Talanta 77 (2009) 1609 1613

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Talanta
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A LCMS method all owing the ana lysis of HMX and RDX present at the picog ram level in natu ral aqueous samples without a concent ration step
Olivier Vigne au, Xavier Ma churon-Manda rd
Commissariat lEnergie At omi que, Cent re DAM Ile-de-France,

Bruy res-le-Ch tel, 91297 Arpajon Cedex, France

a r t i c l e

i n f o

a b s t r a c t
The int roduction of chlo roform into the nebulising gas of a LC/MS elect rosp ray inter face (ESI), in a per fect ly cont roll ed way, leads to the formation of intense adducts ([M+Cl] ) when a mobile phase containing HMX (1,3,5,7- tetranit ro- 1,3,5,7- tetrazac yclooctane or oc togen) and RDX (1,3,5-trint ro- 1,3,5triazac ycloh exane or hexogen) is elu ted. This LC/MS method all ows the di rect ana lysis of aqueous samples containing HMX and RDX at the pic tog ram level without a concent ration step. The method is us ed to determine HMX and RDX concent rations in ground water samples from a milita ry site. 2008 Els evier B.V. All rights reser ved.

Article his tory: Rece ived 13 June 2008 Rece ived in revis ed form 19 Se pt em ber 2008 Acce pted 29 Se pt em ber 2008 Available online 1 N em ber 2008 ov Keywords: LC/MS ESI Explos ive HMX RDX

1. Int roduction In the group of organic explos ives, HMX and RDX are, among the milita ry explos ives, the mo st wide ly us ed (Fig. 1). Bec ause of their toxicity (RDX is a possible human carcino gen [1]), the moni toring of the explos ives releas ed into the envi ronment is an important topic. Those compounds can be detected in soils or in natu ral waters [2]. So, the use of suitable and efcient ana lytical methods devot ed to trace ana lysis is the refo re requi red. The thermal lability of HMX and RDX (nam ed as nit ramines) preferably leads to the use of liquid chroma tography (LC) instead of gas chroma tography (GC) [3]. Usually, as descri be d in the US EPA Method 8330, the ana lysis of aqueous samples containing explosives is carri ed out by LC/UV after a concent ration step [4]. A sample volume equal to 1 L is necessa ry. This method is tedious, time-consuming and UV detection leads to a poor sensit ivity (1 10 g L1 ). The 2004 edition of the drinking water standa rds and health advisories of the US Environmental Prot ection Agency (US EPA) recommends a maximum of 400 g L1 for HMX and a maximum of 2 g L1 for RDX in drinking water for a life-time exposu re [1]. The estima ted quantitation limits of the EPA Method 8330,

Cor responding autho r. Tel.: +33 16926 5127. E-mail add resses: olivie r.vigne au@cea.fr (O. Vigne au), xavie r.m chu ron-m a anda rd@ cea.fr (X. M chu ron-M a anda rd).

when no concent ration step is achieved, are equal to 13 g L1 for HMX and 14 g L1 for RDX [5]. Owing to its poor sensit ivit y, this method is not fully fulll ed for direct ana lysis of aqueous sam- ples containing traces of RDX. Than ks to its good sensit ivit y, mass spect rom etry all ows the detection of a specic ion at traces level. The development of atmospheric pressu re ionisation (API) interface in mass spect rometry all ows di rect detection of traces of explo- sives by LC/MS, without a concent ration step [68]. The compl ete sepa ration of different explos ives having very different physical properties (su ch as nit ramines, nitric esters and nit roa romatics) in a same run cann ot be easi ly realis ed on conventional reverse phase columns. LC sepa ration with porous graphitic carbon (PGC) column foll owed by MS detection with ne gative adduct formation all ows the ana lysis of nit ramines, nitric esters and nit roaromatics in a single method [9,10]. Tachon et al. compa re the ana lysis by LC/MS of explos ives with an atmospheric pressu re chemical ioni- sation (APCI) interface for forensic investigations [9]. The descri bed LC/APCI-MS method claims good performances in terms of select iv- ity, sensit ivity and robu stness. However, the limits of detection for HMX and RDX are respect ively equals to 3.6 ng and 2.2 ng. Holm- gren et al. us ed the same method (LC sepa ration with PGC column foll owed by APCI/MS) and obtain ed the qui te similar limits of detec- tion for HMX (0.7 ng) and RDX (3.2 ng) [10]. The main drawba ck of APCI interface is that it requi res a high tempe ratu re (typical ly a minimum of 300 C) to ensu re the co mpl ete vaporisation of the mobile phase leading to partial loss of heat labile compounds. For exa mple, the thermal deco mposition values of HMX and RDX are

0039- 9140/$ see front m ter 2008 Els evier B.V. All rights reser ved. at doi: 10.1016/j.talanta.2 008.09. 060

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O. Vigneau, X. Ma churon-Manda rd / Talanta 77 (2009) 1609 1613

liquid addit ive into the nebulising gas of the elect rosp ray ionisation (ESI) interface of a mass spect rometer, in a per fect ly cont roll ed way, leading to limits of detection for HMX and RDX at pic tog ram level. 2. Experimental 2.1. Chemicals RDX and HMX standa rds are provid ed by Supelco (Saint Quentin Fall avie r, France) as 1 mg mL1 solutions in acetonitrile. An interm edia te solution containing 1 mg L1 of each compound in acetonitrile is prepa red. This solution is us ed to prepa re cal- ibrated aqueous solutions at different concent rations. Methanol and acetonitrile are both HPLC grade and are provid ed by VWR (Fontenay-sous-Bois, France). HPLC grade chlo roform is purchas ed from SigmaAldri ch (Saint Quentin Fall avie r, France). Ammonium chloride salt is from Merck (Fon ten ay-sous-Bois, France). Ult ra- pu re water used for the prepa ration of mobile phases and standa rd solutions is obtain ed from a Millipo re MilliQ- 10 system. Standa rd solutions and samples are l tered with syrin ge l ters befo re anal- ysis (PALL Acrodisc CR 25 mm Syrin ge l ter with 0.45 m PTFE memb rane) provid ed by VWR (Fontenay-sous-Bois, France). 2.2. Standa rd LC/MS apparatus A VARIAN 1200L triple quadrupole mass spect rom eter, equipp ed with an ESI interface, coupl ed with a VARIAN liquid chro- ma tog raph is used. In the ne gative mode, synth etic air is empl oyed as nebulising gas and nit rogen is us ed as drying gas. The liquid chroma tographic system is compos ed of two VARIAN 210 pumps, a vacuum memb rane de gasser and a VARIAN 410 MIDAS autosam- ple r. This congu ration is us ed when the addit ive is add ed to the mobile phase. In this case, the chroma tog raphic sepa ration is achi eved with a VARIAN column (Pursuit C18 revers ed-phase type, 25 cm 2 mm 5 m) using an isoc ratic mobile phase made of water and methanol (50/50, v/v) at a ow-rate of 0.2 mL min 1 . Ammonium chloride is empl oyed as an addit ive and disso lved in the aqueous mobile phase. It is int roduc ed in the aqueous phase at low concent ration (C = 1 103 mol L1 in water) leading to an ammonium chloride concent ration in the mobile phase equal to 0.5 103 mol L1 . The sample injection volume is 100 L. 2.3. Modied LC/MS apparatus Our pa ten ted system [17], all owing the int roduction of the addit ive into the nebulising gas of the ESI interface, ne eds the mod- ication of the gas feed. A syrin ge-pu mp, a syrin ge and a stainless steel tee are connec ted to the nebulising gas supp ly of the ESI interface (Fig. 2). The liquid ionisable addit ive is int roduc ed in the syrin ge and its ow rate is cont roll ed by the syrin ge pump. A timer trig gered by the autosa mpler cont rols the time and the length of the addit ives int roduction. This int roduction occurs during a short time wind ow cent red around the retention time of the ana lyte. The addit ive is sprayed by the nebulising gas and is carri ed to the ESI needle. At the tip of the ne edle, the ana lytes carri ed by the LC eluent and the addit ive are ionis ed leading to the formation of the adduct ions (e.g. [M+X] in a negative mode). Then, those ions are trans ferred to the ana lyser of the mass spect rometer. Note that in principle, any mass spect rometer equipp ed with an ESI interface can be used. The LC eluent is di rec ted to the waste for the r st 6 min and after 17 min of ana lysis via a six-port valve. The fraction of elu- ent

Fig. 1. Structu ral formulae

of HMX and RDX.

respect ively equals to 280 C and 213 C [11]. So, in APCI part of the RDX molecules deco mpose yielding NO 2 species whi ch leads to adduct formation with a second RDX molecule producing abun- dant [M+NO 2 ] clu ster ions [6]. This pa ram eter could ha mper the ana lysis of environmental samples containing ult ratraces of such explos ives. In the case of nit ramines ana lysis, ESI is the mo st suitable interface [8]. The drawba ck of the use of ESI instead of APCI is that nit roa romatics are mo re di fcult to ionise. In ESI, mo st of the explos ives are detected as adduct-species with the exception of nit roa romatics (su ch as trinit rot oluene, dinit robenzene, trinitrobenzene) [12,13]. Inde ed, the use of addit ives can enhance the ana lyte ionisation efciency leading to higher mass spect rometer sensit ivit y. In ESI, HMX and RDX are di fcult to ionise due to a lack of acidic hyd rogen and because their mass spect ra are compos ed of multiple ions [6,14]. To enhance the sensit ivity and to all ow a better identication of the explos ives, some papers report the use of addit ives such as ammonium salts or organic acids [6, 7,12,14,15]. In the presence of those salts, the mass spect ra of HMX and RDX are co mpos ed of intens ive ions of [M+X] (X = Cl, CH 3 COO, NO 3 or HCOO). Mathis and McCo rd present a comprehens ive method to all ow the screening of a panel of high explos ives (HMX, RDX, EGDN, NG and TNT) [12]. Di fferent ammonium salts (nit rate, forma te, acetate and chloride) are add ed to the mobile phase in order to perform multiple ne gative adduct formation of the different ene r- getic compounds ana lysed. By adding those salts di rect ly to the chroma tographic mobile phase additional specicity and select iv- ity is obtain ed. Inde ed, information relating to the relat ive extent of adduct formation (bas ed on intensity ratios) in addition to adduct stability is us ed to provide a multipl exed detection scheme. No information on the limits of detection obtain ed is given. A sensit ive method for HMX quantication in envi ronmental samples using LC/ESI-MS is repor ted by Pan et al. [14]. A detection limit of 0.78 pg for HMX is obtain ed (low est detection limit obtain ed to da te) by adding a small amount of acetic acid to the mobile phase and ope rating at relat ively low hea ted capilla ry tempe ratu re. The int roduction of the addit ives could be done in various points of the LC/MS appa ratus. Inde ed, it could be int roduc ed in the aque- ous mobile phase, by a post-column system or into the gas feed of the API interface of the mass spect rom eter (in the case of a volatile addit ive). In this pape r, a LC/MS method all owing the direct ana lysis of natu ral water samples contamina ted by HMX or RDX at the pictog ram level without prepa ration of the sample is descri bed. The ana lysis of HMX and RDX is conduc ted because these compounds respond poo rly when ana lysed by GC/MS. Nit roaromatics can be easi ly ana lysed by GC/MS so this work on ly focuses on the ana lysis of nit ramines. The inuence of the addit ive int roduction either in the mobile phase of the LC part or in to the nebulising gas of the elect rosp ray (ESI) interface is compa red. The post-column process is not consid- ered he re bec ause the int roduction of the addit ive by a post-column appa ratus dilu tes the ana lyte and spreads the chroma tog raphic pea ks [16]. A system is descri bed, all owing the int roduction of a

O. Vigneau, X. Ma churon-Manda rd / Talanta 77 (2009) 1609 1613

75

between 6 min and 17 min is int roduc ed in the ESI ne edle of the mass spect rometer. In this case, the chroma tog raphic sepa ration is achi eved with a VARIAN column (Pursuit C18 revers ed-phase type,

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O. Vigneau, X. Ma churon-Manda rd / Talanta 77 (2009) 1609 1613

Fig. 2. Int roduction

of a liquid ionisable

addit ive in the nebulising

gas of the ESI inter face.

25 cm 2 mm 5 m) using an isoc ratic mobile phase of water and methanol (70/30, v/v) at a ow-rate of 0.2 mL min 1 . The sample injection volume is 100 L. 2.4. Sample preparation All water samples are kept at 4 C in the da rk during shipment and storage. Prior to their ana lysis, the samples are l tered through syrin ge l ters. Ea ch extract volume is plac ed into an autosa mpler vial (2 mL) for LC/MS ana lysis. 2.5. Identication and quantication of HMX and RDX

3.2. Introduction of an additive into the nebulising gas of the ESI interface As mention ed above, to int roduce an addit ive into the nebulising gas of the elect rosp ray inter face, it is better to use an ionisable volatile organic compound. In the case of the ana lysis of ethylene glycol dinit rate (EGDN), Lawrence [22] us ed diffusion tubes to dope the gas stream of an ion mobility spect rometer with di chlo rom ethane and to produce chloride adducts. However, this process does not permit careful cont rol of the dopant concent ration. So, in our experiments the int roduction of the vapors of ionisable addit ives (su ch as chlo roform, methylene chloride, nit romethane . . .), in a per fect ly cont roll ed way, is realis ed into the gas feed of an elect rosp ray interface of a mass spect rom eter to improve the ana lyte ionisation efciency. This process needs a slight modication of the ESI interface (Fig. 2). In the presence of chloride ions and in negative mode, HMX and RDX are kn own to give the cor responding adducts [HMX+Cl] and [RDX+Cl] [15,23]. In our system, the chloride ions are produc ed by ionisation of chloroform (CHCl 3 ) [24] or methylene chloride (CH 2 Cl 2 ) [25] at the ESI needle. This is conrm ed by the presence of two abundant ions at m/z values equal to 35 Th and 37 Th with a 3:1 ratio in mass abundance. Fig. 4 demon strates the ne ed to cont rol the ow rate of additive injec ted in the nebulising gas. The chroma tographic peak areas

HMX and RDX are identi ed, in selec ted ion moni toring (SIM) mode, by their retention time, the presence in their mass spect ra of the ions [HMX+ 35 Cl] (m/z = 331 Th), [HMX+ 37 Cl] (m/z = 333 Th), [RDX+35 Cl] (m/z = 257 Th) and [RDX+ 37 Cl] (m/z = 259 Th) and the abundance ratio of the ions [M+ 35 Cl] and [M+ 37 Cl] for whi ch a ratio of 3 to 1 is expec ted. The quantication of HMX and RDX is achieved, also in SIM mode, by measuring the area of the chroma tog raphic peak cor responding to the ions [HMX+ 35 Cl] (m/z = 331 Th) and [RDX+ 35 Cl] (m/z = 257 Th). 3. Results and discussion 3.1. Introduction of an additive in the mobile phase In our r st experiments, ammonium chloride is used as an addit ive. The continuous int roduction of this mobile phase in the elect rosp ray interface has del eterious effects [1820] . The use of salt in the LC eluent clogs up the mass spect rom eter, since the evap- o ration of solvent leads to a deposit of the salt involving a dec rease in sensit ivit y. After 24 h of con stant int roduction of the LC eluent in the elect rosp ray interface, the sensit ivity of the mass spect rom eter dec reases dramatical ly. Inde ed, as can be seen in Fig. 3, the sig- nal response of ions cor responding to [HMX+Cl] and [RDX+Cl] is reduc ed by practical ly 50% after 24 h of repea ted ana lyses of a water sample containing traces of HMX and RDX (2 g L1 of each). So, the liquid ionisable addit ives (su ch as chlo roform or methy- lene chloride) with a low boiling point are mo re suitable because of their volatilit y. Those addit ives can be int roduc ed in the LC elu- ent [16,21] but their concent ration could not be modi ed easi ly and some addit ives are not fully miscible with the aqueous mobile phase. So, the int roduction of the addit ive is realis ed by another way.

Fig. 3. Dec rease of the signal response in LC/ESI-MS after 24 h of repea ted ana lyses of an aqueous m ixtu re of HMX and RDX (2 g L1 of ea ch), w ith a chrom tog raphic m a obile phase containing am onium m chloride (0.5 103 m ol L1 ). The values are calcula ted as the ratio of signal values at di fferent tim to the e signal value obtain ed at the beginning of the study (t = 0 h).

Table 1 LOD and LOQ obtain ed for HMX and RDX co mpa red to EPA recom endations. m Product HMX RDX LOD ( g L1 ) 0.02 0.02 LOQ ( g L1 ) 0.05 0.05 US EPA recommendations 400 2 ( g L1 )

Fig. 4. Inuence of the ow rate of chlo roform int roduc ed into the gas feed of the ESI inter face on the chrom tog raphic peak areas at di fferent concent rations of a HMX and RDX (1 100 g L1 of each).

of ana lytes at different concent rations (1 100 g L1 ) are studied when inc reasing ow rates of chlo roform int roduc ed in the nebulising gas. When the chloride concent ration inc reases (by inc reasing the ow rate of chlo roform), the f ractions of [HMX+Cl] and [RDX+Cl] raise, whi ch results in an inc rease of peak areas of each ana lyte. Mo reover, as can be seen, a minimum of ow rate is necessa ry to ensu re a maximum response signal for HMX and RDX. The ow rate mu st be ada pt ed to a ran ge of concent ration of the ana lytes to obtain a stable signal even if a slight variation of the ow rate of addit ive occurs. The minimum ow rate of chlo roform for the concent ration ran ge 0.02 g L1 to 5 g L1 in nit ramines (HMX and RDX) is equal to 2 L min1 . For the 520 g L1 concent ration range of nit ramines, a minimum of 10 L min1 is requi red. The minimum ow rate of chlo roform requi red for the concent ration range of HMX and RDX between 20 and 100 g L1 is 20 L min1 . 3.3. Detection limit (LOD) and quantication and RDX limit (LOQ) for HMX

RDX (4 replica tes for each calib ration level) are d etermin ed. Those solutions are ana lysed with an injection of chlo roform in the nebulising gas, between the 6th and 17th minu te of the run, at a ow rate of 2 L min1 . As shown in Fig. 5, a good linearity is achieved (R2 = 0.999) for both compounds. The limit of detection and the limit of quantication are obtain ed from a stati stical ana lysis of the signal response. For a di stribution of values presum ed to be G aussian, for a risk value of 5%, the LOD value is equal to 3.29 sB . For the purpose of simplication, the con- ventional value is set to 3 sB . Usually, the LOQ value is set to 10 sB [26]. In LC, the blank standa rd deviation is not direct ly available [26] the refo re the weigh ted linear reg ression m ethod is us ed to estima te the LOD and the LOQ for HMX and RDX. The values obtain ed are summaris ed in Table 1. A good cor relation is obser ved with experimental results obtain ed with standa rd solutions containing the ana lytes at the LOD and LOQ concent ration levels. The two chro- ma tog rams shown on Fig. 6 illu strate the response in LC/MS when standa rd solutions containing HMX and RDX, both at 0.02 g L1 and 0.05 g L1 are injec ted. With a sample loop volume of 100 L, the LOD obtain ed for HMX and RDX equals an injec ted mass of 2 pg. The system all owing addit ive int roduction into the gas feed of an ESI inter face could have a great interest to enhance the sensit iv- ity in liquid chroma tography/atmospheric pressu re ionisatinon by anion atta chment me chanism. Our process, could be us ed instead of salts or po st-column der ivation to improve the detection and the identication of various compounds [27,28] , such as monosac cha- rides [29], oli gosac charides [30], chlorina ted pa rafns [16], lipids [31] or penicillins [32] with an ESI interface. 3.4. Application of the method for natural ground water samples

In all the foll owing experiments, chlo roform is used as addit ive and is int roduc ed into the gas feed of the ESI than ks to our sys- tem. Calib ration standa rds at six concent rations (0.15 g L1 ) are prepa red by diluting the interm edia te standa rd solution in water. The limits of detection and the limits of quantication for HMX and

As mention ed befo re, because of their pot ential to cause ad verse health effects on people expos ed to them, the moni toring of explo- sives releas ed into the environment is an important topic. At this time, the re is a lack of Eu ropean regulations and guidelines regard- ing explos ives. Health advisories have been establish ed for HMX and RDX by US EPA and those are us ed as reference.

Fig. 5. Calib ration cur ves for HMX and RDX w hen nebulising gas ( ow rate = 2 L min1 ).

chlo roform is injec ted in the Fig. 6. Ch rom tog rams obtain ed after injection a of 100 L of solutions containing

0.02

g L1 and 0.05

g L1 of HMX and RDX.

Table 2 HMX and RDX concent rations m easu red in three natu ral ground water sa mples of am ilita ry si te (six replica tes for ea ch sa mple). The data are subjec ted to st ati stical ana lysis by the Student Fisher test, w ith a 95% condence level and Student Fisher con stant of 2. 57. Results are presen ted as m ean er ror. Sa mple Ground water sa mple 1 Ground water sa mple 2 Ground water sa mple 3 [HMX] ( g L1 ) 1.3 0.1 0. 41 0.02 0. 24 0. 01 [RDX] ( g L1 ) 0.42 0.02 <0.02 a 0.02 < x < 0.05 b

mode, acco rding to the addit ive used. The system is also compatible with an atmospheric pressu re chemical ionisation (APCI) interface. References
[1] EPA, 2004 Edition of the Drinking Water Standa rds and Health Advisories, Uni ted States Envi ronm ental Prot ection Agenc y, EPA 822- R-04- 005, 2004. [2] A. Halasz, C. Groom, E. Zhou, L. Paquet, C. Be aulieu, S. Des cha mps, A. Corr iveau, S. Thibou tot, G. Amplem an, C. Dubois, J. Hawari, J. Ch rom tog r. A 963 (2002) a 411. [3] J. Yinon, J.E. McClellan, R.A. Yost, Rapid Com mun. M ass Spect rom 11 (1997) . 1961. [4] EPA, Nit roa rom atics and Nit ram ines by High Performance Li quid Ch rom toga rap hy (HPLC), Uni ted States Envi ronm ental Prot ection Agenc y, Ofce of Solid Waste, Method 8330 SW-846 Upda te III, 1997. [5] EPA, Method 8330: Determination of Concent ration of Nit roa romatics and Nit ramines by High- Performance Li quid Ch roma tog raphy (HPLC), US Ar my Corps of Engineers, Cold Regions Resea rch & Engineering Labo ratory. [6] A. Gape ev, M. Sigman, J. Yinon, Rapid Com un. M m ass Spect rom 17 (2003) . 943. [7] C.S. Evans, R. Sleeman, J. Lu ke, B.J. Kee ly, Rapid Com un. M m ass Spect rom 16 . (2002) 1883. [8] D.A. Cassada, S.J. M onson, D.D. Sn ow, R.F. Spalding, J. Ch rom tog r. A 844 (1999) a 87. [9] R. Tachon, V. Pi chon, M. Bar be-Le Bo rgne, J.-J. Min et, J. Ch rom tog r. A 1154 a (2007) 174. [10] E. Holmg ren, H. Ca rlsson, P. Go ede, C. Crescenzi, J. Ch rom tog r. A 1099 (2005) a 127. [11] J. Akh avan, Chemi stry of Explos ives, 2nd ed., Royal Soci ety of Chemi stry, Do rch- ester, 2004. [12] J.A. Mathis, B.R. McCo rd, Rapid Commun. Mass Spect rom. 19 (2005) 99. [13] A. Schrei ber, J. Efer, W. En ge wald, J. Ch rom tog r. A 869 (200 0) 411. a [14] X. Pan, K. Tian, L.E. Jones, G.P. Cobb, Talanta 70 (2006) 455. [15] J. Yinon, S. Zitrin, M odern Methods and Applications in Ana lysis of Explos ives, 1st ed., John W ey & Sons, Ea stbourne, 1996. il [16] Z. Zenca k, M. Oehme, Rapid Commun. M ass Spect rom. 18 (2004) 22 35. [17] X. M chu ron-M a anda rd, O. Vigne au, Fren ch Patent 0650209, 2006. [18] T.L. Con stan topoulos, G.S. Jackson, C.G. En ke, J. Am. Soc. M ass Spect rom 10 . (1999) 625. [19] S. Kromidas, Mo re Pratical Problem So lving in HPLC, 1st ed., W ey-VCH, 2005. il [20] R. King, R. Bonglio, C. Fernandez-M etzle r, C. Mille r-Stein, T. Olah, J. Am. Soc. M ass Spect rom. 11 (200 0) 942. [21] A. Colo rado, Varian Inc. LC/MS Application Note 18, 2004. [22] A.H. Lawrence, P. Neudor, Anal. Chem. 60 (1988) 104. [23] I. Cotte-Rodriguez, Z. Takats, N. Talat y, H. Chen, R.G. Cooks, Anal. Chem. 77 (2005) 6755. [24] J.M. Warm an, M.C. Sauer Jr., Int. J. Radiat. Phys. Chem. 3 (1971) 237. [25] S. M tejci k, G. Senn, A. Kiendle r, A. Stam tovic, T.D. Ma rk, J. Chem. Phys. 107 a a (1997) 8955. [26] J. Vial, A. Jardy, Anal. Chem 71 (1999) 2672. . [27] S. Gao, Z.-P. Zhang, H.T. Karnes, J. Ch rom tog r. B 825 (2005) 98. a [28] X. Zhao, J. Yinon, J. Ch rom tog r. A 977 (2 002) 59. a [29] H.R. Liang, T. Taka gaki, R.L. Foltz, P. Benn ett, J. Ch rom tog r. B 824 (2 005) 36. a [30] J. Zhu, R.B. Cole, J. Am. Soc. M ass Spect rom 12 (2 001) 1193. . [31] X. Han, H. Cheng, J. Lipid Res. 46 (2005) 163. [32] S. Horim oto, T. Mayumi, K. Aoe, N. Nishimu ra, T. Sa to, J. Pharm. Biom ed. Anal. 30 (2002 1093). [33] H. Abadin, J.J. Liccione, Toxicological Prole for HMX, Agency for Toxic Substances and Disease Regi stry, 1997. [34] C. Smith-Simon, S. Goldha ber, Toxicological Prole for RDX, Agency for Toxic Sub stances and Disease Regi stry, 1995. [35] J. Yinon, Forensic and Envi ronm ental Detection of Explos ives, 1st ed., John W ey il & Sons, Chi che ster, 1999. [36] F.W. Dubois, J.F. Baytos, Weathering of Explos ives for Twenty Years, Los Alamos National Labo ratory, LA 11931, 1991.

a RDX is not detec ted in the sa mple, if RDX is present its concent ration is low er than LOD. b

RDX concent ration is between LOD and LOQ.

The ana lysis of three natu ral ground water samples, obtain ed from a milita ry site is realis ed. All samples are ana lysed six times. The quantitat ive data (six replica tes by sample) are subjec ted to stati stical ana lysis by the Student Fisher test, with a level of significance of 0.05 (i.e. at the 95% condence level with a Student Fisher con stant of 2.57). Results, repor ted in Table 2, are presen ted as mean er ror. Ult ra-t races of HMX and RDX in those ground water samples have been measu red. The actual milita ry site is loca ted on a Second World War battleeld. HMX and RDX are kn own to be present in lea chate from various types of soils and are likely to mig rate into ground water [33,34] despi te a low water solubility [35] but the re is also evidence of their persi stence in soil for several years [36]. Inde ed, a study carri ed out at the Los Alamos National Laboratory, over a period of 20 years, estima tes the half-li fe of HMX and RDX at 39 years and 36 years, respect ively [36]. So, the absence of signicant deg radation, a lea ching and a transport from soil to ground water may explain the presence of traces of HMX and RDX in the samples. 4. Conclusion A sensit ive method, all owing the di rect ana lysis by LC/MS of ground water samples containing HMX and RDX at concent rations as low as 0.02 g L1 , is descri bed. The greatest advanta ge of our sensit ive method is that any concent ration step is necessa ry to be fulll ed with the US EPA recommendations. It can also be useful in forensic ana lysis because after a bomb atta ck, the po st-bla st residues left on the bombing site are present only at trace level. The anion atta chment me chanism is important in the elect rospray process. Negative ion adduct formation could be us ed to improve the sensit ivity of detection for some ana lytes whi ch do not have sites with permanent cha rge or ionisable groups. Elect ron decient atoms, whi ch bind to elect rone gative atoms such as the hydrogen present on hydroxyl groups, could offer binding sites for anions. Our patented system could be of great interest to enhance the identication and the sensit ivity to various compounds ionis ed with an ESI interface. It also could be used in ne gative or in posit ive