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FORENSIC TOXICOLOGY - Bio Medical Forensics

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K26 The Distribution of Gamma-Hydroxybutyric<br />

Acid (GHB) in Both Ante and Postmortem<br />

Specimens, of a Single Fatality After<br />

Long-Term Storage<br />

Laureen J. Marinetti, MS*, Bradford R. Hepler, PhD, Daniel S.<br />

Isenschmid, PhD, and Sawait Kanluen, MD, Wayne County <strong>Medical</strong><br />

Examiner’s Office, 1300 East Warren Avenue, Detroit, MI<br />

The objective is to document GHB concentrations in various<br />

specimens collected from a single GHB fatality at four different times of<br />

specimen collection and after long-term storage.<br />

Content: A 15-year-old female victim allegedly consumed an<br />

unknown amount of a drink laced with a mixture of GHB/GBL on 1/16/99<br />

at approximately 11:30 p.m. She became violently ill and her friends<br />

decided to take her to the hospital when she stopped breathing, arriving on<br />

1/17/99 at approximately 5:00 a.m. In the course of her treatment, several<br />

specimens were collected from her at three different times. Upon<br />

admission to the hospital, five tubes of blood with the following color tops<br />

were collected: red, yellow, green, blue, and purple. Also an admission<br />

urine specimen was collected. At 11:00 a.m. a spinal fluid specimen was<br />

collected and between 12:35 and 12:55 p.m., four more tubes of blood<br />

were collected with the following color tops: yellow, green, purple, and<br />

gray. At 7:34 p.m. on 1/17/99 she was pronounced dead, having never<br />

regained consciousness. The cause of death was GHB intoxication with<br />

no other drugs present including ethanol. At autopsy the next day<br />

(1/18/99) the following specimens were collected: urine, heart blood,<br />

femoral blood, vitreous fluid and bile. All of the specimens were analyzed<br />

either soon after the autopsy or after long-term freezer storage. The<br />

original specimens analyzed were the 5:00 a.m. red top blood, the hospital<br />

urine, the postmortem heart blood, and the postmortem urine. These specimens<br />

were sent out to a reference laboratory for analysis on 1/19/99. The<br />

remaining specimens were stored in the refrigerator until March 6, 1999,<br />

at which time they were moved to the freezer where they remained until<br />

July of 2001. At this time the specimens were analyzed in duplicate at the<br />

Wayne County <strong>Medical</strong> Examiner’s Office using a modified version of the<br />

United Chemical Technologies GHB solid phase extraction method which<br />

derivatizes GHB with no conversion to GBL. This method directly<br />

measures the GHB di-TMS derivative by EI GC/MS with no conversion<br />

to GBL and utilizes d-6 GHB as the internal standard with a specimen size<br />

of 200 microliters. The method has a linearity range from 2.5 to 100<br />

mcg/mL, and a recovery between 35% and 60% (specimen type<br />

dependent). Variation in concentration between analytical runs of both<br />

internal and external controls and specimens was no greater than 15%,<br />

n=54. Table 1 shows the results of analysis of the antemortem specimens<br />

and Table 2 shows the results for the postmortem specimens.<br />

Table One - Antemortem<br />

Specimen GHB in mcg/ml<br />

original analysis re-analysis<br />

Red Top Bld 5:00a.m. 510 466<br />

Yellow Top Bld 5:00a.m. NP 566<br />

Yellow Top Bld 12:55p.m. NP 187<br />

Green Top Bld 5:00a.m. NP QNS<br />

Green Top Bld 12:35p.m. NP 232<br />

Purple Top Bld 5:00a.m. NP QNS<br />

Purple Top Bld 12:55p.m. NP 242<br />

Blue Top Bld 5:00a.m. NP 418<br />

Gray Top Bld 12:55p.m. NP 93<br />

Spinal Fluid 11:00a.m. NP 220<br />

Urine 5:00m<br />

NP = test not performed<br />

2300 QNS<br />

QNS = quantity not sufficient for analysis<br />

Table Two – Postmortem<br />

Specimen GHB in mcg/ml Endogenous GHB<br />

Original analysis Re-analysis in postmortem specimens*<br />

Heart Blood (NaFl) 15 12 1.6 – 36 Average = 12<br />

Femoral Blood NP 18 1.7 – 48 Average = 11<br />

Urine 150 102 0 – 14 Average = 4.6<br />

Urine at pH =<br />

14 for 30 min.<br />

NP 167 NA<br />

Bile NP 48 Unknown<br />

Vitreous Humor NP 127 Less than 10 mcg/ml<br />

*Anderson, D.T., Kuwahara, T., “Endogenous Gamma Hydroxybutyrate (GHB)<br />

levels in Postmortem Specimens”, Abstract CAT/NWAFS/SWAFS/SAT combined<br />

meeting, Las Vegas, Nevada, Nov. 7, 1997.<br />

The concentrations of GHB at the various collection times would<br />

indicate a reduced clearance of GHB in a patient who is alive but severely<br />

compromised by respiratory and metabolic acidosis (admission blood<br />

pH=6.8) from the respiratory depression caused by the high dose of GHB.<br />

The lower GHB concentration in the 12:55 p.m. gray top tube blood as<br />

compared to the yellow, green, and purple top tubes collected at around the<br />

same time cannot be explained at this time. The apparent half-life of GHB<br />

calculated in narcoleptic patients is 53 +/- 19 minutes for two 3gram doses<br />

(Scharf et al., 1998) but this is dose dependent. Even after 14 hours there<br />

is still evidence of past GHB ingestion in the postmortem urine and<br />

vitreous fluid and perhaps even the bile. More bile specimens should be<br />

analyzed to determine a postmortem GHB range in this specimen type.<br />

Table 3 has vitreous humor results from two other GHB fatality cases, both<br />

of these cases involved deaths with less survival time and no hospitalization<br />

with both victims being found dead some 5 to 7 hours after they<br />

were last seen alive. Also it is apparent that GHB conversion to the<br />

lactone does occur over time especially in a postmortem urine with a low<br />

pH such as in this case. The conversion back to GHB was accomplished<br />

because any GBL in the urine should have been from GHB excretion.<br />

Ingested GBL is rapidly converted enzymatically in the blood and liver to<br />

GHB by a lactonase enzyme. The converted GBL is then excreted as<br />

GHB. However, due to the pH dependent chemical equilibrium that exists<br />

between GHB and GBL, the low pH (4.2) of the postmortem urine would<br />

facilitate a shift in the equilibrium over time such that some of the GHB is<br />

converted to its lactone GBL. GBL analysis was not performed on any of<br />

these specimens.<br />

Table 3 – Two Additional GHB Fatality Cases<br />

Postmortem Specimen Case 1 GHB in mcg/mL Ethanol in g/dL<br />

Heart Blood 66 ND<br />

Femoral Blood 77 ND<br />

Vitreous Humor 85 ND<br />

Urine 1260 ND<br />

Postmortem Specimen Case 2<br />

Blood 400 .22<br />

Vitreous Humor 212 .12<br />

Specimen Storage, GHB Distribution, Ante and Postmortem<br />

Specimens<br />

K27 Zolpidem Intoxication<br />

Nancy B. Wu Chen PhD*, Edmund R. Donoghue MD, Aldo J. Fusaro, DO,<br />

Nancy L. Jones, MD, Barry D. Lifschultz, MD, Jennifer L. Jakalski, BS,<br />

Kathleen A. Mittel, BS, and James R. Reich Jr., MLT, Office of the <strong>Medical</strong><br />

Examiner, Cook County, 2121 West Harrison Street, Chicago, IL<br />

The participant will learn the tissue distribution of zolpidem in<br />

three cases.<br />

Zolpidem (Ambien® ) is a hypnotic agent. Case histories and<br />

toxicological findings from three zolpidem intoxication cases are<br />

271 * Presenting Author

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