusar, D. Vajtrr, K. Kotaskal, M.

Bunesoval, R. Kizek3. ]Charles Universiý 2nd Medical School and

Un ivers ih, Hospital Motol, Prague, Czech Republic, 2 Institute for Clinical

and Experimental Medicine, Prague, Czech Republic, 3Mendel Universiý

ofAgriculture and Forestry, Brno, Czech Republic,

InÚoduction. one ofthe substances used for the treatment ofosteoporosis is strontium

ranelate (SrR). It was shown that it lowers risk of fractues and it increases bone

mineral density. Toxic symptoms due to overdosing have not been reported in man.

However, data on Sr concenlrations at tÍeated patients or effects ofstrontium on other

metabolic pathways remain mexplored properly.

Objective. Our goal was to investigate changes of strontium concentration and free

thiols (metallothionein, reduced glutathion, cysteine, and homocysteine)level inplasma, serum, and urine samples ofpatients tÍeated with strontium ranelate.

Materials and methods. Blood md 24 hours urine samples were collected during

routine biochemical examination from 28 patients with osteoporosis before SrR

administration (Z e/day) and at 30th day of the therapy. Differential pulse

voltammetry (Brdicka reaction) coupled with an adsorptive transfer stripping

technique was used for metallothionein (MT) detection in sem using Autolab

instment (EcoChemie, Netherlmd). For detection of reduced glutathione (GSH),

cystein (C), and homocystein (HC) was used high perfomance liquid

chromato$aphy with electrochemical detection (ESA, USA). Strontium

concentration in plasma was measured by atomic absorption spectrometry (Varian,

Australia).Results. Strontium concentration in plasma was 55+5 pgll- (mean+standard enor)

before and 1046311376 yglL at 30th day of SrR administration, respectively.Moreover, we found signiírcant conelation between stlontium levels in plasma and

serum (R=0.98). Mean uine concenhation of strontium was22,2+3.3 mýL and mean

daily uine strontium output was 46.9+6.7 mg/24 hours. Levels of metallothionein in

serum ranged from 0.l4 to the híghest 6.43 pmol/L (nomal values < 1 pmol/L). The

mean serum concentration of cystein was 2 times higher (542+77 pmol/L) thmphysiological one. Serum level of HC wm increased (22.5Í6.2 pmol/L; nomal values

5-12 pmoVl). We found GSH concentration 5.7+0.8 pmol/L in patients aftertreatment with SrR.

Conclusions. Plasma strontium is extremely elevated (2,000 times) at patients treatedu'ith stontium ranelate' We proved very good coÍrelation between plasma and serumle\ els of strontium. Metallothionein and other thiols are also highly elevated as direct

chelating molecules to eliminate potential toxicity at patients treated with strontiumranelate.

Analt'tes Recovery from deproteinized serum for HPLC

Z. K. Shihabi. Wake Foresí UniversiÍt, Baptist Medical Center,, I|inslon-

Sa/err. .\-C

Serum deproteinization is often med as a rapid method for malysis of drugs and small

molecules in serum in HPLC. Acetonitrile is a common method for deproteinization

not just tor HPLC but also in capil lary electrophoresis. The objective of this study is

to demonsrrate that acetonitrile present in the sample limits the amount of small

molecules injected on the HPLC column and also to offer other altematives. When therr'corcn ofcompounds in serum is calculated based on peak height ofaqueousstandards rreated also by the acetonitrile similar to s.'rum. a high recovery is obtained.

Houc'rcr. \\hen these drugs treated with the acrrtrnitrile are calculated based onaqu&rus srandards diluted in the mobile phase rpump soh'ent) a low recovery is

obl3in.'d. Diilťrenl tests (caffeine, voriconazole. lamtrtrigine. and pentobarbital) with

L2 CLINICAL CHEMISTRY, Vol. 54 No. 6, Supplement, 2008