Toxicology

Thin Layer Chromatography

Yoknapatawpha County Forensic Laboratory

DATE ANALYSIS PERFORMED:
5/5/2021

Madeline Holland, Ph.D.
555 Jackson Avenue
Oxford, MS 38655
662-234-XXXX (FAX 662-234-XXXX)

Analysis: Toxicological Analysis

 

REPORT OF FORENSIC TOXICOLOGICAL ANALYSIS

Case Description:
 Barbara Dubois Homicide

Case #
 001995-01E-2021

Investigative Agency:
 Yoknapatawpha County Sheriff's Department

Investigating Officer(s):
 Det. S. Murphy, Det. T. Armstrong

EVIDENCE ANALYZED

  • 001995-06 and 001995-07: Two (2) samples from possible vomit pool in hallway outside YCCC Room 273
  • 001995-22: One (1) medium-sized plastic bag labeled "YCCC Dry Cleaning"
    • Contents: semi-liquid substance visually consistent with vomit

Evidence items were extracted with 5% methanol/chloroform and the organic layer was evaporated to afford a residue, which was subjected to thin-layer chromatography (TLC). Control samples included emetine (an active emetic alkaloid found in Psychotria ipecacuanha) and Syrup of Ipecac, USP, from Roxane Laboratories (an extract of Psychotria ipecacuanha used for poison control to induce vomiting).

Thin Layer Chromatography of Evidence Items:

  Evidence Item(s) Analysis Thin Layer Chromatogram
 1 

 A: 001995-06

 B: 001995-07

 C: Emetine (control)

 D: 001995-22

 E: Syrup of Ipecac (control)

Normal Phase thin layer chromatography; mobile phase = 10% Methanol/Chloroform; Anisaldehyde/sulfuric acid developed Anisaldehyde/sulfuric acid developed
 2 

 A: 001995-06

 B: 001995-07

 C: Emetine (control)

 D: 001995-22

 E: Syrup of Ipecac (control)

Normal Phase thin layer chromatography; mobile phase = 10% Methanol/Chloroform; Dragendorff developed Dragendorff developed

Results:

Two methods for developing the chromatograms were used to visualize different chemical compounds. Anisaldehyde is a general reagent used to develop many different types of compounds, while Dragendorff's Reagent is specific for alkaloids.

From the above chromatograms, evidence items 001995-06, 001995-07 and 001995-22 are consistent with the Syrup of Ipecac control.

As well, evidence items 001995-06, 001995-07 and 001995-22 are all positive for an alkaloid consistent with emetine.

Thin layer chromatography was used as a presumptive test, and the presence of emetine in the evidence items has been confirmed using HPLC-MS.

HPLC-MS

Yoknapatawpha County Forensic Laboratory

DATE ANALYSIS PERFORMED:
5/6/2021

Madeline Holland, Ph.D.
555 Jackson Avenue
Oxford, MS 38655
662-234-XXXX (FAX 662-234-XXXX)

Analysis: Toxicological Analysis

 

REPORT OF FORENSIC TOXICOLOGICAL ANALYSIS

Case Description:
 Barbara Dubois Homicide

Case #
 001995-01E-2021

Investigative Agency:
 Yoknapatawpha County Sheriff's Department

Investigating Officer(s):
 Det. S. Murphy, Det. T. Armstrong

EVIDENCE ANALYZED

  • 001995-06 and 001995-07: Two (2) samples from possible vomit pool in hallway outside YCCC Room #273
  • 001995-22: One (1) medium-sized plastic bag labeled "YCCC Dry Cleaning"
    • Contents: semi-liquid substance visually consistent with vomit
  • 001995-34: Chocolates found in Barbara Dubois' YCCC hotel room (#203)
  • 001995-38: Chocolates found in Ingrid Freeman's YCCC hotel room (#206)
  • 001995-39-013: 1-fl-oz bottle ipecac syrup found in Rachel Webb's YCCC hotel room (#212)
  • 001995-40: Chocolates found in Rachel Webb's YCCC hotel room (#212)
  • 001995-41 through 001995-43: Chocolates found in Sam Tuttle's YCCC hotel room (#217)
  • 001995-45: Chocolates found in Mary and Billie Jo Jones' YCCC hotel room (#247)
  • 001995-46: Urine sample from deceased
  • 001995-47: Blood sample from deceased

Evidence items were extracted with 5% methanol/chloroform and the organic layer was evaporated to afford a residue, which was subjected to high-performance liquid chromatography (HPLC) using a reversed-phase column coupled with an electron-ionization mass spectrometer. A control sample of emetine was used for comparison.

HPLC-MS of Evidence Items:

Evidence #

Brief Description

Analysis Results

 001995-06  vomit sample (from hallway)  Peak with retention time of 5.2 min. (molecular ion = 481); 5.2mg
 001995-07  vomit sample (from hallway)  Peak with retention time of 5.2 min. (molecular ion = 481); 6.3mg
 001995-22  vomit sample (from bag)  Peak with retention time of 5.2 min. (molecular ion = 481); 4.2mg
 001995-34  chocolates (Barbara Dubois)  Peak with retention time of 5.2 min. (molecular ion = 481); 25mg
 001995-38  chocolates (Ingrid Freeman)  No peak (molecular ion = 481)
 001995-39-013  ipecac syrup (Rachel Webb)  Peak with retention time of 5.2 min. (molecular ion = 481); 1.3mg/ml
 001995-40  chocolates (Rachel Webb)  Peak with retention time of 5.2 min. (molecular ion = 481); 22mg
 001995-41  chocolates (Sam Tuttle)  No peak (molecular ion = 481)
 001995-42  chocolates (Sam Tuttle)  No peak (molecular ion = 481)
 001995-43  chocolates (Sam Tuttle)  No peak (molecular ion = 481)
 001995-45  chocolates (Mary & Billie Jo Jones)  No peak (molecular ion = 481)
 001995-46  deceased's urine  Peak with retention time of 5.2 min. (molecular ion = 481); 0.006mg
 001995-47  deceased's blood  Peak with retention time of 5.2 min. (molecular ion = 481); 15.3ng/ml
 Emetine  control  Retention time = 5.2 min., molecular ion = 481

Results:

Based on the positive presumptive test for emetine (thin layer chromatography), the above evidence items were analyzed using HPLC-MS to confirm the presence of emetine.

Evidence items 001995-06, 001995-07, 001995-22, 001995-30, 001995-34-013, 001995-35, 001995-41 and 001995-42 are positive for emetine, an emetic alkaloid known to occur in Syrup of Ipecac.

 


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