Thin Layer Chromatography
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Yoknapatawpha County Forensic Laboratory |
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REPORT DATE: Madeline Holland, Ph.D. |
Analysis: Toxicological Analysis
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REPORT OF FORENSIC TOXICOLOGICAL ANALYSIS |
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Case Description: |
Case # |
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Investigative Agency: |
Investigating Officer(s): |
EVIDENCE ANALYZED
- 003093-06 and 003093-07: Two (2) samples from possible vomit pool in hallway outside YCCC Room 273
- 003093-22: 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: 003093-06 B: 003093-07 C: Emetine (control) D: 003093-22 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: 003093-06 B: 003093-07 C: Emetine (control) D: 003093-22 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 003093-06, 003093-07 and 003093-22 are consistent with the Syrup of Ipecac control.
As well, evidence items 003093-06, 003093-07 and 003093-22 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
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Yoknapatawpha County Forensic Laboratory |
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REPORT DATE: Madeline Holland, Ph.D. |
Analysis: Toxicological Analysis
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REPORT OF FORENSIC TOXICOLOGICAL ANALYSIS |
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|
Case Description: |
Case # |
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Investigative Agency: |
Investigating Officer(s): |
EVIDENCE ANALYZED
- 003093-06 and 003093-07: Two (2) samples from possible vomit pool in hallway outside YCCC Room #273
- 003093-22: One (1) medium-sized plastic bag labeled "YCCC Dry Cleaning"
- Contents: semi-liquid substance visually consistent with vomit
- 003093-34: Chocolates found in Barbara Dubois' YCCC hotel room (#203)
- 003093-38: Chocolates found in Ingrid Freeman's YCCC hotel room (#206)
- 003093-39-013: 1-fl-oz bottle ipecac syrup found in Rachel Webb's YCCC hotel room (#212)
- 003093-40: Chocolates found in Rachel Webb's YCCC hotel room (#212)
- 003093-41 through 003093-43: Chocolates found in Sam Tuttle's YCCC hotel room (#217)
- 003093-45: Chocolates found in Mary and Billie Jo Jones' YCCC hotel room (#247)
- 003093-46: Urine sample from deceased
- 003093-47: 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 |
| 003093-06 | vomit sample (from hallway) | Peak with retention time of 5.2 min. (molecular ion = 481); 5.2mg |
| 003093-07 | vomit sample (from hallway) | Peak with retention time of 5.2 min. (molecular ion = 481); 6.3mg |
| 003093-22 | vomit sample (from bag) | Peak with retention time of 5.2 min. (molecular ion = 481); 4.2mg |
| 003093-34 | chocolates (Barbara Dubois) | Peak with retention time of 5.2 min. (molecular ion = 481); 25mg |
| 003093-38 | chocolates (Ingrid Freeman) | No peak (molecular ion = 481) |
| 003093-39-013 | ipecac syrup (Rachel Webb) | Peak with retention time of 5.2 min. (molecular ion = 481); 1.3mg/ml |
| 003093-40 | chocolates (Rachel Webb) | Peak with retention time of 5.2 min. (molecular ion = 481); 22mg |
| 003093-41 | chocolates (Sam Tuttle) | No peak (molecular ion = 481) |
| 003093-42 | chocolates (Sam Tuttle) | No peak (molecular ion = 481) |
| 003093-43 | chocolates (Sam Tuttle) | No peak (molecular ion = 481) |
| 003093-45 | chocolates (Mary & Billie Jo Jones) | No peak (molecular ion = 481) |
| 003093-46 | deceased's urine | Peak with retention time of 5.2 min. (molecular ion = 481); 0.006mg |
| 003093-47 | 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 003093-06, 003093-07, 003093-22, 003093-30, 003093-34-013, 003093-35, 003093-41 and 003093-42 are positive for emetine, an emetic alkaloid known to occur in Syrup of Ipecac.
