Characterization of Fetterbush 

Lyonia lucida Liquid Extractions

Roderquita K. Moore

Doreen Mann

Frederick Matt

Mark Dietenberger

David Weise

United States Department of Agriculture

Forest

Service

Forest Products

Laboratory

Research Note

FPL–RN–0385

September

2020

First dimension (s)

Second dimension (s)
September 2020

Moore, Roderquita K.; Mann, Doreen; Matt, Frederick; Dietenberger, 

Mark; Weise, David. 2020. Characterization of fetterbush Lyonia lucida

liquid extractions. Research Note FPL-RN-0385. Madison, WI: 

U.S. Department of Agriculture, Forest Service, Forest Products 

Laboratory. 8 p.

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Abstract

Fetterbush (Lyonia lucida) grows in the southern states of 

the United States. The chemical components in fetterbush 

are highly concentrated with phenolic compounds. These 

compounds contribute to the toxicity of the plant and 

could possibly be igniters for wildfire. In this study, liquid 

extractions were obtained from fresh Fetterbush samples. 

Two-dimensional gas chromatography and time of flight 

mass spectrometry (GC×GC-TOFMS 4D) were used 

to characterize the chemical compositions in the liquid 

extractions. GC×GC technology has existed for more than 

30 years. This technique opens the peak capacity in which 

trace and hidden chemicals can be identified. GC×GC 

detected 1,899 and 916 chemicals from the solvent and 

bead bug extractions, respectively. There were 72 chemicals 

shared by both methods identified with similarity match 

>900.

Keywords: GC×GC; extractives; bushes, Lyonia lucida; 

liquid extraction

Contents

Introduction..........................................................................1

Experimental Methods.........................................................1

Results and Discussion ........................................................2

Conclusion ...........................................................................8

Acknowledgments................................................................8

References............................................................................8
Characterization of Fetterbush Lyonia 

lucida Liquid Extractions

Roderquita K. Moore, Research Chemist

Doreen Mann, Physical Science Technician

Frederick Matt, Chemist

Mark Dietenberger, Research General Engineer

USDA Forest Service, Forest Products Laboratory, Madison, Wisconsin, USA

David Weise, Research Forester

USDA Forest Service, Pacific Southwest Research Station, Fire and Fuels Programs, Riverside, California, USA

Introduction

Fetterbush (Lyonia lucida) is a shrub native to the southern 

United States. This bush, along with many other species, is 

being investigated to determine its contribution in the severe 

wildfires that take place every year. Pyrolysis modeling 

is important to understanding wildland fire. Amini and 

others (2019) used slow pyrolysis to identify more than 200 

compounds in tar from dead and live vegetation. Countless 

forest fires occur yearly and sometimes grow uncontrollably 

and rapidly. The chemicals from the various bushes found 

on the floor of the forest are known for contributing fuel 

to these fires. Fetterbush is one of the vegetations being 

used for pyrolysis modeling. Fetterbush is highly toxic, 

possibly fatal if eaten, and produces allelic compounds that 

deter plant growth in its vicinity (Fisher 1987). The poison 

parts of the bush are the leaves and nectar from the flowers. 

The chemicals from this plant are of extreme interest for 

medicinal, herbicidal, and potential fuel source purposes. In 

this study, the primary chemicals of fetterbush leaves were 

extracted by two solvent methods. Liquid extractions are 

known for separating chemicals into various fractionation 

using different solvents. (Moore and others 2015a, 2015b, 

2017a, 2017b). Both methods used solvents, but one 

method was shaken and the other method used steel beads 

to remove the extracts from the leaves. Comprehensive 

two-dimensional gas chromatography and time of flight 

mass spectrometry (GC×GC TOFMS) were used to identify 

the isolated extractives from both extraction methods. 

This comparison determined which method isolated the 

most chemicals from the leaves. The chemical fingerprint 

from the GC×GC chromatograms will assist in identifying 

chemical groups that could possibly be hazardous fuel 

sources in wildland fires.

Experimental Methods

Materials

Fetterbush plants were purchased from a nursery. The 

moisture content of the plants was kept at 30%. Solvents 

were purchased from Thermo Fisher Scientific (Waltham, 

Massachusetts, USA). Tetracosane 99% is the calibration 

standard and was purchased from Sigma-Aldrich (St. Louis, 

Missouri, USA).

Solvent Extraction

Fetterbush leaves were extracted using solvent extraction 

(SE). The leaves were picked and frozen until ready to be 

extracted. The leaves were slightly thawed and weighed 

and then placed in a 250-mL Erlenmeyer flask with 100 mL 

of solvent solution. The first solvent solution (fb-mle) was 

made of acetone and water (90:10), and the second solvent 

solution (fb-c2) was made of hexane and isopropyl alcohol 

(50:50). In both solvent extractions, the solvent solutions 

were filtered, and fresh solvent was poured over the leaves 

every 24 hours. The leaves in the solution were shaken 

(orbital shaker) for a total of 48 hours. Solvent solutions 

were evaporated using a rotary evaporator, and then the 

extracts were transferred to preweighed vials, sealed, and 

stored in the freezer.

Bead Bug Extraction

Bead Bug (BB) (Benchmark Scientific, Edison, New Jersey, 

USA) was also used for extraction of fetterbush leaves. 

The BB uses a bead mill vial containing steel beads. The 

BB vials were filled with 1:1 hexane/isopropanol solution 

(fb-c2) and were centrifuged for 2 min at 400 rpm. After 

this cycle, the vials were placed in a desktop centrifuge at 

12,000 rpm for 1 to 2 min. The supernatant was decanted 

into a 10-mL glass syringe and filtered with a tarred nylon 

0.45-μm luer lock into a 50-mL flask. This was repeated 

until the leaves were a brownish tan color and all the 

green was gone from them. For the last extraction, a hand 

vortex was used for 50 seconds to remove any remaining 

extractives in the leaves. Four replicates were performed 

using 9:1 acetone and water (fb-mle) and were collected 

in different 50-mL volumetric flasks. At this point in the 

procedure, the tissue was completely pulverized.
Research Note FPL–RN–0385

2

Instrumentation Operations

Table 1 shows the GC×GC instrument parameters. The oven 

programming temperature for the secondary column was 

5 °C higher than the primary column. The transfer line and 

injection temperatures remain the same for every run. The 

split ratio varied based on the concentration of the complex 

mixture. The modulation temperature was 15 °C higher 

than the secondary oven. The retention time for the second 

dimension was less than 5 s for each sample.

Results and Discussion

Two solvent extractions were performed for each method 

(SE and BB), but only the first extraction from each method 

was used for the GC×GC analysis. The first extraction from 

the SE method was the acetone extraction (fb-mle), and 

the first extraction from the BB method was the hexane 

extraction (fb-c2). The extractions were run in triplicates. 

NIST and Wiley libraries were used to establish spectral 

similarity between the chemicals identified in the fetterbush 

complex extract mixtures. The minimum similarity for 

matches was 700. The number of chemicals identified from 

the library databases were 1,899 and 916 chemicals in the 

acetone (fb-mle) and hexane (fb-c2) extracts, respectively.

Table 2 lists the number of chemicals detected at various 

similarity matches. There were 252 and 96 unknown 

chemicals detected for the fb-mle and fb-c2 extracts, 

respectively.

Table 1—Gas chromatography and two-dimensional 

gas chromatography-time of flight mass spectroscopy 

(GC×GC-TOFMS) (Pegasus BT 4D) instrument 

parameters

Gas chromatograph agilent 7890B with LECO dual stage quad jet 

modulator and LPAL 3 autosampler

Injection 1 uL, split 20–150:1 at 250 °C

Carrier gas He at 1.4 mL/min

Primary column: Rxi-5 ms, 30-m by 0.25-mm i.d. by 

0.25-μm coating (Restek, Bellefonte, 

PA, USA)

Secondary column Rxi-17SilMS, 0.60-m by 0.25-mm by 

0.25-μm coating (Restek, Bellefonte, 

PA, USA)

Oven programming Primary column starting temperature 

50 °C, ramped 10 °C/min to 

320 °C, held 5 min. Secondary column 

temperature parallel ramp offset by 

+5 °C (total run time 32 min).

Modulation 3–5 s with temperature maintained 

+15 °C relative to second oven

Transfer line 280 °C

Mass spectrometer LECO Pegasus BT 4D

Ion source temperature 250 °C

Mass range 50–650 m/z

Acquisition rate 150 spectra/s (2D)

Table 2—Number of chemicals detected by twodimensional gas chromatography-time of flight mass 

spectrometry (GC×GC-TOFMS) and identified by NIST 

and Wiley libraries

Solvent 

solution

Total 

>700

Peaks 

unknown

Peaks 

known

>800 

known

>800 

knowna

fb-mle

(SE method)

1,899 252 1,647 798 245

fb-c2

(BB method)

916 96 820 530 104

a

Number of known chemicals after removing shared and isomers.

Chemicals with isomers and duplicates caused by high 

concentrations were removed from the list. The modified 

number of chemicals detected was 245 in fb-mle and 104 in 

fb-c2 with greater than 800 similarity matches. Seventy-two 

chemicals were shared between both extracts. These were 

not included in the total for each extract.

The lists of chemicals in Tables 3 and 4 are not duplicates 

and are specific to their solvent and extraction method. The 

chemicals listed are with spectral matches greater than 900. 

The percentage peak area was calculated using the sum 

of all chemicals with matches greater than 800 similarity. 

Several ketones and furanones were identified from the fbmle extracts. These chemicals are known for various flavors 

and scents. Shikimic acid is used in antiviral chemicals such 

as that in Tamiflu (Genentech, Inc., South San Francisco, 

California, USA). The chemicals in Tables 3 and 4 did 

not yield any significantly high percentage peak areas 

with spectral matches at 900 similarity. However, there 

were chemicals below the 900 spectral matches with high 

percentage peak areas such as 1,2,3,4,5,6-cyclohexanehexol 

11.97% and 1-ethyl-5-methylcyclopentene 9.71% in the 

fb-mle extracts and 2-hexanol 14.19% and (+)lactic acid 

11.63% in the fb-c2 extracts.

GC×GC analysis identified 72 chemicals shared between 

both solvent methods. Sugars and phenolic compounds 

had high percentage peak areas. Tables 5 and 6 show 

chemicals with greater than 900 similarity matches. Fewer 

chemicals were identified in the fb-c2 extracts. This could 

have been caused by the concentration of the extracts. 

Fb-mle chemicals with high percentage peak area were 

β-D-glucopyranose; 1,6- anhydro- 23.8%; (pyro)-catechol 

12.6%; and 5-hydroxymethylfurfural 7.65%. Glucose 

10.9% was high, but it was below the 900 similarity match 

threshold. Sugars were highly concentrated in the fb-mle 

extracts.

Figures 1 and 2 show different ways to visually demonstrate 

the GC×GC chromatograms of the fb-mle (SE method)

extract. Figure 1 is a surface plot, and Figure 2 is a contour 

plot. The retention time of the chemicals eluted in the 

first dimension is based on volatility, and in the second 

dimension, it is according to polarity. Any ripple in the blue 

surface is an indication of a chemical being detected. The 

black dots on the contour plot are chemicals detected. The 

peaks that are most intense or highly concentrated are red.
Characterization of Fetterbush Lyonia lucida Liquid Extractions

3

Table 3—Two-dimensional gas chromatography-time of flight mass spectometry (GC×GC-TOFMS) list of 

compounds specifically identified in fetterbush extracts (fb-mle, SE method) >900 similarity

Number Chemical identified Formula

Peak area 

(%)

Retention time

(s) Similarity

1 4-Hydroxy-4-methylpentane-2-one C6H12O2 4.359 203.001 0.989 960

2 Maleic anhydride C4H2O3 0.012 209.004 1.459 931

3 2-Furanmethanol C5H6O2 1.822 212.005 1.080 944

4 5-O-Methoxymethylbacillariolide I C22H32O4 0.391 215.006 0.972 918

5 Malonic acid C3H4O4 0.018 230.012 0.489 904

6 Acetic acid C2H4O2 0.276 233.014 0.605 909

7 2,3-Butanedione C4H6O2 0.148 242.017 0.316 962

8 1-Hydroxy-3-penten-2-one C5H8O2 1.489 251.021 1.198 951

9 2(5H)-Furanone C4H4O2 0.777 257.024 1.735 962

10 1,2-Propadiene-1,3-dione C3O2 0.141 266.027 1.390 928

11 5-Methyl furfural C6H6O2 3.144 296.04 1.383 916

12 3-Hydroxy-2(5H)-furanone C4H4O3 0.088 299.041 1.550 927

13 Methyl 6-formyl-hept-6-eneoate C9H14O3 0.007 317.048 0.841 943

14 β-Myrcene C10H16 0.470 317.048 0.908 941

15 2-Hydroxy-gamma-butyrolactone C4H6O3 0.981 317.048 1.838 905

16 2H-Pyran-2,6(3H)-dione C5H4O3 0.102 320.05 1.502 909

17 Ethanol, 2-(2-ethoxyethoxy)- C6H14O3 0.298 329.053 1.219 933

18 Anhydromevalonolactone C6H8O2 0.008 362.067 2.112 938

19 1,3,6-Octatriene, 3,7-dimethyl-, (E)- C10H16 0.355 368.07 0.957 937

20 3.,22(S),29-Trihydroxy-22,25-epoxylanost-9(11)-ene C30H50O4 0.071 371.071 1.456 940

21 2,5-Dimethyl-4-hydroxy-3(2H)-furanone C6H8O3 1.044 374.072 1.366 914

22 5-Hydroxy-6-methyl-2,3-dihydropyran-4-one C6H8O3 8.548 404.084 1.532 916

23 Benzene, 1-methyl-4-(1-methylethenyl)- C10H12 0.041 407.086 1.159 900

24 Undecane C11H24 0.124 410.087 0.818 932

25 Benzeneethanol C8H10O 0.294 428.094 1.468 932

26 3-Allyloxypentane C8H16O 0.059 443.101 1.354 902

27 2,3-Dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one C6H8O4 1.730 452.104 1.512 924

28 2,2-Dimethyl-4-oxidanyl-hexan-3-one C8H16O2 0.025 470.112 0.639 906

29 5-(Hydroxymethyl)dihydro-2(3H)-furanone C5H8O3 0.090 491.121 2.142 911

30 FURAN C4H4O 0.080 506.127 1.862 929

31 3-Butenal-2-one C4H4O2 0.011 512.129 2.218 929

32 3-Cyclohexene-1-acetaldehyde, α,4-dimethyl- C10H16O 0.368 518.132 1.285 904

33 6-Octen-1-ol, 3,7-dimethyl- C10H20O 0.098 524.134 1.102 901

34 Cyclopentanol, 2-ethenyl-2-methoxy-, trans- C8H14O2 0.029 524.134 1.712 999

35 2-Decenal, (Z)- C10H18O 0.046 551.145 1.145 910

36 Benzoic acid, 2-(acetyloxy)- C9H8O4 0.128 581.158 1.348 909

37 Ketene C2H2O 0.025 599.165 0.551 941

38 (5S)-5-Ethyl-2-oxolanone C6H10O2 0.005 611.17 1.110 921

39 2H-Pyran-3,4,5-triol, tetrahydro-2-methoxy-6-methyl- C7H14O5 0.461 626.176 1.641 933

40 1,6-Anhydro-β-d-talopyranose C6H10O5 0.749 638.181 2.087 904

41 (3-Propyl-2-oxiranyl)methanol C6H12O2 0.136 653.188 1.734 918

42 Benzeneethanol, 4-hydroxy- C8H10O2 0.509 683.2 1.813 920

43 2-Heptanoylfuran C11H16O2 0.021 695.205 0.631 907

44 2-Methoxy-4-(n-propyl)phenol C10H14O2 0.026 713.212 1.788 905

45 p-Hydroxybenzoic Acid C7H6O3 0.524 737.222 1.761 903

46 2-Propanone, 1-hydroxy- C3H6O2 10.719 740.224 0.538 942

47 α-Selinene C15H24 0.010 746.226 1.149 912
Research Note FPL–RN–0385

4

Table 3—Two-dimensional gas chromatography-time of flight mass spectometry (GC×GC-TOFMS) list of 

compounds specifically identified in fetterbush extracts (fb-mle, SE method) >900 similarity—con.

Number Chemical identified Formula

Peak area 

(%)

Retention time

(s) Similarity

48 3,5-Diethylhex-5-en-3-ol C10H20O 0.029 749.227 0.701 904

49 Oct-7-en-1-ol C8H16O 0.018 770.236 0.594 906

50 Benzaldehyde, 3,4-dihydroxy- C7H6O3 0.160 797.247 1.914 904

51 3,4-Dihydroxyphenyl-2-propanone C9H10O3 0.161 863.275 1.963 904

52 Dodecyl acrylate C15H28O2 0.044 872.278 1.064 921

53 D-Allose C6H12O6 3.675 920.298 1.162 910

54 Shikimic acid C7H10O5 2.453 926.301 2.120 934

55 Phytol C20H40O 0.619 1,124.38 1.091 938

56 Docosane C22H46 0.046 1,169.4 0.939 938

57 4,8,12,16-Tetramethylheptadecan-4-olide C21H40O2 0.025 1,253.44 1.312 923

58 3-Eicosene, (E)- C20H40 0.007 1,313.46 1.179 918

59 Pentacosane C25H52 0.156 1,361.48 1.030 947

60 Hentriacontane C31H64 0.153 1,490.53 1.159 940

61 Methyl 5-(4-methoxyphenyl)-2,2,4,4-tetramethyl-3,5-

dioxopentanoate

C17H22O5 0.060 1,577.57 0.607 912

62 Octacosanol C28H58O 0.035 1,733.63 2.780 901

63 (3aS,6S,6aS)-6,8,8-Trimethyl-3,3a,4,5,6,6a,7,9-

octahydroazuleno[4,5-c]furan-1-one

C15H22O2 0.012 1,760.65 1.789 904

64 Friedelan-3-one C30H50O 0.180 1,790.66 2.671 907

Table 4—Two-dimensional gas chromatography-time of flight mass spectroscopy (GC×GC TOFMS) list of 

compounds specifically identified in fetterbush extracts (fb-c2 BB method) >900 similarity

Number Chemical identified Formula

Peak area

(%)

Retention time

(s) Similarity

1 4,5-Bis(3’,4’-methylenedioxyphenyl)benzo[15]crown-5 C34H44O13 0.36 212.005 0.083 978

2 2-Propanone 1-(acetyloxy)- C5H8O3 0.27 218.007 1.199 939

3 Isopropyl alcohol C3H8O 0.73 224.01 0.383 900

4 2,5-Hexanedione C6H10O2 0.26 266.027 1.361 950

5 9-Oxa-cis(1,8),cis(2,7),trans(7,8)-tricyclo[6.3.0,0(2,7)]

undeca-3,5,10-triene

C14H14O 0.58 299.041 1.170 922

6 1,2,3-Propanetriol C3H8O3 0.06 314.047 1.366 905

7 2,3-Dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one C6H8O4 0.09 452.104 1.511 937

8 Undecane 2-methyl- C12H26 0.07 467.111 0.822 938

9 Cyclobutanone C4H6O 1.18 473.113 2.427 949

10 Undecane 2,6-dimethyl- C13H28 0.26 509.128 0.823 907

11 Benzaldehyde 3-hydroxy-4-methoxy- C8H8O3 0.13 665.193 1.860 931

12 Mequinol C7H8O2 0.09 758.231 0.703 900

13 Scyllo-inositol C6H12O6 2.47 818.256 2.117 937

14 Pentadecane 2,6,10-trimethyl- C18H38 0.04 842.266 0.859 902

15 Neophytadiene C20H38 0.37 965.317 0.939 931

16 1,2,4,5-Tetrakis(4-phenylphenyl)benzene C54H38 0.49 1,889.7 0.382 992
Characterization of Fetterbush Lyonia lucida Liquid Extractions

5

Table 5—Two-dimensional gas chromatography-time of flight mass spectrometry (GC×GC-TOFMS) list of 

shared compounds identified in fetterbush extracts (fb-mle SE method) >900 similarity

Number Shared chemicals identified Formula

Peak area 

(%)

Retention time 

(s) Similarity

1 Benzene (1-methylethyl)- C9H12 0.517 266.027 0.980 907

2 Glycerin C3H8O3 0.273 296.04 1.459 933

3 2,4-Dihydroxy-2,5-dimethyl-3(2H)-furan-3-one C6H8O4 0.958 308.045 1.128 918

4 Phenol C6H6O 2.480 308.045 1.252 931

5 (Octahydro-7a-methylbenzofuran-6-yl) Formate C10H14O3 0.306 350.062 0.572 909

6 Limonene C10H16 0.575 353.063 0.963 915

7 Benzenemethanol C7H8O 1.474 356.065 1.431 939

8 Nonanal C9H18O 0.062 416.089 1.064 916

9 (Pyro)-catechol C6H6O2 12.630 497.123 1.481 924

10 Benzofuran 2,3-dihydro- C8H8O 2.904 515.13 1.471 912

11 5-Hydroxymethylfurfural C6H6O3 7.652 521.133 1.895 924

12 1,2-Benzenediol 4-methyl- C7H8O2 1.000 572.154 1.515 901

13 2-Methoxy-4-vinylphenol C9H10O2 0.629 599.165 1.499 924

14 2,6-Octadienoic acid 3,7-dimethyl- (E)- C10H16O2 4.962 623.175 1.220 912

15 Phenol 2-methoxy-3-(2-propenyl)- or guaiacol 6-allyl- C10H12O2 2.789 632.179 1.446 910

16 Benzaldehyde 4-hydroxy- C7H6O2 0.065 632.179 1.760 908

17 1,4-Benzenediol 2-methoxy- C7H8O3 0.283 662.191 1.812 900

18 Hydroxychavicol C9H10O2 1.655 719.215 1.573 907

19 β-D-Glucopyranose 1,6-anhydro- C6H10O5 22.217 725.217 2.245 927

20 β-D-Glucopyranoside methyl C7H14O6 5.290 791.245 1.916 944

21 Benzenepropanol 4-hydroxy-3-methoxy- C10H14O3 0.788 848.268 1.763 912

22 3,7,11,15-Tetramethyl-2-hexadecen-1-ol C20H40O 0.070 992.328 0.952 905

23 n-Hexadecanoic acid C16H32O2 0.162 1,037.35 1.110 923

24 Eicosane C20H42 0.046 1,115.38 0.926 916

25 Octadecanoic acid C18H36O2 0.035 1,148.39 1.141 903

26 (20R,22R,24S)-20,22,24,25-Tetrahydroxy-6β-methoxy3α,5-cyclo-5-α-cholestaneC28H48O5 0.214 1,151.39 0.492 905

27 Triacontane C30H62 0.124 1,220.42 0.954 917

28 (1,2-Dihydroxyethyl)oxirane C4H8O3 0.069 1,322.46 0.504 967

29 Arbutin C12H16O7 0.562 1,337.47 0.104 904

30 Heptacosane C27H56 0.154 1,448.52 1.107 920

31 Squalene C30H50 0.104 1,463.52 1.393 910

32 Vitamin E C29H50O2 0.309 1,589.58 2.220 910

33 Hexadecanal C16H32O 0.053 1,622.59 1.830 900

34 1-Docosene C22H44 0.042 1,685.62 2.337 907

35 7-Hydroxy-7-phenyl-3,9-diisopropyl-2,10-

dioxadispiro[3.3.3.1]dodecan-1,11-dione

C22H28O5 0.599 1,781.65 0.140 968

36 Furan 2,3-dihydro- C4H6O 0.709 1,898.7 0.084 999
Research Note FPL–RN–0385

6

Table 6—Two-dimensional gas chromatography-time of flight mass spectroscopy (GC×GC TOFMS) 

list of shared compounds identified of fetterbush extracts (fb-c2, BB method) >900 similarity

Number Shared chemical identified Formula

Peak area 

(%)

Retention time 

(s) Similarity

1 (20R,22R,24S)-20,22,24,25-Tetrahydroxy-6βmethoxy-3α,5-cyclo-5-α-cholestaneC28H48O5 19.380 203.001 0.994 957

2 7H-Isobenzofuro[4,5-b][1,4]benzodioxepin-11-

carboxaldehyde, 1-(acetyloxy)-5-[(acetyloxy)

methyl]-1,3-dihydro-4,10-dihydroxy-8-

C22H16O12 0.181 227.011 0.098 973

3 2-Propanone C3H6O 1.089 260.025 0.251 975

4 Benzene, (1-methylethyl)- C9H12 0.425 266.027 0.986 931

5 2-Propenal, 3-(2-furanyl)- C7H6O2 1.361 269.029 0.580 956

6 Benzenemethanol C7H8O 0.494 356.065 1.429 926

7 Nonanal C9H18O 0.073 416.089 1.062 943

8 (Pyro)-catechol C6H6O2 2.461 494.122 1.501 925

9 (Octahydro-7a-methylbenzofuran-6-yl) Formate C10H14O3 3.941 548.144 0.541 915

10 2-Methoxy-4-vinylphenol C9H10O2 1.216 596.164 1.523 925

11 2,6-Octadienoic acid, 3,7-dimethyl-, (E)- C10H16O2 6.048 623.175 1.211 937

12 Benzaldehyde, 4-hydroxy- C7H6O2 0.050 629.178 1.787 927

13 Tetradecane C14H30 0.447 671.195 0.846 900

14 Phenol, 2-methoxy-4-(1-propenyl)- or eugenol C10H12O2 0.328 704.209 1.525 919

15 Hydroxychavicol C9H10O2 1.494 719.215 1.577 910

16 β-D-Glucopyranose, 1,6-anhydro- C6H10O5 23.794 719.215 2.263 931

17 β-D-Glucopyranoside, methyl C7H14O6 0.408 779.24 1.983 924

18 Benzenepropanol, 4-hydroxy-3-methoxy- C10H14O3 0.710 848.268 1.773 909

19 p-Coumaric acid C9H8O3 0.168 926.301 2.016 911

20 3,7,11,15-Tetramethyl-2-hexadecen-1-ol C20H40O 0.126 992.328 0.954 915

21 n-Hexadecanoic acid C16H32O2 0.233 1,037.35 1.114 923

22 2-Hexadecen-1-ol, 3,7,11,15-tetramethyl-, 

[R-[R*,R*-(E)]]-

C20H40O 0.433 1,124.38 1.094 934

23 9,12,15-Octadecatrienoic acid, (Z,Z,Z)- C18H30O2 0.206 1,139.39 1.295 900

24 1-Docosene C22H44 0.207 1,313.46 1.178 933

25 2,6,10,14,18,22-Tetracosahexaene, 

2,6,10,15,19,23-hexamethylC30H50 0.743 1,463.52 1.393 933

26 1-Cyclohexene-1-carboxaldehyde, 

2-(3,3-dimethyl-1-butenyl)-, (E)-

C13H20O 0.138 1,517.55 0.618 933

27 γ-Tocopherol C28H48O2 0.529 1,559.56 2.056 901

28 β-Sitosterol C29H50O 0.553 1,673.61 0.107 927

29 7-Hydroxy-7-phenyl-3,9-diisopropyl-2,10-

dioxadispiro[3.3.3.1]dodecan-1,11-dione

C22H28O5 0.665 1,694.62 0.349 970
Characterization of Fetterbush Lyonia lucida Liquid Extractions

7

Figure 1. Two-dimensional gas chromatography (GC×GC) chromatogram surface plot for fetterbush extracts 

(fb-mle, SE method).

Figure 2. Two-dimensional gas chromatography (GC×GC) chromatogram contour plot for fetterbush extracts 

(fb-mle, SE method).

First dimension (s)

Second dimension (s)

First dimension (s)

Second dimension (s)
Research Note FPL–RN–0385

8

A GC×GC chromatogram is known for displaying different 

regions for various classes of chemicals found in complex 

mixtures. This is called molecular chemical mapping 

(MCM). The formatting notice in these chromatograms is 

similar to the periodic table. The periodic table is grouped 

according to chemical families and physical properties. 

GC×GC chromatograms can be used to create a fingerprint 

for various complex mixtures. This is one of the best ways 

to visually and chemically identify unknown chemicals in 

complex mixtures.

On the bottom left side of the chromatograms were long 

chain alcohols, aldehydes, and alkenes. In this region, there 

were two unknown peaks at 1018 and 1046. Because of the 

position of the chemicals and the spectrum of the unknown, 

we used the known chemicals around the unknown chemical 

with its spectrum to establish the chemical name. Each 

unknown has a spectrum that can be used to determine the 

structure.

The chemical composition of fb-mle extracts contained 

several sugars, and they were extremely concentrated. In the 

middle top of the chromatogram, there were several sugars 

visually detected. There is also an unknown peak (470)

detected in this region. Visual assessment of the position of 

the peak on the chromatogram could assist with identifying 

the chemical name from the mass spectrum.

Identifying regions of classes of chemical (RCC) on the 

chromatograms is important when characterizing the 

chemicals found in complex mixtures. Because of the 

resolving power and the visual view of MCM on a GC×GC 

chromatogram, we can identify chemicals in extracts. This 

technique will help with future characterization of chemical 

contributors to wildfires in our forests.

Conclusion

The GC×GC detected 1,899 chemicals from the first 

extraction using the SE method (fb-mle) and 912 from the 

first extraction using the BB method (fb-c2). These results 

could be based on the polarities of the different solvents 

used in the extractions. However, there were 72 shared 

chemicals identified in both extractions. It can be assumed 

but not concluded that there are some similarities in the 

polarity of both solvent systems.

We believe the concentration played a role in the number 

of chemicals detected in the fb-c2 extract. There were 

several unknown chemicals detected. The spectra of these 

chemicals will be important in characterizing them. There 

is not much literature on the chemical components of 

fetterbush. Shikimic acid was identified in fetterbush. This 

chemical is typically found in sweetgum trees. Fetterbush 

could be another source for isolating chemicals with 

antiviral properties. Further characterization of the second 

extractions from each method will be investigated. Which 

chemicals or chemical groups could be the contributing 

igniters in wildfires is still under review. GC×GC TOFMS 

is a great analytical technique to further the investigation 

on identifying these chemicals. This study is also the first 

to identify chemicals in fetterbush leaves using GC×GC 

TOFMS.

Acknowledgments

This research was funded by the DOD/EPA/DOE Strategic 

Environmental Research and Development Program Project 

RC-2640, funded through contract number 16-JV-11272167-

024, which was administered by the USDA Forest Service, 

Pacific Southwest Research Station. The USDA Forest 

Service, Forest Product Laboratory, and the University of 

Wisconsin-Madison provided research support. Zinan Yu 

assisted with GC×GC sorting and data collection.

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