Brandon Doss

Dept of Chemistry & Biochemistry

Introduction
Ponderosa pine resin samples will be analyzed
by GC-MS to determine whether attack by bark
beetle alters the relative terpene composition of
the resin. Seven terpenes will be monitored: !-
pinene, "-pinene, myrcene, 3-carene, limonene,
terpinolene, and longifolene. Fenchone is used
as an internal standard.

Sample Preparation
Resin samples are prepared as 0.25g/mL
solutions in dichloromethane. Dichloromethane
was chosen as a solvent because of its strong
dissolving power towards the resins, and
because it has a very low boiling point, which
makes it a good choice for gas
chromatography. The samples were further
diluted in dichloromethane to a resin
concentration of 2.5e-3g/mL (2,500ppm) prior to
analysis by GC-MS. Each sample was spiked
with 9.5e-5g/mL (95ppm) of fenchone as an
internal standard.

Terpene Identification
A stock terpene solution was prepared to
establish the retention order and identification
of the compounds by GC-MS. 10uL of each
terpene was added to 930uL of hexane. This
was further diluted by a factor of 100 in
dichloromethane prior to GC-MS analysis.
Figure 1 shows the chromatogram of the stock
solution.

All terpenes are well resolved from each other
and from the internal standard with the
exception of limonene and terpinolene. After
comparison of the Kovats retention index
1(NIST)

and analysis of an extracted ion chromatogram
for m/z=136, Figure 2, it was determined that
these two components are co-eluting.



Pine Resin Terpene Analysis
by GC-MS.

Instrumentation
Varian Saturn 3900 GC
Varian Saturn 2100T Ion-Trap MS


Figure 1. Terpene standard solution by GC-MS.



Figure 2. Extracted ion chromatogram of
terpene standard, m/z=136.

The peak at 14.5min is nearly twice the height and
area of its counterparts, suggesting that two
components are eluting at the same time. Notice the
absence of fenchone (m/z=152) and the substantial
decrease of the longifolene signal (m/z=204).





Data Analysis
There will be no absolute measurement of
terpene concentration in this study, rather the
concentration of individual terpenes will be
measured against the internal standard
component, fenchone. Although the absolute
terpene concentrations will be unknown, the
concentration of fenchone is well specified and
thus provides a point of reference for a relative
interpretation of the data. We will monitor the
ratio of terpene signal to fenchone signal in
each sample. For example, the ratio of a-
pinene to fenchone in BSC3 is 5.02, Figure 3.
We will monitor this ratio in BSB3 to see how it
changes.


Figure 3.