Moss species of the family Brachytheciaceae, Kindbergia praelonga (Hedw.) Ochyra and Kindbergia stokesii (Turn.) Ochyra, were preliminarily analysed for their fatty acid composition with the aim of studying the chemical relationship of these two entities. Fatty acid methyl esters were examined by GC and GC MS in their methanol extracts. Thirteen fatty acids were identified. It is likely that the mosses are chemically distinguishable and should be treated as separate entities. However, additional chemical constituents of various moss samples, such as phenolic acids, their derivatives and flavonoids, must be also analyzed in order to support the re-examination of the relationship between these two species.Preliminarno je ispitivan sastav viših masnih kiselina dve mahovine iz familije Brachytheciaceae, Kindbergia praelonga (Hedw.) Ochyra i Kindbergia stokesii (Turn.) Ochyra, sa hemotaksonomskim ciljem. Ukupno je identifikovano 13 viših masnih kiselina GC i GC-MS analizom. Na osnovu dobijenih eksperimentalnih rezultata se može zaključiti da se navedene biljne vrste značajno hemijski razlikuju i da bi se mogle smatrati zasebnim entitetima ukoliko se to potvrdi i dodatnim analizama

Pejin, Boris

Sabovljevic, Aneta

Sabovljevic, Marko

Tešević, Vele

Vajs, Vlatka

Vujisić, Ljubodrag V.

Journal of the Serbian Chemical Society

English

Sabovljević, Marko

Faculty of Chemistry Repository - Cherry

 J. Serb. Chem. Soc. 75 (12) 1637–1640 (2010) UDC Kindbergia+631.445.1:582.32: JSCS–4083 581.184+543  Short communication 1637 SHORT COMMUNICATION Preliminary analysis of fatty acid chemistry of Kindbergia praelonga and Kindbergia stokesii (Brachytheciaceae) BORIS PEJIN1*#, LJUBODRAG VUJISIĆ2#, MARKO SABOVLJEVIĆ3, ANETA SABOVLJEVIĆ3, VELE TEŠEVIĆ1# and VLATKA VAJS2# 1Department of Organic Chemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 16, 11000 Belgrade 2Center of Chemistry, Institute of Chemistry, Technology and Metallurgy, Njegoševa 12, 11000 Belgrade and 3Institute of Botany and Garden, Faculty of Biology, University of Belgrade, Takovska 43, 11000 Belgrade, Serbia (Received 9 February, revised 5 September 2010) Abstract: Moss species of the family Brachytheciaceae, Kindbergia praelonga (Hedw.) Ochyra and Kindbergia stokesii (Turn.) Ochyra, were preliminarily analysed for their fatty acid composition with the aim of studying the chemical relationship of these two entities. Fatty acid methyl esters were examined by GC and GC–MS in their methanol extracts. Thirteen fatty acids were identified. It is likely that the mosses are chemically distinguishable and should be treated as separate entities. However, additional chemical constituents of various moss samples, such as phenolic acids, their derivatives and flavonoids, must be also analyzed in order to support the re-examination of the relationship between these two species. Keywords: bryophytes; mosses; Kindbergia; fatty acids; chemotaxonomy. INTRODUCTION Kindbergia praelonga (Hedw.) Ochyra (typified by Hypnum praelongum Hedw.) and Kindbergia stokesii (Turn.) Ochyra (typified by Hypnum stokesii Turn.) belong to the subgenus Oxyrrhynchium. The latter species is now usually considered as a synonym for or variety of K. praelonga. The genus Kindbergia Ochyra was recently treated as a separate genus from the Eurhynchium Schimp.1,2 According to Hill et al.,1 in Europe only one species is present within the genus: K. praelonga (Hedw.) Ochyra (syn. Eurhynchium praelongum (Hedw.) Schimp.). The same authors classified Eurhynchium praelongum var. stokesii (Turner) Dixon and Eurhynchium stokesii (Turner) Schimp. as a syno-                                                                                                                    * Corresponding author. E-mail: borispejin@yahoo.com # Serbian Chemical Society member. doi: 10.2298/JSC100209129P 2010 Copyright (CC) SCS_________________________________________________________________________________________________________________________Available online at www.shd.org.rs/JSCS/1638 PEJIN et al. nym for K. praelonga. These two entities were separated into the genus Sto-kesiella (Kindb.) H. Rob., which was later considered as an incorrect homonym of the algal generic name Stokesiella Lemmerm. Ochyra renamed the genus Kin-dbergia Ochyra. Now molecular results support this generic independence.3 Worldwide, eleven taxa found at various locations are known, namely K. af-ricana (Herz.) Ochyra, K. altaica Ignatov, K. arbuscula (Broth.) Ochyra, K. brit-toniae (Grout) Ochyra, K. dumosa Mitt., K. kenyae (Dix. Ex Tosco & Piovano) O’Schea et Ochyra, K. oedogonium (C. Müll.) Ochyra, K. oregana (Sull.) Ochy-ra, K. praelonga, K. stokesii and K. squarrifolia Broth. ex Iishiba. The main reason for the repeated nomenclatural confusion and the resulting complications surrounding Oxyrrhynchium is that the name E. praelongum was widely used with two different meanings. Schimper employed it for E. hians (syn. Hypnum hians, Oxyrrhynchium hians (Hedw.) Loeske) including the con-specific E. swartzii Turn. (syn. Hypnum swartzii Turn.) and several closely re-lated elements) for varietal synonymy, whereas his Eurhynchium stokesii corres-ponded to E. praelongum, or following the recent nomenclature K. praelonga. Although many bryologists knew of the problem, frequently the use of the name E. praelongum, instead of E. hians, continued.4 Thus, K. stokesii was often easily overlooked. Analyses were performed to test if chemotaxonomy can be helpful in the as-signment of the two entity species, K. praelonga and K. stokesii. EXPERIMENTAL Both moss species, which were available as fresh material, were collected in Germany in December 2007: Kindbergia praelonga (Hedw.) Ochyra (BEOU4701) in Kologne and Kind-bergia stokesii (Turn.) Ochyra (BEOU4703) in the surroundings of Bonn. Voucher specimens were deposited in the Herbarium of the Institute of Botany, University of Belgrade, Serbia (bryophyte collection – BEOU). The moss samples were carefully selected and cleaned from soil and other contaminants. The gametophyte tips were used for the extraction. Air-dried parts of both mosses were ground (1 g) and extracted 3 times with 90 % MeOH for 1 h at room temperature. The extracts were evaporated to dryness and were further transesterified with 5 % H2SO4 in MeOH (v/v) for 4 h at 80 °C. The resulting methyl esters of the fatty acids were analysed by comparing their GC-FID chromatograms with the chromatogram of a standard mixture (Supelco 37) ob-tained under the same conditions, and/or by analysis of GC–MS data using NIST 5 and Wiley 7 libraries. The GC analyses were performed on an Agilent 7890A GC system equipped with a 5975C MSD and an FID, using a DB-23 column (30 m×0.25 mm×0.25 μm). The injection vo-lume was 1 μL and injector temperature was 220 °C with 10:1 split ratio. The carrier gas was He at a flow rate 0.9 ml min-1, while the column temperature was linearly programmed in the range of 150–240 °C at a rate of 4 °C min-1 and held at 240 °C for 10 min. The transfer line was maintained at 240 °C. The FID detector temperature was 300 °C. The EI mass spectra (70 eV) were acquired in the m/z range 40–500. 2010 Copyright (CC) SCS_________________________________________________________________________________________________________________________Available online at www.shd.org.rs/JSCS/ ANALYSIS OF FATTY ACID OF Kindbergia SPECIES 1639 RESULTS For K. stokesii, 13 fatty acids were identified (Fig. 1): palmitic acid (C16:0, 25.04 %), arachidonic acid (C20:4,n-6, 18.29 %), linolelaidic acid (C18:2,n-6t, 14.57 %), α-linolenic acid (C18:3,n-3, 11.13 %), cis-5,8,11,14,17-eicosapentae-noic acid (C20:3,n-6, 8.94 %), elaidic acid (C18:1,n-9t, 6.32 %), behenic acid (C22:0, 3.04 %), lignoceric acid (C24:0, 2.90 %), palmitoleic acid (C16:1, 2.76 %), stearic acid (C18:0, 2.31 %), myristic acid (C14:0, 1.31 %), oleic acid (C18:1,n- -9c, 1.30 %) and arachidic acid (C20:0, 1.28 %). K. praelongum showed less variety in these constituents: only two fatty acid constituents were found (Fig. 2), palmitic acid (C16:0, 88.58 %) and stearic acid (C18:0, 11.42 %).  Fig. 1. Fatty acid methyl ester chromatogram for K. stokesii; myristic acid (RT 10.399 min); palmitic acid (RT 13.150 min); palmitoleic acid (RT 13.581 min); stearic acid (RT 16.198 min); elaidic acid (RT 16.669 min); oleic acid (RT 16.798 min); linolelaidic acid (RT 17.523 min); α-linolenic acid (RT 18.570 min); arachidic acid (RT 19.273 min); arachidonic acid (RT 21.495 min); behenic acid (RT 22.234 min); cis-5,8,11,14,17-eicosapentaenoic acid (RT 22.543 min) and lignoceric acid (RT 25.294 min).  Fig. 2. Fatty acid methyl ester chromatogram for K. praelongum; palmitic acid (RT 13.167 min) and stearic acid (RT 16.212 min). 2010 Copyright (CC) SCS_________________________________________________________________________________________________________________________Available online at www.shd.org.rs/JSCS/1640 PEJIN et al. CONCLUSIONS The fatty acid composition of the two related and often synonymized Kind-bergia species strongly suggested that they are chemically distinguishable and, thus, could be treated as separate entities. This, however, has still to be confirmed by the analyses of additional chemical constituents, such as phenolic acids and their derivatives as well as flavonoids,5 of various moss samples in order to support a re-examination of the relationship between them. For a general consi-deration of the quality of fatty acid profiling for chemotaxonomy, more replicates and additional species would have to be included in a follow-up study. Acknowledgment. This work was supported by the Ministry of Science and Techno-logical Development of the Republic of Serbia (Research grants No. 142053 and 143015). И З В О Д  ПРЕЛИМИНАРНА АНАЛИЗА ХЕМИЈЕ МАСНИХ КИСЕЛИНА ВРСТА Kindbergia praelonga И Kindbergia stokesii (BRACHYTHECIACEAE) БОРИС ПЕЈИН1, ЉУБОДРАГ ВУЈИСИЋ2, МАРКО САБОВЉЕВИЋ3, АНЕТА САБОВЉЕВИЋ3, ВЕЛЕ ТЕШЕВИЋ1 и ВЛАТКА ВАЈС2 1Katedra za organsku hemiju, Hemijski fakultet, Univerzitet u Beogradu, Studentski trg 16, 11000 Beograd, 2Centar za hemiju, Institut za hemiju, tehnologiju i metalurgiju, Wego{eva 12, 11000 Beograd i 3Institut za botaniku i botani~ka ba{ta, Biolo{ki fakultet, Univerzitet u Beogradu, Takovska 43, 11000 Beograd Прелиминарно је испитиван састав виших масних киселина две маховине из фамилије Brachytheciaceae, Kindbergia praelonga (Hedw.) Ochyra и Kindbergia stokesii (Turn.) Ochyra, са хемотаксономским циљем. Укупно је идентификовано 13 виших масних киселина GC и GC– –MS анализом. На основу добијених експерименталних резултата се може закључити да се наведене биљне врсте значајно хемијски разликују и да би се могле сматрати засебним ен-титетима уколико се то потврди и додатним анализама. (Примљено 9. фебруара, ревидирано 5. септембра 2010) REFERENCES 1. M. O. Hill, N. Bell, M. A. Bruggemann-Nannenga, M. Brugues, M. J. Cano, J. Enroth, K. I. Flatberg, J.-P. Frahm, M. T. Gallego, R. Garilleti, J. Guerra, L. Hedenäs, D. T. Ho-lyoak, J. Hyvönen, M. S. Ignatov, F. Lara, V. Mazimpaka, J. Munoz, L. Söderström, J. Bryol. 28 (2006) 198  2. M. Sabovljević, R. Natcheva, G. Dihoru, E. Tsakiri, S. Dragićević, A. Erdag, B. Papp, Phytol. Balcan. 14 (2008) 159 3. M. S. Ignatov, S. Huttunen, Arctoa 11 (2002) 245 4. M. Ignatov, P. Isoviita, Taxon 52 (2003) 352 5. N. Jocković, P. B. Andrade, P. Valentao, M. Sabovljević, J. Serb. Chem. Soc. 73 (2008) 1161. 2010 Copyright (CC) SCS_________________________________________________________________________________________________________________________Available online at www.shd.org.rs/JSCS/

Preliminarna analiza hemije masnih kiselina vrsta Kindbergia praelonga i Kindbergia stokesii (Brachytheciaceae)

 CER - Central Repository of the Institute of Chemistry;  Technology and Metallurgy

 
J. Serb. Chem. Soc. 75 (12) 1637–1640 (2010) UDC Kindbergia+631.445.1:582.32: 
JSCS–4083 581.184+543 
 Short communication 
1637 
SHORT COMMUNICATION 
Preliminary analysis of fatty acid chemistry of Kindbergia 
praelonga and Kindbergia stokesii (Brachytheciaceae) 
BORIS PEJIN1*#, LJUBODRAG VUJISIĆ2#, MARKO SABOVLJEVIĆ3, 
ANETA SABOVLJEVIĆ3, VELE TEŠEVIĆ1# and VLATKA VAJS2# 
1Department of Organic Chemistry, Faculty of Chemistry, University of Belgrade, Studentski 
trg 16, 11000 Belgrade 2Center of Chemistry, Institute of Chemistry, Technology and 
Metallurgy, Njegoševa 12, 11000 Belgrade and 3Institute of Botany and Garden, 
Faculty of Biology, University of Belgrade, Takovska 43, 11000 Belgrade, Serbia 
(Received 9 February, revised 5 September 2010) 
Abstract: Moss species of the family Brachytheciaceae, Kindbergia praelonga 
(Hedw.) Ochyra and Kindbergia stokesii (Turn.) Ochyra, were preliminarily 
analysed for their fatty acid composition with the aim of studying the chemical 
relationship of these two entities. Fatty acid methyl esters were examined by 
GC and GC–MS in their methanol extracts. Thirteen fatty acids were identified. 
It is likely that the mosses are chemically distinguishable and should be treated 
as separate entities. However, additional chemical constituents of various moss 
samples, such as phenolic acids, their derivatives and flavonoids, must be also 
analyzed in order to support the re-examination of the relationship between 
these two species. 
Keywords: bryophytes; mosses; Kindbergia; fatty acids; chemotaxonomy. 
INTRODUCTION 
Kindbergia praelonga (Hedw.) Ochyra (typified by Hypnum praelongum 
Hedw.) and Kindbergia stokesii (Turn.) Ochyra (typified by Hypnum stokesii 
Turn.) belong to the subgenus Oxyrrhynchium. The latter species is now usually 
considered as a synonym for or variety of K. praelonga. The genus Kindbergia 
Ochyra was recently treated as a separate genus from the Eurhynchium 
Schimp.1,2 According to Hill et al.,1 in Europe only one species is present within 
the genus: K. praelonga (Hedw.) Ochyra (syn. Eurhynchium praelongum 
(Hedw.) Schimp.). The same authors classified Eurhynchium praelongum var. 
stokesii (Turner) Dixon and Eurhynchium stokesii (Turner) Schimp. as a syno-
                                                                                                                    
* Corresponding author. E-mail: borispejin@yahoo.com 
# Serbian Chemical Society member. 
doi: 10.2298/JSC100209129P 
2010 Copyright (CC) SCS
_________________________________________________________________________________________________________________________
Available online at www.shd.org.rs/JSCS/
1638 PEJIN et al. 
nym for K. praelonga. These two entities were separated into the genus Sto-
kesiella (Kindb.) H. Rob., which was later considered as an incorrect homonym 
of the algal generic name Stokesiella Lemmerm. Ochyra renamed the genus Kin-
dbergia Ochyra. Now molecular results support this generic independence.3 
Worldwide, eleven taxa found at various locations are known, namely K. af-
ricana (Herz.) Ochyra, K. altaica Ignatov, K. arbuscula (Broth.) Ochyra, K. brit-
toniae (Grout) Ochyra, K. dumosa Mitt., K. kenyae (Dix. Ex Tosco & Piovano) 
O’Schea et Ochyra, K. oedogonium (C. Müll.) Ochyra, K. oregana (Sull.) Ochy-
ra, K. praelonga, K. stokesii and K. squarrifolia Broth. ex Iishiba. 
The main reason for the repeated nomenclatural confusion and the resulting 
complications surrounding Oxyrrhynchium is that the name E. praelongum was 
widely used with two different meanings. Schimper employed it for E. hians 
(syn. Hypnum hians, Oxyrrhynchium hians (Hedw.) Loeske) including the con-
specific E. swartzii Turn. (syn. Hypnum swartzii Turn.) and several closely re-
lated elements) for varietal synonymy, whereas his Eurhynchium stokesii corres-
ponded to E. praelongum, or following the recent nomenclature K. praelonga. 
Although many bryologists knew of the problem, frequently the use of the name 
E. praelongum, instead of E. hians, continued.4 Thus, K. stokesii was often easily 
overlooked. 
Analyses were performed to test if chemotaxonomy can be helpful in the as-
signment of the two entity species, K. praelonga and K. stokesii. 
EXPERIMENTAL 
Both moss species, which were available as fresh material, were collected in Germany in 
December 2007: Kindbergia praelonga (Hedw.) Ochyra (BEOU4701) in Kologne and Kind-
bergia stokesii (Turn.) Ochyra (BEOU4703) in the surroundings of Bonn. Voucher specimens 
were deposited in the Herbarium of the Institute of Botany, University of Belgrade, Serbia 
(bryophyte collection – BEOU). 
The moss samples were carefully selected and cleaned from soil and other contaminants. 
The gametophyte tips were used for the extraction. Air-dried parts of both mosses were 
ground (1 g) and extracted 3 times with 90 % MeOH for 1 h at room temperature. The extracts 
were evaporated to dryness and were further transesterified with 5 % H2SO4 in MeOH (v/v) 
for 4 h at 80 °C. The resulting methyl esters of the fatty acids were analysed by comparing 
their GC-FID chromatograms with the chromatogram of a standard mixture (Supelco 37) ob-
tained under the same conditions, and/or by analysis of GC–MS data using NIST 5 and Wiley 
7 libraries. 
The GC analyses were performed on an Agilent 7890A GC system equipped with a 
5975C MSD and an FID, using a DB-23 column (30 m×0.25 mm×0.25 μm). The injection vo-
lume was 1 μL and injector temperature was 220 °C with 10:1 split ratio. The carrier gas was 
He at a flow rate 0.9 ml min-1, while the column temperature was linearly programmed in the 
range of 150–240 °C at a rate of 4 °C min-1 and held at 240 °C for 10 min. The transfer line 
was maintained at 240 °C. The FID detector temperature was 300 °C. The EI mass spectra (70 
eV) were acquired in the m/z range 40–500. 
2010 Copyright (CC) SCS
_________________________________________________________________________________________________________________________
Available online at www.shd.org.rs/JSCS/
 ANALYSIS OF FATTY ACID OF Kindbergia SPECIES 1639 
RESULTS 
For K. stokesii, 13 fatty acids were identified (Fig. 1): palmitic acid (C16:0, 
25.04 %), arachidonic acid (C20:4,n-6, 18.29 %), linolelaidic acid (C18:2,n-6t, 
14.57 %), α-linolenic acid (C18:3,n-3, 11.13 %), cis-5,8,11,14,17-eicosapentae-
noic acid (C20:3,n-6, 8.94 %), elaidic acid (C18:1,n-9t, 6.32 %), behenic acid 
(C22:0, 3.04 %), lignoceric acid (C24:0, 2.90 %), palmitoleic acid (C16:1, 2.76 %), 
stearic acid (C18:0, 2.31 %), myristic acid (C14:0, 1.31 %), oleic acid (C18:1,n- 
-9c, 1.30 %) and arachidic acid (C20:0, 1.28 %). K. praelongum showed less 
variety in these constituents: only two fatty acid constituents were found (Fig. 2), 
palmitic acid (C16:0, 88.58 %) and stearic acid (C18:0, 11.42 %). 
 
Fig. 1. Fatty acid methyl ester chromatogram for K. stokesii; myristic acid (RT 10.399 min); 
palmitic acid (RT 13.150 min); palmitoleic acid (RT 13.581 min); stearic acid (RT 16.198 
min); elaidic acid (RT 16.669 min); oleic acid (RT 16.798 min); linolelaidic acid (RT 17.523 
min); α-linolenic acid (RT 18.570 min); arachidic acid (RT 19.273 min); arachidonic acid 
(RT 21.495 min); behenic acid (RT 22.234 min); cis-5,8,11,14,17-eicosapentaenoic acid 
(RT 22.543 min) and lignoceric acid (RT 25.294 min). 
 
Fig. 2. Fatty acid methyl ester chromatogram for K. praelongum; palmitic acid 
(RT 13.167 min) and stearic acid (RT 16.212 min). 
2010 Copyright (CC) SCS
_________________________________________________________________________________________________________________________
Available online at www.shd.org.rs/JSCS/
1640 PEJIN et al. 
CONCLUSIONS 
The fatty acid composition of the two related and often synonymized Kind-
bergia species strongly suggested that they are chemically distinguishable and, 
thus, could be treated as separate entities. This, however, has still to be confirmed 
by the analyses of additional chemical constituents, such as phenolic acids and 
their derivatives as well as flavonoids,5 of various moss samples in order to 
support a re-examination of the relationship between them. For a general consi-
deration of the quality of fatty acid profiling for chemotaxonomy, more replicates 
and additional species would have to be included in a follow-up study. 
Acknowledgment. This work was supported by the Ministry of Science and Techno-
logical Development of the Republic of Serbia (Research grants No. 142053 and 143015). 
И З В О Д  
ПРЕЛИМИНАРНА АНАЛИЗА ХЕМИЈЕ МАСНИХ КИСЕЛИНА ВРСТА Kindbergia 
praelonga И Kindbergia stokesii (BRACHYTHECIACEAE) 
БОРИС ПЕЈИН1, ЉУБОДРАГ ВУЈИСИЋ2, МАРКО САБОВЉЕВИЋ3, АНЕТА САБОВЉЕВИЋ3, 
ВЕЛЕ ТЕШЕВИЋ1 и ВЛАТКА ВАЈС2 
1Katedra za organsku hemiju, Hemijski fakultet, Univerzitet u Beogradu, Studentski trg 16, 11000 
Beograd, 2Centar za hemiju, Institut za hemiju, tehnologiju i metalurgiju, Wego{eva 12, 11000 Beograd 
i 3Institut za botaniku i botani~ka ba{ta, Biolo{ki fakultet, 
Univerzitet u Beogradu, Takovska 43, 11000 Beograd 
Прелиминарно је испитиван састав виших масних киселина две маховине из фамилије 
Brachytheciaceae, Kindbergia praelonga (Hedw.) Ochyra и Kindbergia stokesii (Turn.) Ochyra, са 
хемотаксономским циљем. Укупно је идентификовано 13 виших масних киселина GC и GC– 
–MS анализом. На основу добијених експерименталних резултата се може закључити да се 
наведене биљне врсте значајно хемијски разликују и да би се могле сматрати засебним ен-
титетима уколико се то потврди и додатним анализама. 
(Примљено 9. фебруара, ревидирано 5. септембра 2010) 
REFERENCES 
1. M. O. Hill, N. Bell, M. A. Bruggemann-Nannenga, M. Brugues, M. J. Cano, J. Enroth, K. 
I. Flatberg, J.-P. Frahm, M. T. Gallego, R. Garilleti, J. Guerra, L. Hedenäs, D. T. Ho-
lyoak, J. Hyvönen, M. S. Ignatov, F. Lara, V. Mazimpaka, J. Munoz, L. Söderström, J. 
Bryol. 28 (2006) 198  
2. M. Sabovljević, R. Natcheva, G. Dihoru, E. Tsakiri, S. Dragićević, A. Erdag, B. Papp, 
Phytol. Balcan. 14 (2008) 159 
3. M. S. Ignatov, S. Huttunen, Arctoa 11 (2002) 245 
4. M. Ignatov, P. Isoviita, Taxon 52 (2003) 352 
5. N. Jocković, P. B. Andrade, P. Valentao, M. Sabovljević, J. Serb. Chem. Soc. 73 (2008) 
1161. 
2010 Copyright (CC) SCS
_________________________________________________________________________________________________________________________
Available online at www.shd.org.rs/JSCS/


Moss species of the family Brachytheciaceae, Kindbergia praelonga (Hedw.) Ochyra and Kindbergia stokesii (Turn.) Ochyra, were preliminarily analysed for their fatty acid composition with the aim of studying the chemical relationship of these two entities. Fatty acid methyl esters were examined by GC and GC–MS in their methanol extracts. Thirteen fatty acids were identified. It is likely that the mosses are chemically distinguishable and should be treated as separate entities. However, additional chemical constituents of various moss samples, such as phenolic acids, their derivatives and flavonoids, must be also analyzed in order to support the re-examination of the relationship between these two species

BORIS PEJIN

LJUBODRAG VUJISIĆ

MARKO SABOVLJEVIĆ

ANETA SABOVLJEVIĆ

VELE TEŠEVIĆ

VLATKA VAJS

Directory of Open Access Journals

 J. Serb. Chem. Soc. 75 (12) 1637–1640 (2010)  UDC Kindbergia+631.445.1:582.32: JSCS–4083 581.184+543  Short  communication 1637 SHORT COMMUNICATION Preliminary analysis of fatty acid chemistry of Kindbergia praelonga and Kindbergia stokesii (Brachytheciaceae) BORIS PEJIN1*#, LJUBODRAG VUJISIĆ2#, MARKO SABOVLJEVIĆ3, ANETA SABOVLJEVIĆ3, VELE TEŠEVIĆ1# and VLATKA VAJS2# 1Department of Organic Chemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 16, 11000 Belgrade 2Center of Chemistry, Institute of Chemistry, Technology and Metallurgy, Njegoševa 12, 11000 Belgrade and 3Institute of Botany and Garden, Faculty of Biology, University of Belgrade, Takovska 43, 11000 Belgrade, Serbia (Received 9 February, revised 5 September 2010) Abstract: Moss species of the family Brachytheciaceae, Kindbergia praelonga (Hedw.) Ochyra and Kindbergia stokesii (Turn.) Ochyra, were preliminarily analysed for their fatty acid composition with the aim of studying the chemical relationship of these two entities. Fatty acid methyl esters were examined by GC and GC–MS in their methanol extracts. Thirteen fatty acids were identified. It is likely that the mosses are chemically distinguishable and should be treated as separate entities. However, additional chemical constituents of various moss samples, such as phenolic acids, their derivatives and flavonoids, must be also analyzed in order to support the re-examination of the relationship between these two species. Keywords: bryophytes; mosses; Kindbergia; fatty acids; chemotaxonomy. INTRODUCTION Kindbergia praelonga (Hedw.) Ochyra  (typified by Hypnum praelongum Hedw.) and Kindbergia stokesii (Turn.) Ochyra (typified by Hypnum stokesii Turn.) belong to the subgenus Oxyrrhynchium. The latter species is now usually considered as a synonym for or variety of K. praelonga. The genus Kindbergia Ochyra was recently treated as a separate genus from the Eurhynchium Schimp.1,2 According to Hill et al.,1 in Europe only one species is present within the genus: K. praelonga (Hedw.) Ochyra (syn. Eurhynchium praelongum (Hedw.) Schimp.). The same authors classified Eurhynchium praelongum var. stokesii (Turner) Dixon and Eurhynchium stokesii (Turner) Schimp. as a syno-                                                                                                                    * Corresponding author. E-mail: borispejin@yahoo.com # Serbian Chemical Society member. doi: 10.2298/JSC100209129P 2010 Copyright (CC) SCS_________________________________________________________________________________________________________________________ Available online at www.shd.org.rs/JSCS/1638 PEJIN  et al. nym for K. praelonga. These two entities were separated into the genus Sto-kesiella (Kindb.) H. Rob., which was later considered as an incorrect homonym of the algal generic name Stokesiella Lemmerm. Ochyra renamed the genus Kin-dbergia Ochyra. Now molecular results support this generic independence.3 Worldwide, eleven taxa found at various locations are known, namely K. af-ricana (Herz.) Ochyra, K. altaica Ignatov, K. arbuscula (Broth.) Ochyra, K. brit-toniae (Grout) Ochyra, K. dumosa Mitt., K. kenyae (Dix. Ex Tosco & Piovano) O’Schea et Ochyra, K. oedogonium (C. Müll.) Ochyra, K. oregana (Sull.) Ochy-ra, K. praelonga, K. stokesii and K. squarrifolia Broth. ex Iishiba. The main reason for the repeated nomenclatural confusion and the resulting complications surrounding Oxyrrhynchium is that the name E. praelongum was widely used with two different meanings. Schimper employed it for E. hians (syn. Hypnum hians, Oxyrrhynchium hians (Hedw.) Loeske) including the con-specific E. swartzii Turn. (syn. Hypnum swartzii Turn.) and several closely re-lated elements) for varietal synonymy, whereas his Eurhynchium stokesii corres-ponded to E. praelongum, or following the recent nomenclature K. praelonga. Although many bryologists knew of the problem, frequently the use of the name E. praelongum, instead of E. hians, continued.4 Thus, K. stokesii was often easily overlooked. Analyses were performed to test if chemotaxonomy can be helpful in the as-signment of the two entity species, K. praelonga and K. stokesii. EXPERIMENTAL Both moss species, which were available as fresh material, were collected in Germany in December 2007: Kindbergia praelonga (Hedw.) Ochyra (BEOU4701) in Kologne and Kind-bergia stokesii (Turn.) Ochyra (BEOU4703) in the surroundings of Bonn. Voucher specimens were deposited in the Herbarium of the Institute of Botany, University of Belgrade, Serbia (bryophyte collection – BEOU). The moss samples were carefully selected and cleaned from soil and other contaminants. The gametophyte tips were used for the extraction. Air-dried parts of both mosses were ground (1 g) and extracted 3 times with 90 % MeOH for 1 h at room temperature. The extracts were evaporated to dryness and were further transesterified with 5 % H2SO4 in MeOH (v/v) for 4 h at 80 °C. The resulting methyl esters of the fatty acids were analysed by comparing their GC-FID chromatograms with the chromatogram of a standard mixture (Supelco 37) ob-tained under the same conditions, and/or by analysis of GC–MS data using NIST 5 and Wiley 7 libraries. The GC analyses were performed on an Agilent 7890A GC system equipped with a 5975C MSD and an FID, using a DB-23 column (30 m×0.25 mm×0.25 μm). The injection vo-lume was 1 μL and injector temperature was 220 °C with 10:1 split ratio. The carrier gas was He at a flow rate 0.9 ml min-1, while the column temperature was linearly programmed in the range of 150–240 °C at a rate of 4 °C min-1 and held at 240 °C for 10 min. The transfer line was maintained at 240 °C. The FID detector temperature was 300 °C. The EI mass spectra (70 eV) were acquired in the m/z range 40–500. 2010 Copyright (CC) SCS_________________________________________________________________________________________________________________________ Available online at www.shd.org.rs/JSCS/  ANALYSIS OF FATTY ACID OF Kindbergia SPECIES  1639 RESULTS For K. stokesii, 13 fatty acids were identified (Fig. 1): palmitic acid (C16:0, 25.04 %), arachidonic acid (C20:4,n-6, 18.29 %), linolelaidic acid (C18:2,n-6t, 14.57 %), α-linolenic acid (C18:3,n-3, 11.13 %), cis-5,8,11,14,17-eicosapentae-noic acid (C20:3,n-6, 8.94 %), elaidic acid (C18:1,n-9t, 6.32 %), behenic acid (C22:0, 3.04 %), lignoceric acid (C24:0, 2.90 %), palmitoleic acid (C16:1, 2.76 %), stearic acid (C18:0, 2.31 %), myristic acid (C14:0, 1.31 %), oleic acid (C18:1,n- -9c, 1.30 %) and arachidic acid (C20:0, 1.28 %). K. praelongum showed less variety in these constituents: only two fatty acid constituents were found (Fig. 2), palmitic acid (C16:0, 88.58 %) and stearic acid (C18:0, 11.42 %).  Fig. 1. Fatty acid methyl ester chromatogram for K. stokesii; myristic acid (RT 10.399 min); palmitic acid (RT 13.150 min); palmitoleic acid (RT 13.581 min); stearic acid (RT 16.198 min); elaidic acid (RT 16.669 min); oleic acid (RT 16.798 min); linolelaidic acid (RT 17.523 min); α-linolenic acid (RT 18.570 min); arachidic acid (RT 19.273 min); arachidonic acid (RT 21.495 min); behenic acid (RT 22.234 min); cis-5,8,11,14,17-eicosapentaenoic acid (RT 22.543 min) and lignoceric acid (RT 25.294 min).  Fig. 2. Fatty acid methyl ester chromatogram for K. praelongum; palmitic acid (RT 13.167 min) and stearic acid (RT 16.212 min). 2010 Copyright (CC) SCS_________________________________________________________________________________________________________________________ Available online at www.shd.org.rs/JSCS/1640 PEJIN  et al. CONCLUSIONS The fatty acid composition of the two related and often synonymized Kind-bergia species strongly suggested that they are chemically distinguishable and, thus, could be treated as separate entities. This, however, has still to be confirmed by the analyses of additional chemical constituents, such as phenolic acids and their derivatives as well as flavonoids,5 of various moss samples in order to support a re-examination of the relationship between them. For a general consi-deration of the quality of fatty acid profiling for chemotaxonomy, more replicates and additional species would have to be included in a follow-up study. Acknowledgment. This work was supported by the Ministry of Science and Techno-logical Development of the Republic of Serbia (Research grants No. 142053 and 143015). ИЗВОД ПРЕЛИМИНАРНА АНАЛИЗА ХЕМИЈЕ МАСНИХ КИСЕЛИНА ВРСТА Kindbergia praelonga И Kindbergia stokesii (BRACHYTHECIACEAE) БОРИС ПЕЈИН1, ЉУБОДРАГ ВУЈИСИЋ2, МАРКО САБОВЉЕВИЋ3, АНЕТА САБОВЉЕВИЋ3, ВЕЛЕ ТЕШЕВИЋ1 и ВЛАТКА ВАЈС2 1Katedra za organsku hemiju, Hemijski fakultet, Univerzitet u Beogradu, Studentski trg 16, 11000 Beograd, 2Centar za hemiju, Institut za hemiju, tehnologiju i metalurgiju, Wego{eva 12, 11000 Beograd i 3Institut za botaniku i botani~ka ba{ta, Biolo{ki fakultet, Univerzitet u Beogradu, Takovska 43, 11000 Beograd Прелиминарно је испитиван састав виших масних киселина две маховине из фамилије Brachytheciaceae, Kindbergia praelonga (Hedw.) Ochyra и Kindbergia stokesii (Turn.) Ochyra, са хемотаксономским циљем. Укупно је идентификовано 13 виших масних киселина GC и GC– –MS анализом. На основу добијених експерименталних резултата се може закључити да се наведене биљне врсте значајно хемијски разликују и да би се могле сматрати засебним ен-титетима уколико се то потврди и додатним анализама. (Примљено 9. фебруара, ревидирано 5. септембра 2010) REFERENCES 1.  M. O. Hill, N. Bell, M. A. Bruggemann-Nannenga, M. Brugues, M. J. Cano, J. Enroth, K. I. Flatberg, J.-P. Frahm, M. T. Gallego, R. Garilleti, J. Guerra, L. Hedenäs, D. T. Ho-lyoak, J. Hyvönen, M. S. Ignatov, F. Lara, V. Mazimpaka, J. Munoz, L. Söderström, J. Bryol. 28 (2006) 198  2.  M. Sabovljević, R. Natcheva, G. Dihoru, E. Tsakiri, S. Dragićević, A. Erdag, B. Papp, Phytol. Balcan. 14 (2008) 159 3.  M. S. Ignatov, S. Huttunen, Arctoa 11 (2002) 245 4.  M. Ignatov, P. Isoviita, Taxon 52 (2003) 352 5.  N. Jocković, P. B. Andrade, P. Valentao, M. Sabovljević, J. Serb. Chem. Soc. 73 (2008) 1161. 2010 Copyright (CC) SCS_________________________________________________________________________________________________________________________ Available online at www.shd.org.rs/JSCS/

Preliminary analysis of fatty acid chemistry of Kindbergia praelonga and Kindbergia stokesii (Brachytheciaceae)

Moss species of the family Brachytheciaceae, Kindbergia praelonga (Hedw.)
   Ochyra and Kindbergia stokesii (Turn.) Ochyra, were preliminary analysed for
   their fatty acid composition with the aim of studying the chemical
   relationship of these two entities. Fatty acid methyl esters were examined by
   GC and GC-MS in their methanol extracts. Thirteen fatty acids were
   identified. It is likely that the mosses are chemically distinguishable and
   should be treated as separate entities. However, additional chemical
   constituents of various moss samples, such as phenolic acids, their
   derivatives and flavonoids, must be also analyzed in order to support the
   re-examination of the relationship between these two species.</jats:p

Boris Pejin

Ljubodrag Vujisic

Marko Sabovljevic

Aneta Sabovljevic

Vele Tesevic

Vlatka Vajs

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Central Repository of the Institute of Chemistry, Technology and Metallurgy (CER)

Moss species of the family Brachytheciaceae, Kindbergia praelonga (Hedw.) Ochyra and Kindbergia stokesii (Turn.) Ochyra, were preliminarily analysed for their fatty acid composition with the aim of studying the chemical relationship of these two entities. Fatty acid methyl esters were examined by GC and GC MS in their methanol extracts. Thirteen fatty acids were identified. It is likely that the mosses are chemically distinguishable and should be treated as separate entities. However, additional chemical constituents of various moss samples, such as phenolic acids, their derivatives and flavonoids, must be also analyzed in order to support the re-examination of the relationship between these two species

Preliminarna analiza hemije masnih kiselina vrsta Kindbergia praelonga i Kindbergia stokesii (Brachytheciaceae)

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