Organic Geochemistry 76 (2014) 194–203

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Organic Geochemistry
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Hopanoid hydrocarbons and perylene in Lake Biwa (Japan) sediments:

Environmental control on their abundance and molecular composition
Ryoshi Ishiwatari a,b,⇑, Hitoshi Uemura b,c, Shuichi Yamamoto d
a
Geotec Inc., Takaido-nishi 3-16-11, Suginami-ku, Tokyo 168-0071, Japan
b
Graduate School of Science, Tokyo Metropolitan University, Hachioji, Tokyo 192-03, Japan
c
Chemical Division, Kanagawa Prefectural Institute of Public Health, Shimomachiya 1-3-1, Chigasaki, Kanagawa 253-0087, Japan
d
Faculty of Engineering, Soka University, Hachioji 192-8577, Japan

a r t i c l e i n f o a b s t r a c t

Article history: We examined the vertical distribution of hopanoid hydrocarbons and perylene in sedimentary sections
Received 13 March 2014 from Lake Biwa (Japan) and discuss the results with respect to paleoenvironmental information from
Received in revised form 6 June 2014 around the lake region over the last 120 kyr.
Accepted 11 August 2014
Moretene concentration was high above the sections where fall-out events of tephra (tephra AT)
Available online 23 August 2014
occurred ca. 30 kyr before present (BP). This feature suggests that the tephra fall-out events created O2
deficient conditions, resulting in high activity of methane producing bacteria, and of methanotrophic
Keywords:
moretene-producing bacteria becoming active above the tephra layer. Five sections at 10–20 kyr, ca.
Lake Biwa
Lake sediment
35 kyr, ca. 70 kyr, ca. 90 kyr and ca.110 kyr BP exhibited peaks in the ratio of 17a, 21b-22R-homohopane
Hopanoid hydrocarbons to 17b, 21b-22R-homohopane (hopC31ab/bb). The n-C23/n-C31 alkane ratio in these sections was 0.6 ± 0.1
Moretene (n = 49), very low if compared with typical characteristics of n-alkanes for a Sphagnum peat environment
n-Alkanes (n-C23/n-C31 ca. 3.6; Bingham et al., 2010). We conclude that Sphagnum peat may not have been an impor-
Perylene tant contributor of n-alkanes over the past 120 kyr. Comparison with previous pollen records for the lake
Pollen record region suggests that high hopC31ab/bb values may be caused by production of acidic soil from expansion
of temperate conifer (e.g. Cryptomeria) forest under cold and wet climate conditions. The abundance of
perylene relative to sediment or total organic carbon (TOC) was high in the sections at ca. 20 kyr, ca.
70 kyr and ca. 90 kyr BP. These sections were approximately in accord with those having a maximum
in hopC31ab/bb. This concordance indicates that the climatic conditions suitable for the production of
perylene quinone in soil surrounding the lake coincided with those for a high hopC31ab/bb ratio. Thus,
hopC31ab/bb and perylene/TOC might be useful as proxies of soil acidity, temperature and humidity.
Ó 2014 Elsevier Ltd. All rights reserved.

1. Introduction of associating environmental information with past living organ-

A major target of organic geochemistry is to explore the use of
marker compounds in studies of sedimentary records of environ-
mental change, such as fluctuation in temperature and other cli-
matic conditions. Alkenones and glycerol dialkyl glycerol
tetraether (GDGT) lipids can be quoted as examples of compounds
being successfully developed for such purposes (Schouten et al.,
2013). It is becoming increasingly apparent that a wealth of infor-
mation on paleoenvironmental change is contained in the composi-
tion, concentration and isotopic characteristics of biomarkers in the
sedimentary record. Indeed, many compounds have the possibility
⇑ Tel.: +81 3 5930 7634; fax: +81 3 5930 2329.
E-mail address: ry.ishiwatari@jcom.home.ne.jp (R. Ishiwatari).
isms (Schouten et al., 2013).
Hopanoids (biohopanoids) are produced as membrane lipids by
bacteria of diverse taxonomic groups (Rohmer et al., 1980; Innes
et al., 1998). The occurrence of hopanoids, including 17b, 21b-
22R-hopanes (bb-hopanes), in recent sediments has been reported
frequently (e.g. Rohmer et al., 1980; Simoneit, 1986; Cranwell and
Koul, 1989). Hexafunctionalised biohopanoids were most abun-
dant in lake sediments (Farrimond et al., 2000). Biohopanoids in
lake and marine sediments and the soil environment undergo dia-
genesis, leading to geohopanoids, including hydrocarbons, acids,
hopanols, aldehydes and ketones (Innes et al., 1998).
Abundances of hopanoid hydrocarbons in recent sediments
may reflect the intensity of bacterial activity, combined with favor-
able conditions for organic matter (OM) preservation. A lake sedi-
ment example (Priest Pot, English Lake district 1997) showed that

http://dx.doi.org/10.1016/j.orggeochem.2014.08.003
0146-6380/Ó 2014 Elsevier Ltd. All rights reserved.
 R. Ishiwatari et al. / Organic Geochemistry 76 (2014) 194–203
hopanoic acids and hopanols were abundant (25–48% of total
geohopanoids for the former and 11–27% for the latter). Hop-
17(21)-ene was also abundant (22–44% of total geohopanoids),
while hopanoid hydrocarbons were minor (Innes et al., 1997).
The molecular composition and abundances of geohopanoids in
sediments have the potential to be used as a proxy for past environ-
ment related to bacterial activity. However, there are few examples
of their evaluation and application as paleoclimate proxies. During a
survey of the distribution of hopanoid hydrocarbons in lake
sediments, we recognized that their molecular compositions (e.g.
ab-hopanes, moretene) differed among lakes and with sediment
depth (Uemura and Ishiwatari, 1992, 1995). The ab type of hopanoid
hydrocarbons, frequently seen in subsurface sediments, is generally
oil pollution derived because hopanoids with this configuration are
transformation products formed under thermal stress from biohop-
anoids with the bb configuration (Peters and Moldowan, 1993).
However, hopanoids with the ab configuration have also been found
in acidic peat bogs and forest soil and it has been demonstrated that
ab-hopanes can be formed via acid catalyzed reactions (Quirk et al.,
1984; Crossman et al., 2001; Pancost et al., 2003).
Perylene is also frequently found in marine and freshwater sed-
iments. Its distribution pattern in sediments is different from that
of other combustion-derived polynuclear aromatic hydrocarbons
(PAHs). It is therefore thought to originate by way of natural
in situ formation in sediments. A variety of precursors has been
proposed, such as perylenequinone from terrigenous (Orr and
Grady, 1967; Aizenshtat, 1973; Laflamme and Hites, 1978;
Ishiwatari et al., 1980) and aquatic sources (Wakeham et al.,
1979). The hypothesis that its formation is controlled by in situ
microbial processing of non-specific OM was proposed later by
Silliman et al. (1998, 2000, 2001). Its origin therefore remains
unsolved. We previously reported the abundance and distribution
of perylene in various lake sediments (Lake Biwa, Ishiwatari and
Hanya, 1975; Lake Haruna, Ishiwatari et al., 1980). Based on the
hypothesis that it is generated from perylene quinone pigments
via in situ reduction in lake sediments, we developed a method
for the quantification of perylene quinone in soils and lake sedi-
ments and applied it to several lake sediments. We found up to
10 x the amount of perylene precursor in some lake sediment sam-
ples, including from Lake Biwa (Japan, Ishiwatari and Matsushita,
1986), leading us to think that perylene is generated from perylene
quinone pigments via in situ reduction.
Here we report the findings of the ratio of ab C31 hopane to bb
C31 hopane and perylene abundance to TOC as possible indicators
of acidity of catchment soil, which is related to temperature and
humidity. We determined the abundance of hopanoid hydrocar-
bons and perylene in two sediment cores from Lake Biwa and
examine the factors controlling their temporal variation, such as
molecular characteristics, vegetation history from pollen analysis
and regional paleoclimatic change over the past 120 ky.
2. Study site and previous studies
Lake Biwa is the largest freshwater lake in Japan and is in the
central part of the mainland (Fig. 1). It is situated in a temperate
zone and receives relatively large precipitation with moderate tem-
perature. In both 1971 and 1982–1983, long (200 m and 1400 m)
sediment cores were obtained from almost the same location. The
lake lies at a surface elevation of 84.4 m, and is 63.49 km long
and 22.8 km wide (max. width), covers an area of 674 km2 and
has a maximum water depth of 104 m and average water depth
of 48 m. The watershed area is 3850 km2 (Kuwae, 2004). Currently,
the lake receives a mean annual precipitation of 1570 mm and the
mean annual temperature is 14.3 °C (10 yr average for 1991–2000).
The trophic status is oligotrophic-mesotrophic (Otsuki et al., 1981).
195
Studies of the sediments have revealed diagenetic changes in
various biomarkers such as fatty acids (e.g. Kawamura and
Ishiwatari, 1984), lignin compounds (Ishiwatari and Uzaki, 1987),
lipids and pheopigments (Ogura et al., 1990), sterols (Ogura
et al., 1990), carbohydrates (Uzaki et al., 1991) and humic sub-
stances (Ishiwatari, 1977; Ishiwatari and Kawamura, 1981;
Ishiwatari et al., 1981). These studies showed various aspects of
environmental change responding to global climate change, such
as glacial/interglacial changes in vegetation from fossil pollen anal-
ysis, correlation between lake productivity and climate from fossil
diatom analysis, and millennial-scale oscillation of eolian dust
since the last glacial from quartz analysis (Ishiwatari et al., 2009
and references therein). Recently, Ito and coworkers reported the
abundance and possible source of perylene in a mixed subsurface
sediment, as well as surface sediments from the lake (Itoh and
Hanari, 2010; Itoh et al., 2010, 2012).
3. Experimental
3.1. Sampling site and age model
Two cores (BIW95-5, 15 m long, and the 250 m unit of a 1400 m
core) were used. BIW95-5 was collected near the center
(35°15.000 N, 136°03.00’E; water depth 67 m at ca. 4 km off the lake
shore) in 1995. An additional short gravity core (BIW95-A, 83 cm in
length) was collected at the same site. BIW95-5 is composed of
homogeneous gray clay with 7 volcanic ash layers (Ishiwatari
et al., 2009). It is known that a significant volcanic eruption
occurred at Kyushu island, off Japan mainland (ca. 600 km west
of Lake Biwa), at 29 kyr BP. The thickness of tephra AT in the lake
sediment is ca. 14 cm. In addition, other tephra (Daisen tephra,
Sakate, U-Oki, KAh and Kg) are present (Table 1).
Drilling of the 1400 m core was conducted in the 68 m deep
southern basin of the lake (35°130 0600 N, 136°000 4900 E) during
1982–1983 (Takemura and Yokoyama, 1989; Takemura, 1990).
The upper 250 m unit (T-bed) of the core is essentially composed
of lacustrine deep water bluish gray, non-laminated clays, indicat-
ing that the depositional conditions were under deep water, simi-
lar to the present conditions. The chronostratigraphic scale of the
cores was revised after re-examination of marker tephra layers
(Machida et al., 1991) and various parameters obtained from sed-
imentary sections (Meyers et al., 1993). Ages of the sections were
determined essentially from comparison between the tephra
records in BIW 95-5 and the 1400 m cores and those of well-estab-
lished tephra chronology record for the Takashima-Oki sediment
core (Takemura et al., 2000; Machida et al., 1991; Nagahashi
et al., 2004). Table 1 shows the age control points. The age of each
sampling depth was obtained from linear interpolation between
the control points, assuming uniform sedimentation rate.
3.2. Sample preparation and analytical procedures
The BIW95-5 core was divided into 2.4 cm sections and sub-
samples from each were stored at 20 °C in glass bottles. Samples
from the 1400 m core were essentially kept frozen from the time of
collection until analysis. These samples were freeze-dried and
powdered before analysis.
For the samples from the 1400 m core, hydrocarbons were ana-
lyzed according to the procedure reported by Ishiwatari et al.
(2005). Briefly, a powdered sample was saponified with 0.5 M
KOH in MeOH under reflux and extracted with n-hexane/Et2O
(9:1) for neutral compounds. The neutral compounds were frac-
tionated into aliphatic and aromatic hydrocarbons via silica gel
chromatography. For BIW 95-5, each powdered sample (ca. 3 g)
was placed in a 50 ml Teflon tube and extracted with 30 ml CH2Cl2/
MeOH (3:1) via sonication in a Nihon Seiki apparatus. After ele-
196 R. Ishiwatari et al. / Organic Geochemistry 76 (2014) 194–203

Fig. 1. Location of coring site.

Table 1
Age control points used.

BIW95-5 1400 m core Tephra name and number Age Ref.b

Depth (m) Depth (m) kyr BP
2.11 – Kawagodaira (Kg) 3.1 (1)
3.32 10.0 Kikai-Akahoya (K-Ah) (BB15a) 7.3 (1)
4.18 13.4 Ulreung-Oki (U-Oki) (BB23) 10.7 (1)
6.78 19.2 Sakate (BB37) 17.2 (1)
10.55 – Daisen-Higashi Daisen (DHg) (2)
11.05 24.9 Daisen Sasaganaru (DSs) (BB51) 27.1 (1,2,3)
11.22–11.34 26.0 Aira-Tn (AT) (BB55) 29.0 (1)
– 37.1 BB85 49.0 (1)
– 62.3 BB153 83.5 (1)
– 72.5 Kikai-Tozuhara (K-Tz) (BB173) 91.0 (1)
– 74.0 Ata (BB179) 98.6 (1)
– 88.8 BB207 122.0 (1)
– 131.5 BB323 212.4 (1)
a
Tephra number for 200 m core from Lake Biwa.
b
(1) Nagahashi et al. (2004), (2) Takemura et al. (2000), (3) Machida et al. (1991).

mental S was removed with Cu grains, the extract was applied to a Hopanoid hydrocarbons and perylene were analyzed using GC
silica gel column, eluted first with hexane and then with hexane/ with a HP 5890 series II instrument equipped with flame ionization
benzene (3:1). The second fraction was used for gas chromatogra- detection (FID), splitless injector or on-column injector and
phy (GC) and GC–mass spectrometry (GC–MS) analysis. electric pressure control system. A DB-5 fused silica column
 R. Ishiwatari et al. / Organic Geochemistry 76 (2014) 194–203
(30 m  0.32 mm i.d.; 0.25 lm film thickness) was used with He as
carrier gas. The oven temperature was programmed from 50 °C to
120 °C at 30 °C/min and then to 310 °C at 6 °C/min. Injector and
detector were at 300 °C and 315 °C, respectively. Quantitation was
achieved by integration of GC peak area of individual compounds.
Typically, n-C24D50 alkane was used as a reference (injection) stan-
dard for quantitation and the response factor of the individual com-
pounds relative to it was assumed to be 1.0. Analytical precision for
individual compounds was ± 5% relative standard deviation.
GC–MS was performed using a Finnigan INCOS-50 mass spec-
trometer equipped with a Varian 3400 gas chromatograph. In full
scan mode, the spectrometer was scanned at 1.5 s/decade over
m/z 60 to 650. The ion source was at 180 °C, with 70 eV ionization.
The gas chromatograph was equipped with a DB-5 fused silica col-
umn (30 m  0.32 mm i.d.; 0.25 lm film thickness).
Quantitation for C31bb hopane was achieved by comparison of
GC (FID) peak areas with those of external standards (n-C18 and
n-C28 alkanes). C31bb hopane was alternatively quantified as the
equivalent of 1-docosene or n-C24D50 alkane on a total ion current
basis (Uemura and Ishiwatari, 1992). The other hopanoid hydro-
carbons were quantified by comparing the peak area of C31bb
hopane with the peak area from each hopanoid hydrocarbon on
m/z 191 mass chromatograms, with the assumption of a response
factor of 1.0.
For quantification of moretene, the following procedure was
used since it co-eluted with hopC29bb (GC peak 5 + 6 in Fig. 2;
Uemura and Ishiwatari, 1995). The mass spectrum of GC peak
5 + 6 (Fig. 2) exhibited a combination of (i) fragments from more-
tene (signals at m/z 189, 191 and 410) and (ii) those from hopC29bb
(signals at m/z177, 191 and 398, where m/z 177 was > m/z 191).
The concentration of moretene was estimated by subtracting the
contribution of hopC29bb from peak 5 + 6. The concentration of
hopC29bb was calculated from that of hopC31bb on the assumption
that the ratio of hopC29bb to hopC31bb was 0.68, which was calcu-
lated for the 1400 m core (0.68 ± 0.14, n = 32).
4. Results and discussion
TOC in the BIW 95-5 core ranges from 5–21 mg/g dry sediment
(ds) and C/N (wt. ratio) from 6 to 9 (Table 2; Ishiwatari et al., 2009).
The 1400 m core had TOC from 8–14 mg/g ds and C/N from 6 to 8
(Table 3). These results imply that the bulk OM was essentially
autochthonous. TOC was relatively high in the Holocene [marine
isotope stage (MIS) 1] and MIS 5a, and significantly lower in other
MIS stages. The levels of TOC were essentially the same as those in
Fig. 2. Partial m/z 191 mass chromatogram of hopanoid hydrocarbons from Lake
Biwa core. The sample is no. 33-3 from the 1400 m core (55.5 m depth; 73.2 kyr BP).
Peak numbers: 1, 17b(H)-22,29,30-trisnorhopane, C27b; 2, C30D17(21); 3,
17a(H),21b(H)-hopane, C30ab; 4, C30D13(18); 5, 17b(H),21b(H)-30-norhopane,
C29bb; 6, 17b(H)-moret-22(29)-ene (moretene), C30baD22(29); 7, 17a(H),21b(H)-
22R-homohopane, C31ab; 8, 17b(H),21b(H)-hopane, C30bb; 9, 17b(H),21b(H)-hop-
22(29)-ene (diploptene), C30bbD22(29); 10, 17b(H),21b(H)-22R-homohopane, C31bb;
11, 17b(H),21b(H)-22R-bishomohopane, C32bb.
197
studies for other Lake Biwa cores (e.g. Kawamura and Ishiwatari,
1984; Iwamoto and Inouchi, 2007). The TOC concentration and C/
N were at a similar level to those (TOC 3 to 28 mg TOC/g ds and
C/N 6 to 13) for Lake Baikal Holocene sediments (Ishiwatari
et al., 2005).
4.1. Hopanoid hydrocarbons
A series of hopanoid hydrocarbons was present in the aliphatic
hydrocarbon fractions. Fig. 2 shows a typical partial m/z 191chro-
matogram from the 1400 m core. Tables 2 and 3 list the concentra-
tions of hopanoid hydrocarbons and perylene together with basic
information on depth, age and bulk properties [TOC, total nitrogen
(TN)] for the two cores.
Diploptene was detected (Fig. 3) only in the sections above ca.
4 m (ca. 10 kyr BP). Hop-17(21)-ene and neohop-13(18)-ene
occurred throughout the sections of both cores. The abundance
of hop-17(21)-ene was similar to that of neohop-13(18)-ene for
BIW95-5, whereas the abundance of the former was higher than
the latter in the 1400 m core. Hopenes comprised 5–24% of the
total hopanoid hydrocarbons (avg. 10%, n = 30) in BIW95-5, and
8–46% (avg. 24%, n = 32) in the 1400 m core. The abundance distri-
bution of moretene in the cores was quite wide (0–176 ng/g ds)
and its abundance was remarkably high in the BIW 95-5 core when
compared with that in the 1400 m core (Fig. 3). The characteristics
of the vertical profile of moretene are discussed in the next section.
The composition of hopanoid hydrocarbons could indicate that
transformation of the precursor biohopanoids to geohopanoids
took place at an early diagenetic stage. The presence of diploptene
was limited to the sections younger than ca.10 kyr BP, which might
be reasonable, because it is known to occur in living bacteria
(Rohmer et al., 1980, 1984). It tends to change to more stable hop-
enes, such as hop-17(21)-ene and neohop-13(18)-ene, probably as
a result of microbial and/or chemical isomerization (Ageta et al.,
1987; Brassell et al., 1980; Moldowan et al., 1991; Innes et al.,
1998). The relative abundance of neohop-13(18)-ene and hop-
17(21)-ene was similar in BIW 95-5, and hop-17(21)-ene was more
abundant than neohop-13(18)-ene in the 1400 m core.
C27–C32 hopanes were recognized throughout the cores. Major
ones were the C27b, C29bb, C30bb and C31bb hopanes with C32bb
hopane being a minor contributor. The ratio of C27b, C29bb, C30bb
relative to C31bb was 0.5 ± 0.1, 0.6 ± 0.1, 0.9 ± 0.1 (n = 63) respec-
tively. Hopanes with the ab configuration were also present. The
homohopanes had the 22R configuration characteristic of a biolog-
ical origin, i.e. without thermal stress (Peters and Moldowan,
1993).
4.2. Source of moretene
As described in Section 4.1, a high concentration of moretene
occurred in the BIW95-5 core. In particular, a peak in concentration
normalized to TOC was recognized in the sections at ca. 30 kyr BP
(Fig. 3), where tephra AT and other tephra were present (Table 1).
The following two scenarios giving rise to this vertical profile
could be envisaged: (i) Scenario 1, whereby tephra AT fallout pro-
duced acidic conditions which stimulated the reactions for produc-
ing moretene; or (ii) Scenario 2, whereby the tephra fallout events
may have produced O2 deficient conditions in the sections beneath
the tephra, resulting in promotion of the activity of methane pro-
ducing bacteria. Then, the produced methane may have moved
upward in the sediment. As a result, moretene derived from met-
hanotrophic bacteria would become abundant in the sections
above the tephra AT layer at ca. 30 kyr BP.
We consider the possibility of Scenario 1 to be low, because the
fallout of tephra AT would probably not produce acidic conditions
in the sedimentary sections, given that the pH of water mixed with
198 R. Ishiwatari et al. / Organic Geochemistry 76 (2014) 194–203

Table 2
Hopanoid hydrocarbons and perylene in BIW95-5 core.

Sample Depth Age MIS TOC TN C/N Moretene Total hopenesa Total hopanoidsb C31ab/bb Moretene/C31bb Perylene
(m) (ky BP) stage (mg/gds) (mg/gds) (wt. ratio) (ng/gds) (ng/gds) (ng/gds) (wt. ratio) (wt. ratio) (lg/gds)
A-10 0.11 0.16 1 16.9 2.1 8.1 90 285 821 0.38 0.66 0.07
A-50 0.51 0.75 11.9 1.5 7.7 19 36 166 0.47 0.52 0.28
A-80 0.81 1.14 16.3 2.0 8.1 63 131 496 0.47 0.61 0.26
2-1-14 1.45 2.13 16.1 2.0 8.2 81 144 417 0.34 0.98 0.47
2-2-7 2.16 3.27 20.8 2.4 8.8 132 174 797 0.37 0.80 0.34
2-2-22 2.52 4.52 19.6 2.3 8.7 37 65 317 0.38 0.54 0.34
2-3-12 3.04 6.30 19.8 2.3 8.6 127 213 738 0.36 0.84 0.29
2-3-32 3.52 8.06 14.0 1.8 7.7 39 55 224 0.38 0.82 0.38
2-4-13 4.04 10.13 13.0 1.7 7.7 139 258 869 0.69 0.82 0.47
2-4-32 4.50 11.48 16.7 2.0 8.4 33 57 378 0.71 0.39 0.53
2-5-15 5.04 12.84 2 12.2 1.6 7.6 34 34 366 1.49 0.44 0.74
2-5-35 5.52 14.04 9.3 1.1 8.7 69 84 355 2.33 1.47 0.27
3-1-10 6.02 15.29 9.6 1.4 6.7 34 52 222 1.04 0.90 0.41
3-1-31 6.52 16.55 9.5 1.4 6.9 58 87 356 1.09 0.88 0.62
3-2-10 7.02 17.81 11.3 1.6 7.0 77 109 413 1.26 1.09 0.51
3-2-31 7.52 19.12 9.7 1.4 6.8 81 117 514 0.82 0.78 0.76
3-3-13 8.01 20.38 11.5 1.7 6.6 86 145 538 0.64 0.89 1.01
3-3-34 8.51 21.69 9.1 1.5 6.3 79 126 464 0.43 0.84 0.73
3-4-16 9.02 23.00 8.7 1.3 6.5 98 154 503 0.52 1.04 0.38
3-4-36 9.50 24.25 3 10.1 1.4 7.0 74 74 284 0.71 1.50 0.37
3-5-17 10.04 25.64 9.9 1.6 6.1 48 58 218 0.56 1.11 0.32
3-5-38 10.52 26.88 8.0 1.4 5.6 176 245 632 0.54 2.11 0.31
4-1-13 11.01 28.14 4.6 0.8 6.1 11 11 25 0.50 2.67 0.01
4-1-34 11.51 29.14 7.0 1.2 5.7 102 141 285 0.46 2.40 0.01
4-2-17 12.02 30.46 9.5 1.3 7.5 64 92 313 1.03 1.27 0.34
4-2-37 12.50 31.70 10.0 1.5 6.9 21 38 177 0.55 0.57 0.47
4-3-18 13.02 33.04 11.4 1.5 7.4 57 82 428 0.52 0.62 0.35
4-4-2 13.56 34.42 11.3 1.6 7.3 51 69 398 0.75 0.61 0.50
4-4-21 14.02 35.61 11.1 1.7 6.7 37 56 379 0.54 0.41 0.52
4-5-3 14.51 36.87 11.9 1.9 6.3 38 105 687 0.40 0.23 0.51
4-5-24 15.01 38.17 9.3 1.6 5.8 32 83 486 0.37 0.28 0.64
a
Total hopenes = C30bbD22(29) + C30D17(21) + C30D13(18) + moretene[C30baD22(29)].
b
Total hopanoids = total hopenes + C27b + C29bb + C30bb + C31bb + C30ab + C31ab.

volcanic ash similar to tephra AT (Sakura-jima volcano, Kyushu
island) did not produce strongly acidic conditions (pH 4.5–5.1:
Suenaga and Kusamizu, 1986). In addition, as reported previously
(Uemura and Ishiwatari, 1995), a small amount of authentic stan-
dard diploptene was dispersed and stayed in an aqueous solution
of pH 1 for 7 days and did not change to moretene.
We therefore consider the possibility of Scenario 2 to be strong,
although it may not be conclusive considering that no measure-
ment of d13C values for moretene in this core was carried out.
Moretene has been suspected to be derived from methanotrophic
bacteria by several authors on the basis of its carbon isotopic com-
position (Uemura and Ishiwatari, 1995; Aichner et al., 2009;
Kristen et al., 2010). That is, the d13C value of moretene from Lake
Suigetsu was 60.7‰ (Uemura and Ishiwatari, 1995) and moreten-
e from Lake Koucha (alkaline lake) gave an isotopically depleted
signal (63.4‰ to 50.1‰: Aichner et al., 2009). Moreover, high
concentrations were observed in acidtrophic lake sediments
(Uemura and Ishiwatari, 1995). The abundance ratio of moretene
to diploptene for surface sediments from acidtrophic lakes (pH
3.4–4.7 for lake water) is 6–10. Moretene has been observed in
Black Sea sediment (lacustrine sediment section; Blumenberg
et al., 2009), a hypersaline lake (Lake Tswaing, South Africa;
Kristen et al., 2010) and an alkaline lake (Lake Koucha, Tibetan Pla-
teau; Aichner et al., 2009).
4.3. Environmental factors for high hopC31ab/bb values
The presence of hopC31ab has been reported in moss and lignite
(Miocene age: van Dorsselaer et al., 1977), Sphagnum spp. moss
peat (Quirk et al., 1984) and forest soil (Crossman et al., 2001;
Quirk et al., 1984). Quirk et al. (1984) measured a vertical profile
of its abundance in moss peat (350 cm in depth and younger than
3750 yr) composed predominantly of Sphagnum spp. remains, and
recognized that it began to appear at the early stage of moss decay.
The presence of a hopC31ab hopanoid in modern lake sediments
was confirmed by later studies (Innes et al., 1997). Pancost et al.
(2003) reported that the hopC31bb(ab + bb) ratio in a Bargerveen
peat (Sphagnum) core (ca. 2800 yr ago) was in the range 0.04–
0.10, which is equivalent to hopC31ab/bb of 9–24. High C31ab/bb
values were also reported for sediments from peat environments
(Ries-Kautt and Albrecht, 1989; Dehmer, 1995). These studies indi-
cate clearly that high hopC31ab/bb values are associated with the
presence of Sphagnum moss peat, forest soil or lignite. In addition,
in laboratory decay experiments with Sphagnum cuspidatum under
aerobic and anaerobic conditions, Quirk et al. (1984) observed an
increasing trend in hopC31ab/bb values and claimed that hopC31ab
was of both microbial and chemical origin. Innes et al. (1997) men-
tioned a possibility of an early diagenetic origin for ab hopanoids
involving acid catalyzed isomerization of a 17b(H) precursor, via
a D17(21) intermediate. This acid catalyzed isomerization seems
to be supported by the work of Pancost et al. (2006), where the
change from 17b,21b to 17a,21b hopanoids (C31 and C32 hopanoic
acids) correlated with pH, that is, hopC32ab/bb increased from ca.
0.1 to ca. 0.7 with decreasing pH (from ca. 9 to ca. 2).
For the Lake Biwa cores, hopC31ab/bb values ranged from 0.3 to
3.1, showing variation with depth (Fig. 4E). The values are signifi-
cantly high, although they are considerably lower than those
reported for Sphagnum peat sections (9–24; Pancost et al., 2003).
It therefore appears that the possibility of the presence of a typical
peat environment around Lake Biwa is low. The Biwa hopC31ab/bb
 R. Ishiwatari et al. / Organic Geochemistry 76 (2014) 194–203 199

Table 3
Hopanoid hydrocarbons and perylene in 1400 m core (nd, not determined).

Sample Depth Age MIS TOC TN C/N Moretene Total hopenesa Total hopanoidsb C31ab/bb Moretene/C31bb Perylene
(m) (ky BP) stage (mg/g ds) (mg/g ds) (wt. ratio) (ng/g ds) (ng/g ds) (ng/g ds) (wt ratio) (wt ratio) (lg/gds)
a7-3 16.0 13.6 2 12.5 1.8 6.9 <2 8 54 1.55 < 0.1 1.63
a15-2 28.0 32.4 3 9.7 1.3 7.2 <2 7 32 1.04 < 0.1 0.96
a15-3 28.5 33.3 9.7 1.5 6.4 <2 27 78 1.03 < 0.1 1.08
a17-3 31.0 37.8 11.5 1.7 6.7 <2 41 127 0.52 < 0.1 1.14
a21-3 37.0 48.8 9.5 1.5 6.5 <2 27 65 0.53 < 0.1 1.07
a23-2 40.0 52.9 10.2 1.6 6.3 <2 5 37 0.49 < 0.1 1.13
a23-3 40.5 53.6 10.0 1.6 6.1 <2 7 47 0.47 < 0.1 1.13
a25-2 43.0 57.0 11.3 1.6 7.3 <2 12 55 0.50 < 0.1 0.97
a27-1 46.0 61.0 4 9.7 1.6 6.2 <2 10 43 0.88 < 0.1 1.21
a29-2 49.0 65.1 10.7 1.6 6.7 <2 18 102 1.30 < 0.1 1.12
a31-2 52.0 69.1 7.7 1.4 5.6 4 43 111 1.96 0.3 1.23
a31-3 52.5 69.8 10.1 1.4 7.0 <2 18 108 3.10 < 0.1 1.33
a33-2 55.0 73.2 11.8 1.6 7.2 6 67 309 2.62 0.2 2.14
a33-3 55.5 73.8 5a 12.2 1.7 7.3 11 23 147 1.25 0.4 1.48
a35-1 58.0 77.2 12.9 1.7 7.7 <2 14 135 0.57 < 0.1 1.06
a37-2 61.0 81.2 9.7 1.8 5.5 <2 39 101 0.37 < 0.1 1.11
a39-3 64.0 84.3 11.0 1.9 5.8 0 18 74 0.25 < 0.1 1.18
a41-3 66.5 86.3 5b 10.8 1.8 6.1 <2 28 97 0.42 < 0.1 1.40
a43-2 69.2 88.4 10.1 1.7 5.9 <2 46 105 0.50 < 0.1 1.22
a45-3 70.0 89.1 12.9 1.8 7.1 <2 20 122 0.68 < 0.1 1.53
a47-3 71.7 90.4 11.7 2.0 6.0 <2 16 130 0.44 < 0.1 1.41
a49-3 73.0 93.5 5c 12.7 1.9 6.7 <2 67 188 0.46 < 0.1 1.41
a51-3 73.5 96.1 11.7 2.0 5.8 <2 117 248 0.40 < 0.1 1.06
a55-3 75.3 100.7 11.8 1.9 6.2 <2 26 174 0.29 < 0.1 0.79
a57-3 76.0 101.8 11.7 1.9 6.2 <2 12 89 0.25 < 0.1 0.93
a59-3 77.4 104.0 10.2 1.7 5.9 <2 38 104 0.34 < 0.1 1.29
a61-3 79.3 107.0 5d 12.3 1.8 6.7 <2 56 124 0.36 < 0.1 1.07
a63-3 81.0 109.7 11.2 1.9 5.9 4 52 193 1.30 < 0.1 nd
a65-3 82.1 111.2 12.0 1.9 6.2 4 82 251 1.30 < 0.1 nd
a67-3 83.0 112.8 13.7 1.8 7.7 9 83 206 1.04 0.3 nd
a69-3 84.7 115.5 13.9 2.0 7.1 3 21 137 0.55 < 0.1 nd
a71-3 87.0 119.2 5e 13.7 1.9 7.1 10 120 303 0.62 0.2 nd
a
Total hopenes = C30bbD22(29) + C30D17(21) + C30D13(18) + moretene[C30baD22(29)].
b
Total hopanoids = total hopenes + C27b + C29bb + C30bb + C31bb + C30ab + C31ab.

values exhibited 5 maxima, including both small and large ones,
over the last 120 kyr BP at: ca.110 kyr BP (medium), ca. 90 kyr BP
(small), ca. 70 kyr BP (large), 40–30 kyr BP (small) and 20–10 kyr
BP (large).
4.3.1. Possibility of Sphagnum peat environment as inferred from long
chain n-alkane composition
According to previous studies, a large number of Sphagnum
moss species show an n-alkane predominance at C23 or C25 (Baas
et al., 2000; Nott et al., 2000; Pancost et al., 2002; 2003; Nichols
et al., 2006; Bingham et al., 2010). Bingham et al. (2010) reported
n-C23/n-C31 values for Sphagnum spp. from ombrotrophic bogs to
be 3.6 on average (0.2–28.7, n = 41) and n-C23/n-C25 values 1.6 on
average (0.4–4.5 n = 41). These studies indicate that the n-C23/n-
C31 and n-C25/n-C31 ratios may be used as a clue to the occurrence
of a Sphagnum moss environment in the past.
Fig. 5 shows profiles of the ratios of n-C29, n-C27, n-C25 and n-C23
relative to n-C31 alkanes. The most abundant alkanes were n-C27-,
n-C29- and n-C31, indicating a high contribution from terrigenous
higher plants. The ratios of n-C23 and n-C25 relative to n-C31 in
the sections examined were ca. 0.6, i.e. 0.5 ± 0.1 for n-C23/n-C31
and 0.6 ± 0.1 for n-C25/n-C31 (n = 49) except for those (n = 20) at
30 – 10 kyr BP where they were 0.7 ± 0.2 for n-C23/n-C31 and
0.9 ± 0.2 for n-C25/n-C31. Both n-C23/n-C31 and n-C25/n-C31 values
for the sections at ca. 100 kyr BP, ca. 90 ky BP and ca. 70 kyr BP
were 0.5 ± 0.1 and 0.7 ± 0.2 (n = 5), 0.4 ± 0.0 and 0.6 ± 0.1 (n = 5),
and 0.5 ± 0.0 and 0.7 ± 0.1 (n = 5), respectively. These values are
considerably lower than those reported for Sphagnum spp. cited
above. Consequently, we conclude that Sphagnum peat may not
have been an important contributor of n-alkanes over the past
120 kyr. Although there is a possibility of occurrence of a Sphag-
num peat environment for 30–10 kyr BP, we should raise another
possibility – larger contributions of n-C23 and n-C25 alkanes rela-
tive to n-C29 and n-C31 alkanes from submerged and floating-
leaved plants (Ficken et al., 2000). Further study is needed on this
matter.
4.3.2. Past coniferous forest and possible acidic soil environment
According to Hayashi et al. (2010a,b), Cryptomeria japonica
(temperate conifer: Japanese cedar), Sciadopytys verticillata and
Cupressaceae trees were the dominant vegetation during MIS 5.
In particular, during the second half of MIS 5e–5d (120–110 kyr
BP), MIS 5c–5b (ca. 100 kyr BP), and MIS 5a-4 (ca. 75 kyr BP), C.
japonica was dominant (Fig. 6). For the Lake Biwa cores, maxima
in the abundance of fossil Cryptomeria occurred at 120–110 kyr
BP (MIS 5e–5d), ca. 100 kyr BP (MIS 5c–5b), 80–60 kyr BP (MIS
5a–4), 55–45 kyr BP and 40–30 kyr BP (MIS 3). A similar temporal
profile of fossil Cryptomeria pollen abundance was reported for a
sediment core from the northwest Pacific off central Japan
(IMAGES core MD01 – 2421: Igarashi and Oba, 2006), which may
represent the vegetation history of central Japan.
According to Igarashi and Oba (2006), a high abundance of Cryp-
tomeria fossil pollen indicated that the summer monsoon was
stronger than at present, and the East Asian summer monsoon
was relatively strong during 121–117 kyr BP, 115–89 kyr BP, 80–
70 kyr BP and 4–1 kyr BP. Annual precipitation during these peri-
200 R. Ishiwatari et al. / Organic Geochemistry 76 (2014) 194–203
ods may have exceeded 2000 mm. Igarashi and Oba (2006)
observed that Cryptomeria pollen began to increase with decreas-
ing atmospheric temperature, which was estimated from the pol-
len temperature index (Tp). Here, Tp is defined as [100  Tw/
(Tw + Tc)], where Tc = sum of subalpine taxa and Tw = warm-tem-
perate taxa (Igarashi and Oba, 2006).
It is known that coniferous trees tend to retain their leaves all
year round; in addition, the mineralization of fallen leaves pro-
ceeds slowly and creates a more soil type deposit characterized
by accumulation of raw humus, thereby producing a low base sta-
tus and low pH (Millar, 1974; Miyazawa, 1964). Therefore, this
process could raise the hopC31ab/bb ratio of hopanoid hydrocar-
Fig. 3. Temporal profile of hopenes in Lake Biwa cores.
μ μ
Fig. 4. Temporal profiles of (A) TOC, (B) total hopanoids, (C) perylene, (D) moretene/C31bb and (E) C31ab/bb ratios in Lake Biwa cores.
bons in the catchment area of Lake Biwa in the past. When the
temporal variation in hopC31ab/bb is compared with the relative
abundance (% of total; Hayashi et al., 2010a) of fossil Cryptomeria
pollen, the periods at ca.110 kyr BP, ca. 90 kyr BP and 75–65 kyr
BP, showing maxima in hopC31ab/bb, seem to coincide with the
peaks in the relative abundance of the fossil Cryptomeria pollen
(Figs. 4 and 6). In both the ca. 50 kyr BP and < 20 kyr BP sections,
no positive correlation was apparent between the maxima in Cryp-
tomeria abundance and those in hopC31ab/bb. The maxima in
hopC31ab/bb in the ca. 35 kyr BP and 20–10 kyr BP sections may
be explained by increased acidity of the catchment soil, rather than
by a Sphagnum peat environment.
Consequently, we consider that hopC31ab/bb tended to be high in
the catchment soil under temperate to cold transition with humid
climatic conditions. The highest hopC31ab/bb value was at 75–
65 kyr BP. This maximum may indicate that the environmental con-
ditions were highly suitable for conversion from hopC31bb to
hopC31ab, that is, the conditions were temperate and highly humid.
However, in the periods after 10 kyr BP, the relationship
between Cryptomeria abundance and hopC31ab/bb is not apparent,
as observed for 75–65 kyr BP. HopC31ab/bb was almost constant
during this period (Fig. 4E). The lack of concordance may be
explained by the idea that the rate of isomerization to C31ab in soil
was low, probably because of low soil acidity, although further
study is needed to verify this idea.
4.4. Perylene and its temporal profile in relation to paleoclimate
The abundance of perylene in the BIW 95-5 (a short core) ran-
ged from 0.3 to 1.0 lg/g ds, except for the 11.0–11.5 m section,
where extremely low values occurred because of the dominant
presence of volcanic ash (AT-tephra 29 kyr BP; Tables 1 and 2).
The concentration ranged from 0.8 to 2.1 lg/g ds in the 1400 m
core (Table 3), generally higher than that in the BIW 95-5 core.
We assume that perylene was produced by in situ reduction of
perylene quinone pigments originating from forest soil surrounding
the lake (Section 1). DHPQ (4,9-dihydroxyperylene-3,10-quinone)
from terrestrial organisms (e.g. fungi) has been suggested as an
ββ αβ ββ
 R. Ishiwatari et al. / Organic Geochemistry 76 (2014) 194–203
Fig. 5. Temporal profiles of long chain n-alkanes in Lake Biwa cores.
important precursor of perylene by many authors since the discov-
ery of perylene in sediments (e.g. Orr and Grady, 1967; Aizenshtat,
1973; Ishiwatari et al., 1980). Kumada and Hurst (1967) found soil
humic material (P-type humic acid), to contain a perylene quinone
pigment, and claimed that Cenococcum graniforme (= Cenococcum
geophilum; sclerotia) was a potential precursor of P-type humic
acids. Therefore, a promising scenario exists whereby perylene qui-
none was transported to the lake in hyphae and sclerotia as well as
in P-type humic substances from the watershed forest soil via sur-
rounding rivers. This is partly supported by a recent study by Itoh
et al. (2012), who found sclerotium grains of C. geophilum, together
with the particles containing DHPQ, at different stages of transfor-
mation to perylene in a Lake Biwa sediment sample.
The temporal profile of perylene concentration normalized to
TOC (Fig. 4C) exhibits broad peaks around 20 kyr BP (MIS 2),
70 kyr BP (MIS 4) and 90 kyr BP (MIS 5b). These peaks in abun-
dance imply that (i) perylene precursors (perylene quinones)
became abundant in the catchment forest soil, and/or that (ii)
transportation of the precursors from the catchment soil to the
lake was enhanced during these periods.
With respect to possible environmental controls on the source of
perylene in soil, a study of the temporal change in abundance
(expressed as absorption intensity) of the Pg fraction (perylene qui-
none pigment) of soil humic acid in a tephra soil sequence from
Kyushu (southern island) in Japan is relevant (Watanabe and
Kobayashi, 2001). There, Pg abundance, expressed using the
absorption at 615 nm, was relatively high at MIS stage 1 (< 10 kyr
BP), 3 (60–25 kyr BP) and 5a (90–70 kyr BP), and it was suggested
that the variation in Pg abundance reflected the strength of leaching
controlled by the relative strength of precipitation and evaporation.
The Kyushu soil results match roughly with our results for per-
ylene (Fig. 4C), implying that climatic conditions, such as lower
temperature and greater wetness (higher precipitation) may have
been favorable for the increase in abundance of Pg as a perylene
precursor.
201
αβ/ββ
Fig. 6. Comparison between Cryptomeria pollen records for BIW95-4 core and
Takashima-oki core (same site as BIW95-5 core) reported by Hayashi et al.
(2010a,b) and C31ab/bb ratio. Pollen proportion is based on the sum of the total tree
pollen (Hayashi et al., 2010a).
4.5. Significance of hopanoid hydrocarbons and perylene in the study
of paleoclimate change and potential as paleoclimate proxies
It is noteworthy that the sections at ca. 70 kyr BP and ca. 90 kyr
BP, with high perylene concentration, coincide roughly with those
with peaks in hopC31ab/bb (Fig. 4C and E). Comparison of perylene/
TOC and hopC31ab/bb for the two cores, indicates relatively good
correlation (Fig. 7) for the 1400 m core (R2 0.44, n = 27). This
implies that the enhancement in perylene precursor (perylene qui-
none) production and/or transportation from the catchment was
related to vegetation change and climatic conditions such as tem-
perature and humidity. However, the relationship was poor (R2
0.04, n = 31) for the BIW95-5 core.
Overall, the temporal changes in the Lake Biwa sediment for
hopanoid hydrocarbons and perylene imply the presence of a rela-
tionship with regional climatic change. However, the extent of the
response to climatic change was different between hopanoid
hydrocarbons and perylene, probably reflecting differences in
source, microbial processes, etc. Further study is necessary to
understand these differences before there is any practical use for
these parameters.
5. Conclusion and implications
We examined the vertical distribution of hopanoid hydrocar-
bons and perylene in sedimentary sections of Lake Biwa and dis-
cussed the relationship between the behavior of these
biomarkers and the regional paleoenvironmental conditions:
202 R. Ishiwatari et al. / Organic Geochemistry 76 (2014) 194–203
αβ ββ
Fig. 7. Relationship between C31ab/bb and perylene concentration normalized to
TOC for Lake Biwa cores.
(i) Moretene concentration was high above the sections where
tephra (tephra AT) fallout events occurred at ca. 30 kyr BP.
This occurrence suggests that O2 deficient conditions were
produced beneath the tephra, resulting in high activity of
methane producing bacteria, followed by high activity of
methanotrophic bacteria which produced moretene.
(ii) Five sections at 10–20 kyr BP, ca. 35 kyr BP, ca. 70 kyr BP, ca.
90 kyr BP and ca. 110 kyr BP exhibited peaks in hopC31ab/bb.
However, n-C23/n-C31 alkanes in these sections was 0.6 ± 0.1
(n = 49), which is very low compared with typical character-
istics for n-alkanes for a Sphagnum peat environment (n-C23/
n-C31 ca. 3.6; Bingham et al., 2010). Therefore, we conclude
that Sphagnum peat may not have been an important con-
tributor of n-alkanes over the past 120 kyr. Comparison with
pollen records reported by Hayashi et al. (2010a,b) for the
lake region suggests that high hopC31ab/bb values may be
caused by production of acidic soil from expansion of tem-
perate conifer (e.g. Cryptomeria) forest under cold and wet
climate conditions, rather than from a Sphagnum peat
environment.
(iii) The high abundance of perylene relative to sediment or TOC
in the sections at ca. 20 kyr BP, ca. 70 kyr BP and ca. 90 kyr
BP was roughly in accord with maxima in the hopC31ab/bb
ratio. This indicates that the climate was suitable for the pro-
duction of perylene quinone in the soil surrounding the lake,
that is, cold and wet climate conditions.
The study indicates a possibility for hopC31ab/bb ratio and per-
ylene/TOC being indicators of soil acidity, temperature and humid-
ity, although further investigation is required.
Acknowledgements
Samples from the 1400 m core were provided by the late Prof. S.
Horie, Kyoto University. We are indebted to Profs. M. Torii, K.
Takemura and A. Hayashida for supervising the coring of BIW 95-
5 core and their support with sampling. We thank E. Hoshino, D.
Ashida, K. Negishi and K. Unoki for analytical assistance and R.
Hayashi for providing the pollen data. We would like to thank
P.A. Meyers and J. Nichols for helpful suggestions and constructive
comments, which greatly improved the manuscript.
Associate Editor—B. van Dongen
References
Ageta, H., Shiojima, K., Arai, Y., 1987. Acid-induced rearrangement of triterpenoid
hydrocarbons belonging to the hopane and migrated hopane series. Chemical
and Pharmaceutical Bulletin 35, 2705–2716.
Aichner, B., Wilkes, H., Herzschuh, U., Mischke, S., Zhang, C., 2009. Biomarker and
compound-specific d13C evidence for changing environmental conditions and
carbon limitation at Lake Koucha, eastern Tibetan Plateau. Journal of
Paleolimnology. http://dx.doi.org/10.1007/s10933-00909375-y.
Aizenshtat, Z., 1973. Perylene and its geochemical significance. Geochimica et
Cosmochimica Acta 37, 559–567.
Baas, M., Pancost, R., van Geel, B., Sinninghe Damsté, J.S., 2000. A comparative study
of lipids in Sphagnum species. Organic Geochemistry 31, 535–541.
Bingham, E.M., McClymont, E.L., Valiranta, M., Mauquoy, D., Roberts, Z., Chambers,
F.M., Pancost, R.D., Evershed, R.P., 2010. Conservative composition of n-alkane
biomarkers in Sphagnum species: implications for palaeoclimate reconstruction
in ombrotrohic peat bogs. Organic Geochemistry 41, 214–220.
Blumenberg, M., Seifert, R., Kasten, S., Bahlmann, E., Michaelis, W., 2009. Euphotic
zone bacterioplankton sources major sedimentary bacteriohopanepolyols in the
Holocene Black Sea. Geochimica et Cosmochimica Acta 73, 750–766.
Brassell, S.C., Comet, P.A., Eglinton, G., Isaacson, P.J., McEvoy, J., Maxwell, J.R.,
Thomson, I.D., Tibbetts, P.J.C., Volkman, J.K., 1980. The origin and fate of lipids in
the Japan Trench. In: Douglas, A.G., Maxwell, J.R. (Eds.), Advances in Organic
Geochemistry 1979. Pergamon Press, Oxford, pp. 375–392.
Cranwell, P.A., Koul, V.K., 1989. Sedimentary record of polycyclic aromatic and
aliphatic hydrocarbons in the Windermere catchment. Water Research 23, 275–
283.
Crossman, Z.M., McNamara, N., Parekh, N., Ineson, P., Evershed, R.P., 2001. A new
method for identifying the origins of simple and complex hopanoids in
sedimentary materials using stable isotope labelling with 13CH4 and
compound specific stable isotope analyses. Organic Geochemistry 32, 359–364.
Dehmer, J., 1995. Petrological and organic geochemical investigation of recent peats
with known environments of deposition. International Journal of Coal Geology
28, 111–138.
Farrimond, P., Head, I.M., Innes, H.E., 2000. Environmental influence on the
biohopanoid composition of recent sediments. Geochimica et Cosmochimica
Acta 64, 2985–2992.
Ficken, K.J., Li, B., Swain, D.L., Eglinton, G., 2000. An n-alkane proxy for the
sedimentary input of submerged/floating freshwater aquatic macrophytes.
Organic Geochemistry 31, 745–749.
Hayashi, R., Takahara, H., Hayashida, A., Takemura, K., 2010a. Millennial-scale
vegetation changes during the last 40,000 yr based on a pollen record from Lake
Biwa, Japan. Quaternary Research 74, 91–99.
Hayashi, R., Takahara, H., Yoshikawa, S., Inouchi, Y., 2010b. Orbital-scale vegetation
variability during MIS 6, 5, 4, and 3 based on a pollen record from the
Takashima-oki core in Lake Biwa, western Japan. Japanese Journal of Palynology
56, 5–12.
Igarashi, Y., Oba, T., 2006. Fluctuations in the East Asian monsoon over the last 144
ka in the northwest Pacific based on a high-resolution pollen analysis of
IMAGES core MD01-2421. Quaternary Science Reviews 25, 1447–1459.
Innes, H.E., Bishop, A.N., Head, I.M., Farrimond, P., 1997. Preservation and diagenesis
of hopanoids in recent lacustrine sediments of Priest Pot, England. Organic
Geochemistry 26, 565–576.
Innes, H.E., Bishop, A.N., Fox, P.A., Head, I.M., Farrimond, P., 1998. Early diagenesis of
bacteriohopanoids in recent sediments of Lake Pollen, Norway. Organic
Geochemistry 29, 1285–1295.
Ishiwatari, R., Hanya, T., 1975. Organic geochemistry of a 200 meter core sample
from Lake Biwa. II. Vertical distribution of mono- and di-carboxylic acids and
polynuclear aromatic hydrocarbons. Proceedings of the Japan Academy 51,
436–441.
Ishiwatari, R., 1977. Organic geochemistry of a 200 meter core sample from Lake
Biwa. III. Isolation and characterization of kerogen and humic acids. Proceedings
of the Japan Academy 53(Ser. B), 47–50.
Ishiwatari, R., Ogura, K., Horie, S., 1980. Organic geochemistry of a lacustrine
sediment (Lake Haruna, Japan). Chemical Geology 29, 261–280.
Ishiwatari, R., Kawamura, K., 1981. Abundance of humic compounds in the 200 meter
sediment core sample from Lake Biwa. In: Horie, S. (Ed.), Paleolimnology of Lake
Biwa and the Japanese Pleistocene, vol. 9. Kyoto University, pp. 103–111.
Ishiwatari, R., Ogura, K., Kawamura, K., 1981. Total carbon, total nitrogen and humic
compounds in the 1000 meter sediment core from Lake Biwa. In: Horie, S. (Ed.),
Paleolimnology of Lake Biwa and the Japanese Pleistocene, vol. 9. Kyoto
University, pp. 112–123.
Ishiwatari, R., Matsushita, T., 1986. Determination of perylenequinones in
sediments and soils. Conversion to perylene followed by gas chromatographic
analysis. Proceedings of the Japan Academy 62 (Ser. B), 287–290.
 R. Ishiwatari et al. / Organic Geochemistry 76 (2014) 194–203
Ishiwatari, R., Uzaki, M., 1987. Diagenetic changes of lignin compounds in a more
than 0.6 million-year-old lacustrine sediment (Lake Biwa, Japan). Geochimica et
Cosmochimica Acta 51, 321–328.
Ishiwatari, R., Yamamoto, S., Uemura, H., 2005. Lipid and lignin/cutin compounds in
Lake Baikal sediments over the last 37 kyr: implications for glacial-interglacial
palaeoenvironmental change. Organic Geochemistry 36, 327–347.
Ishiwatari, R., Negishi, K., Yoshikawa, H., Yamamoto, S., 2009. Glacial-interglacial
productivity and environmental changes in Lake Biwa, Japan: a sediment core
study of organic carbon, chlorins and biomarkers. Organic Geochemistry 40,
520–530.
Itoh, N., Hanari, N., 2010. Possible precursor of perylene in sediments of Lake Biwa
elucidated by stable carbon isotope composition. Geochemical Journal 44, 161–
166.
Itoh, N., Tamamura, S., Kumagai, M., 2010. Distribution of polycyclic aromatic
hydrocarbons in a sediment core from the northern basin of Lake Biwa, Japan.
Organic Geochemistry 41, 845–852.
Itoh, N., Sakagami, N., Torimura, M., Watanabe, M., 2012. Perylene in Lake Biwa
sediments originating from Cenococcum geophilum in its catchment area.
Geochimica et Cosmochimica Acta 95, 241–251.
Iwamoto, N., Inouchi, Y., 2007. Reconstruction of millennial-scale variations in the
East Asian summer monsoon over the past 300 ka based on the total carbon
content of sediment from Lake Biwa, Japan. Environmental Geology 52, 1607–
1616.
Kawamura, K., Ishiwatari, R., 1984. Fatty acid geochemistry of a 200 m sediment
core from Lake Biwa, Japan. Early diagenesis and pleoenvironmental
information. Geochimica et Cosmochimica Acta 48, 251–266.
Kristen, H., Wilkes, H., Vieth, A., Zink, K.G., Plessen, B., Thorpe, J., Partridge, T.C.,
Oberhänsli, H., 2010. Biomarker and stable carbon isotope analyses of
sedimentary organic matter from Lake Tswaig: evidence for deglacial wetness
and early Holocene drought from South Africa. Journal of Paleolimnology 44,
143–160.
Kumada, K., Hurst, H.M., 1967. Green humic acid and its possible origin as a fungal
metabolite. Nature 214, 631–633.
Kuwae, M., 2004. Reconstruction of a climate record for the past 140 kyr based on
diatom valve flux data from Lake Biwa, Japan. Journal of Paleolimnology 32, 19–
39.
Laflamme, R.E., Hites, R.A., 1978. The global distribution of polycyclic aromatic
hydrocarbons in recent sediments. Geochimica et Cosmochimica Acta 42, 289–
303.
Machida, H., Arai, F., Yokoyama, T., 1991. Re-examination of marker-tephra layers in
the 200 m Biwa-Lake core. Quaternary Research 30, 439–442.
Meyers, P.A., Takemura, K., Horie, S., 1993. Reinterpretation of Late Quaternary
sediment chronology of Lake Biwa, Japan, from correlation with marine glacial-
interglacial cycles. Quaternary Research 39, 154–162.
Millar, C.S., 1974. Decomposition of coniferous leaf litter. In: Dickinson, C.H., Pugh,
G.J.F. (Eds.), Biology of Plant Litter Decomposition, vol. 1. Academic Press,
London and New York, pp. 105–128.
Miyazawa, K., 1964. Acidification of soils and chemical properties. In: Kanno, I. (Ed.),
Soil Types in Japan. Nobunkyo, Tokyo, pp. 242–264.
Moldowan, J.M., Fago, F.J., Carlson, R.M.K., Young, D.C., van Duyne, G., Clady, J.,
Schoell, M., Pillinger, C.T., Watt, D.S., 1991. Rearranged hopanes in sediments
and petroleum. Geochimica et Cosmochimica Acta 55, 3333–3353.
Nagahashi, Y., Yoshikawa, S., Miyakawa, C., Uchiyama, T., Inouchi, Y., 2004.
Stratigraphy and chronology of widespread tephra layers during the past 430 ky
in the Kinki district and the Yatsugatake mountains: major element composition
of the glass shards using EDS analysis. Quaternary Research 43, 15–35.
Nichols, J.E., Booth, R.K., Jackson, S.T., Pendall, E.G., Huang, Y., 2006. Paleohydrologic
reconstruction based on n-alkane distributions in ombrotrophic peat. Organic
Geochemistry 37, 1505–1513.
Nott, C.J., Xie, S., Avsejs, L.A., Maddy, D., Chanmbers, F.M.t, Evershed, R.P., 2000. N-
alkane distributions in ombrotrophic mires as indicators of vegetation change
related to climatic variation. Organic Geochemistry 31, 231–235.
Ogura, K., Machihara, T., Takada, H., 1990. Diagenesis of biomarkers in Biwa Lake
sediments over 1 million years. Organic Geochemistry 16, 805–813.
Orr, W.L., Grady, J.R., 1967. Perylene in basin sediments off southern California.
Geochimica et Cosmochimica Acta 31, 1201–1209.
203
Otsuki, A., Aizaki, M., Kawai, T., Fukushima, T., 1981. Application of Carlson’s trophic
state index to Japanese lakes. Research Report from National Institute for
Environmental Studies 23, 3–48.
Pancost, R.D., Baas, M., van Geel, B., Sinninghe Damsté, J.S., 2002. Biomarkers as
proxies for plant inputs to peats: an example from a sub-boreal ombrotrophic
bog. Organic Geochemistry 33, 675–690.
Pancost, R., Baas, M., van Geel, B., Sinninghe Damsté, J.S., 2003. Response of an
ombrotrophic bog to a regional climate event revealed by macrofossil molecular
and carbon isotopic data. The Holocene 13, 921–932.
Pancost, R.D., Pressley, S., Coleman, J.M., Talbot, H.M., Kelly, S.P., Farrimond, P.,
Schouten, S., Benning, L., Mountain, B.W., 2006. Composition and implications
of diverse lipids in New Zealand geothermal sinters. Geobiology 4, 71–92.
Peters, K.E., Moldowan, J.M., 1993. The Biomarker Guide. Prentice-Hall, New Jersey.
Quirk, M.M., Wardroper, A.M.K., Wheatley, R.E., Maxwell, J.R., 1984. Extended
hopanoids in peat environments. Chemical Geology 42, 25–43.
Ries-Kautt, M., Albrecht, P., 1989. Hopane-derived triterpenoids in soils. Chemical
Geology 76, 143–151.
Rohmer, M., Dastillung, M., Ourisson, G., 1980. Hopanoids from C30 to C35 in recent
muds. Naturwissenschaften 67, 456–458.
Rohmer, M., Bouvier-Nave, P., Ourisson, G., 1984. Distribution of hopanoid
triterpenes in prokaryotes. Journal of General Microbiology 130, 1137–1150.
Schouten, S., Hopmans, E.C., Sinninghe Damsté, J.S., 2013. The organic geochemistry
of glycerol dialkyl glycerol tetraether lipids: a review. Organic Geochemistry 54,
19–61.
Silliman, J.E., Meyers, P.A., Eadie, B.J., 1998. Perylene: an indicator of alteration
processes or precursor materials? Organic Geochemistry 29, 1737–1744.
Silliman, J.E., Meyers, P.A., Ostrom, P.H., Ostrom, N.E., Eadie, B.J., 2000. Insights into
the origin of perylene from isotopic analyses of sediments from Saanich Inlet,
British Columbia. Organic Geochemistry 31, 1133–1142.
Silliman, J.E., Meyers, P.A., Eadie, B.J., Klump, J.V., 2001. A hypothesis for the origin of
perylene based on its low abundance in sediments of Green Bay, Wisconsin.
Chemical Geology 177, 309–322.
Simoneit, B.R.T., 1986. Cyclic terpenoids of the geosphere (Chapter 2). In: Johns, R.B.
(Ed.), Biological Markers in the Sedimentary Record. Elsevier, Amsterdam, pp.
43–59.
Suenaga, H., Kusamizu, T., 1986. Chemical properties of Sakurajima volcanic ash and
classification according to color. Kyushu Nogyo Kenkyu 48, 118.
Takemura, K., Yokoyama, T., 1989. Sedimentary environments inferred from
lithofacies of the Lake Biwa 1400 m core sample, Japan. Japanese Journal of
Limnology 50, 247–254.
Takemura, K., 1990. Tectonic and climatic record of the Lake Biwa, Japan, region
provided by the sediments deposited since Pliocene times. Palaeogeography,
Plaeoclimatology, Palaeoecology 78, 185–193.
Takemura, K., Hayashida, A., Okamura, M., Matsuoka, H., Ali, M., Kuniko, Y., Torii, M.,
2000. Stratigraphy of multiple piston-core sediments for the last 30,000 years
from Lake Biwa, Japan. Journal of Paleolimnology 23, 185–199.
Uemura, H., Ishiwatari, R., 1992. Aliphatic hydrocarbons and hopanes in Lake
Haruna sediments with special reference to oil pollution. Japanese Journal of
Limnology 53, 293–303.
Uemura, H., Ishiwatari, R., 1995. Identification of unusual 17b(H)-moret-22(29)-ene
in lake sediments. Organic Geochemistry 23, 675–680.
Uzaki, M., Ishiwatari, R., Machihara, T., 1991. Long-term diagenesis of cellulose and
non-cellulose carbohydrates in Lake Biwa sediments (0.6 million years old). In:
Manning, D. (Ed.), Organic Geochemistry, Advances and Application in the
Natural Environment. Manchester University Press, Manchester and New York,
pp. 264–267.
Van Dorsselaer, A., Albrecht, P., Connan, J., 1977. Changes in composition of
polycyclic alkanes by thermal maturation (Yallourn lignite, Australia). In: (Eds.),
Advances in Organic Geochemistry 1975. Enadimsa, Madrid, pp. 53–59.
Wakeham, S.G., Schaffner, C., Giger, W., 1979. Perylene in sediments from the
Namibian Shelf. Geochimica et Cosmochimica Acta 43, 1141–1144.
Watanabe, M., Kobayashi, T., 2001. Existing state and forming environment of
humic acid Pg fraction in tephra soil sequence of southern Kyushu, Japan.
Quaternary Research 40, 19–28.