Sug ars in en vi ron men tal sam ples and sed i men tary rocks – ef fec tive ness in derivatization for GC-MS anal y sis
Justyna SMOLAREK-LACH1, *, Magdalena GORYL2 and Leszek MARYNOWSKI1
1 Uni ver sity of Silesia, Fac ulty of Nat u ral Sci ences, Bêdziñska 60, 41-200 Sosnowiec, Po land
2 Pol ish Acad emy of Sci ences, In sti tute of Geo log i cal Sci ences, Re search Cen tre in Kraków, Senacka 1, 31-002 Kraków, Po land
Smolarek-Lach, J., Goryl, M., Marynowski, L., 2021. Sug ars in en vi ron men tal sam ples and sed i men tary rocks – ef fec tive - ness in derivatization for GC-MS anal y sis. Geo log i cal Quar terly, 2021, 65: 40, doi: 10.7306/gq.
Sac cha rides (sug ars), com mon con stit u ents of the bio- and geosphere, are use ful in de ter min ing the source of or ganic mat - ter (OM) in aero sols and early diagenetic phases. Due to the chem i cal prop er ties and great di ver sity of sug ars in na ture, a wide range of tech niques is used for the iso la tion and quan ti fi ca tion of sac cha rides. Here, gas chro ma tog ra phy-mass spec - trom e try anal y sis, pre ceded by derivatization with N-methyl-N-trimethylsilyl-trifluoroacetamide (MSTFA) and N,O- bis(trimethylsilyl)tri fluoro acetamide (BSTFA), was un der taken. The ef fec tive ness of this derivatization was dem on strated for soils, aero sols, fungi, and in di vid ual sugar stan dards. The ef fi cacy of BSTFA derivatization for in di vid ual sugar stan dards and, sim i larly, for sam ples of sug ars in win ter and sum mer aero sols was ad e quate. In fungi and soil sam ples, the MSTFA re - agent was much more ef fec tive. Dif fer ent mono- and disaccharides were iden ti fied in bi o log i cal sam ples and sed i men tary rocks, show ing not only the great di ver sity of these com pounds but also the spec i fic ity of their or i gin in re cent and an cient OM. Greater vari a tion in the oc cur rence of sug ars in soils com pared to rocks may sug gest dif fer ences in the sta bil ity of some sac cha rides at an early stage of diagenetic trans for ma tion. Com par i son of the oc cur rence of cer tain sug ars in bi o log i cal sam ples and sed i men tary rocks sheds new light on the or i gin and pres er va tion of these biomolecules.
Key words: BSTFA, MSTFA, sac cha rides, par tic u late mat ter, fungi, lig nites.
INTRODUCTION
Sac cha rides (sug ars) rep re sent the pri mary form of photo - synthetically as sim i lated car bon in the bio sphere and can serve as a tool for clar i fy ing the sources and pro cesses of or - ganic ma te ri als in nat u ral en vi ron ments (Cowie and Hedges, 1984). Mono- and disaccharides are com mon biomolecules in plants and an i mals, oc cur ring widely both in ma rine and ter - res trial en vi ron ments (e.g., BeMiller, 1989; Moers et al., 1989;
Jia et al., 2008). Sug ars are the main con stit u ents of soils (e.g., Gleixner et al., 2002) formed in var i ous cli mate con di - tions, i.e. trop i cal (Simoneit et al., 2004) or tem per ate (Mary - no wski et al., 2020); how ever, their or i gin and role in pedo - genic pro cesses are usu ally di verse and com plex (Maryno - wski et al., 2020). More over, they are used as mo lec u lar trac - ers in at mo spheric aero sols (Simoneit et al., 2004). Anhydro - sugars (e.g., levoglu cosan, mannosan, and gala ctosan) are in di ca tors of burn ing of bio mass and lig nite (Fabbri et al., 2009); sugar al co hols (ara bitol and mannitol) are fun gal spore trac ers; mono- and disa ccha rides (fruc tose, glu cose and su -
crose) are in di ca tors of pol len grains (Simoneit et al., 2004;
Bauer et al., 2008). Given fa vour able depositional and diage - netic con di tions, sac cha rides may be pre served in Ce no zoic and even Me so zoic rocks (Maryno wski et al., 2018, 2019;
Rybicki et al., 2020). Sug ars are typ i cal con stit u ents of var i ous types of li chens and fungi (e.g., Bido chka et al., 1990; Nehls et al., 2010), and their oc cur rence in geo log i cal sam ples may in - di cate a plant-based or fun gal or i gin for or ganic mat ter (Mary - no wski et al., 2018, 2019).
The dif fi culty in eval u at ing and com par ing sug ars is due to the va ri ety of tech niques used for their iso la tion and quan ti fi ca - tion (cf. meth ods in Simoneit et al., 2004). The chem i cal prop - er ties of sac cha rides such as high po lar ity, hydrophilicity, and low vol a til ity ne ces si tate their con ver sion into volatilisable and sta ble de riv a tives prior to gas chro ma tog ra phy-mass spec - trom e try (GC-MS) anal y ses (Ruiz-Matute et al., 2011). Deri - vati zation is a chem i cal pro cess that mod i fies pri mary com - pounds and gen er ates new prod ucts with su pe rior chro mato - graphic prop er ties (Schummer et al., 2009). Briefly, the mech - a nism of the most com mon derivatization method, i.e. silyla - tion, is based on the in tro duc tion of a silyl group in place of a la bile hy dro gen atom. This group can be readily re moved with the re in tro duc tion of ac tive hy dro gen (e.g., via hy dro ly sis).
Schummer et al. (2009) enu mer ated the re agents most com - monly used in silylation: trimethylchlorosilane (TMCS), trime - thylsilylimidazole (TMSI), N-(t-buty ldimethylsilyl)- N-methyl tri - fluo roacetamide (MTBSTFA), N- me thyl-N-trimethylsilyl- tri -
* Corresponding author, e-mail: justyna.smolarek-lach@us.edu.pl Received: April 23, 2021; accepted: July 13, 2021; first published online: September 14, 2021
fluoroacetamide (MSTFA), and N,O-bis(trime thy lsilyl)trifluoro - ace ta mide (BSTFA); for ad di tional re agents, see Poole (2013).
In this study, we show the di ver sity of sac cha rides in the at - mo sphere, bio sphere, pedosphere, and litho sphere, as well as the ef fec tive ness of BSTFA and MSTFA in their silylation. Both re agents are known for suc cess ful derivatization in en vi ron - men tal and geo log i cal stud ies (Simoneit et al., 2004; Burshtein et al., 2010; Rushdi et al., 2016; Marynowski et al., 2019, 2020), but there is still lit tle in for ma tion on the per for mance of these re - agents when derivatizing sugar-rich sam ples in bi o log i cal and early diagenetic set tings such as fungi or soils. In this work, we test which re agent is most ap pro pri ate for derivatization of sam - ples with a rich and di verse saccharide dis tri bu tion. Com par ing the oc cur rence of cer tain sug ars in bi o log i cal ma te rial and sed i - men tary rocks sheds new light on the or i gin and pres er va tion of these com pounds.
MATERIALS AND METHODS
MATERIALS
Five types of sam ple were in ves ti gated:
1 – stan dards of in di vid ual sug ars (Ta ble 1), pur chased from Sigma-Aldrich, AMRESCO, Acros Organics, Fluka, Alfa Aesar, and Merc;
2 – ex am ples of soil: arenosol from the B³êdowska Desert (S Po land) and soil de vel oped on gyp sum from Busko- Zdrój (S Po land);
3 – two PM10 sam ples col lected dur ing win ter (Jan u ary) and sum mer (Au gust) in Sosnowiec (S Po land);
4 – wood-de cay fungi (Lenzites betulina, Inonotus obliquus, Fomitiporia robusta, and Phellinus tremulae);
5 – sed i men tary rocks: two fos sil wood sam ples (Doubrava sandpit, Czech Re pub lic, Pleis to cene; Jaroszów mine, SW Po land, Mio cene), a de tri tal lig nite and an over ly ing clay (Jaroszów mine, Mio cene).
METHODS
EXTRACTION AND DERIVATIZATION
Pow dered sam ples ~15 g (arenosols, fungi, sed i men tary rocks) or quartz fi bre fil ters – Whatman QMA, 47 mm (PM10 sam ples) were ex tracted us ing a di chloro methane:meth a nol mix ture (1:1 v:v) with an ul tra sonic bath (30 min utes at 30°C).
The ex tracted aliquots were fil tered us ing Munksjö fil ters (grade 388, Ahlstrom-Munksjö) for the re moval of in sol u ble par ti cles.
The ex cess re agent was re moved un der blowdown with dry ni - tro gen, then the sam ple mix ture was dis solved in an equiv a lent vol ume of n-hex ane. In di vid ual stan dard so lu tions were pre - pared by dis solv ing 10 mg of each stan dard in 25 cm3 of meth a - nol and ana lys ing 100 mL of the so lu tion, which was evap o rated, derivatized, and dis solved in n-hex ane to 1.5 mL. We have used 100 mL of both BSTFA and MSTFA to check their ef fec - tive ness of derivatization.
The first se ries was con verted to trimethylsilyl (TMS) de riv a - tives via re ac tion with 100 mL BSTFA and the sec ond with 100 mL MSTFA. Both re agents con tained 1% trimethylchlo - rosilane (TMCS). Fol low ing the ad di tion of the silylation re agent and 10 mL pyridine, sam ples were heated for 3 hours at 70°C. A blank sam ple (sil ica gel) was ana lysed us ing the same method.
Trace amounts of phthalates, n-fatty ac ids (FAs), and n-alka - nols were found. No sac cha rides were de tected in the blank sam ples.
GAS CHROMATOGRAPHY-MASS SPECTROMETRY
Gas chro ma tog ra phy-mass spec trom e try (GC-MS) anal y - ses were car ried out with an Agilent Tech nol o gies 7890A gas chromatograph and Agilent 5975C Net work mass spec trom e ter with a Tri ple-Axis Detector (MSD) at the Fac ulty of Earth Sci - ences, Sosnowiec, Po land. He lium (6.0 Grade) was used as a car rier gas at a con stant flow of 2.6 ml/min. The sep a ra tion was ob tained us ing a J&W HP-5ms (60 m × 0.32 mm i.d., 0.25 µm film thick ness) fused sil ica cap il lary col umn coated with a chem - i cally bonded phase (5% phenyl, 95% methylsiloxane), for which the GC oven tem per a ture was pro grammed from 45 (1 min) to 100°C at 20°C/min, then to 300°C (held for 60 min) at 3°C/min, with a sol vent de lay of 10 min. The GC col umn out let was con nected di rectly to the ion source of the MSD. The GC-MS in ter face was set at 280°C, while the ion source and quadrupole an a lyzer were set at 230 and 150°C, re spec tively.
Mass spec tra were re corded from 45–550 (0–40 min) and 50–700 da (>40 min). The MS was op er ated in the elec tron im - pact mode, with an ion iza tion en ergy of 70 eV.
QUANTIFICATION AND IDENTIFICATION
For data col lec tion and mass spec tra pro cess ing, Agilent Tech nol o gies MSD ChemStation E.02.01.1177 and Wiley Reg - T a b l e 1
The per cent age num ber of fully derivatized com pounds of in ter nal sugar stan dards
(BSTFA ver sus MSTFA) af ter silylation
In ter nal stan dard BSTFA MSTFA
[% fully derivatized]
Anhydrosaccharides
Levoglucosan 100 100
Galactosan 89 95
Sugar al co hols
Threitol 98 99
Arabitol 100 100
Adonitol 98 100
Pinitol 100 100
Mannitol 95 100
Sorbitol 91 94
myo-inositol 100 100
Monosaccharides
a- + b-arabinose 87 91
a- + b-xylose 92 98
a- + b-fruc tose 80 97
Mannose 93 96
Galactose 81 91
a-glu cose 92 98
Disaccharides
Su crose 100 100
Trehalose (mycose) 100 100
is try of Mass Spec tral Data (9th edi tion) soft ware were used. To re port the val ues of the monosaccharides, which com monly oc - cur as two iso mers (1a- and 1b-con fig u ra tions of OH on the pyrano or furano ring), the two GC peaks were added to gether (Medeiros and Simoneit, 2007). Ethylvanillin ³98% (Sigma- Aldrich) was used as an in ter nal stan dard in bi o log i cal sam ples.
RESULTS AND DISCUSSION
EFFECTIVENESS OF THE DERIVATIZATION OF SACCHARIDES USING BSTFA VERSUS MSTFA
INTERNAL STANDARDS
Based on the sugar con cen tra tions in the bi o log i cal sam - ples, the dom i nant com pounds were se lected for ver i fi ca tion of the ef fec tive ness of BSTFA ver sus MSTFA derivatization (Ta - ble 1). The iden ti fi ca tion of sac cha rides was made based on the com par i son of mass spec tra and re ten tion times with those of com mer cial sugar stan dards. The m/z 73, 147, 191, 204, and 217 frag ments are the key ions for pentoses, hex oses, and hep tos es as TMS ethers; disaccharides have an ad di tional ion
at m/z 361; while the key ions for the re duced sug ars are at m/z 205, 217, 307, and 319 (Simoneit et al., 2004). As in the case of en vi ron men tal sam ples ex am ined by Medeiros and Simoneit (2007), most of the monosaccharides in our sam ples showed two GC peaks (1a- and 1b-con fig u ra tions of OH on the pyrano or furano ring, Fig. 1). The sum of the val ues for iso mers was pre sented as a per cent age of fully derivatized com pounds (e.g., a-glu cose in Ta ble 1). To iden tify sugar com pounds, es pe cially monosaccharides such as TMS, two mass fragmento grams were pri mar ily used. The m/z 204 frag ment ion (e.g., gluco - pyranose as TMS) char ac ter ized the mass spec tra of sac cha - rides for pyrano ring 5C, the m/z 217 ion for furano ring 4C.
Medeiros and Simoneit (2007) noted no iso mers in their chromatograms, for ei ther sugar al co hols or anhydrosugars;
these were also ab sent in the case of mannitol, arabitol, and myo-inositol in fungi and ini tial soil from this study (Fig. 1). Most disaccharides, such as trehalose or su crose, are char ac ter ized as TMS de riv a tives by the m/z 361 key ion (con junctly with m/z 204). Fully derivatized com pounds were iden ti fied and cal cu - lated (Ta ble 1) based on mass spec tra and re ten tion times. The ef fec tive ness of BSTFA and MSTFA in the derivatization of sugar stan dards was high, rang ing from 80 to 100% and 90 to 100%, re spec tively.
Fig. 1. To tal ion cur rent (TIC) chromatogram of arenosols (sandy soils lack ing any sig nif i cant de vel op ment of a soil pro file) from the B³êdowska Desert show ing the dis tri bu tion of sac cha rides
as trimethylsilyl de riv a tives derivatized us ing: MSTFA – A and BSTFA – B
As ter isks de note com pounds not fully derivatized; note a higher num ber of com pounds not fully derivatized in the case of the BSTFA re agent
ARENOSOLS
Sac cha rides con sti tute a ma jor frac tion of soil or ganic mat - ter (Gleixner et al., 2002). In tem per ate cli mates, glu cose, su - crose, and trehalose dom i nate in soils, with inositols, rha - mnose, arabinose, and lyxose pres ent in lesser amounts (Simo neit et al., 2004). The sam ple of ini tial soil from the B³êdowska Desert rep re sents the third stage of pedogenesis, at which a bi o log i cal soil crust is formed by fungi, li chens, al gae, bac te ria and the de tri tus of higher plants (Marynowski et al., 2020). The dom i nant sug ars in this arenosol were disacchari - des – su crose, trehalose, and sugar al co hol (mannitol); oc cur - ring in mi nor amounts were glyc erol, arabitol, b-glu cose, myo- inositol, and glyceryl glucopyranoside (Fig. 1 and Ta ble 2). The num ber of non-fully-derivatized com pounds was much higher for the sam ples derivatized with BSTFA com pared to MSTFA (Fig. 1 and Ta ble 3). More BSTFA in fact can in crease deriva - tization ef fec tive ness. How ever, in this re search we have stud -
ied the use ful ness of par tic u lar derivatization agents us ing the same amounts of BSTFA and MSTFA. Even us ing twice as much BSTFA than MSTFA gives higher con cen tra tions of non-fully derivatized com pounds. The gen er ally lower ef fec tive - ness of derivatization in arenosols may have been con nected with the higher con cen tra tion of sac cha rides in the sam ple;
how ever, in this case, MSTFA was a su pe rior silylation re agent.
All of the sam ples were dried prior to pow der ing; hence dif fer - ences in derivatization ef fi ciency were not due to the sus cep ti - bil ity of the re agents to mois ture.
AEROSOLS
In aero sols, de pend ing on the sea son, the most com mon sac cha rides are anhydrosugars (with levoglucosan be ing dom i - nant), sugar al co hols (arabitol and mannitol), a- and b-glu cose, a- and b-fruc tose, a- and b-arabinose, and trehalose. Other sug ars, in lesser amounts, in clude su crose, myo-inositol, sor -
T a b l e 2 Oc cur rence and dis tri bu tion of sac cha rides in soils, lig nites, clays,
aero sols and wood-rot fungi
Sam ples 1 2 3 4 5 6 7 8 9 10 11
soils lig nites and clays aero sols fungi
Anhydrosaccharides
Levoglucosan 0.1 100 92
Mannosan 17 14
Galactosan 1 4
Sugar al co hols
Glyc erol 5 15 43 7 11 100 6 13
Threitol 0.1
Erythritol 1 9
Arabitol 3 1 5 5 1 100 2 100 4
Adonitol 0.3 1 1 1
Pinitol 1 100 2 6
Mannitol 32 10 100 64 2 77 12 42 25
Sorbitol 3 1 1 6 0.1
scyllo-inositol 0.2 0.2
myo-inositol 3 4 1
Monosaccharides
a- + b-arabinose 0.7 31 14 2 21 22
a- + b-xylose 17 2
a- + b-fruc tose 0.7 3 100 2 17 8 4 26
Mannose + galactose 1.4 1 22 5 15 2 0.2
a- + b-glu cose 3 9 24 6 38 48 8 79 2 2 1
Glyceryl
glucopyranoside 6 1
Disaccharides
Su crose 100 93 2 4 10 32 7 5 1
Trehalose (mycose) 54 100 9 23 6 27 1 37 100 24 100
1 – arenosole from B³êdowska Desert (data from Marynowski et al., 2020); 2 – ini tial soil from Busko Zdrój; 3 – fos - sil wood, Doubrava sandpit; 4 – fos sil wood sam ple, Jaroszów mine (JR14K); 5 – de tri tal lig nite, Jaroszów mine (JR7Z); 6 – clay, Jaroszów mine (JRIL1); 7 – aero sol sam ple, win ter (80114); 8 – aero sol sam ple, sum mer (80725);
9 – Lenzites betulina fungi; 10 – Inonotus obliquus fungi; 11 – Fomitiporia robusta fungi; rel a tive abun dances in %, nor mal ized to ma jor peak – 100
bitol, pinitol, and glyc erol (Ta ble 2). Levoglucosan is a wide - spread anhydrosaccharide and pres ent mostly in win ter aero - sols as a bio mass burn ing tracer (Simoneit et al., 2004). The ef - fi ciency of derivatization with BSTFA com pared to MSTFA was sim i lar. Pos si bly, in the case of lower con cen tra tions of sac cha - rides (con cen tra tion of in di vid ual com pounds not ex ceed ing 2000 ng/m3 of flow ing air, usu ally within a range of 20–100 ng/m3), both re agents are ef fec tive; derivatized sac - cha rides reached a level near 100%.
FUNGI
Arabitol, mannitol, and trehalose, when they oc cur to - gether, are char ac ter is tic of mycorrhizas and mycorrhizal (e.g., Nehls et al., 2010), entomopathogenic (Bidochka et al., 1990), and wood- and root-rot ting spe cies of fungi (Maryno - wski et al., 2019). The lead ing role of these com pounds in the or gan isms enu mer ated is car bo hy drate stor age (e.g., Asie - gbu, 2000). Com pared to in di vid ual sugar stan dards, the ef - fec tive ness of derivatization with BSTFA was much lower in these sam ples (Fig. 2 and Ta ble 3). For ex am ple, mannitol and trehalose were poorly derivatized by BSTFA (rang ing from 13 to 23% and from 2 to 20%, re spec tively); both show higher val ues for TMS de riv a tives with MSTFA: 79–100 and 89–98%, re spec tively (Ta ble 3 and Fig. 3). BSTFA is not as ef - fec tive for mix tures of many dif fer ent or ganic com pounds with dif fer ent po lar i ties and re ac tiv i ties. Thus, ef fi ciency in deriva - tization de pends not only on the type of re agent (Schummer et al., 2009) but also on the com po si tion of sam ples. For ex am - ple, BSTFA is su pe rior to MSTFA in the derivatization of ste - roids (Huang et al., 2010). In more com plex sam ples MSTFA is much more ef fec tive (com pare deriva tization of stan dards ver sus derivatization of fun gal sam ples).
Re cent stud ies sug gest that sac cha rides are use ful in di ca - tors of or ganic mat ter sources and ma tu rity (Medeiros and Simoneit, 2007; Marynowski et al., 2018, 2019). Thus, ac cu rate cal cu la tion ap pears to be cru cial both in quan ti ta tive re search and in palaeo environmental re con struc tion. Omis sions in the in te gra tion of peaks and to tal ing of com pounds that are not fully derivatized may un der es ti mate their con cen tra tions. Com plete deriva tization achieved with the use of MSTFA helps to avoid these prob lems.
OCCURRENCE OF SACCHARIDES IN BIOLOGICAL MATERIAL AND SEDIMENTARY ROCKS
The dis tri bu tion of sac cha rides in soils, rocks, and fungi sam ples are shown in Ta ble 2 and Fig ure 4, and in areosols in Ta ble 2. Whereas fungi in clude only three ma jor and a few mi - nor com pounds, the com po si tion of sac cha rides is the most di - verse in soils and aero sols, con sist ing of var i ous el e ments of or gan isms in clud ing al gae, cyanobacteria, pol len grains, fungi, and li chens and their spores, as well as frag ments of higher plants (Bauer et al., 2008; Marynowski et al., 2020), and is less het er o ge neous in lig nites and clays. More over, some saccharide-rich par ti cles in aero sols orig i nate from bio mass and coal burn ing (Fabbri et al., 2009; Rybicki et al., 2020). Al - though brown coals are also com posed of var i ous or gan isms, some less sta ble sac cha rides may dis ap pear dur ing early diagenetic pro cesses, in which case other neu tral sac cha - rides, such as hemicellulose and cel lu lose, are gen er ated from pre served biopolymers (Marynowski et al., 2018, 2019).
How ever, the or i gin of some sug ars ap pears to be the same in the case of soils, fos sil wood, and aero sols. Pinitol and fruc - tose, which are ma jor com po nents of de tri tal lig nites and one fos sil wood sam ple, are as so ci ated more closely with pol len grains and pri mary wood com po si tion (e.g., Roach et al., 2017). Glu cose was found in all sam ples ana lysed, in clud ing ex tant fungi. This com pound ap pears to be a com mon con stit - u ent of many geo log i cal, en vi ron men tal, and bi o log i cal ma te ri - als. Many sugar com pounds have been iden ti fied in soils (Simoneit et al., 2004; Marynowski et al., 2020), aero sols (Medeiros and Simoneit, 2007; Bauer et al., 2008), and sed i - men tary OM (Marynowski et al., 2018, 2019), but more stud ies are re quired in or der to un der stand their role and or i gin.
CONCLUSIONS
Sac cha rides are com mon and di verse con stit u ents of soils, aero sols and low-rank coals. Based on this study we rec om mend the ap pli ca tion of MSTFA as a derivatization re - agent in the case of bi o log i cal and en vi ron men tal sam ples, such as fungi and soils, where sac cha rides are the dom i nant com pounds. To avoid un der es ti ma tion of saccharide con -
T a b l e 3 The per cent age of fully BSTFA ver sus MSTFA derivatized com pounds
of bi o log i cal sam ples
Sam ple Fungi spe cies/soil type
L-(-)-arabitol Mannitol Trehalose
[% fully derivatized]
BSTFA MSTFA BSTFA MSTFA BSTFA MSTFA
Fungi15 Lenzites betulina 85 100 15 90 14 91
Fungi18 Inonotus obliquus 68 100 13 79 14 89
Fungi48 Fomitiporia robusta 52 84 14 81 2 93
Fungi70 Phellinus tremulae 27 100 5 100 5 98
Fungi94 Ischnoderma resinosum 80 94 23 91 20 97
OR2 Arenosol 100 100 44 77 72 97
Fig. 2. TIC chromatogram of Phellinus tremulae fungi show ing the dis tri bu tion of sac cha rides as trimethylsilyl de riv a tives derivatized us ing:
BSTFA – A and MSTFA – B
As ter isks de note com pounds not fully derivatized;
note the lower ef fi ciency of BSTFA derivatization
Fig. 3. Mass spec tra of TMS de riv a tives of arabitol (A), mannitol (B) and trehalose (C), the most com mon fungi sug ars de tected in wood-rot fungi
and sed i men tary rocks
cen tra tions in sugar-rich nat u ral sam ples, the use of MSTFA would be cru cial. MSTFA en ables near-full derivatization of the sac cha rides. Even though BSTFA is known for suc cess - ful deriva tization in en vi ron men tal and geo log i cal stud ies, the com po si tion of bi o log i cal sam ples may play a role in the ef - fec tive ness of deriva tization.
Com par ing the oc cur rence of cer tain sug ars in bi o log i cal ma te rial and sed i men tary rocks sheds new light on the or i gin and pres er va tion of these com pounds. The or i gin of arabitol,
mannitol, and trehalose in sed i men tary rocks is as so ci ated with fun gal spores and wood-de grad ing fungi ac tiv ity. Fruc tose and pinitol are more typ i cal of pol len spores as well as of her ba - ceous plants and trees, whereas glu cose or i gin is het er o ge - neous.
Ac knowl edge ments. We thank S. Kurkiewicz for his tech - ni cal as sis tance. This study was funded by the Na tional Sci - ence Cen tre, Po land (grant 2015/19/B/ST10/00925 to LM).
REFERENCES
Asiegbu, F.O., 2000. Ad he sion and de vel op ment of the root rot fun - gus (Heterobasidion annosum) on co ni fer tis sues: ef fects of spore and host sur face con stit u ents. FEMS Mi cro bi ol ogy Ecol - ogy, 33: 101–110.
Bauer, H., Claeys, M., Vermeylen, R., Schueller, E., Weinke, G., Berger, A., Puxbaum, H., 2008. Arabitol and mannitol as trac - ers for the quan ti fi ca tion of air borne fun gal spores. At mo spheric En vi ron ment, 42: 588–593.
BeMiller, J., 1989. Car bo hy drates. In: Nat u ral Prod ucts of Woody Plants: Chem i cals Ex tra ne ous to the Ligno cellulosic Cell Wall (eds. W.E. Hillis and J.W. Rowe): 155–178. Springer Se ries in Wood Sci ence. Springer, Berlin.
Bidochka, M.J., Low, N.H., Khachatourians, G.G., 1990. Car bo - hy drate stor age in the entomopathogenic fun gus Beauveria bassiana. Ap plied and En vi ron men tal Mi cro bi ol ogy, 56:
3186–3190.
Fig. 4. Summed mass chromatogram (m/z 204 + 217 + 361) show ing the dis tri bu tion of mono- and disaccharides of: A – soil de vel oped on gyp sum from Busko-Zdrój, derivatized us ing MSTFA; B – Mio cene xylite (fos sil wood) from the Jaroszów mine, derivatized us ing BSTFA; and C – wood rot fungi Fomitiporia robusta, derivatized us ing MSTFA
Burshtein, N., Yona, N.L., Rudich, Y., 2010. Er gos terol, arabitol and mannitol as trac ers for biogenic aero sols in the East ern Med i ter ra nean. At mo spheric Chem is try Phys ics Dis cus sion, 10:
27725–27758.
Cowie, G.L., Hedges, J.I., 1984. Car bo hy drate sources in a coastal ma rine en vi ron ment. Geochimica et Cosmo chi mica Acta, 48:
2075–2087.
Fabbri, D., Torri, C., Simoneit, B.R.T., Marynowski, L., Rushdi, A.I., Fabiañska, M.J., 2009. Levoglucosan and other cel lu lose and lignin mark ers in emis sions from burn ing of Mio cene lig - nites. At mo spheric En vi ron ment, 43: 2286–2295.
Gleixner, G., Poirier, N., Bol, R., Balesdent, J., 2002. Mo lec u lar dy nam ics of or ganic mat ter in a cul ti vated soil. Or ganic Geo - chem is try, 33: 357–366.
Huang, B., Pan, X., Liu, J., Fang, K., Wang, Y., Gao, J., 2010. New dis cov er ies of heat ing ef fect on trimethylsilyl derivatization for si mul ta neous de ter mi na tion of ste roid en do crine dis rupt ing chem i cals by GC–MS. Chromato graphia, 71: 149–153.
Jia, G., Dungait, J.A.J., Bingham, E.M., Valiranta, M., Korhola, A., Evershed, R.P., 2008. Neu tral mono saccha rides as biomarker prox ies for bog-form ing plants for ap pli ca tion to palaeovegetation re con struc tion in ombro trophic peat de pos its.
Or ganic Geo chem is try, 39: 1790–1799.
Marynowski, L., Bucha, M., Smolarek, J., Wendorff, M., Simoneit, B.R.T., 2018. Oc cur rence and sig nif i cance of mono-, di- and anhydrosaccharide biomolecules in Me so zoic and Ce - no zoic lig nites and fos sil wood. Or ganic Geo chem is try, 116:
13–22.
Marynowski, L., Goryl, M., Bucha, M., Smolarek-Lach, J., Det - man, A., Sikora, A., Chojnacka, A., Simoneit, B.R.T., 2019.
Trehalose, mannitol and arabitol as in di ca tors of fun gal me tab o - lism in Late Cre ta ceous and Mio cene de pos its. In ter na tional Jour nal of Coal Ge ol ogy, 201: 51–61.
Marynowski, L., Rahmonov, O., Smolarek-Lach, J., Rybicki, M., Simoneit, B.R.T., 2020. Or i gin and sig nif i cance of sac cha rides dur ing ini tial pedogenesis in a tem per ate cli mate re gion. Geo - derma, 361: 114064.
Medeiros, P.M., Simoneit, B.R.T., 2007. Anal y sis of sug ars in en vi - ron men tal sam ples by gas chro ma tog ra phy–mass spec trom e - try. Jour nal of Chro ma tog ra phy A, 1141: 271–278.
Moers, M.E.C., Boon, J.J., de Leeuw, J.W., Baas, M., Schenck, P.A., 1989. Car bo hy drate speciation and Py- MS map ping of peat sam ples from a sub trop i cal open marsh en vi ron ment.
Geochimica et Cosmochimica Acta, 53: 2011–2021.
Nehls, U., Göhringer, F., Wittulsky, S., Dietz, S., 2010. Fun gal car - bo hy drate sup port in the ectomycorrhizal sym bi o sis: a re view:
Fun gal car bo hy drates in the ecto my corrhizal sym bi o sis. Plant Bi ol ogy, 12: 292–301.
Poole, C.F., 2013. Alkylsilyl de riv a tives for gas chro ma tog ra phy.
Jour nal of Chro ma tog ra phy A, 1296: 2–14.
Roach, M., Arrivault, S., Mahboubi, A., Krohn, N., Sul pice, R., Stitt, M., Niittylä, T., 2017. Spa tially re solved met a bolic anal y - sis re veals a cen tral role for transcriptio nal con trol in car bon al - lo ca tion to wood. Jour nal of Ex per i men tal Bot any, 68:
3529–3539.
Ruiz-Matute, A.I., Hernández-Hernández, O., Rodríguez- Sán - chez, S., Sanz, M.L., Martínez-Cas tro, I., 2011. Derivatization of car bo hy drates for GC and GC–MS anal y ses. Jour nal of Chro - ma tog ra phy B, 879: 1226–1240.
Rushdi, A.I., Oros, D.R., Al-Mutlaq, K.F., He, D., Mede iros, P.M., Simoneit, B.R.T., 2016. Lipid, sterol and saccharide sources and dy nam ics in sur face soils dur ing an an nual cy cle in a tem - per ate cli mate re gion. Ap plied Geo chem is try, 66: 1–13.
Rybicki, M., Marynowski, L., Simoneit, B.R.T., 2020. Com po si tion of or ganic com pounds from low-tem per a ture burn ing of lig nite and their ap pli ca tion as trac ers in am bi ent air. Chemosphere, 249: 126087.
Schummer, C., Delhomme, O., Appenzeller, B., Wennig, R., Mil - let, M., 2009. Com par i son of MTBSTFA and BSTFA in derivatization re ac tions of po lar com pounds prior to GC/MS anal y sis. Talanta, 77: 1473–1482.
Simoneit, B.R.T., Elias, V.O., Kobayashi, M., Kawamura, K., Rushdi, A.I., Medeiros, P.M., Rogge, W.F., Didyk, B.M., 2004.
Sug ars dom i nant wa ter-sol u ble or ganic com pounds in soils and char ac ter iza tion as trac ers in at mo spheric par tic u late mat ter.
En vi ron men tal Sci ence and Tech nol ogy, 38: 5939–5949.
