The author(s) shown below used Federal funds provided by the U.S. Department of Justice and prepared the following final report: Document Title: Development of Advanced Raman Spectroscopy Methods and Databases For The Evaluation of Trace Evidence and The Examination of Questioned Documents (Phase I) Author: John R. Lombardi, Marco Leona, Tuan Vo-Dinh, Philip Antoci Document No.: 227843 Date Received: August 2009 Award Number: 2006-DN-BX-K034 This report has not been published by the U.S. Department of Justice. To provide better customer service, NCJRS has made this Federally- funded grant final report available electronically in addition to traditional paper copies. Opinions or points of view expressed are those of the author(s) and do not necessarily reflect the official position or policies of the U.S. Department of Justice. This document is a research report submitted to the U.S. Department of Justice. This report has not been published by the Department. Opinions or points of view expressed are those of the author(s) and do not necessarily reflect the official position or policies of the U.S. Department of Justice. Report Title: Development of Advanced Raman Spectroscopy Methods and Databases For The Evaluation of Trace Evidence and The Examination of Questioned Documents (Phase I) Authors: John R. Lombardi, Center for Analysis of Structures and Interfaces, City College of New York (CCNY); Graduate Faculty, Forensic Sciences, John Jay College of Criminal Justice Marco Leona, The Metropolitan Museum of Art (MMA), New York Tuan Vo-Dinh, Director of the Fitzpatrick Institute for Photonics and Professor of Biomedical Engineering, Duke University Philip Antoci, NYPD Crime Laboratory and John Jay College of Criminal Justice Award Number: 2006-DN-BX-K034 Award Number 2006-DN-BX-K034 1 of 146 This document is a research report submitted to the U.S. Department of Justice. This report has not been published by the Department. Opinions or points of view expressed are those of the author(s) and do not necessarily reflect the official position or policies of the U.S. Department of Justice. (blank) Award Number 2006-DN-BX-K034 2 of 146 This document is a research report submitted to the U.S. Department of Justice. This report has not been published by the Department. Opinions or points of view expressed are those of the author(s) and do not necessarily reflect the official position or policies of the U.S. Department of Justice. Abstract: Researchers from the City University of New York, Fitzpatrick Institute of Photonics at Duke University, the New York Police Department Crime Lab, and The Metropolitan Museum of Art formed a multidisciplinary collaborative to further the application of Raman spectroscopy and non-destructive surface enhanced Raman scattering (SERS) analysis techniques as applied to the evaluation of trace evidence and the examination of questioned documents. Through the sharing of instrumentation (e.g. the only multiwavelength open architecture Raman/FT-Raman microscope currently available), reference materials (e.g. a collection of natural and synthetic pigments and inks dating to the first half of the century), and their respective research expertise the team developed a much needed reference database of spectra for forensic applications and cross-validated newly developed techniques for non-destructive SERS analysis. Raman spectroscopy is an established and increasingly utilized technique for the rapid and non-destructive analysis of paints, inks, fibers, mineral residues, pharmaceuticals, and controlled substances. The technique is applicable to a variety of substances, but analytes of great forensic interest such as some natural and synthetic dyes found in textiles, inks, and paints display an excessive fluorescent background, which limits Raman efficiency in most situations, leading to poor analytical results. The team's goal is to further explore the application of Raman spectroscopy to the evaluation of trace evidence and the examination of questioned documents, with the aim of solving the problems that have so far limited the applicability of this technique to the identification of organic colorants and other materials in trace amounts. This project is divided into two phases. Phase I methods and results are covered in this report. Based on the success of Phase, funding for Phase II is currently being requested. The work in Phase I of the project successfully demonstrated that surface enhanced Raman scattering (SERS) can be applied to the identification of organic colorants present in inks, paints, and textile fibers. The techniques we developed Award Number 2006-DN-BX-K034 3 of 146 This document is a research report submitted to the U.S. Department of Justice. This report has not been published by the Department. Opinions or points of view expressed are those of the author(s) and do not necessarily reflect the official position or policies of the U.S. Department of Justice. are especially suited for handling microscopic samples: textile dyes were successfully identified from samples as small as a one-millimeter section of a single silk fibril of fifty-micrometer diameter and even from textiles severely degraded by burial. The research conducted in Phase I also demonstrated that SERS is a valuable technique for the identification of organic colorants used in inks and for the dyeing of textile fibers, as well as for trace analysis of pharmaceuticals and of drugs of abuse. Analytical procedures for SERS of a number of representative dyes were developed, the core of a high quality spectral database was assembled as a proof of concept experiment, and innovative non-destructive approaches were investigated. To date our colorants database includes approximately 50 natural and synthetic dyes, and it is, to our knowledge, the first SERS spectral database ever assembled (in the course of the work, a normal Raman database of approximately 100 spectra of organic colorants was also assembled). The database is both chemically inclusive (as dyes belonging to the principal classes are represented) and spectrally comprehensive, since both SERS spectra obtained at different wavelengths (488, 633, 785 nm) and normal dispersive Raman (488, 633, 785 nm), and FT-Raman (1064 nm) are included. Scientific articles describing this first phase of the work were accepted for publication in the Journal of Forensic Sciences. Award Number 2006-DN-BX-K034 4 of 146 This document is a research report submitted to the U.S. Department of Justice. This report has not been published by the Department. Opinions or points of view expressed are those of the author(s) and do not necessarily reflect the official position or policies of the U.S. Department of Justice. Table of Contents TABLE OF CONTENTS ..................................................................................... 5 EXECUTIVE SUMMARY .................................................................................. 6 I. INTRODUCTION.......................................................................................... 10 1. STATEMENT OF THE PROBLEM ................................................................... 10 1.1 Surface Enhanced RAMAN Spectroscopy (SERS) .............................. 10 1.2 SERS and RAMAN Spectroscopy in Forensic Science ....................... 11 2. STATEMENT OF HYPOTHESIS OR RATIONALE FOR THE RESEARCH ............. 14 2.1 Objectives............................................................................................ 14 2.2 Development of Analytical Protocols and Creation of a SERS Database ................................................................................................... 14 2.3 Non-Destructive Analysis with SERS Nanoprobes ............................. 15 2.4 Non-Destructive Analysis with Matrix Tranfer SERS......................... 16 2.5 Sensitivity Enhancement with Immobilized Colloids .......................... 16 II. METHODS..................................................................................................... 17 1. PREPARATION OF AG COLLOIDS ................................................................ 17 2. RAMAN SPECTRA ....................................................................................... 17 3. MATRIX TRANSFER SERS ......................................................................... 18 4. IMMOBILIZED COLLOIDS ............................................................................ 19 III. RESULTS..................................................................................................... 20 1. STATEMENT OF RESULTS ........................................................................... 20 1.1 Results from the Metropolitan Museum of New York Site-................. 20 1.2 Results from the Duke University site................................................. 23 2. TABLES AND FIGURES ................................................................................ 26 IV. CONCLUSION.......................................................................................... 133 1. DISCUSSIONS OF FINDINGS....................................................................... 133 2. IMPLICATIONS FOR POLICY AND PRACTICE .............................................. 136 3. IMPLICATIONS FOR FURTHER RESEARCH ................................................. 137 V. REFERENCES............................................................................................ 141 VI. DISSEMINATION OF RESEARCH FINDINGS.................................. 143 1. PRESENTATIONS AND INFORMATION