Vol. 19 No. 5 October/November 2007 SPECTROSCOPY europe /—— Midday Davis fur deh) Suaeoussouy UU/yis Siuutiss of Pouap five aii) EhsnGéutues MME of Diebeieissude dyes lat es ®WI LEY I WY TRiblicationsay ae VARIAN Overtake your competition with accurate ICP-OES analysis that's so fast it's practically unbelievable. The mumber one supper of HOES instuents rng ou the Varian 720/70-65 Seis. he | sero ep Consumables work!’ best and fastest ICP-OES platform. An innovative custom-designed CCD detector gives you | YORU true simultaneous measurement for unrivalled productivity timate precision andlawestever detection | Ds Systems limits. Custornize or upgrade your 720/730-ES with a range of performance and productivity enhancing | Supper & Training options, ensuring the best posible soltion for your most demanding rasds - both now and in the future. you havea need for speed and proven performance, arian has the ICP-OES solution fr you. Another productivity-enhancing soliition from Varian View free webinar at www.varianine,com/ICPwebinar/ ab as ce eN Ane RF OU GNI Fe Raman Fonscnce*Okstucons AW tums sina “Dat Sptm VARIAN FASTLINK / CIRCLE 0011 FOR FURTHER INFORMATION 2 SPECTROSCOPYEUROPE www.spectroscopyeurope.comVOL. 19 NO. 5 (2007) jc.“ "18 ‘Nell Macleod, Charlo:te Eliasson and Pavel Matousek descibe new Raman spectroscopy techniques for analysis through turbid mecie, They describe Spatially Offset Raman Spectroscopy (ORS) and transmission Reman, and applications of them for detection of concealed explosives, counterfeit drugs and quality control of pharmaceutical Products tis possible, using these tech Tiques, to detect liquid explosives and other substances inside white plastic ‘AnnaStina Jaskelainen and Tina ita investigete the use of LV/is reflectance spectroscopy to study the reactions caus ing colour changes in paper We are used to the yellowing of ole paper, and itis important for the production of quality paper to identity the reactions causing this, especially since some take place shortly efter the papermaking process. Understanding the ageing will also help in determining the correct storage condi tions for historical documents. ur third article, by Berd Diehl, Frank Malzand Ulike Holzgrabe, describes the use of quantitative NMR in the pharma- ceutical industry, an area where it hes not been widely applied in the past Capabilities of gNMIR for drug quality evalation are highlighted, end show that NMR should be used more often when chromatographic methods are not work ing effectively Tony (A.N.) Davies’ topic for this monits columns the use of Adobe POF for archiving and storage of analytical deta, As most other publications, we use POF extensively: all advertisements are provided in PDF, proofs ave sent out as POF and, finaly, articles and columns are posted on the web as PDF. The current functionality of Adobe Acrobat means hat it can succesful be used in many areos including anahticol ercivng ond storege applications. Peter Jenks gets political in his RM Columa! Recent changes in UK govern ment departments have brought a cher isto the role of overseeing UK cherical metrology and have brought all aspects of metrology together which offers the opportunity for more logical manage ment. The second of our interviews with senior people in instument compa: nies is with Chris van ingen of Agilent Technologies. This i particualy timely, because we get Chis’ views before he teres atthe end of October Quatrry Wirnour Co WHY SETI WHEN THE a eS UE FOR L EST COSTS NO MORE? POU eee orgy ere re eee PURE: 99.99% & 99,999% — Caisse fluxes will not contaminate ferkeruresigelnreitbsonoorept eileen Ce Re Road CLAISSE yr 350, rue Fanquet, Suit 4, Sainte-Foy (Quebec) Canada GP 43 Tel: 418.656.6453 Fax: 618.656.1169. sopportecaisse.com www.claisse.com ‘PASTLINK / CIRCLE 002 FOR FURTHER INFORMATION ‘www. sgectroscopyeurope.cam ‘SPECTROSCOPYEUROPE 3SPECTROSCOPY re ee spect Cg lan Michael (pas ETS Pe rane ee on ey Tee ca) nue eee eee Peo pts bebe es coer peers et aT oe) Cee ead Beto ISSN 0966-0941 CONTENTS Editorial Hidden depths? New techniques for sub-surface spectroscopy Nell A, Macleod, Charlotte Eliasson and Pavel Matousek UV/vis reflectance spectroscopy reveals the changes in fibre chemistry during ageing Anna-Stiina J8askelainen and Tiina Lita Quantitative NMR spectroscopy in the quality evaluation of active gredients and Bernd W.K. Diell, Frank Malz and Ulrike Holzgrabe Tony Davies Column Adobe PDF use in analytical information storage and archiving RM Column UK chemical metrology: a new opportunity? News New Products Advertisers Index Interview Chris van Ingen of Agilent Technologies Diary Specroscopy Europe isa contvaied culation journal, published seven times a yeor and avalible free-of-charge to qualyng individuals in Europe. Others can subscribe atthe rte {of €120 (Europe), £83 (UK), S16D (ROW, postage incided) forthe sever ssues published in 2005. Al paid subscipton enuries should be addressed to: Spectroscopy Europe, John Wiley & Sons Ltd, Joumals Administration Department, The Atrium, Souther Gate, Chichester, West Susex POTS 850, Uk. Copyright 2007 John Wiley & Sons Ltd and IM Publications Al ihtsresecved. 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A dedicated team of scientists, backed by the resources of a company that’s 30,000 people strong, Stand behind our spectrometers Afully upgradeable, routine and edvanced research bench focused on power and Join the many satisfied customers who rely on Thermo sity, Scientific products they know and trust, like Nicolet FFAR. Visit wwvw:thermo.com/spectroscopy today to begin planning for your lab’s next spectrometer. Tel: 1-800-532-4752 / Email: analyze@thermofisher.com FASTLNA/ CIRCLE 003 FOR FURTHER INFORMATION Partof Therma Fisher Scientiio SCIENTIFICi SHIMADZU OLUELONS FOT SCLN Ce itVoL. 19 NO. 5 (2007) Hidden depths? New techniques for sub-surface spectroscopy Neil A, Macleod, Charlotte Eliasson* and Pavel Matousek* Central Laser Facility, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Hanwell Science and Innovation Campus, Didcot, Oxfordshire OX11 OQX, United Kingdom. E-mail: PMatousek@rlacuk “present address: Peek Oseberg South, Hydro, Sandsliveien 90, postboks 7190, 5020 Bergen, Nonway Introduction several hundred micrometes. Deeper A number of analytical appiaions in leyers cannot be readiy resolved and, _ ; the area of security screning, medical typically, are overwhelmed by Raman ~S : diagnosis, drug authentication and quak and fluorescence signals emanating from. a aD ity conte often require non-iwasive the surface layer F probing of difusely scattering turbid) : media in oder‘o obtain chemical cher. — Deep probing Raman Tani g LEE actefsaton ofeep-yng sampleregons. techniques : Eamples include non-invasive isease Recent research efforts by Everall tol? Figure 1. scheracilustatono he dlagnosis, the detection of concealed into time-resolved Ramen photon migr-_ransmssion Raman (fi) and SORS ‘explosives and ilicit material, te iden- tion in turbid media, in particular, have concepts (igh). tification of counterfeit drugs and quality stimulated substantial progress in the contol applications in the pharmaceuti-- area af deep probing of turbid samples cAlindustry, and led to the development of effective suppresses interfering Raman and fluo- Raman spectioscopy holds particu- spatial variants requiring only continu- rescence signals from the surface layers lat promise inthis area due to its inher ous wave laser beams. The methods ely and provides Raman spectra contain- ently high chemical specifiy [exceeding onthe ciffuse component of ight Two ing a varying degree of surface and sub- that of near inftared (IR) absorption variants in particular have emerged as surface content at diferent spatial fel; spectoscopy and comparable with mid+ promising practical tools: Spatially Offset complete separation of the spectrum of inreted end THz methods], the ality to. Raman Spectroscopy (SORS)* in which each layer can be accomplished using probe samples in the presence of water the Raman specta of individual sub- numerical methods.* This isin contrast (the Raman scattering cross-section of layers within a complex multilayered with conventional backscattering Raman waters very low) and its high penetra- system are isolated and transmission spectroscopy, where the solitary Raman tion depth into turbid non-absorbing Raman spectroscopy which provides spectrum avaiable is typically dominated ‘or weakly absorbing samples. On the information on buik content? (see Figure by intense surface layer components downside, the technique is rested to). Since the fst experimental dernonsta- ‘samples that do not exhibit strong fluo- tion of the SORS concept on powders,* rescence emission although this poblen SORS. the technique has been used in numer- «an, in the majority of cases, be avoided The SORS method is based on the ous applications including the demon- by using NIR excitation.’ Until recently, collection of Raman signals from spatial __ stration of Raman tomography in turbid Raman techriques have general been regions offset (As) from the point of media, now imasive Raman spectoscopy confined to applications involvingsurface _ilumination on the sample surface. The of bones in the pharmaceutical indus- layess of tubid media due to limitations laterally offset spectra contain differ- ty” and in security screening applica- imposed by the backscattering colection ent relative contributions from sample _tions.* geometry common to the majority of layers located at different depths due commerce Raman probes In pinciple, to the wider spread of photons emerg- ‘Transmission Raman confocal Raman microscopy can poten- ing from deeper layers on the semple Inthe transmission Raman concept, the tially resolve objects to depths of up to surface. Consequently, SORS effectively laser beam is incident on the sample ‘wn spectroscopyeurope.com ‘SPECTROSCOPYEUROPE 7VOL. 18.NO. 5 (20071 AN op AS concealed H:02 Me Dk Wavenuarbes(" Figure 2. Detection of lqué explosives. Convertonal backscattering Raman (CR) and processed SORS Raman specta ofa white past jar with e thickness af 1.2mm filled vith H,0, (30% aqueous solution) The bottom trace i the Raman spect of the empty jar tsof and is essentially identical othe conventional Raman spectrum ofthe iat containing 11,0, wth no absous tace of the Rarran signature of H,0, The SORS specu, onthe ether hand, shows cleat the HO, ‘Raman marker band. [Reprinted with permission flor C. Elasson, NA ‘Macleod and P, Matouses, ‘Non-inasive Detection af Concealed tiqus Explosives using Raman Specvescopy’, Anal. Cher. 2007, in press. Copyright (2007) Arserican Chemical Socety} from one side and the Reman signal collected from the opposite side (Figure 1). This concept can be considered as 2 special case of SORS with the illumi- ration and collection points situated at extreme displacement} Although the transmission Raman technique was demonstrated in the very eatly days of Raman spectroscopy! ts benefits forthe ‘nom invasive probing of the bulk content of turbid samples have not been previ- ously recognised. In particular, these include the removal of the so-called ssub-sampling problem (oversensiity to the surface layers of the probed medium) and the effective suppression of fluorescence components emanating from surface layers." (SORS) 8 SPECTROSCOPYEUROPE ‘The two techniques and their appli= tions are reviewed in Reference 3. In this article we present a brief snapshot of potential applications in security and in the pharmaceutical industy. Non-invasive detection of concealed explosives The recent heightened terrorist threa underlines the importance of robust security screening techniques wi chemical specificity. Recently, we have ddemoastated the applicabiity of SORS to the detection of concealed liquid explosives Uinougl tubid plastic packag ing’ In this work, hydrogen peroxide, @ citcal constituent of home-made explo- | SoRS coed h oe fac lh am ei how Wovetunbes (or) Figure 3, Detection of counteret drugs Nom invasive Ramen spectra ‘of paracetamol tablets measured through 9 white, dfsy scattering 1.7mm thick plastic container. Canvenional Raman and SORS raw data are shown together wit the reference Raman spectrum of paacete ‘mal, The processed SORS spectrum matches well that of paracetamol, ‘while the conventional Raman spectum is dominated by Rarnan signals criginaing rom the batle wall (Adapted with perission for Casson enc P. Metousek Anal Cherm. 79, 1696-170) (2007) Copyright 2007) American Chemical Society sives used recently in several terrorist bombings across Europe, was detected through various packaging materials. Figure 2 shows the results of probing 2 small white plastic jar containing 1,0, (30% aqueous solution). Such plastic jars are cormmonly used by travelers to nsfer a smaller ammount of moisturising ream or other products for te purposes of travel to save space and meet current travel regulations in cary-on luggage. The high containers highly ifusely scattering and presented an insurmountable challenge {to conventional Raman spectroscopy; the signature Raman band of HO; at 76cm” is completely overwhelmed by Raman signals originating from the container wail. in contrast, SORS, after ‘ynwnw.spectroscopyeurope.comVOL. 19ND. 5 (2007) = “blind” automated data processing using spectra obtained at two diferent spatial ‘offsets, clearly revealed the H,0, Raman marker band, The experiments were performed using a continuous wave diode laser (830m, 250 mW); an acgui sition time of 1s was sufficient to obtain good quality spectra, Non-invasive detection of counterfeit. driigs (SORS) ‘Another hot topic is the non-invasive detection of counterfeit drugs through plastic bottles and blister packs. These drugs present an increasingly severe threat to health and life in our society. Recently we have demonstrated” that ‘SORS can provide a chemical signature ofthe intemal content through unopened plastic containers with higher sensitiv- ity than that available with conventional Raman spectroscopy. This is demon- strated in ure 3 for peracetero! tables held inside white plastic pharmaceu- tical bottles. The conventional Raman approach, dominated by the Raman signal originating rom the container wall, is dearly nettectve. In contast the SORS approach, following the scaled sublrac: tion of two SORS spe-tra obtained at different spatial offsets, provides a clean Raman spectium of the tablets held inside the bottle. The experiments were performed using a ccntinuous diode laser (827nm, SOM) with an acquis tion time of 105. A pradcal use of SORS in the identification of “authentic” fake tablets was very recently demonstrated on ant-malarias by Ricci et at As the SORS concept can be easily incorpo- tated into existing commercial hand-held Raman instruments it holds great prom- ise for more accurate ard sensitive iden- tification of drugs at each stage of the supply drain, Bulk probing of pharmaceutical products In quality control (transmission Raman) In a number of pharmaceutical process analytical technology (FAT applications itis essential o monitor the bulk content cof pharmaceutical products. Ideally, this shoul be accomplished quick and non- invasively with high chemical specificity wan. szectroscopyeurope.com © z 3 z inputty 1100 1200-1300 14001500 160017001800 Weverumbers (er) z z 2 N00 1200 130014001500 600 «17001800, \Wavenumber (cm Figure 4, Removal of sub-sampling rablem in quality contol of pharmaceutical tablets. The Reman spectre obtained from a tw layersarple (e paracetara tablet and a 2mm thick ras stibene “impurity” layer using @) conventional backscattering geometry and (b) ansrission ‘geometry. The meesurements ate performed at two sarpe oiertations, with paracetamol at the top and bottom of the wans-stibene col as indicated. The top and bottom races are refer cence spectra of paracetamol and wons-stbene, respectively, obtained in separate expeiments ‘The vansmisson Raman spectra ate notable forthe absence ofthe sub-sampling problem, Le over sensitivity to the surface yer. Legend: P: paracetamol, T:rans-sibene, R: Rar ih, L? lsser beam. (Reprinted with permission ftom P Matousek and ALN, Parker, Appl Spectrosc 60, 1385-1357 (2006), Copright (2006) The Socety for Applied Spectroscopy.) -Athough NI absorption spectroscopy is _The experimental demonstration of the ‘widely used inthis area it provides insuf- elimination ofthe sub-sampling problem ficient chemical specicity in @ number was performed on a standard paraceta- ‘of applications. The conventional Raman mol tablet of 3.9mm thickness with a approach, on the other hand, suffers simulated impurity layer of a 2mm thick- from the sub-sampling problem, which ness of ors-stibene powder. The impur precludes the characterisation of deep ity layer was located ether at the font sample components, Research in this of the back of the tablet; conventional area has recently demonstrated the Raman spectroscopy is beset by the sub- removal ofthis effect with the tansmis- sampling problem and yields only the sion Ramen approach,® thus widening Raman signature ofthe surface layer in the prospects for the quantification of both possible orientations [Figue 4(@)] the bulk content of undisturbed tablets In contrast, the transmission geometry and capsules. [Figure 4(b)| provides a Raman spec ‘SPECTROSCOPYEUROPE 9VOL. 18.NO. 5 (2007) Cue acu ee Tt See eS eee) sensitivity for Confocal Raman eres unas cane ated with the Petey elas Erno Pen eee ae Tee oan eee of minutes, Confocal . Fluorescence Raman . SNOM . AFM Bie ‘FASTLINA/ CIRCLE O05, FOR FURTHER INFORMATION 10 SPECTROSCOPY Ral ‘rum comprising of a misture ofthe tablet and the "impurity; the similarity of the spectra in both sample ariontaions is expected for a tecinique that does not suffer from the sub-sampling issue. ‘The experiments were performed using 2 continuous 5 (627rm, BOW) and acquisition times ranging ‘The transmission Raman geometry appears to be very w suited to the requirements of pharmaceutical production lines, Lndetining the potential ofthis method to displace NIR absorp- tion spectroscopy in applications where higher chernical specificity is desited, Further studies are needed to establsh the technique's sensitivity lirits and validate its potent ‘accurate quantitative information on the composition of the probed sample, We are presently watking on establishing these pints in collaboration with major pharmaceutical companies. Our ate that the quantification of active pharma ceutical ingredients with errors comparable to established meth ods (eg. 1 ransmission Raman approach is indeed feasible. oreliminary data Conclusions The advert of SORS and the rensissance of transmission Ramen spectroscopy have stimulated the development of numerous new methods for probing tissue anc powders at previously ible depths, Many new exciting practical applications are ‘already looming on the horizon including the diagnosis of bone se, breast cancer detection, accurate quality control and rtication of pharmaceutical products, a5 well as the detec and liquid explosives through packaging Feferen 1. MJ, Pelletier, Analytical Application Blackwell Science, Oxford (1999). 2. N. Everall, 7. Hahn, P. Matousek, A: a .c 55, 1701 (2001) 3. P. Matousek, Chem, Soc. Rev. 36, 1292 (2007). 4, P. Metousek, LP. Clark, E.R.C. Draper, M.D. Mortis, Goodship, N. Everall, M. Towne, WF. Sinney and ANN. Patker, Parker and M. Tow, Spectrosc. 59, 395 (2005), P, Matousek and A.W. Parker, Appl Spectrosc. 60, 1353 (2006). 6. MY, Schulmerich, KA. Dooley, TM. Vanasse, S.A. Goldsteir and M.D. Monts, Appl. Spectrasc. 61, 871 (2007) 7. C.Ellasson and P. Matousek, Anal. Chem. 78, 1696 (2007). 8 C. Eliasson, N.A. Macleod and P. Matousek, "Non-inva sive Detection of Concealed Liquid Explosives using La! Spectroscopy’, Anal. Chem. (2007), in press. 9, B, Schwader and G. Bergmann, Zeitsch Chemie Freseniv 20 (1967) 10.5, Johansson, S. Pettersson and S. Folestad, . Pharmaceut Biomed. 39, 510 (2005) 1. P. Matousek and AWW, Parker, J. Rama 2007) 12, R. Mukhopadhyay, Anal. Chem, 79, 2522 (2007). 13. C Ricci, €. Eliasson, NA, Madleod, P. Newton, P.Matousek and SG. Kazarian, Anal. Bioanal. Chern, (2007), in press. jpectrose 38, 563 www.spectroscopyeurope.comVOL. 19 NO. 5 (2007) UV/vis reflectance spectroscopy reveals the changes in fibre chemistry during ageing Anna-Stiina Jadiskelainen* and Tiina Liitia? “TKK Helsinki University of Technology, Laboratory of Forest Products Chemistry, PO Box 6300, 02015 Espoo, Finland. E-mail: tina jaaskelainen@tkk i °KCL Science and Consulting, PC Box, 02150 Espoo, Finland. E-mail: tina litia@kel.fi Introduction Papers are susceptible to colour charges during storage. This is easily seer as 2 yellowish or brownish colour of old printed material, such as old books and Newspapers tis important to identify the reactions causing yellowing, as some of those take place quicky after papermak- ing and that way decrease the value of the material. Furthermore, understanding the reactions and mechanisms of ageing ‘would help in choosing the conect stor age conditions for historical documents. Pulp, the main raw material for paper, «an be produced by two different meth: ‘ds: mechanical or chemical pulping in mecharicel pulping, wood fibres are rst ‘separated mechanically and then the fibres are bieached if @ high whiteness level is targetec. Mechanical pulp is 2rla- tively cheap raw material for paper andit is used mainly in low grade papers, eg, newsprint. The chemical compositor of mechanical pulp is cose to the compo sition of wood. Hence, itis composed ‘mainly of polysaccharides (cellulose end hhemicelluioses) and aromatic lignin. Papers with better optical propertes, such as offce papers, are made from ‘chemical pulp. In chemical pulping, the wood fibres are separated chemically by depolymerising and dissolving ligrin in kraft pulping, which nowadays is the ‘most common pulping process, sodium lhydronide and sodium sulphide are used a lgnindissohing chemicals. After pup- ‘www. spectrascopyeurope.com ing, wood is defibrated, but the colour of the pulp has become dark brown, like @ cartonboard, Hence, the remain- ing lignin is removed by other chemi: cals, such as onygen, chlorine dioxide, provide and ozone. Because this entire proces targets the removal of lignin, the bleached chemical pulp is composed mainly of polysaccharides. However, some of the polysaccharides undergo chemical reactions during pulping and therefore their structure does not entirely correspond to the structure of native polysaccharides. Because mechanical and chemical pulps differ in their chemical composi- tion, their behaviour during ageingis also different! Yellowing of mechanical pulp is mainly @ photochemical reaction, Of course, other factors such as tempera- ture, humidty, onygen and pH also infl- ence the rate and the mechanisms of yellowing, but ight exposure isthe most critical parameter that initiates yellowing reactions, Light is absorbed by aromatic moieties in lignin. Ths is followed by radical reactions that result in formation of highly coloured structures, chromo- phores. These reactions are fest and Under direct sunlight mechenicel pulp turns yellow within 2 few hours, as can be observed visually when newsprint is left in sunshine, The understanding of yellowing reactions requies the identi- fication of the chromophores formed. However, their identification is challeng- ing. as ther concentration is low and they 2 bound in the fibre matric Yellowing of chemical pulp is @ thermochemical process, whereas light irradiation has only limited impact on brightness loss. In chemical pulps, the formed chromophores originate mainly ‘rom polysaccharides, while the traces ‘of ignin may aso undergo reactions that result inthe formation of coloured struc tures. Like the chromophores in mechan ical pulp those in chemical pups are also very low in concentration and bound to the fibres. This makes the analysis of chromophores difficult and only a few techniques are avaiable to characterise them Artificial aging The brightness stabiity of pulps can be tested by artificially ageing pulp hand- sheets under strictly controlled condi tions. For mechanical pulps, ageing is ‘most commonly performed by expos- ing the sempies to light that has a wave length range similar to sunight.On the ‘other hand, chemical pups are aged by heat treatment. There are several stand arcised ageing treatments induding both dry and humid heat treatments. Ageing in moist conditions [eg 80°C, 6546 rele tive huricty (RH), 48 hours), however, probably simulates more accurately the Conditions during pulp drying and stor age? ‘SPECTROSCOPYEUROPE 11VOL. 19 NO. 5 (2007) ‘he change in pulp brightness during which increases during light exposure. 300m, whereas remarkable changes light or heat exposure eflects the quan- On the other hand, the reflectance band in the 300-500nm spectral range can tity of chromophore formation during at 420-600nm decreases during light__be observed, The shoulder at c 350m ‘ageing As a standard measure, pulp irradiation, explaining the reduced bright’ reduced upon ageing, probably ilustiat brightness is equal tothe reflectance of | _ness ofthe sample, ing the degredation of conifer aldehyde anindefinely thick semple atthe wave- ‘The conversion of a reflection spec structures present in lignin. The absorp- lergts of blue light mainly at 457m. trum into an absorbance spectrum tions close to 315m and above 400m This peremeteris one of the mostimpor by applying the Kubelka-Munk equa- increased during light irradiation, indicat- tartin determining pulp quay, and typi tion (1) improves the applicability of ing the formation of conjugated cromo- caly mechanical pulps and chemical _UVAis spectroscopy for pulp and paper pores, probably arcarbory! structures pulps have brightness of 70-85% and samples. Paper's a ciffusively scatter- and quinones. 88-92%, respectively. During ageing the ing media, and reflectance is affected by Chemical pulps ae composed aimost bightness may decrease several units both absorption and scattering proper- entirely of cellulose and hemiceliloses and this reduces the veluc of the pulp. tes, as deserbed by the Kubela-Munk (ryan and glacamannan). Therefore the Even i rightness is an important quaty equation (7), where R,, stands for the brightness reversion takes pace in carbo~ parameter, it does not give information reflectance of an infinitely thick sample, _hyate movetes. The most heatsensitve fon chomophori structures contrbutg forthe light absorption coelident ands structures in these crbobydaiesouctures tothe pulp colour forte light scattering coefcient. are hexenuroric acid groups-conjugated k_0-RF side-gioups formed 1 altaline puping As UV/Vis spectroscopy as ss ()athermal degradation product of hexen: a tool to observe pulp 5 Re tronic acid, 5-formyl-2-furancarboxylic chromophores Ifthe light scattering coefficient in acid (FFA) is forme, which reacts further Uv/vis reflectance spectroscopy is a paper samples is considered constant, _and forms highly coloured small-molec- practical method to study the chemistry the k/s is directly proportional to light ular components.‘ It is noteworthy that ‘of pulp yellowing? The visible spectral absorbance and thereby pulp chromo- bth hexenuronic zcid and the primary region and reflection spectra are sensi- hore content. During ageing, the degradation product FFA are colourless, tive to the changes in coloured pulp scallering properties of paper remain but due to ther thermolablty hey both components determining the brightness, unchanged, end k/s can be used as have significant roles in brightness ever Absorption maxima in non-linear reflec such without resolving k and s separately. _ sion of chemical pulps. ticn spectra are not always easy to iden- When performing the conversion for the ‘The thermal degradation of hexen- ‘ify, but structural information on the reflectance spectrum, it is possible to. uronic acid can easily be followed by ‘chromophores can be obtained from the gather information on the light absorb-_UV/vis spectroscopy. Figure 3 shows & ‘spectrum, where the reflectance of an jing species, both in the UV and visible UV/vis absorbance spectrum of kraft “ity thick somple i plotted aginst ranges. Figure 2 shows the &/s spee pulp with a high hexenuronic acid wavelength, as is shown for a mechanical trum of mechanical pulps before and content. The strong band at 240nm_ pulp sample before and after light irad- after light exposure. t can be observed —_arises from hexentronic acid. After heat Sten fore hours (Fue 1). The mein that not many chenges occurred in treatment, the band hes decreased Giference oceus ate 340-400nm, the UV range between 200nm and in intensity, whie the intensity of wore ager oe a Son rear 3D i asengh or) reg) Figure 1. Refiectance specta of mechanical pulp before and atterlght ‘Figure 2. Absorbance (b/s) spectra of mechanical pulp before and ‘exposure for fve hous after ight exposure fo ve hous. 12 SPECTROSCOPYEUROPE wwww.spectroscopyeurope.comVOL. 19.NO. 5(2007) provides a very fast and simple method = HOA 240 nn) degeder dung case to study the changes in the concentre- tions of certain important pup compo: rents during ageing. In addition, the Degudtonyott ea 205-0) concentrations of these components unto athanss bans een) have been studied also in other pulp processes, such as mechanical and 3 1s chemical pulp bleaching References as 1. L Forsskahl, in Forest Products Chemisty, Ed by P. Stenius. Fapet, ° Helsinki, pp. 277-832 (2000), ee oe = 00 eee ro oe 2. T. Liitid and T. Tamminen, Proc. oe 5rd international Colioquim on Figure 3. Absbance is) specta of bleached cher pulp belo and afer heat reatment Fucolptus Kot Pulp. Belo Horizonte, R0VC, 65% Re and 48 hours Brazil (2007). 3. Schmidt and C Heiney in Advances band arising tom FFA at 2850m has Summary in Lgnocelulesics Characterization, increased, Simuleneoush, the absorb- __UV/is election spectroscopy isa prac Ed by DS. Argyropouios. TAI Press, ance at 320-500nm has increased. tical method to investigate pulp ageing, ‘lant, pp. 179-199 (1993). When heatireatment is prolonged, especialy when reflectance spectra are 4, O. Sevestyanova, J. Li and G the band at 285nm decreases and the converted to absorbance (k/s) spectra. Gellerstedt, Nordic Pulp Pop. Res. J. absorption inthe visible range increases Even if detalled reaction paths cannot 21, 188 2006). consequently. be solved with this technique alone, it cee ee eee eer eae or) Creat + Spectral respon z ete ree reeatis ”For the discerning collector with an eye for data quality the ECS-400 completes the set From 300MHz to 930MHz CISC A MULL ORR Ce eC * Compact NMR system The new ECS 400 NMR Spectrometer High Performance / Cost ratio www.jeoluk.com IPN ME MeL Nel ROL etaVoL, 18 NO. 5 (2007) Quantitative NMR spectroscopy in the quality evaluation of active pharmaceutical ingredients and excipients Bernd W.K. Diehl," Frank Malz and Ulrike Holzgrabe™ 'Specvalservice, Emil-Hoffmann-Str, 33, 50996 Kéln, Germany Federal institute for Materials Research and Testing (BAM), Richard-WillstétterSt. 11, 12489 Berlin, Germany Snsttute of Pharmacy and Food Chemistry, University of Wurzburg Am Hubland, $7074 Wurzburg, Germany Intreduction Quantitative (q) methods in nuclear rmegneticresonance (NMR) spectroscopy have been used successfully for many years, However there is a lack of general acceptance of gNMR in the pharma- ceutical industry, whereas chromato- arephic methods are well established and documented in analytical stend- ads, pharmacopoeia and drug master files. However, NMR spectroscopy can be considered as a quentilative primary ratio method of measurement, since the ratio of substances in a mixture can be determined direct from the NMR spectral measurement without referencing to another substance. The absolute amount of substances can be determined by using simple reference substances. ‘The purpose of this short review arti- cle is to highlight some capabilities of GNM spectroscopic methods in drug qualty eveluation, indicating that NMR spectroscopy should be more often applied when chromatographicméthods ‘are not working effectively. NMR spectroscopy can be used in diferent lds ofthe quality evaluation of drugs? W to identify a drug to determine the level of impurities and elucidate their structure, and 10 ‘wwn.spectroscopyeurape.com observe the course of decomposi- ton 1 to evaluste the content of residual sohents fm to determine the isomeric compos tion: the ratio of diastereomers and the enantiomeric excess (ee) by rears of chiral adltves to determine molar ratios of (proto: nated) basic drugs and (deprote- nated) organic acids in respective salts, Basic gNMR The most important fundamental rel tion for qNWIR is that the signal response: 4, (signal area) in a spectrum is directly proportional tothe number of nucle! Ny generating the coresponding resonance ine: by = KN oO with k; being a spectrometer constant. ‘The prerequisite for quantitative meas- Uutements is that femains constant for all resonance lines within @ NMR spec trum. & number of criteria have to be fulfled for ensuring this (1) The pulse excitation must be uniform for the entire spectral width (GW) of interest, which requires short pulses (typically 10s). In this case, 'H NMR spectre can be acquired quantit tively, However, spectra of heavier nuclei (CF, "P etc) with larger cherical shift ranges may suffer intensity distortion, pattculerly if measured at very high magnetic fields“ Q) The repetition time + (orrecycing time) depends on the longest longitu- inal relaxation time 7, ofall signals of interest. The T, relaxation is described br M,=mfi-e4| @ with 4, and being the magnetisation along the zavs (response factor) after waiting time + and at thermal equilid- rium, respectively. Routinely, measure- ments are done by choosing r= 5x T, Hence, 99.3% of the equilibrium magnetisation (signal) is measured.* For nuclei with very long M, values, e.g %C, Si, "18 etc, paremagnetic relax ation reagents such as chromium) acetylaceionate [Cr(acac),] are added in order to shorten the relaxation times, ) Heteronudeer NMR experiments of X nuclei (%C, "F,..) with simultaneous ‘ti broadbend decoupling cause inher ent intensity distortions by the nuclear Overhauser effect (ne). This nde effect ‘onthe spectrometer constant; can be described? by: ‘SPECTROSCOPYEUROPE 15,VOL. 19 NO. 5 (2007) kg=hy-(140):| 2 —} sina 1-cosa-e% @ where fois constant instrumental factor, 1 the nuclear Overhauser enhance- ant factor and ais the flip angle of the exctation pulse. n orderto suppress possible intensity distortions below 196, the following rules must be fulfiled: () "decoupling is apped only during the signal acquisition time (inverse gated technique) in order to minimise ny Gi) the repetition must be set to 5..7%Ty° ard (i) 90° pulse should be used for the excitation, ‘Additional parameters may affect the ‘measurement uncertainty (wiz. accuracy ‘ard precision) of the signal intensities in NuIR spectra: (4) The required acquisition time fay depends on the smallest line width in the spectrum, and truncation ofthe NMR signal in the time domain free induction decay (FID)] must be avoided. tf trunca- tion occurs, signal forms with (substan- tial) “wiggles” appear in the spectra, and, in combination with FID baseline corection modes, wrong intensities will result” Usually, the signel should decay completely half way through the acquis tion period. aso ensures that enough data points describe the NMR lines (at least five date points above the halt- height) such thet the integration proce dure does not cause artifical distortions (tensity error less than 146) °° (©) The accuracy of the integration procedure depends on the signal-to- roise-rtio (S/N). Fora precise integra- ‘ion a maximum S/N is tequited, For & precision better than 19, S/N of 250: 1 (CH), 300:1 (°F) and 600:1 €"P) have been proposed in the literature.” It should be noted tat there fs no eason forthese citferent numbers; one and the same S/N should yield the same preci sion, iespective of the NMR frequency (nuceus), Nevertheless, a detailed validation of QNMR using single pulse excitation of ‘HNMR spectra proved that the method is robust, if acquisition times, repettion tines and the signal-to-noise-etios are set according to rules mentioned above? 1 SPECTROSCOPYEUROPE Sample preparation Easy and fast sample preparation is possible for qNMIR purposes. For relative measurements, the anal is dssolved and diluted in an appropriate deuterated solvent. In contrast, for absolute measure- ‘ments, 2 one-point calibration has to be Cored out aking 2 sander of ran assay gravimetrically such that the inten- Sty ato ofthe ana signals of erst rite tarda ae nest ecu ame Specil requirements forthe standards have been described elsewhere!®'> Spectra evaluation integration) The operator is most commonly the main error source of NMR measure- ments for various reasons Baseline and phase correction must be performed with very high predéson to ensure accur rate results. It was found that auto- matic routines could not solve this problem in general; therefore the opera- tor’s experience determines the quality of the results.? Furthermore, the inte- eration mits forthe NMR lines have to be set according to the full width at half height (FWHH) of each signal. Extending the integration over a frequency range of 64 times the FWHH value ensures that 99% ofthe entre signal intensity (Lorentzian) is obtened. Quantitative NVR spectroscopy Flelative method The determination of ratios is the easi- est way to obtain quantitative results by NMR. The molar ratio n,/ny of two compounds X and ¥ can be calculated streigntowardly using Pe be Me yh My @) In this relation ks cancels provided itis, constant (@ necessary prerequisite, see above) for all lines. Consequently, the ‘amount fraction of a compound Xin a rmisture of rm components is given by: fu LiNe y00% 6) La Lay inespectve of the solvent signal in which ‘the mixture is dissolved. Furthermore, aNMR is a most impor tant method for quantifying the ratios of isomers, diastereomers and enenti- comers, because of its excellent selec tivity for structural analysis combined with the easy and fast quentfiation procedure. Even the knowledge of the molar messes ofthe components i not requited. I should be noted, that enan- tiomers, although having identical spec ta, can be dfferertiated by NMI too, i chiral solverts or complexing agerts'®"” are used Absolute method NMR offers two different methods for quantifying absolute values such 2s content concentration oF assay. {() fallimpurties show up in the NMR specttum, and ifthey can be assigned structurally and if they can be meas- tured quantilatively, the assay is simply the difference from the 100% value 60 called 100% method). This approach is limited for partially overlapping signals, and its impossible for impurties not containing the observed nucleus [e.g inorganicirmpurties (NaC) in case of 'H NWR (id The rein component P, can be caleulated directy from the NMR using ‘a standard of known assay Py be Nag, Meas ig 1H Mag with Mand being the molar masses of analyte and standatd,m and gy the "weights of the sample and stant, ‘and By and Pag the assays of analyie end standard, respectively. Furthermore, other procedures for ‘quantifying concentrations or assays exist using NMR. The use of an intemal standard is the most popular. If contarn- ination of the analyte by the chosen standard must be avoided, external standards Fave to be used. This can be achieved either by puting a capillary, filed withthe dissolved standard, inside ‘the analyte NMR tube or by using two (MR tubes one filled with the analyte solution, the other fled withthe stand- ard solation. In both cases analyte and standard should be dissolved in the same solvent and the volumes of the tubes (capillary) have to be calibrated wunw. spectroscopyeurope.comVOL. 18.NO. 5 (2007) [U0 == before. Capillaries are commonly used for the determination of deuterated assays or the water content of solvents. The standard addition method isa third possibilty of absolute value determina- tions. If known amounts of the active ‘compound are added to the solution in several steps the content can be cau lated without the knowledge of the molar mass of the analyte. lin 1998 Akoks and co-workers! Published a new technique called ERETIC (Electronic Reference To access 1n vivo Concentrations). Here, an elec: tronic reference signal is fed in the resonance circuit of the probe during the acquisition time using a tree coil in the probe (heteronuclear chan ne). Amplitude, FWHH and frequency (chemical shift position) can be set by the opetator. The acvantage is that this electronic signal can be shifted to free spectral range avoiding superposition with the analyte signals. Salecuvity The analyte NMR signal used for the quantification procedure must be assigned unambiguously in terms of the molecular structure (molecular sgf0ups). It must be ensured that this, ‘monitor signal belongs exclusively to the analyte without any contibu- tions from an impurity. If it neverthe- less happens, one can (i) change the temperature, (i) the pH value, (ii) add specific sift reagents, (jv) use another solvent or different concentration in order to separate such overlapping signals ifthese attempts fail, one may search for another (resolved) signal of the same impurity. If the specific impurity resonances can be assigned unambigu- ‘ously and determined quantitatively, the hidden impurity signal can be subtracted ‘rom the total signel area, ‘Another serious problem may arise ifthe impurity signals cannot by distin. guished from the analyte at all. This stuation was met in a purity determi- nation of atrazine containing simazine and propazine as impurities.”® The authors used elevated temperatures and homodecoupling to tackle the problem, \www.spectroscopyeurope.com Applicetions in internetional pharmacopoeias Due 1o the broad range of possibli- ties for application intemational phar- macopoeias such as the European Pharmacopoeia (EP, PhEur 6" edition), the Britis Pharmacopoeia (BP) and the United States Pharmacopeia USP29, cite use of NMR spectroscopy for iden- tification purposes and quantitative NMR spectroscopy for evaluation of the composition of polymers, mainly excipients and impurities in drugs. The PhEur 6" edition describes the method ‘of NMR spectroscopy only in principle using continuous wave (no longer in use nowadays) and pulsed spectiom- etry. In contrast, beside the descrip- tion of the physical background of MIR spectroscopy, the apparatus, the general method and the interpreta- tion of @ spectrum, the USP 29 gives detailed information about procedures to be applied for both qualitative and ‘quantitative purposes. For quantitative applications an absolute method utils- ing an internal stendaré and a relative method is given. A special working group on NMR spectroscopy in the European Directorate for Quality of Medicine has been commissioned which will adopt the general monograph on NMR spectroscopy in the EP to modern applications utlising 1D and 2D meth- ‘ods, cfferent methods of quantification, and liquid and solid phase measure- ments Currently the following applications can be found in the three aforemen- tioned pharmacopoeias, Identity PhEur 5.4: Buserelin (1H), Goserelin CO), Tobramycin ('H); BP 1998: Hydrocortisone Sodium Phosphate (1H); USP2S: Aminitite isomers (1H); PhEur 5.4: Heparins low-molecularmass (°C), vaccines such as Haemophilus Type b conjugate vaccine, Meningococcal Group ‘C Conjugate Vaccine and Pneumococcal Polysaccharide Conjugete Vaccine ‘(@dsorbed) (1H), and Salmon oil farmed {distbution of (8(2)-acyl of fatty acids CO). Tests DAB 9: Gentamicin ('H); Pheu 5.4 Poloxemer ratio of axypopylene/onyeth- lene; Hydroxypropylbetadex: molar subsituiion (H); USP2S: Orphenadrine ciate: meta/para isomer (H). Assay [USP29: Amyinitrte isomers (1H), For identification purposes the phar- macopoeias use NMR spectroscopy in a similar way to IR spectroscopy, ie. by @ comparison of the spectra of the chemical reference substance and the substance being examined. Thus, NMR specttoscopy is mainly applied to complex molecules such as, for exam- ple, hydrocortisone, goserelin, buserelin, ‘obramycin and different low-molecu- lar mass heparins such es delteparin, cenoxaparin, pamaparin, certoparin and nadroparn, Recently H VMR was intro- duced for charecterisatior of conjugated \accines inthe EP in addon to the das- sical methods Drug and active pharmaceutical ingredient analysis by gNMR Drug analysis is probably the most important appication for qNMR. In one step itis possible to measure several parameters for drug acceptance or stor age stability, NMR spectroscopy provides both identification and quantification of the active ingredient, of by-products and degradation products, of excip- ents and solvent residues. Applicaton of NMR for characterisation of reference compounds derived from natural mater- is wel established; thet relat was confirmed in round-obin sts Several ‘examples for drug analysis are reported in the literature, eg the quantification of inydroquinone in wound ue which is complicated for chromatogrephic meth- ods due to the reactivity ofthe acrylate system. In this short atic the potential of gNMR analysis willbe illusated on the basis of only afew spe examples. Forthe analyss ofa medial formulation containing 2 platinum complex the inter nal standard 3-G34-dinycroxyphery)-- dlanin (OHPA) was used. "he oxaliplatin as wellas the formation cd lactase can ‘SPECTROSCOPYEUROPE 17VOL. 18 NO. 5 (2007) sed as a metker to distinguish between raw materials. ny the green part of be | vera produces citicand iscitic acd, the latter is partly converted to the lactone, Ln if both molecules can be detected in an i no Aloe vera product, the sample is there fore either a whole leaf material or itis contaminated with pats ofthe cortex. Using the intemal standard method Intemel Standard | with cotnis acd amide (NSA) the char DHRA Oxsltan acterisation of each Aloe vera sample can be performed quantitatively or at least ssemi-quantatvely. Malic acid as well as, ‘glucose and the polymer Aloverose are ‘the main constituents of fresh Aloe vero gel and are quantified routinely by gNMR. - “ ‘Additional signals originatinge from WL | 96 888072 6456 ABAD BZ 24 16 OB OO |e ae quand the same nay Figure 1. qNiMR of an cxalziatin formulation using the internal standard DHPA (S00MH2). (Figure 2). in addition to the main constituents chemical and bacterial degradation prod- be quantified in an aquecus solution ple, the NMR specta of Aloe vera inner uc, additives (eg. preservatives) and containing added D,0 as the releence gel (CEL) and Aloe vere whole leaf (WL) _adulterating agents can be quentified in signal lock at O ppm) (see Figue 1). are discussed as examples for qualita the same NMR analysis Hetero nucle: containing drugs are tive and quantitative analysis to dstim- also targets for gNWR. Phosphorous _guish between different plant sources —_EXCipients containing molecules such as chloctonic and different qualities of extracts. For Besides the active molecules in drugs, or ethideonic acid have been analysed econamical reasons itis necessary to many excipients, including polymers, by routine qNMR for many years.” characterise whethera product was taken can be analysed qualitaely and often Quentitication and simultaneous dever- for the inner gel of Ale vera, which & _uantitatvely. Emulsifies are often a rmination of enantiomencexcess in the more expensive then producs from the mixture of similar molecules (isomers case of cyclophosphamide and related whole leaf. The compesiton of fruit acids or homologues, e.g, pareisononyiphe- substances using diferent cyclodextrin species hevo been reported. Meny — drugs and pesticides contain the hetero ruclei fiurine or phosphorous atoms. A 100% natural abundance of "Fand *P predestines NMR spectroscopy as the selective and sensitive analytical tool for {quantification of these chemicals, even, Fermulation ad | derived from the civic acid cycle can be Lectose in complex matices. The only require- ee | Malic Arr sr ment is the availability of 2 suitable “ * oN internal standard and probably also a valid exraction method, Hetero nudear tnt Si, NA teodiic acd NMR is an especially powerful tool in the analysis of by-products in amounts ‘of 0.19 and less, The structural simila rity of active compounds and their by- products results in an almost identical relaxation behaviour thus, by knowledge of the respective molecular structures no specific standard is necessary for quent= fication, Phytopharmaceutics are another oe domain of qNMR applications. For exarn-FAgre 2. "H-NMR spectum of Aloe vevo whole leaf (OO MH2) ‘18 SPECTROSCOPYEUROPE www.spectroscopyeurope.com9 8B FT 6 5 Figure 3. '¥ Nui specrum of NPE (SOMME). nol fatty acid mistues). By reaction of these basic materials (alcohols, amines, fatty acids, tiglycerides, carbohydrates, phenols et.) with ethylene oxide and/ or propylene oxide, polymeric srucures ate formed with @ wide variety of molee uler weights. The quantitation of emul- siflers by qNMR based on the ratio of ‘oxyethylene and ompropylene contain- ing copolymers is desenbed in EPS and USP29 (see above). similar procedure is used for characterisation of polylectid/ polygycolid copolymers. Furthermore, IC GNMR enables the determination of polyoryethylene chain length in PEG or ‘modified PEG ab initio by comparison of the integral areas of the terminal meth- lene groups with the sum of the meth- veene chain* (One representative "H-NMR applica- tion is the analysis of nonyiphenol poly- ‘ethylene glycol (NPE). The functional ‘groups of these mixtures cluster to sepa- rated signal areas in an NMR spectrum as shown in Figure 3, By normalisation othe integral areas 10 10 fora singe, proton the mean ethyt- ‘ene gycol (EO) distribution and the alkyl chain lengths are directly determinable. ‘The mean number of EO units (-CH,— CH1,-0-) in the respective sample is: = (19:954350)/40=9.25 www. spectroscopyeurope.com, ‘The integral value of the alkyl chain between §=05 and 20pm confirms ‘the presence of 19 protons in the alkyl part. Quantfication of NPE ‘rom complex formulations is possible by standard addition or using the specific integrals 3 5=6.8ppm (b) or at =4.1ppm (0) However, for calculation the aromatic rotons at '=6.8ppm are most suit able. Beside NPE, there are more complex emulsifiers based on alylated phenols. Differences in their molecu- ler structure can be identified by NMR spectroscopy. The terminal pars of the respective emulsifier motecules, eg, the ‘aromatic protons enable the calculation Of the polyethylene and polypropylene slycol (0) chain lengths and the mean ‘molecular weight Though these few examples the value and versatlty of aNMR spectroscopy to the quality and compositional analyses ‘of active pherraceutical ingredients ‘and organic excipients has been dearly demonstrated, and it is expected that its use in these areas of application will continue to increase significantly. References 1. Ti. Quinn, Metrologia 34, 61-65 (1997), 2. F Malz and H. Jancke, J. Pharm, Biomed, Anal, 38, 813-823 (2005). VoL. 19 NO. (2007) 3. U, Holegrabe, R. Detbner, ¢ Scholiayer and B. Waibel Phorm. Biomed, Anal. 38, 806-812 (2005) Freeman, Handbook of Nuclear ‘Magnetic Resonance. Addison Wesley Longman, Einburgh (1987), 5. F.ElShahed, K. Doerffel and R Radegla, Prat Chem. 821, 859- 864 (1979) 6. JR Mooney, in Ancijtical NMR, Ed by LD. Feld and S. Stemtell. John Wiley & Sons, Chichester (1989). 7 DIL Rabenstein, KK. Mill and EJ Strauss, Anal. Chem. 60, 1580A~ 13914 (1988). 8. FG. Hening and PS. Philips, 1 Mogn. Reson. 62,19 (1985). 9. J.P, Grivel, Signal Treatment and Signal Analysis in NMR. Elsevier, “Amsterdam (1996). 10. K McLeod and MB. Conisarow, J ‘Magn. Reson. 84, 490-500 (1989). 11. TS. Al Deen, Dissertation, Jrversity ‘oF South Wales, Syne, usta 12. L Grifiths and A.M. Irving, Analyst 123, 1061-1068 (1996), 1. CK. Latve, D.Jayaickrama and L Orfi, Appl. Spectose. 51, 1531-1536 997). 14. DL. Rabenstein and DA. Ki, Pract. Spectrosc. W, 325-369 (1991), 15. R Deubner and U. Holegrabe, Magn Reson. Chem. 40, 762-766 (2002). 16. GM, Hanna, Enantiomer 5, 308-512 2000). 17JS, Salsbury and PX. Isbester, Magn. Reson, Chem. 43, 919 (2005), 18S. Akoka, 8. Barentin and M. Tiemvele, Anal. Chem, 71, 2554— 2557 (1999). 19. GS, Remaué, V. Silvests and 5. ‘oka, Acted Quel. ASsus, 10, 415— 420 (2005). 20.F. Maz and H. Jancke,Anol Bioanal Chem. 385, 760-765 (206). 21.U. Holegrabe, R. Deubner, C. Schollmayer and B. Wael. harm. ‘Biomed, Ana. 38, 806-812 (2008). 22. BWK Diehl, Round-obin test, unpu- bished resus. 23.U. Holagrabe, |. Wawer and B. Diehl, NiR Spectroscopy in Drug Development and Anais. Wiley ‘VCH, Weinheim (1999), 24, Spectral Senvce, informaton avai tebe, » ‘SPECTROSCOPYEUROPE 1SVeb. 19 NO. 5 2007) pTONY DAVIES COLCONN Adobe PDF use in analytical information storage and archiving ALN. Davies. External Professor, University of Glamorgan, UK, Director, ALIS Ltd and ALIS Gmbli—Analyticel Laboratory Informatics Solutions Background There are more things in Heaven and Earth Horatio, Thon ore dreamt of in your philosophy Yes, the children are now studying Hamlet at school ané this seemed an appropriate quote to stat ofthis edition, When we set up ALIS GmbH one of the first major “discoveries” was probably the most embarrassing for ‘me. Having worked on analytical data standards for so long, I seem to have successfully generated a blind spot or the developments which have taken place in the structure, standardisa- tion and functionality of the Portable Document Format (POF)? Maybe its due to a subconscious aversion to what | hed for a long time seen as 2 simplistic “get out" solution for those too lazy to convert data into a long: term, stable, vendor neutral format How often have we heard the “wel. we just print to POF" as an excuse for ‘not having in place # property thought through analytical data storage and archiving policy teking no account of the future use to which that data may well be put within an organisation. Anyway, it has been prety difficult ‘or me to adit that my knowledge of the available functionality lay somewhere back in the eatty 1990s (see Figure 1) but hope in this column to make some amends! ‘We wil look atthe POF fle format and stendarcistion by ntemationa sanders bodies. On the functional side we will look at: 20 SPECTROSCOPYEUROPE Figure 1, Adobe historical company development leading to ISO standardisation of PDF/A fot ‘Document Archiving How the PDF document can work in the Analytical Laboratory environ- ‘ment not only asa stable document format but also communicating with ther computer systems such as 3 UMS or ERP software suite such 35 ‘SAP, essentially making the POF form into 2 database data erry and report ing tool. Integrating spectroscopic and other analytical data into @ PDF document. Wi The use of so-called rights enabled PDF documents within the free Adobe Reader software pack age providing additional function- ality such as editing ond saving content. in addition | wil riety outine how a PDF document can workin an anal ‘allaboratory environment puling and pushing data like 2 database front-end system. 1 To finish off| will very briefly touch on document security and digital signa- tures. History The "PDF format" is actually a series of formats which have buit on each other with increasing (and sometimes spect ically limiting) functionality (see text bon. The POF format was originally intended to provide a platform end generating software for independent documenta tion presentation. So whatewer compu- ter operating system is your personal favourite from Apple Macs, Microsoft ‘Windows or UNDCbased etc, you can receive, view and print dacuments fom wwa.spectroscopyeurope.comThe world leader in IR spectroscopy with a big reputation eae ee ee a ae SSC Mae age eu aa eee oe ea a eo ee nt er oad eee Te en cen nea ee eh cet Peas SM ce acces ean MCE ee eclataceroren GN Unread a eae Ronee MN er he at each eee Die i Rec eee Tac ee cir art From our flagship product the Golden Gato™ diamond ATR, through to our latest SID 1000 Portable Eee ENS eS Arr ee tte ees ed e.@®2,5\/ TD \g www.specac.comYUL 19 NO. 5 (2007) Formal agreed ISO PDF Standards PDF/X for the prirting and grephic ars. The guidelines for developing the PDF/X standards were puoished as Graphic technology-Prepress digital dota exchange Guidelines and principles forthe development of PDF/X standards 150 15929:2002 and a succession of PDF/X standerds were agreed under the general ISO 15930 ‘numbering wit increesing levels of functionality and allowed graphical types and handing options. 1 {50 15950-1:2001-Part 1: Complete exchange using OMYK data (POF/X-1 and POF/ia) 1m 150 15930-3:2002-Part 3: Complete exchange suitable for colour managed workflows (PDFX3) 1m 150 15930~4:2003~Part 4: Complete exchange of CMYK and spot colour pint- ing data using POF 1.4 (PDF/X-10) 1 ISO 15930-5:2003-Part 5: Partial exchange of printing data using POF 14 (PDF/ x2) 1m 150 15930-6:2003-Complete exchange of printing data suitable for colour- ‘managed workdows using POF 1.4 (POE/-3) PDF/A for archiving in corporate, govemment library etc. environments as ISO 19005~1:2005 published on October 1, 2005, Document ManagementElectrnic document file format for long term preservation-Part 1: Use of POF 14 (PDF/A-1). ‘Akey element to tis reproducbilyis the requirement for POF/A documents to be 100% seltcortained.$ 1m Audio and video content are forbidden. Javascript end executable file launches are prohibited 1 All fonts must ke embedded and also must be leglly embeddable for unimited, Universal rendering. This also applies to the so-called PostScriet standard fonts such 2s Times ot Helvetica 1§ Colourspaces svectied in a device independent manner 18 Encryption is dsallowed. Use of standards based metadata is mandated. Additional PDF standardisation under development targeting specific application areas: PDF/E currently under development for exchange of engineering drawings PDF/UA aizo under development for universally accessible PDF fle. PDF/H (PDF fr Heath Care Indust). Under development forthe sorage and tans- port of patient care data such as: tm Diagnoses: 1m Lab reports Pidures 1 X-Ray and Computer Tomography data ECG end EEG information et: colleagues who prefer to work in one ofA number of versions of the PDF the other emizonments without worrying format have been and continue to be whether the document you are viewing adopted as ISO standards (see text bos). is actualy showing the same image as Adobe published the first PDF stand- the original author intended. ard in 1993 and since then the stand- ‘Although the POFformatis proprietary it ard has been enhanced with edcitional is ako open and well documented allon- functionality with almost every major ing other software vendors to work with release of the Adobe Acrobat softwar. PDF formatted documents without resort The organisation coordinating these to licensing the format from Adobe. 150 efforts is called AlIM and was orig- 22 SPECTROSOOPYEUROPE inally founded in 1943 as the National Micofiim Associaton. later became the Association for Information and image Management (AIK) before mutating in 2 similar manner to ASTM by dropping its orginal ttle in favour ofthe infials end 's now known as ‘Al—The Enterprise Cantent Management Association".” AIM is accreditec by ANSI (American National Standards institute) as a stand ards development oganisation and holds the Secretariat forthe ISO (International Organization forStadarcization) commit- tee focused on Information Management Compliance issues, C1713 (OK, so POF is now an intemationally recognised documentation standard, but how can that help us in the analytical field? Well lets fook atthe internal wosk- ings of the PDF fie PDF structure What surprised me most wen learning about how far PDF formats had come while | wasn't wotching, was to lear about the inteligence underlying the Visible document which we are used to seeing | could wre for several hundred pages on the structure of the POF docu ment but asa picure paints a thousand ‘words you will be spared! Figure 2 shows a schematic representation of how a modern POF fle is built ‘The presentation layer is essentially what we see either on the computer screen or when printed out This may be ‘only fraction ofthe information actully stored within the POF fle down at the XML level asthe display is controled in part by speaficbusiness rules. tn an analytic laboratory this may well mean that a single POF form contains data entry fields for all the possible information to be captured for all the different types of analyses carried out in the company. A very simple POF presents itself to the user when a new work orders fist opened. say just enough information for the User to identify chemsehes and their group. This information can be used to Control the presentation layer restict- ing the available felds to those relevant to the actual scr. A user registering @ sample for analysis in the NMR labore tory would, for example, only see the www.spectroscopyeurope.com