Robert Rothchild
Science Department An Organic Chemistry Experiment for
John Jay College of Criminal Justice
City University of New York
445 West 59th Street
Forensic Science Majors
New York, NY 10019
I
Teaching organic chemistry within a program for Forensic
Science majors presents a special challenge; the course is a n
essential part of a satisfactory background for future required
courses in biochemistrv a n d forensic science. M a n -v a .. ~olica-
tions are frequently encountered by t h e practicing forensic
science orofessional who must have a n understanding of or-
ganic renrtim m w h a n ~ s m tosupport
s variuu.icharacteri7ation
tests :and analyticd results, particularly if these resulti are
heing prest.nrtd n i part of s o m . courtroom proceedings. To
some extent, the organir chemistry course r a n he "slanted"
toward the firld of specialirntiun with no sacrifice of course
rontent and, i~idet,d,with enhancement oistudent motivation.
Eiferrive mpthods for doinr" this would inrlude freouent use
-
of compounds of special interest t o forensic scientists, such
as drugs, controlled substances, toxic materials a n d reagents
used for specific purposes i n modern criminalistics laborato-
ries.
Of particular concern i n presenting courtroom evidence is
t h e degree of certainty t o he ascribed t o a n experimental
conclusion. Has t h e Identity of a sample been confirmed
"hevond a reasonable doubt"? A wav of introducine this
concept in a numerical sense (hut n o t a philosophic one!) has
develooed a s a simole and reasonable offshoot of one of t h e
organiE chemistry Lboratory experiments. T h e basic exper-
iment is one which is oarticularlv. a .. ~ ~ r .o o r i a for
t e future
criminologists. I t is t h e familiar use of thin-layer chroma-
tography' (tlc) for qualitative analysis, specifically theiden.
tification of d r u ~ sWhile
. student analysis of suhstances such
~~~
.~
~ , - - he e s ~ e c i a l.l vaD-
as mornhine.. heroin. cocaine.. etc... might
propriate for eliciting student interest, there are real problems
- mixture; difficulties of using mixed solvent systems should be dis-
associated with access t o and inventow control of such strictly
regulated materials. I n practice, student response has
.been encountered during analyses of various commercial
over-the-counter analgesic formulations, including assorted
headache, pain, a n d cold tablets.
Specllics
Ire w~uldurge some mdification* in the performance of this rx-
prrimmt ,>r it 1% wmetimwdelrrit,cd in lnhorarury manuals. Com-
mon solvents such as benzene, chloroform, and carbon tetra-
chloride a r e classified as cancer suspect agents and should not
be used for extraction of drugs from sample tablets, for solution of
known reference standards or for development of the tlc chromato-
grams. We have managed almost totally to phase out the use of these
solvents in our organic chemistry laboratory by use of substitutes such
as hexane, toluene, or methylene c h l ~ r i d eAnother
.~ modiGcation that
we suggest is the use of commercial Silica Gel tle plates impregnated
with fluorescent indicator. This allows multiple visualization tech-
niques to be applied for more effective sample characterization. Vi-
sualization of spots can be performed with both shortwave and
longwave radiation and students can note plate appearance, spot
fluorescence and color while separately observing with 254 nm and
360 nm light sources. Students are often quite surprised to note the
behavior of different sample spots and readily appreciate the added
information for sample identification that can be provided in this way.
Students should be cautioned against direct viewing of the ultraviolet
light sources? Visualization with iodine (in a hood) should be applied
after use of the ultraviolet lamp.
To expand the experiment with development of concepts of po-
larity, solvent strength, and eluotropic series, students should run
plates with low and high polarity solvents, such as methylene chloride
and acetone, and then prepare solvent mixtures of intermediate po-
larity. Of special interest would be the use of the newly available re-
versed-phase tlc plates4 in which octyl- or oetadecylsilyl groups are
chemically bonded to silica gel. With these plates, solvent strength
(eluting power) will generally increase with decreasing solvent po-
larity; methanol will be a more powerful solvent than water. Use of
reversed-phase tlc plates is especially relevant because of the pre-
ponderance of analyses and separations being carried out on re-
versed-phase columns with high performance liquid chromatography
(HPLC). Applications of this technique in'forensic (and other) labs
is growing very rapidly. While HPLC has not yet become common in
undergraduate organic chemistry courses because of cost, etc., the
concepts of reversed-phase chromatography which is so useful in
HPLC may be cheaply and easily introduced by using the corre-
sponding tlc plates.
Finally, students can perform a very elementary statistical treat-
ment of their tlc results if they carry out replicate runs of their known
reference materials and the unknowns. This is useful to evaluate re-
producibility of student technique. Comparisons of methods involving
side-by-side spotting across asingle tlc plate with spatting on different
plates and with superimposed spotting of two different materials
should be made and discussed from the viewpoint of achieving max-
imum confidence in results.
For forensic science laboratory results to lead to effectiveprobative
evidence. it could be useful to have a auantitative sense of haw sie-
~ ~ ~~ ~ ~ ~ ~~ ~~~
hnckpmnd d~srurrmnoi~tntirti&,studens can drtrrmmr standard
.
nificant or believable the data mav h.without reauirine an extensite ~
drr i;nionr for the ti, valut-s determined tn,m replienre sputrlnoi on
a single and on different tlc plates. If different brands of commercial
from a single individual with valuesbbtained from Rrvalues deter-
mined by different students. Reproducibility using a single solvent
for development should he compared with results from a solvent
cussed, including inaccuracies in mixture preparation by different
individuals and selective solvent evaporation from the mixture.
Students can determine whether Rrvalues are susceptibleto matrix
effects.Thus, a student might plot average Rrtslues determined from
solutions of pure known reference compounds against values obtained
after extraction from a commercial tablet containine several com-
pounds HS wrll as an excipient (tinder, zurh a\ <larch.Lktrrmmnlion
of the rorrrlatlm cwfficwnr fr,r the straight linr 1,1<~t through this
swim dpointrshould he w r y hieh in thc nhiencr of interfering mntrix
effects. The points corresponding to the averaged Rr values from
replicate determinations can he used to define the best straight line
fit. Invariably, some student has a calculator that the class can use
far the task of establishing this line and the corresponding correlation
coefficients as welL5
Finally. when Rrvaluesformainajor Imisfor nrmpc,und idmtif:.
m r m , dtudrnts shuuid determine the rmfidencc limit; that michr
be SPI tvarrd on their replicate dctermlnntion.. Srudenti quwkl :sin
1 One good treatment of the topic appears in: Perry, S. G., Amos,
R.. and Brewer. P. I.. "Practical Liauid
' Chromatop;raphy."
. .~ Plenum
~ i e s sNew
, ~ & k 1973,
, Chapt. 6.
2 RVPII
.... short.
~~
t e ~ mexnosum should he avoided. Verv relevant for
.... ~~~~
7 ~~ ~~
student laboratories is the maximum allow~blc5 p.pm t,rlrhorr fw
15 minutes fg,r brnrrne. See: Green. M.E., and'l'urk, A . "Safety in
Work~nr:with Chrmirals." The M~crn~llan Puhlishinr Co.. Inc. X e w
York, 1978,p. 84.
3 Lamp model UVSL-25 has been used satisfactorily; Ultraviolet
products. Inc., San Gabriel, CA.
4 Available from Kontes, Vineland, N.J. or from Whatman Inc.,
Clifton, NJ.
5 See, for example, Publication LCB-2510, "Calculating Better
Decisions," Texas Instruments, 1977, especially pp. 2-14 to 2-18.
6A useful basic statistics text is: Weinberg, George H., and Schu-
maker, John A,, "Statistics: An Intuitive Approach," 3rd Ed.,
BrooksICole Publishing Ca., Manterey, CA, 1974.
7 Sherma, J., and Beim, M., HRC CC, J. High Resolut. Chromatogr.
Chromotogr. Cornrnun., 1,309 (1978);CA, 90 110065 (1979).
Volume 56, Number 11, November 1979 1 757
 an appreciation of the desirability and necessity of being able to
quantitate this concept and to be able to say, for example, that a
compound's Rrvalue has been shown to be within specified limits from
a particular value with, e.g., 95 or 99% confidence. The desirability
-
of estahlishine narrower limits or hieher confidence can he weiahed
against the time required for additional determinations.
These simple statistical analysesbpplied to this particular ex-
__3
758 1 Journal of Chemical Education
periment greatly enhance its value and interest for the forensicscience
major and should surely do the same for other students as well.
Note Added in Proof. Reversed-phase tlc plates are also available
frllm Analtech.
~-~~~~~~~
~~~,Inc.., Newark.. Delaware. For a discussion of the de-
~
termination of caffeine in APC tablets using reversed-phase tlc plates,
see Sherrna and Beim."