55th SEMI-ANNUAL SEMINAR (Spring 1980)
CALIFORNIA ASSOCIATION OF CRIMINALISTS
MAY 7-10, 1980
SANTA BARBARA, CALIFORNIA

ADVENTURES IN THE APPLICATION OF EXISTING ELECTROPHORETIC EQUIPMENT TO THE SUB-TYPING OF PGM
James M. White

Sub-typing of PGM provides enhanced discrimination. Commercial iso-electric focusing equipment is expensive. Home-made focusing devices and non-focusing methods are described in the literature. These were tried by the author; Carbon-rod isoelectric focusing. Gels prepared as suggested in the forensic literature. Carbon rods 5 X 150mm, emission spec grade used as electrodes, samples run at 1050V after a pre run at constant current of 7mA for 1 hour. Initial focusing runs showed great promise; high resolution and good sharp bands. Results then deteriorated, apparently due to deterioration of the stock acrylamide. Acid starch gel. Separation of the subtypes has been achieved with 0.1M tris, 0.1M maleic acid, 0.01M EDTA, 0.01M MgCl2, adjusted to pH5.6 with NaOH in a 10% starch gel.


PHOSPHOGLUCOMUTASE SUBTYPING BY CONVENTIONAL ELEGTROPHORESIS
Brian Wraxall and Joan Provost

Subtyping of Phosphoglucomutase is a valuable discriminator largely unused by crime laboratories because of the expense of iso-electric focusing and the complexity of interpretation of focused gels.

A method is proposed which utilizes conventional electrophoresis equipment and supplies available in most laboratories to sub-type PGM without iso-electric focusing.

PGM subtyping of blood, vaginal secretions and semen by this conventional method produces well separated and resolved isozyme bands which are easily interpreted in both fresh samples and older stains.

It is felt that PGM subtyping by this procedure may offer crime laboratories an alternative to the expense and complexity-of iso-electric focusing.


THE ADDITION OF 6PGD DETECTION TO A 4 HOUR EAP, EsD, AK, ADA SYSTEM
James M. White

The simultaneous determination of AK, ADA and 6PGD has been suggested by Brinkmann. Its inclusion into a pH.5.9 system for EAP was suggested by Polesky. It was tested on the Wraxall pH5.5 system as modified in this laboratory to a 4 hour run time. Electrophoresis on 10% starch gels. Citrate/ Phosphate buffer pH5.5, Gel buffer diluted 1:80 15V/cm, 4 hours. Compact banding of the PGD allowed sufficient resolution of the A and dimeric AC bands. A homozygous C has not been encountered. This detection is at the sacrifice of the anodal AK band, which may require second runs to resolve AK 2-1 or 2 questions.

6PGD has been conveniently added to a 4 hour EAP detection system. Casework experience so far has successful typing in almost all cases in which AK and ADA types were identified.


RATES OF DRYING OF VAGINAL SWABS
George F. Sensabaugh, Jan Bashinski and Edward T. Blake

It is probable that most of the degradative processes affecting markers on vaginal swabs occur while the swab is still wet. Prerequisite to investigating the extent of these degradative processes we have sought to determine how long swabs remain wet while they are.drying. The major determinant in the drying process is the air flow around the swab. Swabs in still air require 3-5 hours for complete drying depending upon how much fluid is taken up initially. Swabs in closed containers require an estimated 150-250 hours for drying. Swabs in tubes with breathing holes dry in 10-20 hours. Swabs in moving air can dry within an hour.


THE RECOGNITION OF THE RED CELL ACID PHOSPHATASE (AGP) R ALLELE, WITH SPECIAL REFERENCE TO ITS DETECTION FOLLOWING GEL STAINING FOR EsD
James M. White

The R allele of EAP has an incidence in the Black population such that it should be encountered occasionally in most laboratories. Detection is impaired by the comparatively low activity of the enzyme and that the anodic band is in the-region of "storage" activity.

It has been further suggested that prior staining for esterase in the anodic area of the gel impairs detection of this allele.

Method; Electrophoresis on 10% starch gels. Citrate/Phosphate buffer pH5.5, Gel buffer diluted 1:80. 15v/cm, 4 hours.

Although the anodic R band is in the area of "storage", there is sufficient resolution between the anodic A and B bands to clearly resolve the cathodic R band.

Resolving the cathodic R band between the Anodic A and B bands is proposed as a more selective method for recognition of this variant, and overcomes the problems encountered with storage bands and prior esterase staining.


CARBONIC ANHYDRASE TYPE II PHENOTYPING
Rodney Andrus

Problem Under Investigation: Simple and rapid method for typing of CA-II in fresh blood and bloodstains.

Methods Used: Cellulose Acetate Membrane

Results: Achieves good separation in relatively short analysis time.

The method presented provides a rapid and effective means for phenotyping of CA-II polymorphisms in fresh blood and bloostains in as little as 30 minutes of electrophoresis.


THE NATION DECISION AND THE PRESERVATION OF EVIDENCE
Duane Mauzey

The recent decision of the California Supreme Court in reference People vs Nation has necessitated a reexamination of the way physiological fluid evidence is handled by the investigator and laboratory.

The literature dealing with the stability of various genetic markers of forensic interest was reviewed.

The literature review suggested a procedure to be used that would comply with Nation. With the aid of the Santa Barbara District Attorneys Office, this procedure has been implemented.

The procedure is to request submission of all evidence involving physiological fluids to the laboratory within 48 hours. Upon receipt, the evidence is examined for the presence of stains. Stains that are determined to be from physiological fluids are sampled. Samples of each stain are divided. Part of each stain is either subjected to immediate further examination or temporarily stored in a refrigerator for several days until it is convenient for a complete analysis. The remainder of the stain, now thoroughly dry, is placed in a properly marked coin envelope and stored in a -20°C freezer. Exemplar fluids from victims and suspects are put into the form of stains on cloth, and also frozen.


BLOOD "SPLATTER" PATTERN CAUSED BY FLIES
Robert G. Cranston

A crime scene had a blood splatter pattern resembling sprayed blood. However, the presence of such a pattern did not fit the circumstances.

Analysis of the pattern, the circumstances and the presence of large numbers of flies indicated flies were the source. Experimentation resulted in similar patterns.

Under favorable conditions (such as large numbers of flies, ample blood, time, confined areas) flies can produce blood patterns resembling sprayed blood.


EFFECTIVENESS OF LONG TERM FREEZER STORAGE OF BLOODSTAINS
Jan Bashinski

For several years, the Oakland Police Department Crime Laboratory has routinely stored frozen lysates and whole blood dried stains as standard samples. We have also routinely preserved bloodstain evidence awaiting analysis and subsequent to analysis by freezing. The present study was undertaken to investigate the viability of dried bloodstains stored frozen for long periods.

Laboratory-prepared dried stain samples stored frozen up to five years were screened for typable anti A and anti B, hemoglobin, PGM, Es-D, EAP, AK, and ADA. Comparisons were also made between frozen dried stains, frozen lysates, and samples of the same stains stored at room temperature.

Typable genetic markers were detected regularly in dried stains stored frozen for up to two years and, in many instances, for much longer periods.

Long term freezer storage of dried bloodstains is an effective means of preserving genetic markers in typable condition.


STABILITY OF PHENCYGLIDINE IN STORED BLOOD
Darrell Glardy and J.L. Ragle

Does Phencyclidine break down on storage? Does the concentration of phencyclidine increase on storage? 45 Bloods which had been stored for up to 18 months using sodium fluoride and potassium oxalate as the pre-servative. They were reanalyzed using a GC/MS procedure with PCPd5 as the internal standard.

There is no loss of phencyclidine upon storage for up to 18 months. Blood which is preserved with sodium fluoride and potassium oxalate will not affect the concentration of PCP upon storage in or out of refrigeration for up to 18 months.


TITRES IN ABH BLOOD GROUP SUBSTANCES IN HUMAN SEMEN
George F. Sensabaugh, Jan Bashinski and Edward T. Blake

A recent study by the Home Office Central Research Establishment reports that in semen'the titres of H substance generally exceed the titres of A and/or B substances; they used an automated technique in their study. This observation is inconsistent with earlier observations. We have reinvestigated the question using a standard macroscopic hemagglutination technique. Two sets of semen samples have been studied.

A large collection ( n ~ 100) of semen from donors of unknown blood and secretor type have been titred; this parallels the sample set used in the HOGRE study.

A smaller collection of semen samples from donors of known blood and secretor type; for these, saliva ABH titres have also been determined.

The results for both indicate that A and/or B titres generally exceed H substance titres; this contradicts the HOGRE findings.


PHYSICAL EVIDENCE LEADS TO IDENTITY OF ORIGIN OF A CORPSE
Talib ul Haq and Duane Lovass

Problem Under Investigation: Establish the possible identity of a corpse

Knowledge of calligraphy, the Muslim religion, language structure and the metallic composition of a pendant were used.

Possible place of origin of the person whose corpse was found in Yosemite National Park was established to be Iran.

Conclusions:

  1. Eclectic nature of Criminalistics
  2. Information potential of criminalistics.

SURVIVAL OF MARKERS IN PRESERVED SEMEN SAMPLES
George F. Sensabaugh, Jan Bashinski and Edward T. Blake

The survival of acid phosphatase activity, ABH blood group substance titre, and PGM activity has been investigated in seminal plasma samples stored frozen (-10°C ) for periods of up to seven years. The acid phosphatase activity levels in these samples have declined at a rate of about 5% per year; the maximum activity lost in any sample was 75%. ABH titres show no systematic pattern of decline over time. PGM typing could not be done on samples stored before 1975. The proportion of samples that could be typed increased in each succeeding year since 1975 and almost all the samples stored in 1979 could be easily typed. Preliminary results indicate that samples stored dried and frozen exhibit better survival properties.


POTENTIAL AND LIMITATIONS OF FORENSIC BLOOD AND BLOODSTAIN ANALYSIS
Benjamin W. Grunbaum

Problem Under Investigation; To simplify technical operations for phenotyping blood and bloodstains, to generate useful data for statistical interpretation of results and to recognize possible sources of error in reading genetic patterns, especially with regard to bloodstains.

Methods Used: Cooperation of experts in biochemistry, immunology, blood grouping, and statistics in developing frequency data; cooperation with crime laboratories in developing and adapting methodology.

Useful statistical data has been generated. Rapid, reliable and comparatively simple technology now permits phenotyping of more than 25 genetic markers.

A handbook has been prepared for use as reference for those concerned with identification and investigation of blood in civil and criminal justice. Financially assisted through a federal grant from the Law Enforcement Assistance Administration (LEAA) and the California Office of Criminal Justice (OGJP).


THE INDIVIDUALIZATION OF A VISE-GRIP AS THE INSTRUMENT RESPONSIBLE FOR TOOLMARKS IN SKIN; PART I
N. Wall is and R.L. Taylor

A murder victim was found to have toolmarks in her skin. A suspect tool - specifically a Vise-grip - was found and submitted for comparison. The actual specimens as well as silicone impressions of the specimens were examined under the stereoscopic and scanning electron microscopes. A total of 83 new and used Vise-grips were also examined for comparison purposes. Many individualizing features were found in the skin impression and in the impressions made from the suspect tool. Using the techniques of professional film animators, a 16 mm film was prepared to demonstrate the comparison to the jury.


THE INDIVIDUALIZATION OF A VISE-GRIP AS THE INSTRUMENT RESPONSIBLE FOR TOOLMARKS IN SKIN; PART II
R.L. Taylor and N. Wallis

A murder victim was found to have toolmarks in her skin. A suspect tool - specifically a Vise-grip - was found and submitted for comparison. The actual specimens as well as silicone impressions of the specimens were examined under the stereoscopic and scanning-electron microscopes. A total of 83 new and used Vise-grips were also examined for comparison purposes. Many individualizing features were found in the skin impression and in the impressions made from the suspect tool. Using the techniques of professional film animators, a 16 mm film was prepared to demonstrate the comparison to the jury.


DISCRIMINATION AMONG SMOKELESS GUNPOWDER BY MEANS OF COLOR TESTS
Ralph Maloney and John Thornton

To discriminate among smokeless gunpowders through the use of spot tests, and to detect the presence of diphenylamine added to smokeless gunpowders as a stabilizer.

To discriminate among smokeless gunpowders, a battery of sulfuric acid reagents (Marquis, Mecke, etc.) was used. To detect diphenylamine,.conc. HNO3 was used to convert diphenylamine to diphenylnitrosamine; this then reacts with conc. H2SO4 to give a blue color.

Smokeless gunpowder may be characterized by means of H2SO4 -containing alkaloidal reagents; those powders containing diphenylamine can be identified by means of a two-step reaction;

  1. conc. HNO3, followed by
  2. conc. H2SO4

A sensitive test for diphenylamine in gunpowder or gunpowder residues is inherently more attractive from a forensic standpoint then is a test for ubiquitous nitrates. The tandem HNO3 -H2SO4 test appears to achieve this aim.


PROBABILITY AND HAIR COMPARISON
Robert R. Ogle, Jr. and Peter D. Barnett

A series of articles by Gaudette purport to establish certain probabilities regarding the individualization of human hairs. The authors have reviewed the data and conclusions in these articles. The authors conclude that these articles misstate the probabilities involved in hair comparisons. Using the data given in the articles, the authors conclude that Gaudette, et al., grossly over estimate the ability of criminalists to individualize hair. The authors feel that the practice of expert witnesses in citing these articles when giving testimony is misleading to the jury.


RECENT ADVANCES OF HIGH PRESSURE LIQUID CHROMATOGEAPHY IN THE FORENSIC SCIENCES
Matthew Go

Due to its number of diverse applications, high pressure liquid chromatography (HPLC) has been the fastest growing analytical technique for the past 8 years in a row. Recently, HPLC has found its way more and more into the forensic laboratory.

The areas that HPLG offers to the forensic chemist greatest advantage are, (1) screening of poisons and their metabolites in physiological fluids, (2) characterization of illicit drugs and (3) comparative analysis of a variety of materials such as paint, glass, soil, ink, etc.

Toxicology and drug screening in-overdose cases are an important function performed by the forensic chemist, As a very rapid technique, HPLC offers a quick screen for drugs of abuse and their metabolites in blood and urine. Typical analysis times are under 15 minutes.

With increased drug traffic, the analysis of street drugs becomes more important. By using HPLG to "fingerprint" a particular cache, its quality and even possibly, origin, may be determined.

To link possible evidence back to the scene of the crime, comparative analysis involving trace analysis of paint, explosives, ink, soil, etc. is important. Advances in HPLC detector technology has lowered the sensitivity of the technique to low nanogram and in some cases, picogram levels.


FIREARMS EVIDENCE NOMENCLATURE
Lowell W. Bradford

A discussion of approaches to succinct descriptive language in notes and reports involving firearms evidence, dealing with various types of weapons and cartridge components. A model system will be proposed.


THE SEPARATION AND IDENTIFICATION OF ATROPINE AND SCOPOLAMINE FROM SPECIES OF DATURA
Lucien Haag and Margaret Shoumaker

The plants Datura meteloides and Datura Stramonium both grow wild in California and Arizona and have had a history of sacramental use by certain Indian groups. Datura has currently emerged as an infrequent but ready source of the CNS active drugs atropine and scopolamine by young drug abusers occasionally with fatal consequences. The ability to recognize these plants and their seeds and a means to separate and identify their alkaloids was deemed desirable after several instances of poisoning in the Phoenix area. The seeds of Datura sp. are milled with Celite, packed in a column and defatted with petroleum ether. The alkaloids are then extracted with aimnoniacal chloroform. Thin layer chromatography and HPLC are employed to identify the active drug(s) present.

The two most common species of Datura can be discriminated by means of their fruit and seeds. Atropine and scopolamine can be conveniently removed by solvent extraction and subsequently identified by TLC and HPLC.

The symptoms of Datura intoxication are remarkable and the seeds of each species are characteristic and re-cognizable. Both can be extracted in a special Celite column, then analyzed by TLC and HPLC to identify the atropine and scopolamine present.


WIDMARK'S HYPOTHESIS
David W. Sanchez

Widmark's Hypothesis is defined. Its varying mathematical forms are derived from the original expression. The dynamics of alcohol absorption distribution, and elimination are briefly explained and related to the use of Widmark's Hypothesis on forensic problems.


DELAYED ANALYSIS OF BREATH-ALCOHOL
Kurt M. Dubowski, PhD., and Natalie A. Essary, CLS(NCA)

Collection of specimens for breath-alcohol analysis for subsequent analysis is necessary or convenient for certain law enforcement, survey, or research applications. Stored-specimen capability is also a necessity for later analysis by the defense if the result of an official breath-alcohol analysis is to be offered as evidence in the prosecution of alcohol-related traffic offenses, in several jurisdictions, under expanding legal doctrine.

We, therefore, developed and validated methods for breath-alcohol analysis by automated gas chromatography with intervening trapping of the (ethyl) alcohol from measured breath specimens on solid sorbents. Some key findings of extensive in-vitro and in-vivo studies comparing the results of breath-alcohol analysis with and without trapping follow. We conclude that no significant systematic difference exists between the results of direct and delayed breath-alcohol analyses by the specified methods.


PLASMA EMISSION SPECTROSCOPY - A DIFFERENT CONCEPT FOR ELEMENTAL ANALYSIS IN FORENSIC SCIENCE
Cecil L. Hider

A technique to perform elemental analysis on various forensic samples, plasma emission spectroscopy, is presented. Parameters such as cost, sampling techniques, advantages and limitations are presented.

The technique of plasma emission spectroscopy is a viable alternative to EDX and AA when the need for elemental analysis is encountered in the crime laboratory.


MIGROGOULOMETRIC ANALYSIS OF GASOLINE
Bill Johnston

Premium, regular and low lead grade gasoline samples from five separate service stations were analyzed by a microcoulometric technique.

Two different titration cells were required for the determination of relative amounts of sulfur and halogens using an instrument from the Dohrmann Division of Envirotech Corporation. Five microliters of sample were used per analysis. Analysis time was approximately six minutes per sample for each determination.

A digital display on the instrument gave relative amounts of either sulfur or halogen depending on the cell employed. Data provided a means for distinguishing between all but two of the samples analyzed


ACOUSTIC MICROSCOPY
Robert C. Bray and G.F. Quate

The acoustic microscope using microwave frequency sound is now capable of resolution comparable to the best optical microscopes. The acoustic microscope will be described and representative images of a variety of samples (materials, electronic devices, biological) will be presented and compared with optical and electron microscope images.

The acoustic microscope, since it is sensitive to the mechanical properties of the sample under consideration, offers information which is complementary to the optical images. In some cases, the acoustic images reveal information unobtainable by optical imaging.