Investigating the composition of the PPSQ-50 ... - Shimadzu · Shimadzu’s lab4you student ......

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Mineral oil compounds in paper and cardboard packaging

Investigating the composition of the aromatic mineral oil fraction

New generation of protein sequencer

PPSQ-50 with increased sensitivityand FDA compliance

The game changer redefines productivity

GCMS-QP2020 opens new horizons

Clinical

Chemical, Petrochemical,Biofuel and Energy

Higher performance with AtomicBooster – WizAArd software andAtomic Booster optimize AA-7000Fsensitivity 4

The key to success – Confirmed by headspace GC-MS: formation ofICN by lactoperoxidase 6

Tiny ivory chess piece? –Identification of ivory using infraredspectroscopy 10

Simple method development for SFC – Robust, reliable alternative to conventional LC 12

Colorful light for good mood …Emission measurements of glow sticks in party bracelets 14

APPLICATION

New application for the TOC parameter – Testing of plastic packa-ging in the pharmaceutical industry 9

Wanted: Clever Minds! Shimadzu’s lab4you student program 21

Mineral oil compounds in paper and cardboard packaging –Investigating the composition of the aromatic mineral oil fraction 22

LATEST NEWS

MARKETS

The game changer redefines productivity – GCMS-QP2020 opensnew horizons 2

New solution for the UV-VIS-NIRrange – ISR-1503 series 8

Hazardous phthalic acid esters detected quickly with Py-Screener 16

Newborn screening – NeonatalSolution software 18

New generation of protein sequenc er – PPSQ-50 with increasedsensitivity and FDA compliance 20

PRODUCTS

SHIMADZU NEWS 1/2016

CONTENT

W ith the introduction ofthe QP2010 series inearly 2000, Shimadzu

made a big step in the GC-MS

market and has within a few yearsreached a main player position inthe mass spectrometry segment.Now, after a series of innovativeproducts and technologies (pleasesee separate box), Shimadzu intro-duces the new GCMS-QP2020combining all outstanding GC-MSfeatures and opening new hori-zons for the users.

Three choices of carrier gas

With Fast GC-MS technologiesembedded, it is already commonto use hydrogen as carrier gas onShimadzu’s GC-MS. The QP2020now offers a third choice withnitrogen in addition to standardhelium and hydrogen.

GCMS-QP2020 opens new horizons

The game changer redefinesproductivity

Food, Beverages, Agriculture

Pharmaceutical

Environment

Plastics and Rubber

Figure 4: LabSolutions Insight environment

PRODUCTS

3SHIMADZU NEWS 1/2016

SmartSIM reduces developmenttime and improves sensitivity

Users of GCMS triple quadru -poles from Shimadzu are familiarwith SmartMRM technologywhich greatly simplifies the cre-ation of analytical methods. Thenew GCMS-QP2020 gives accessto the same SmartSIM feature,

Shimadzu’s GC-MS product lines in brief

Fully compatible GC-MS systems

Shimadzu was the first to introducethese instruments for both normaland Fast GC-MS analysis supporting0.32 / 0.25 / 0.15 / 0.1 mm innerdiameter capillary columns withoutany hardware change. In collabora-tion with Professor Luigi Mondello’steam at University of Messina (Italy),this lead to a compendium on FastGC showing critical parameters on asystem to adjust in order to performgood Fast GC analysis.

GCxGC-qMS known as comprehensive GC-MS

Introducing the fast scanning GC-MSwith 10,000 amu/s and later 20,000amu/s with no skewing in spectra,

Shimadzu made a new breakthrough.Together with Chromsquare softwarededicated to GCxGC data processing,complex samples such as petroleumextracts, flavor and fragrances sam-ples and environmental or food sam-ples can be screened easily to detectdifferent classes of compounds oridentify new compounds in a complexnatural extract.

MDGCMS multidimensional technology

This unique tool allows food and fla-vors companies to check for adulter-ation of the raw materials they buy.

MDGCMS is an exceptional systemfor working on flavors, e.g. in wineand coffee, when coupled with snif-

fers in the first and second dimen-sion. MDGCMS with two ovens and adean switch tool avoids any retentiontime shift when selecting a windowof time of the chromatogram to bediverted to the second dimension forhigher separation.

Twin Line MS

Thanks to the split dual stage turbo-molecular design, this configurationallows installation of two differentpolarity columns on only one GC-MSsystem, thus reducing pumpdownneeded to exchange columns whenco-elutions have to be resolved, orwhen different types of samplesrequiring either different columns orinjection systems are analyzed on thesame machine.

Powerful technologies andaccessories

Design of high performance quadru -poles, unique configuration of the ionsource with shield plates and dual fil-ament as well as overdrive lens, con-tribute to the most flexible GC-MSplatform available today.

Figure 1: GCxGC-qMS (ZX2-2010) Figure 2: MDGCMS

bottleneck. To sustain the increasein throughput which is demandedby most routine labs nowadays,the LabSolutions Insight platformaccelerates quantitative mass spec-trometry data review while open-ing a new approach in exception-based reporting. The intuitivedesign makes it simple to use andeasy to adapt to individual work-

Further information

on this article

• Application Handbook

Fast GC/GCMS

Figure 3: LabSolutions Insight

reducing development time andimproving the sensitivity of thechosen meth od by optimizingscheduled windows for SIM analy-sis and increas ing the potentialnumber of compounds that can beanalyzed in the same run.

LabSolutions Insight platformaccelerates quantitative MSdata review

Today, laboratory efficiency isdriven by highly automated massspectrometry platforms, deliveringlarge volumes of high quality data.However, manual data review canlimit sample turnaround times andreduce productivity, resulting in a

flows in the lab. A collection ofconfigurable flag criteria based onindustry-standard quality rulescan quickly identify peaks re -quired for data review. A click ofa button automatically showspeaks for review.

The LabSolutions Insight platformsupports data review over a net-work, creating new opportunitiesin remote data review, and enablesmultiple pane display for twomonitors.

APPLICATION

4 SHIMADZU NEWS 1/2016

F lame AAS is one of the fast -est methods in elementalanalysis. However, sensitivi-

ty of the measuring method oftenplays a decisive role. How cansensitivity be optimized? Thisarticle presents an overview ofvarious optimization approachesusing the AA-7000F atomic ab -sorption spectrophotometer.

In order to optimize sensitivity,the flame gas composition as wellas flame observation height can beadjusted. These instrument settingparameters are adjusted individu-ally as part of an automated opti-mization process using the associ-ated WizAArd software.

Fast method creation using the WizAArd software

The WizAArd software providescomplete instrument control.Basic parameters for the respectiveelements are also stored here. Inthis way, new methods can be cre-ated very effectively and requirevery little time.

Changer). This motor is also usedto adjust the burner height in theflame mode when not using thegraphite tube furnace. Flame gascomposition can also be adjustedfully automatically via the soft-ware.

Optimization of the burner height

In flame AAS, the liquid sample isfirst nebulized and subsequentlydesolvated by the thermal energy

Because of the flexibility of theanalytical instrument, it is particu-larly easy to optimize the burnerheight. It is possible to carry outmeasurements not only in theflame mode but also in the muchmore sensitive graphite furnacemode. This option can also beimplemented afterwards.

A software-controlled motor en -ables hands-free switching be -tween flame and furnace mode(AAC: Automatic Atomizer

(temperature) of the flame, andthe elements (present in the flameas ionic or molecular species) arefurther atomized into free atoms.Only in this way the measuringprinciple does fully apply.

Element-specific light is passedthrough the flame. The flame actsas a measuring cell, similar to thecuvette used in UV-VIS spectros -copy. The elements present in theflame can now absorb the ele-ment-specific light. The moreatoms are present, the stronger theattenuation of the light. In addi-tion to qualitative element identi-fication, quantitative measure-ments can also be carried out.

Depending on the characteristicsof the element being investigated,the atoms are present at differentheights (temperature zones) in theflame. If the flame observationheight (height within the flamewhere absorbance is measured) istoo low, the elements have not yetbeen atomized and cannot be de -tected correctly. If the flame obser -vation height is too high, the ele-ments may be present in theirexcited/ionized state, wherebythey are no longer detected.

The sample matrix can have anadditional effect on the flame. If, for instance, the sample con-tains organic compounds such asethanol or methanol, the flame isgenerally hotter. This means thatthe flame must be observed at alower height, or that the flow rateof the combustion gas must beminimized.

Example cesium

The element cesium (Cs) is usedto optimize the burner height.

WizAArd software and Atomic Booster optimize AA-7000F sensitivity

Higher performance withAtomic Booster

Height [mm] Height [mm]Flow [L/min]

Abs.

Abs.

Abs.

c)b)a)

0.000

0.025

0.050

0.000

0.025

0.050

0.000

0.250

0.500

0.7500.66630.075

0.075

0.100

5.0 12.0 2.41.5 2.5 0.0 13.1 17.0

Figure 1: a) Cs – Optimization of burner height; b) Cs – Optimization of flame gas composition; c) Cd Atomic Booster – Optimization of

burner height


APPLICATION

Cs is quite sensitive to atomicabsorption with limits of detec-tion in the double-digit ppb range.In this way, it outperforms ICP-OES (single-digit ppm range). The determination of even lowerCs levels is possible using graphitefurnace AAS (double-digit pptrange).

To optimize the burner height, aninterval is selected within whichvarious burner heights are testedunder simultaneous nebulizationof a Cs standard solution. Measur -ing values are subsequently deter-mined (figure 1a). In this case, thelowest observation height is theoptimal choice because Cs is ther-mally very unstable. Cs atomiza-tion already takes place very closeto the burner head.

Optimization of the flame gas composition

Two different gas mixtures aremainly used for the flame. In bothcases, acetylene (C2H2) is theflame gas. Different so-called oxi-dants are used: air for low flametemperatures and nitrous oxide(N2O) for higher flame tempera-tures. For most elements, air willbe sufficient to generate freeatoms in the flame. For refractoryelements with high dissociationenergies, higher flame tempera-tures are required (for instancealuminum or titanium).

Air-acetylene flames are selectedfor Cs. This is also stored in thesoftware. Analogous to the opti-mization of the burner height, aninterval is selected for the flamegas flow (acetylene). The resultingplot of absorbance versus flow rateshows a signal maximum at a flowrate of 2.4 L/min (figure 1). Thisparameter is now stored as theoptimum flow rate in the method.

Figure 2: Calibration of cesium and cadmium before and after individual optimizations. Details are provided in table 1

(Page 6)

Effects on the detection limit:cesium

Not only the sensitivity, i.e. thehighest measuring signal, but also

signal fluctuations have an effecton the detection limit. For thisreason, improvement of the detec-tion limit is determined via the linearity of the calibration (DIN32645). Comparing the values(table 1, page 6), it becomes clearthat the detection limit is alreadysignificantly improved by adjust-ing the flame gas composition.Adjustment of the burner height

Conc [ppm]

Conc [ppm]

Abs.

Abs.

a)

d)

0.00

0.01

0.02

0.03

0.04

0.05

0.06

0.07

0.08

0.09

0.10

0.11Abs.b)

0.00

0.01

0.02

0.03

0.04

0.05

0.06

0.07

0.08

0.09

0.10

0.11Abs.c)

0.00

0.01

0.02

0.03

0.04

0.05

0.06

0.07

0.08

0.09

0.10

0.11

0.00

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

0.50Abs.

e)

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

0.50Abs.

f )

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

0.50

0.00 0.05 0.10 0.15 0.20 0.25 0.30

Conc [ppm]0.00 0.05 0.10 0.15 0.20 0.25 0.30

Conc [ppm]0.00 0.05 0.10 0.15 0.20 0.25 0.30

0.50 1.00 1.50 2.00

Conc [ppm]0.00 0.50 1.00 1.50 2.00

Conc [ppm]0.00 0.50 1.00 1.50 2.00

Figure 3: Principle of the Atomic Booster – Dwell time of the atoms in the optical axis is

increased

brought about further improve-ment of the detection limit so thatit was possible in standard flameoperation to detect concentrationsas low as 0.011 mg/L. The effectsof the individual steps on the cali-bration of cesium are shown infigure 1 (a - c).

Atomic Booster – optimizationfor cadmium

As an alternative to cesium, Cad -mium was used for optimization.Cadmium is a heavy metal and istoxicologically very harmful. It isalready considered poisonous atlow concentrations and has beenproven to be carcinogenic as wellas mutagenic and teratogenic. Forhumans it is not required physio-logically (non-essential element).This is why trace analysis of thiselement is of critical importance.

To improve the detection limit ofthe AA-7000F for this element,the flame gas composition and theburner height were optimized inthe same way as for cesium, lead-

ing to calibrations with differingsensitivities (see figure 2 d, e). The detection limit for cadmiumcould also be lowered using theseoptimization steps. Instead of 1.5 µg/L, a detection limit of 1.0 µg/L could be achieved (table1). To further increase the detec-tion sensitivity for this toxicologi-cally relevant element, the AtomicBooster is used.

The Atomic Booster is essentiallya quartz tube that is positioned inthe optical path over the exit slotof the burner. The tube has twoslit-shaped openings opposite eachother that are different in length.The flame enters the quartz tubethrough the longer (larger) open-ing. Parts of the flame can subse-quently leave the quartz tubethrough the shorter opening.However, since the exit opening issmaller, a portion of the flame isheld within the optical path orleaves the quartz tube throughboth open ends (figure 3). �

APPLICATION


A s part of the innate immunesystem, the heme proteinlactoperoxidase (LPO) is

essential for defense againstpathogens in mucous membranesand bodily secretions (for instancesaliva, tear fluid, milk and respira-tory secretions).

This enzyme oxidizes thiocyanate(SCN-) and various small sub-strates (see box) in the presence ofhydrogen peroxide [1]. The result-ing hypothiocyanite (HOSCN/-OSCN) is bacteriostatic, passesthrough cell membranes and spe -cifically oxidizes thiols and sele -nols. Consequently, these func-tional groups efficiently activateintracellular proteins [2].

The catalytic and antimicrobialproperties of LPO are also usedby manufacturers of foods, cos-metics and oral care products,where LPO is used together witha hydrogen peroxide generating

system and (pseudo)halogens aspreservatives [3, 4]. In some ofthese systems, iodine is used asthe halogen because the resultinghypoiodous acid (HIO) and otherreactive iodine species such as I2,I3- and I2OH- offer a broaderspectrum of activity thanHOSCN: thioether groups arealso oxidized and iodine is addedto tyrosine residues.

In contrast to the LPO-H2O2-SCN- systems, the I--containingpreparation is also characterizedby bactericidal and antiviral prop-erties [5]. This raised the questionwhether thus far unknown reac-tive products of LPO were gener-ated. And indeed, the reactionproducts of LPO-H2O2-SCN-/Isystems predicted from the resultsof 13C nuclear magnetic resonancespectroscopy could be confirmedusing headspace gas chromatogra-phy-mass spectrometry (head-space GC-MS).

Confirmed by headspace GC-MS: formation of ICN by lactoperoxidase

The key to success

The lactoperoxidase reaction mechanism

LPO belongs to the group of heme peroxidases which oxidize various organicand inorganic substrates in the presence of H2O2. Their enzymatic activitiesare referred to as the halogenation (two-electron oxidation) and peroxidasecycle (two-electron oxidation).

During the halogenation cycle (reactions 1 and 2), the native enzyme is oxi-dized by H2O2 to the reaction complex I, which in turn oxidizes SCN- and I-.The myeloperoxidase present in neutrophilic granulocytes was also able tooxidize Cl- and Br-. In the case of the peroxidase cycle (reactions 1, 3 and 4),numerous organic substrates, for instance epicathechin present in green tea,are oxidized to their corresponding radicals.

software enable automated opti-mization of burner height andflame gas composition. Detectionlimits of 1 µg/L (for cadmium)can be achieved in this way.Cesium is also very sensitive witha detection limit of 11 µg/L.

Using the Atomic Booster, furthersensitivity increases can beachieved. The principle is quiteclear and the results speak forthemselves: the detection limit ishalved and at 520 ng/L and lieswithin the ppt range.

In this way, the dwell time of theatoms in the optical path as wellas the ‘thickness’ of the flame canbe increased.

To achieve a maximum energythroughput of the element-specificlight (good positioning within theoptical axis), the burner heightmust also be adjusted correctly.The lamp mode is set to emissionand the maximum burner height isdetermined (figure 1c). Measure -ments using the Atomic Boostertake place at a burner height of 13 mm.

By focusing the flame in the opti-cal axis, sensitivity increases sig-nificantly, as can be seen in the

Conclusion

Using different approaches, theinstrument sensitivity of the AA-7000F can be adjusted quick-ly and easily and optimized to thesample matrix at hand. The sup-porting functions of the WizAArd

calibration plot (figure 2f) in com -parison to measurements withoutAtomic Booster (figure 2 d, e).This increase in sensitivity lowersin turn the detection limit by afactor of 2, whereby the de tectionlimit for flame AAS lies within theppt range (0.52 µg/L).

Standard operationAfter optimized flame gascomposition After optimized burnerheightWith Atomic Booster

0.018 mg/L (fig. 2a)

0.013 mg/L (fig. 2b)

0.011 mg/L (fig. 2c)

–––

0.00151 mg/L (fig. 2d)

0.00096 mg/L (fig. 2e)

0.00052 mg/L (fig. 2f)

Cesium Cadmium

Table 1: AA-7000F – Limits of detection of cesium and cadmium before and after individual

optimizations (calculated according to DIN 32645). The corresponding calibrations are shown

in figure 2.


APPLICATION

Method

In the first step, the compoundsof the basic systems consisting ofLPO-S13CN- or I- were mixed inphosphate buffer pH 7.0, the reac-tion was initiated by the additionof H2O2 and the resulting reac-tion products were analyzed using13C-NMR. The second step in -volved the combined use of SCN-

and I- in various concentrationratios. It was found that with anexcess of iodine, a new unknownreaction product for LPO wasformed. Additional analyses re -vealed that iodide is first oxidized,this oxidized product subsequent-ly reacts with thiocyanate yieldingcyanogen iodide (ICN) [6].

Headspace analysis for the detection of ICN

To confirm the 13C-NMR results,a second detection technique wasapplied. As ICN is volatile, sam-ple analysis using GC-MS wassuitable. Due to the high volatilityof ICN and the solvent exchangeneeded from water to an organicsolvent, liquid injection was un -suitable for this investigationHeadspace analysis was thereforeapplied for the detection of ICN.

This method is an elegant alterna-tive to conventional GC-MS since,

analogous to NMR, no furtherworkup of the reaction mixturewas required. Another advantagein the analysis of this reactionwith unstable products was theshort amount of time needed tointroduce the analytes into thesystem.

The samples were analyzed usingShimadzu’s GCMS-QP2010 Ultraand the HS-20 headspace auto -sampler. By heating the reactionvessel to 60 °C, the analytes couldbe transferred directly into theheadspace and a GC-MS analysiscould subsequently be carried out.

Conclusion

It could be shown that a ratio of1: 2 of I- to SCN- will lead to theproduction of ICN (figure 1a),while an equimolar use of bothsubstrates did not result in theformation of a product (notshown). This is probably due tothe fact that the value of the rateconstant for the oxidation of I- byreaction complex I of the LPO isonly half of that compared to oxi-dation by SCN-. Therefore, therequired oxidation is not cata-lyzed.

The formation of ICN at an ex -cess of iodine could be confirmedby comparison with a reference

standard regarding retention timeand mass spectrum (not shown).Furthermore, S13CN- (figure 1b)and S13C15N- (figure 1c) in thesystem mentioned were measuredand supported the identificationof ICN. In the underlying spectrafor the molecular ion ICN++,shifts of the m/z values of 159.9 tom/z 153.9 and m/z 154.9 corre-sponding to the 1 u- and 2 u-mass increase of the marked ionswere detected.

In addition, the doubly chargedmolecular ion at m/z 76.5 (figure1a), m/z 77.0 (figure 1b) and m/z77.5 (figure 1c) was identified.Likewise, the corresponding frag-ments were detected. To beginwith, I+ with m/z 127.0 wasdetected in all spectra. Next, frag-mentation within the triple bondled to the formation of IC+ withm/z 139.0 (SCN-, figure 1a) andm/z 140.0 (S13CN- and S13C15N-,figure 1a and c).

The headspace GC-MS measure-ments confirmed that the com-bined use of SCN- and I- in anH2O2 activated LPO system con-tributes to the formation of a newproduct. How this product com-plements the described bacterio-static or bactericidal properties ofHOSCN or HOI/I2 must be clar-ified in further investigations.

AuthorsDenise Schlorke a, Jörg Flemmig a,

Claudia Birkemeyer b, Jürgen Arnhold a

a) Institute for Medical Physics and

Biophysics, Medical Faculty, University

of Leipzig

b) Institute of Analytical Chemistry, Faculty

of Chemistry and Mineralogy, University of

Leipzig.

Literature[1] J. Flemmig, J. Gau, D. Schlorke, J. Arnhold,

Exp. Opion. Therap. Targets 20 (2015)

11 -15.

[2] O. Skaff, D.I. Pattison, M.J. Davies,

Biochem J. 422 (2009) 111 -117.

[3] K.D. Kussendrager, A.C.M. van Hooijdonk,

Br J Nutr. 84 (2000) 19 -25.

[4] E. Seifu, E.M. Buys, E.F. Donkin, Trends

Food Sci Technol. 16 (2005) 137 -154.

[5] F. Bafort, O. Parisi, J.P. Perraudin,

M.H. Jijakli, Enzyme Res. 2014 (2014)

517164.

[6] D. Schlorke, J. Flemmig, C. Birkemeyer,

J. Arnhold, J Inorg Biochem. 154 (2015)

35 - 41.

[7] P.G. Furtmüller, W. Jantschko,

G. Regelsberger, C. Jakopitsch, J. Arnhold,

C. Obinger, Biochemistry. 41 (2002)

11895 -11900.

Time (min)

Relative intensity

a)

m/z

b)

c)

0.5

0.5

1.0

1.5

2.0

0.5

1.0

1.5

2.0

0.5

1.0

1.5

2.0

1.0

1.5

2.0

2.5

3.0

3.5

4.0

2.75

75 80 85 90 95 100 105 110 115 120 125 130 135 140 145 150 155

3.00 3.25 3.50 3.75 4.00 4.25 4.50 4.75

(Total ion current x 100,000)

(Ion current x 100,000)

Headspace GC-MS analysis of the reaction product of the LPO-H2O2-SCN-/I- system. In addition to the chromatograms,

mass spectra are shown for the reaction of 4 µM LPO with 10 mM H2O2 and 40 mM I- mixed with (a) 80 mM SCN-, (b) 80 mM S13CN-

and (c) 80 mM S13C15N-.

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PublisherShimadzu Europa GmbHAlbert-Hahn-Str. 6 -10 · D-47269 DuisburgPhone: +49 - 203 - 76 87-0 Fax: +49 - 203 - 76 66 [email protected]

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PRODUCTS

water. The near-infrared rangedoes not exhibit any ab sorptionof water, as can occur in bariumsulfate measurements in thismeasuring range, when the sam-ple is not embedded in BaSO4.

The ISR-1503/1503F includes aphotomultiplier for the ultravioletand visible range as well as anInGaAs detector and a cooled PbSdetector for the near-infraredrange.

High measurementstability

The 150 mm inte-grating sphere isexceptionally well

suited for samples with stronglight scattering and uneven andrough surfaces. The large spherecollects the strongly reflected lightand generates high measurementstability without being too strong-ly influenced by surface struc-tures.

Due to the design of the sphere, it is possible to place the sample

onto the sphere horizontally fortransmittance measurements. Forreflectance measurements underan angle of incidence of 0° or 8°,the sample can also be put along-side of the sphere (vertically) orunder the sphere (horizontally).This way, different physical prop-erties can be measured, such astotal transmittance, relative direct-ed reflectance and diffuse reflec -tance of the sample under investi-gation.

Properties:

• Wavelength range: 250 to 2300 nm

• Integrating sphere: 150 mminternal diameter

• Detector: integrated PMT,InGaAs and PbS detectors

• Sample positioning: transmit-tance, 0° reflectance – horizon-tal; 8° reflectance – vertical

• Aperture ratio: reflectance meas-urement (0°) 3 %

• Angle of incidence: 0°/8°• Material of the sphere:

ISR-1503: BaSO4ISR-1503F: Spectralon (Teflon)

Solar glass for photovoltaicthermal solar collectors

Reflectance spectra of solid mate-rials like powders, paper and tex-tiles can be measured using theISR-1503 series. Transmittancemeasurements of liquids andtrans parent solid materials can becarried out.

For solar glass, structures are ap -plied onto the glass surface. Inone method, specially designedmoulding rollers are used tostamp patterns on both sides ofthe glass surface. The structureson the surface must ensure a highsolar transmittance factor for per-pendicular irradiation.

In addition, a minimum loss ofreflectance for angles of incidenceof sunlight that are different fromthe perpendicular is required. Thisis specified as the angular factor.A high angular factor yields agood energy production balance.

Shown here is the measurement of a double-sided structured glasswith a layer thickness of approxi-mately 5 mm. The measurementwas carried out in the transmit-tance mode (figure 2). The trans-mittance baseline is at 92 % trans-mittance. This corresponds to theluminous efficiency that is meas-ured with flat glass in direct trans-mittance mode. Flat glass losesapproximately 4 % transmittanceon each surface under the condi-tion of direct transmittance.

Conclusion

Using the 150 mm-diameter inte-grating sphere, the determinationof the characteristics of a struc-tured glass sample is simple andfast. Combining three detectorswithin the integrating sphere al -lows measurements up to the NIRrange and also allows for the de -termination of heat permeability.Various transmittance and reflec -tance parameters can be deter-mined using the ISR-1503.Theaperture ratio allows for stablemeasurements of the structuredsurfaces.


New solution for the UV-VIS-NIR rangeISR-1503 series with large integrating sphere and threedetectors in two white standard versions

I nspired by the successful con-cept of the triple-detector ana-lytics for the UV-VIS-NIR

range, the large integrating sphereof the Shimadzu UV-3600 Plusseries has been redesigned. It has a diameter of 150 mm, and it ismade of either BaSO4 or Spec -tralon®. It integrates three detec-tors that can be jointly controlled

over the entire measuring range: a PMT (photomultiplier tube), anInGaAs detector and a PbS detec-tor.

In comparison with the predeces-sor model, its design has a morespherical shape in order to realizean aperture ratio of less than 3 %at 0° reflectance, even in the pres-ence of a third detector. This way,the sphere meets the requirementsfor applications with a low aper-ture ratio. This parameter is speci-fied by national and internationalstandard methods such as EN,ASTM and JISZ8722 “Methods ofcolor measurement – Reflectingand transmitting objects”.

Two versions of the ISR-1503 are available:

• the ISR-1503 coated with BaSO4

• the ISR-1503F coated withSpectralon (Teflon), which doesnot take up any moisture. Thisis an advantage for the NIRrange, which sensitively reactsto the presence of surface-bound

nm.

T %

101.112

50.000

0.000

-8.279250.00 500.00 1,000.00 1,500.00 2,000.00 2,500.00

Figure 2: UV-VIS-NIR spectrum of a structured glass measured with the ISR-1503F

Figure 1: View of the ISR-1503


LATEST NEWS

I n the pharmaceutical industryplastic packaging is used invarious forms – for example

for intravenous bags, bottles, car-tridges or pre-filled syringes. Thepackaging must be tested for suit-ability for these particular uses.

The United States Pharmacopeiahas published two new chapters tothis effect (661.1 and 661.2), whichwill be valid from May 2016.

• Chapter 661.1 describes thecharacterization and testing ofthe individual plastic materialsused in the manufacture of theplastic packaging.

• Chapter 661.2 deals with therequired testing of the finalpackaging system since packag-ing often consists of more thanone plastic material.

Characterization takes place byidentifying and determining thebio-compatibility, physio-chemi-cal characteristics and extractablemetals.

TOC – determination

The TOC parameter as an indica-tor for extractable organic materi-al is part of the physio-chemicalcharacteristics that must be deter-mined. For this purpose, the plas-tic material used is weighed,mixed with ultra-pure water andsubsequently heated. The amountused and the extraction tempera-ture employed depend on theplastic to be tested. The TOC ofthe ultra-pure water is subtractedfrom the measured value of theextraction solution. The resultantTOC value must not exceed 5mg/L.

For testing the packaging system,it is filled with ultra-pure water,

TOC-V series – wet-chemicaloxidation

The key technology of the TOC-Vseries is the powerful oxidationthrough the combination of sodi-um persulphate and UV oxidationat 80 °C. Since a persulphate solu-tion is used for the determination,it is important that it does notcontain any impurities that couldfalsify the actual measured value.For this purpose, the TOC-VWPhas an automatic reagent prepara-tion which removes potential im -purities from the persulphate solu -tion. This ensures that the TOCvalue recorded actually comesfrom the measuring sample – andnot from the reagents solutionused. Together with the largeinjection volumes (up to 20.4 mL)and the highly-sensitive NDIRdetector, this results in an ex -tremely low detection limit andoutstanding reproducibility in thelower ppb range. For this reason,the TOC-VWP/WS is particularlysuited to TOC determination onan ultra-trace level.

Summary

Both types of device with theirvarious oxidation methods aresuitable for TOC determinationaccording to the American Phar -macopoeia, USP <643> and USP661.1/661.2. Both include the re -quired measuring ranges: thelower range (0.5 mg/L) as well asup to 20 mg/L. The measuringranges are achieved by adjustingthe injection volume and then cal-ibrating accordingly. No furtheradjustments are necessary.

LiteratureSource: www.usp.org

able for TOC determination inpharmaceutical applications.

Two TOC systems for pharmaceutics

Shimadzu offers two systemswhich are perfectly suited for usein the pharmaceutical industry:• TOC-VWP/WS applies wet-

chemical oxidation• TOC-LCPH works with the

catalytic oxidation method at680 °C.

With their large measurementranges from 0.5 µg/L to 30,000mg/L, they support every applica-tion – from ultra-pure water tohighly-polluted water (e.g. fromcleaning validation to extractionsolutions to waste water).

TOC-L series – catalytic oxidation at 680 °C

The ISP module (IntegratedSample Pretreatment) for theTOC-L series significantly re -duces workload, since it carriesout dilution, acidification andsparging. The measuring range isexpanded by automatic dilutionfrom 4 µg/L to 30,000 mg/L.

Additionally, the combustiontechnology can be coupled withthe TNM-L module so that thetotal combined nitrogen (simulta-neous TOC/TNb determination)is recorded with just one injec-tion. Here it is worth referring tothe EN standard determinationconcerning chemiluminescencedetection. Catalytic combustionoccurs here at 720 °C. Simultane -ous TOC/TNb determination isparticular interesting for cleaningvalidation since a differentiatedconsideration between the clean-ing substance and the product ispotentially possible.

sealed and heated in an autoclave.The temperature and dwell timedepend on the plastic used. Inorder to determine the blank val -ue, ultra-pure water is poured intoa glass flask and is heated to thesame temperature. The TOC ofboth solutions is determined. Thedifference between the two meas-ured TOC values should not ex -ceed 8 mg/L.

TOC determination according to USP <643>

The general TOC determination isdescribed in USP <643>. The TOCsystems must be able to differenti-ate between inorganic and organicplastic; this can be achieved byremoving the inorganic carbon(NPOC methods) or by separatedetermination (difference meth -od). The detection limit is 0.05 mg/L. The suitability of the systemmust be verified in a system suit-ability test.

In the controlled ultra-pure water(purified water and water for in -jection purposes), the TOC mustnot exceed a value of 0.5 mg/L. In the actual application, the TOCvalue may be higher. Thus, a lin-ear range of 0.2 to 20 mg/L TOCis required.

TOC determination in pharma-ceutical applications

In TOC analytics, two oxidationtechniques have gained accept-ance: catalytic combustion andwet-chemical oxidation. Catalyticcombustion converts the carboncompounds into CO2 using hightemperature and a catalyst; it isthen detected with an NDIRdetector. Wet-chemical oxidationuses the combination of UV radia-tion and persulphate to achieveoxidation. Both methods are suit-

New application for the TOC parameter Testing of plastic packaging in the pharmaceutical industry

http://www.usp.org

F rom 1895 until 1910, a sugarmill named ‘Oriente’ existedon the Caribbean island of

Puerto Rico. After the mill closed,many heirloom pieces passed intothe hands of the next generationand are considered to be antiques.

Now in the third generation, thequestion arose as to the value ofthe objects. Their authenticity hasbeen confirmed. They are madeeither of natural materials whichare now banned (ivory for in -stance, under the so-called ‘Wash -ington Convention’, the Conven -tion on International Trade inEndangered Species of Wild Faunaand Flora) or of materials thathave been replaced by less flam-mable, cheaper or possibly moredurable substances.

Age and authenticity are criteriaused to classify these objects asantiquities. For age the followingapplies: the object must be at least100 years old. Occasionally, morethan 50 years has been specified.

Natural material or imitation?

The use of historical materials canbe a criterion for classifying ob -jects as genuine antiques and de -

was made by the company Aitken& Co. Aitken, as well as MirrlessWatson from Glasgow, suppliedmachinery to sugar mills in theUnited States around 1900. In thisway the folding ruler travelledfrom Scotland via New York toArecibo, Puerto Rico to the Ori -en te sugar mill and later on, afterthe mill was sold, to Bremen,Germany.

To find out if this folding rulerconsisted of celluloid, a diamond-

from nitrocellulose. They werenot as highly flammable and wereused in the production of manyconsumer goods or as substitutematerials (imitations) in the manu-facture of objects that would oth-erwise be made of ivory, amber,horn or mother-of-pearl.

Celluloid or mother-of-pearl or?

The small folding ruler (with met-ric and inch scale) shown in figure 1

ter mining that they are not animitation. Table 1 provides a listof ‘plastic’ materials from the past.Natural materials such as rubber,linseed oil or casein have beenused. Cellulose and nitric acidwere used to produce the firstartificial silk (nitrocellulose). Thistype of silk was also known asChardonnet silk.

By the addition of camphor as asolvent, it was possible to producethe first thermoplastics (celluloid)

APPLICATION


Tiny ivory chess piece?Identification of ivory using infrared spectroscopy

Figure 1: A folding ruler made around 1900

cm-1

Abs.

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

4,000 3,600 3,200 2,800 2,400 2,000 1,800 1,600 1,400 1,200 1,000 800 600 400

Figure 2: Infrared reflectance spectrum of an ATR measurement of the surface of a folding

ruler made of nitrocellulose

cm-1

cm-1

Abs.

Abs.

0.00

0.25

0.50

0.75

1.00

0.00

0.25

0.50

0.75

1.00

4,000 3,000 2,000 1,500 1,000 900 800 700 600 500 400

4,000 3,000 2,000 1,500 1,000 900 800 700 600 500 400

Figure 3: Search result for the surface of the folding ruler – nitrocellulose was identified


APPLICATION

ATR unit was used to record aninfrared spectrum of the ruler’ssurface (figure 2). The foldingruler was clamped against theATR unit and the surface waspressed onto the diamond win-dow. Figure 1 shows the ruler andfigure 2 the spectrum of its surfacemeasurement.

Via a library search, the spectrumcould unequivocally be assignedto nitrocellulose (figure 3).

Horn, tooth or bone?

Another interesting topic is ivory,which was in the past often usedto make jewelry.

Sources of ivory were not limitedto elephant tusks and rhinoceros

This noise effect is due to thesample not covering the measur-ing window completely.

The infrared library search led toapatite. This result is correct, asteeth or bone contain apatite. Ac -cording to the current literature,the apatite here is hydroxyl apa -tite, a calcium phosphate mineralthat, depending on functionality,

horns, as the long tusks of aquaticanimals such as walrus were usedas well. The Puerto Rico inheri-tance also included a tiny chesspiece made of walrus tusk (figure 4,photographed next to a one Euro -cent coin for size comparison).

A fragment was removed from thebottom of the chess piece and thissample was examined using ATRspectroscopy. The infrared spec-trum obtained exhibited inorganicsignals. Because ivory is a hardmaterial, the ATR spectrum alsoexhibited noise in the absorptionrange of diamond.

This amplifies the CO2 signalfrom air at 2,300 cm-1 and thenoise from water vapor in the airat 3,500 and 1,600 cm-1.

Figure 4: Mini chess figure made of walrus tusk, shown with a one Eurocent piece to visualize

the size of the figure

Table 1: Chronological assignment of the ‘early’ plastics

Figure 6: Result of the library search, infrared spectrum of apatite (1430, 860 cm-1 carbonate bands, 1050 phosphate bands) with traces

of protein deposits (1660, 1550 cm-1)

cm-1

Abs.

0.00

0.02

0.04

0.06

0.08

0.10

0.12

0.14

4,000 3,600 3,200 2,800 2,400 2,000 1,800 1,600 1,400 1,200 1,000 800 600 400

Figure 5: ATR infrared spectrum of walrus tusk ivory, recorded using a diamond

single-reflectance unit

is found in varying concentrationsin bones, hooves, horns or teeth.Figure 7 shows the spectra of dif-ferent sources (equine hoof, whalejaw bone, apatite mineral and thechess piece).

The spectra shown do not repre-sent pure substances, but as theyare natural compounds, their spec-tra arise from a sum of varioussubstances.

A collagen spectrum may be seenin the apatite spectrum as a fur-ther natural substance. Both theapatite region (1,200 – 800 cm-1) as well as the protein region (1,700– 1,400 cm-1) can be used for morespecific determination of the natu-ral material. It helps to distinguishbetween horn, teeth and bone andto use specific signals for agedetermination. [1] �

cm-1

cm-1

Abs.

Abs.

0.00

0.25

0.50

0.75

1.00

0.00

0.25

0.50

0.75

1.00

4,000 3,000 2,000 1,500 1,000 900 800 700 600 500 400

4,000 3,000 2,000 1,500 1,000 900 800 700 600 500 400

1839186018651885189719101913

Year

RubberLinoleumCelluloidChardonnet SilkGalalithBakelitePVC

Name

Natural rubber vulcanized with sulfurLinseed oilNitrocellulose and CamphorCellulose and Nitric acidCasein and formaldehydePhenol and formaldehydeVinyl chloride

Starting materials

APPLICATION


The measurements were car-ried out using Shimadzu’sIRTracer-100 and a Specac Questdiamond unit. The sample stage of the diamond window has adiameter of about 2 mm.

Summary

Infrared spectroscopy can be usedfor the direct analysis of naturaland synthetic materials. Usingspectral library searches, identifi-cation can be achieved quickly.

In this way, it is possible to deter-mine the material characteristicsof antique objects within a veryshort time. For ivory and wood, it is possible to determine the ageof the object.

cm-1

Abs.

1,700 1,600 1,500 1,400 1,300 1,200 1,100 1,000 900 800

Literature[1] „Die Anwendung der spektroskopischen

Analyse auf die Datierung von organis-

chen Materialien ist patentiert.“

It. Patent Nr. 01266808 – G. Matthaes,

1993

Thanks to Bernhard Westphäling for the

interesting topic, his consultation and his

participation as co-author.

Figure 7: Infrared spectra of bone, teeth and hooves of various origins and an apatite

infrared spectrum. In the walrus tusk spectrum, several signals are marked that are relevant

for identification.

Simple method development for SFC Robust, reliable alternative to conventional LC

A lready in the late 18th cen-tury, it was discovered thatheavy non-volatile organic

compounds could dissolve in cer-tain inorganic gases when thesegases are present above their criti-cal point, i.e. in their supercriticalstate. The first ideas to use thesegases as mobile phases in chro-matography were launched in1957. In the early 1990’s it wasshown that the addition of polarsolvents could achieve retentiontime control and this marked thebirth of modern supercritical fluidchromatography (SFC).

In supercritical fluid chromatogra-phy, so-called ‘supercritical’ car-

Fast, efficient chromatographicseparation

By using ‘su per critical’ CO2, chro-matographic separation becomes

supercritical state is non-polar andits solvent strength is often in -creased by addition of a polar mod -ifier. As soon as an ad ditionalorganic solvent is used, the mobile

bon dioxide is used as the mobilephase. Supercritical CO2 is de -scribed as a fluid state of carbondioxide, whereby it must be heldabove its critical temperature of31.1 °C and above its critical pres-sure of 73.8 bar. In this supercriti-cal state, the density of the medi-um is vastly influenced by changesin temperature and pressure, andits physical and thermal propertieslie between those of the gas andthe liquid phase.

The compressibility and the diffu-sion coefficient of the supercriticalsolvent are higher, while the vis-cosity is reduced in comparison tothe pure liquid.

Figure 1: Schematic representation of the Nexera UC method scouting system

Figure 2: GUI of the method scouting solution of the Nexera UC

phase is not truly ‘su percritical’ butthe terminology is used regard less.Many solvents are miscible withCO2 (for instance MeOH, EtOH[hydrogen bonds], ACN [dipoles])and offer additional interactions.

faster, more efficient, cheaper andmore environmentally friend lythan with the use of toxic organicsolvents such as hexane, heptaneor chloroform, as is customary innormal-phase HPLC. CO2 in the

APPLICATION


In this way, they induce controlledchanges in retention and provide avaluable tool to improve the selec-tivity of a separation.

Method Scouting for automatedmethod development

Method development is greatlysimplified using the new NexeraUC system with the meth odscout ing option. This option en -ables rapid testing of combinationsof up to twelve separation col -umns with up to four modifiers in addition to supercritical CO2(schematic representation figure 1).

The associated method scoutingsolution platform creates all scout -ing methods automatically in ac -cordance with predefined meth odparameters (figure 2). The differ-ent combinations of modifierswith various columns can also betested in combination with differ-ent gradients.

Separation of chiral compounds

A very good application exampleis the separation of chiral com-pounds, which are often best sepa-rated using SFC. For two newlydeveloped substances (PS 101 andPS 132) each having a chiral center,a good separation could be ob -tained quickly and easily using theNexera UC method developmentscouting system.

For this purpose, two modifiersmethanol and methanol containing0.1 % formic acid have each beentested at a column temperature of40 °C, a flow rate of 2.0 mL/minand an applied pressure of 150 barbehind the column and the detec-tor for different columns. Six dif-ferent chiral separation columnshave been tested: Lux Amylose-1and -2, Lux Cel lulose-1 to -4, allwith column dimensions 250 x 4.6 mm and a particle size of 5 µm.The individual chromatograms forsample PS 101 are shown in figure3. The Lux Cellulose-3 columnachieves by far the best separationof both enantiomers.

A very similar result in terms ofbest separation was achieved forsample PS 132 on the Lux Cellu -lose-3 column as shown in figure 4.For both separations, the simplestand best option is an isocratic sol-

vent composition with 30 %MeOH + 0.1 % formic acid overthe entire run.

Detection can be achieved using aphotodiode array detector (PDA/DAD) as well as using a massspectrometer (LC-MS).

Conclusion

A fast and simple method forscreening and separation of chiral

compounds on different columnshas been developed in a shortamount of time. A system wasused which allows testing of up to twelve columns in combinationwith four modifiers as additives toCO2. The method is optimized interms of separation and sensitivity.Simultaneous recording of a UVor a PDA signal together with amass signal via an LC-MS detectoris possible within one run. A se -lectivity of > 1.5 with an RSD <

2 % for the retention time wasachieved for all compounds shown.

With this system design, the de -velopment of an SFC separationmethod is very similar to that ofHPLC method development. Asshown in this example, supercriti-cal fluid chromatography offers a robust, reliable and simple alter-native to the conventional LC inuse for decades.

Figure 3: Chromatograms of PS 101 on different columns

Figure 4: Chromatograms of PS 132 on different columns

Luck

y Ly

nda

/flic

kr.c

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hoto

s/ol

sena

rt/

version process known as ‘peroxy-oxalate chemiluminescence’ takesplace. An example is the reactionbetween diphenyl oxalate and hy -drogen peroxide, as shown in fig-ure 3.

The oxalate ester reacts with thehydrogen peroxide to produce phe -nol (2ROH) and the short-livedand high-energy reaction interme-diate 1,2-dioxetanedione, which inturn reacts to CO2 in the presenceof the fluorescent dye. At the sametime, an electron in the fluorescentdye molecule is promoted to thenext unoccupied molecular orbital(excited state). This process is pos-sible because the electron hits theπ-electron cloud of the dye mole-cule (table 1, dyes).

Luminescence occurs when anelectron within the electron cloudis promoted into an energeticallyhigher unoccupied orbital level(excited singlet state). From thisexcited state, the electron fallsback to the ground state under theemission of photons (h*√) [1].This luminescence effect can beanalyzed using emission measure-ments in the visible spectral range.Table 1 lists some of the fluores-cent dyes used in this application.

APPLICATION


G low sticks are increasinglypopular at major eventslike concerts, carnival ses-

sions or New Year's Eve parties –even birthday parties. But they arealso used in high-performance ap -plications, for instance for illumi-nation purposes in professionaldiving processes. Or in general, asa light source when no other lightsources are available. They createcolorful lights and bring aboutgood cheer; they light up continu-ously or flicker, their luminositycan last for a short time or longer(figure 1).

Glow sticks have in common thatthey light up due to the power ofchemiluminescence and that theydo not require any external energysources (power outlet). Chemilu -minescence is caused by a chemicalreaction between certain substanc -es that results in the emission oflight (luminescence). Simple lightsticks (glow sticks) typically con-sist of a polymer tube filled with adye and a solvent. Inside the tube,there is a glass vial, which containsadditional chemicals that are need-ed to catalyze the chemical reaction.

Which substances and chemicalsare present in these glow sticks?What are their chemical proper-ties? Are they hazardous whenthey get into the hands of chil-dren?

Reaction fluids identified using FTIR spectroscopy

FTIR spectroscopy allows, with aminimum of effort, the determina-tion of the polymer tube material.An infrared library search identi-fied polyethylene as the best hitfor the polymer tube spectrum(left spectrum). To identify the re -action fluids in a glow stick aftercompletion of the chemilumines-cence reaction, the resulting yel-lowish, oily fluid was placed ontothe ATR measurement accessory

and measured using a single-reflec -tance unit. The result is seen in fig -ure 2 (right spectrum), an infra redlibrary search identified di meth ylphthalate as the best hit for this oil.

Phthalic acid esters (for instancedimethyl phthalate) are used assolvents for esters, like butyl ben-zoate. This is consistent with theFTIR measurement and identifica-tion of the oily fluid. According tothe packaging specifications, theglow sticks investigated containthe substance butyl benzoate. Thissubstance is an ester of butyl ben-zoic acid and is one of the chemi-cals in the chemiluminescence pro -cess discussed here. Lumines cenceis very intense and therefore onlya small amount of this substance isneeded for luminescence to occur.

What happens inside the glow sticks?

According to the instructions, thepolymer stick must be bent. Theresulting snapping noise is causedby the glass vial inside the polymertube breaking. The polymer sticksubsequently emits an intense glow. Depending on the fluorescentchemicals used, different colors oflight are emitted. A chemical con-

Colorful light for good mood ... Emission measurements of glow sticks in party bracelets

Figure 1: Glow sticks used for party bracelets

cm-1

Abs. Abs.

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

4500

Outside polymer tube of glow stick, Quest Yellow glow stick, oil, Quest

3000 2000 1000 500

cm-1

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

0.50

0.55

0.60

0.65

0.70

0.75

4500 3000 2000 1000 500

Figure 2: Infrared spectra of the polymer tube and the oily reaction fluid.

Substance

APPLICATION


How is chemiluminescencedetected?

Chemiluminescence emissionsfrom chemical reactions can bemeasured with Shimadzu’s RF-6000 spectrofluorophotometer.Luminescence qualification as wellas time-dependent observation ispossible.

For this purpose, a shutter wasplaced in front of the excitationlight inlet window of the optics of

the RF-6000 to prevent excitationlight from entering the samplecom partment. The second opticsfor emission measurement can nowbe implemented for measuring thelight emission of the glow stick.As glow sticks are very light inten-sive, the emission optics are attenu-ated using optical filters (mesh fil-ters) to reduce the amount of lightreaching the detector. Because theglow sticks used in this applicationare longer than the sample com-partment, the small lid inside the

sample compartment cover wasremoved to insert the sticks intothe sample compartment. To pre-vent external light from enteringthe sample compartment duringmeasurement, the opening wascovered by a blackout curtain.

All five bracelets contained in thepackage in the colors blue, yellow,green, red and pink-purple weremeasured consecutively. The as -signment of color is subjective. Itis therefore advisable to processthe obtained spectra from the visi-ble spectral region using color de -

termination software. The colorpink-purple can then be more un -equivocally assigned, as assessedaccording to standardized colorscales.

The emission spectra of the fiveglow sticks are shown in figure 4.The displayed colors of the spectracorrespond to the appearance ofthe luminescence.

Luminescence decay curve of a glow stick

As the light emission duration wasadvertised to be approximatelyeight hours, a corresponding ki -net ic study was started. For thispurpose, a glow stick was placedin the sample compartment formeasurement immediately afteractivation of the luminescence atknown luminescence wavelength(629 nm for red). It was found thatthe highest luminescence intensitywas achieved within the first sec-onds and minutes.

The emission initially decreasesstrongly and then turns into a faintglow, which continuously decreas-es over time to a constant value

Figure 3: Chemical reaction of the substances in a glow stick

nm.

Intensity

314,577.0300,000.0

200,000.0

100,000.0

0.0

300.0 400.0 500.0 600.0 700.0 800.0

Figure 4: Emission spectra of five party bracelets. The colors of the spectra correspond

to the glow stick colors.

Structure Color

9,10-Bis(phenylethynyl)anthracene

Rubren (5,6,11,12-Tetraphenyl -naphthacene)

Rhodamine B

Green

Yellow

Red

Table 1: Fluorescent dyes –

examples for red, green and yellow,

the ring systems and the multiple bonds,

are sources for high-energy π electron

clouds

Time (min)

Intensity721.511

200.000

400.000

600.000

0.000

0.0 100.0 200.0 300.0 400.0 480.0

Figure 5: Kinetics of the luminescence of a red glow stick over a time period of eight hours.

The curve shows the decrease of intensity versus time.

after about 170 minutes (figure 5,total time eight hours).

Conclusion

Glow sticks, which among otherthings are used as party bracelets,should not fall into the hands ofchildren. The polyethylene tubescontain chemicals that smellstrongly and can stain textiles. Inaddition, after bending the glowstick, the polymer tube containsglass splinters. The polymer tubeitself is quite thick but it can becut easily with scissors.

Under normal conditions, every-thing stays in the tube and can bedisposed of after use.

The chemiluminescent colors canbe determined using Shimadzu’sRF-6000 spectrofluorophotometerand the luminescence time is inagreement with the times indicatedon the glow stick packaging: atime period of about eight hoursin which the in tensity, as shownby the time curve measurement –the kinetics – de creases significant-ly within the first few minutes.

Literature[1] C.V. Stevani, S.M. Silva, W.J. Baader,

Eur. J. Org. Chem., 2000, 4037

[2] Shimadzu LAAN-A-RF-E003, application

News No. 493, „Measuring Peroxyoxalate

Chemiluminescence Using a Spectrofluoro -

photometer”

Further information

on this article

• Application: Measuring

Peroxyoxalate Chemi-

luminescence Using a

Spec tro fluoro photometer

PRODUCTS

E ach year, millions of tonnesof plasticizers are processedworldwide in order to make

brittle materials soft, flexible andmoldable. The most importantfield of application by far is plas-tics, in particular products madeof PVC, whereby the largest partis used in the manufacture of filmsand cables.

The industry uses a broad rangeof substances as plasticizers, thatare chemically very different. Interms of quantity, esters of phthal-ic acid (identified as phthalates)are currently still predominant.

What are phthalates used for?

Apart from use as a plasticizer inplastics such as PVC, nitrocellu-lose or synthetic rubber, phtha-lates are applied in many otherareas of daily life. They serve as acarrier substance for fragrances inperfumes, deodorants and otherpersonal hygiene products. Theyare components in nail varnishesand hair sprays. They are used asa formulation agents in pesticides,as industrial solvents and lubri-cants and as additives in the textileindustry. They can be found invarious other products such astoys or pharmaceuticals, e.g. as acoating for film tablets or as plas-ticisers in gelatine capsules(Deutsche Apotheker-Zeitung[German Pharmacist Paper]).

Phthalates are used for externalplasticization, meaning that theplasticizer is not bonded covalent-ly into the polymer, but instead

formation of certain tumours isalso being discussed.

Phthalate bans in Europe

In Europe, there are already manybans on phthalates. According to the EU chemicals regulationREACH (2009), plasticizersDEHP, DBP and BBP which arecurrently classified as toxic toreproduction, must not be presentin concentrations greater than 0.1mass percent in toys and childcarearticles for children under threeyears. This limit also applies toother phthalates (DINP, DIDP,DNOP), the use of which is pro-hibited as far as possible in toysand childcare articles which chil-dren could put in their mouths.Furthermore, some phthalate plas-ticizers must not be included inmixtures for private consumers orin cosmetic products. The use ofphthalates in plastics for foodpackaging has been restrictedthroughout the EU.

Finally, due to REACH regula-tions, companies producing orusing the phthalates DEHP, BBP,DBP and DIBP as of 21.02.2015may only do so with an excep-tional permission (approval) thatis difficult to obtain. In addition,the European Commission hasincluded these four phthalates inthe list of substances whose use isrestricted by the RoHS II Direc -tive (Directive on the Restrictionof the use of certain HazardousSubstances in electrical and elec-tronic equipment).

Also in this case, a maximum per-missible concentration of 0.1 masspercent applies per substance. Theenvisaged transitional period ex -pires for most device groups on22.07.2019. For medical devices as

phthalate (DBP), benzyl butylphthalate (BBP) and diisobutylphthalate (DIBP) are suspected tohave an endocrinologic (hormone-like) effect. These substances, alsoknown as endocrine disruptors,disturb the hormone balance andcan cause health problems if theyenter a human or animal in a suffi-cient concentration.

At present, the effect of substan -ces, in particular those whichinterfere with thesexual hormonesystem, are beingdiscussed.

It is assumed thatthey may impairfertility and thedevelopment ofchildren in thewomb or at cer-tain developmen-tal stages, such aspuberty. Theirinfluence in the

interacts with the polymer onlyvia its polar groups, increasingmobility between the polymerchains. Due to the lack of chemi-cal bonding, the phthalates can beextracted relatively easily from theplastic or can migrate out gradual-ly. In this way they then escapeinto the environment, and, fromvarious products, enter householddust or food.

Health risks

Phthalates have for some timebeen making negative headlinesdue to their potential health risk.However, it must be consideredthat the risks posed by individualcompounds varies widely. Theyare differentiated into low or highmolecular compounds accordingto the length of the esterified alco-hol chains.

The low molecular phthalates,including for example di-(2-ethyl-hexyl)-phthalate (DEHP), dibutyl


Hazardous phthalic acidesters detected quickly with Py-ScreenerEU guideline RoHS II will ban the use of four phthalic acid esters in electrical and electronic devices from July 2019

Figure 1: Pyrolysis GCMS-QP2020

Figure 2: Tool kit for sample preparation


PRODUCTS

well as control and measuring in -struments, a longer period applies– until 2021.

Fast detection of phthalates in polymer material

Current legislation requires quickand easy identification of phtha-lates in polymers. Existing stan-dard methods (e.g. EN 14372, ISO 8124-8, ISO 14389 – Analysisof phthalates in toys, childcarearticles and textiles) are based on aliquid sample preparation, mean-ing that the sample is extractedwith an organic solvent over sev-eral hours and subsequently ana -lysed with a coupled gas chroma -tograph-mass spectrometer (GC-MS). The ‘Py-Screener’ presentedhere is a screening method forphthalates which does not re quiresolvent extraction and offers excel -lent selectivity and the extensionof the method to new target mole-cules.

The Py-Screener system

Screening is carried out with acoupled pyrolysis GC-MS system.For this purpose, an aliquot of thepolymer sample (approx. 500 µg)is placed directly into the pyroly-sis furnace. The semi-volatilephthalates are extracted from thepolymer with a special heatingprogram. The desorbed phthalatesare transported by the inert carriergas into the gas chromatographwhere they are separated on ananalytical column and subsequent-

ly detected with a mass spectrom-eter. The screening system in -cludes an in-depth video tutorialin preparation of the analyticalstandards and polymer samples.

Figure 4a and 4b: Examples of the Maintenance Navigator Window

Since liquid extraction is omittedin the case of pyrolysis GC-MS, asmall piece of the polymer sampleis cut off, placed directly into asample cup and weighed. Thesoftware specially developed forscreening of phthalates providesall necessary, already optimizedmethod and instrument parame-ters as well as sequences for sam-ple measurement and the completequantification method including areport template.

Color-coding (Flagging) ofnoticeable concentrations

When an autosampler is part ofthe system, samples can be meas-ured easily overnight. The data

obtained are displayed in a tabularand graphical overview for quickevaluation, in which the valuesexceeding defined limits are color-coded. Analytical standards need-

ed to quantify the phthalates andcheck system per formance can beconveniently and easily punchedout of the standard material in a defined size using the sampletoolkit’s micro-puncher.

The so-called ‘MaintenanceNavigator’ supports regular main-tenance or trouble shooting proce-dures with detailed descriptions,comprehensive illustrations and avideo.

If required, the ‘Py-Screener’ sys-tem can also be expanded to coverthe other compounds already reg-ulated under the RoHS Directive,PBB (polybrominated biphenyls)and PBDE (polybrominated di -

phenyl ethers), which can be ana-lyzed with GC-MS. The opti-mized methods are already part of the package.

Summary

The ‘Py-Screener’ screening sys-tem provides a comprehensivepackage consisting of a toolkit forsample preparation, standards forthe analysis of phthalic acid estersand special evaluation software.Videos describing the accuratepreparation of standards and testsamples as well as regular mainte-nance of the pyrolyzer and GC-MS round off the system. Withthe complete Py-Screener method,even inexperienced users canquickly and autonomously learnhow to identify phthalates inpoly mers.

LiteratureOfficial Journal of the European Union

04.06.2015, Commission Delegated Regu lation

(EU) 2015/836 from 31st March 2015 amend-

ing Annex II of the Directive 2011/65/ EU from

the European Parliament and of the Council

regarding the list of substances subject to

restrictions.

Figure 3: Special evaluation software: For faster optical identification, differing concentrations of indicator substances are color-coded

Further information on this article• Brochure: Py-Screener • Application:

Analysis of Phthalate Esters using the Py-Screener

• Technical Report: Comparison ofScree ning Method (Py-GC/MS) and Quan -titative Method (Solvent Extrac tion-GC-MS) for Phthalate Esters Analysis

List of the target diseases in Germany

• Congenital adrenal hyperplasia(CAH)

• Maple syrup urine disease (MSUD)

• Biotinidase deficiency

• Carnitine metabolism deficiencies

• Carnitine palmitoyltransferase-Ideficiency (CPT-I)

• Carnitine palmitoyltransferase-IIdeficiency (CPT-II)

• Carnitine-acylcarnitine translo-case deficiency

• Galactosemia

• Glutaric acidemia type 1 (GA1)

• Hypothyroidism

• Isovaleric acidemia

• Long-chain 3-hydroxyacyl-CoAdehydrogenase deficiency (LCHAD)

• Medium-chain acyl-CoA dehydroge-nase deficiency (MCAD)

• Very long-chain acyl-CoA dehydro-genase deficiency (VLCAD)

• Phenylketonuria (PKU) and hyper-phenylalaninemia (HPA)

All tested metabolic disorders can beattributed to congenital enzymedefects.

Failing or incorrectly formed enzymeslead to degradation disorders in theorganism or rather an accumulationof toxic intermediate metabolic prod-ucts, small organic acids that causepoisoning and in turn lead to irrever -sible organ damage.

One of the most frequently occurringcongenital metabolic disorders isphenylketonuria, an amino acidmetabolism disorder that, if undetect-ed, leads to physical and mentaldevelopment disorders.

With a timely begun low-protein diet, however, the symptoms can beprevented.

Extended screening by tandemmass spectrometry

The decisive turning point in thisissue (please see box on page 19)was the introduction of tandemmass spectrometry in routine anal -ysis. Using this technology, it ispossible to simultaneously identi-fy a large number of disorders ofthe amino acid metabolism, themetabolism of organic acids andfatty acid degradation within oneanalytical run. In this way, notonly the number of marker sub-stances (metabolites) investigatedwas increased, but also many morenewborns with metabolic disor-ders could be tested and treated.

PRODUCTS

N ewborn Screening (NBS) isavailable in many Euro -pean countries. The blood

of newborns is tested for raremetabolic disorders within a fewdays following birth. When suchdiseases remain undetected, thiscan lead to massive health damageor even to death in infancy. Someof these tests are carried out usingtandem mass spectrometry (MS/MS). The ‘Neonatal Solution’software package supports rapidand routine evaluation of themeasurement results.

be treated very well with a specialdiet, vitamins or hormones.

Inborn Errors of Metabolism(IEM) of newborns as early aspossible. European countriesscreen for different types of disor-ders. In Germany for example,this comprehensive screening testwas included since 2005 in thenational screening program andsince then every newborn child inGermany is currently tested fortwelve disorders (see box) afterwritten parental consent.

These disorders include aminoacid metabolism disorders such asphenylketonuria (PKU) or maplesyrup urine disease, as well asfatty acid decomposition deficien-cies in which the transport andoxidation of fatty acids in mito-chondria is impaired (medium-chain acyl-CoA dehydrogenasedeficiency (MCAD) and verylong-chain acyl-CoA dehydroge-nase deficiency (VLCAD); carni-tine cycle defects). The screeningalso includes diseases such as thefrequently occurring hyperthy-roidism, an excessive productionof thyroid hormone, or the veryrare galactose mia in which toomuch galactose is present in theblood. Most of these diseases can

Early screening of metabolicdisorders

Newborn screening is a preventivemedical measure to detect andeffectively treat some important


Newborn screening – just a few drops of blood for healthydevelopmentNeonatal Solution software for rapid evaluation of screening data

Figure 1: The LCMS-8050 tandem mass spectrometer

Figure 2: Start window of the Neonatal

Solution software


PRODUCTS

tion of conspicuous concentra-tions it is possible to define limitvalues for each marker substance.When the limit value is exceeded,the deviating values are flaggedusing a color-code.

Summary

Through newborn screening, dis-orders that are clinically not yetapparent can be diagnosed and inmost cases treated at an earlystage. Even when potential diseaseconsequences cannot be preventedfully in all cases, timely treatmentwill often ensure a largely normaldevelopment.

Screening laboratories must meethigh demands in terms of qualityand speed, as the samples must beanalyzed on the day of receipt.Currently, eight of the total oftwelve tested disorders can bescreened using LC-MS/MS analyt-ics. The Neonatal Solution soft-ware with its functions custo -mized to the evaluation of screen-ing data enables rapid and effec-tive processing of large amountsof data.

For Research Use Only. Not for use in

diagnostic procedures. Not available in USA,

Canada and China.

At the same time, the number offalse-positive test results de -creased significantly. Already in2002, the screening committee ofthe German Society of Pediatricsand Adolescent Medicine requiredthe use of mass spectrometry asstandard method in newbornscreening.

The Neonatal Solution software simplifies routine data evaluation

On the third day of its life, bloodis drawn from the newborn’s heel,applied onto paper filter cards andsent to the respective screeninglaboratory on the same day.

Due to the tandem mass spectro -meter’s high specificity, more than20 metabolites, such as aminoacids and acylcarnitine, can bedetermined within the shortestpossible time. Because of continu-ously improving instrument sensi-tivity, time-consuming samplepreparation, such as derivatiza-tion, is now no longer necessary.Chromatographic separation ofthe marker substances is also nolonger required due to the selec-tivity of the MS/MS technology.With an analysis time of less thantwo minutes, a high daily samplethroughput is no longer a prob-lem.

The amount of data acquired isenormous and a clear and effectivedata management is urgently re -quired. With the Neonatal Solu -tion data evaluation software,extremely large quantities of datacan be rapidly and easily proces -sed. In this way, users can selectcertain marker substances and cre-ate evaluation methods selectivelyfor these substances only.

Concentrations, peak areas as wellas concentration ratios for indica-tor substances can be calculatedeasily from the LC-MS/MS analy-sis data. For fast visual identifica-

History

Already in 1934, the Norwegian physician Ivar Asbjørn Følling discovered anincreased excretion of phenylpyruvic acid in the urine of mentally disabledpatients, which could be detected using iron (III) chloride (Fölling’s test).

It took almost 20 years until the German pediatrician Horst Bickel couldprove that the severe developmental disorder, which was later known as thedisease phenylketonuria, could be prevented with a diet low in phenylala-nine. As was quickly shown, the long-term result of this dietary treatmentdecisively depends on starting the diet before the onset of clinical symptoms.

In the early 1960’s, the American microbiologist Robert Guthrie developed an easy to perform bacteriological test for phenylalanine. Since 1969/70, allnewborns in Germany have been comprehensibly tested for elevated pheny-lalanine blood levels using the test named after Guthrie. Another congenitalmetabolic disorder, galactosemia, was included in the screening program.

Over the years, additional tests like thyrotropin screening for congenitalhypothyroidism and screening for 17-OH-progesterone (AGS) were includedin the screening program. The treatment successes in positively tested chil-dren speak for themselves, the demand grew and it turned out that more andmore diseases could, in principal, be treated presymptomatically, i.e. beforeonset of the disease, although effective screening methods for early determi-nation were still not available.

Further information

on this article

• Brochures: Neonatal

Solution

• Application: Simulta-

neous Analysis of Amino Acids and

Acylcarnitines in DBS (Dried Blood

Spot) with LCMS-8040

Figure 3: Neonatal Solution – for rapid, visual identification, deviating concentrations of indicator substances are color-coded

Display data file names

If a component exceeds the permitted

range, a color will be assigned to the

displayed data file name.

Color assigned depending on permit-

ted range

Red: If the warning value is exceeded

Yellow: If the caution value is exceeded

LiteratureGudrun Heyn: Ein lebensrettender Test;

Phar mazeutische Zeitung 23/2009,

Erik Harms, Bernhard Olgemöller:

Neugebore nenscreening auf Stoffwechsel -

erkrankungen und Endokrinopathien;

Deutsches Ärzteblatt, 2011, Jg. 108,

Heft 1-2, 11 ff

PRODUCTS

Protein sequencing via Edmandegradation is a proven andstill very popular method in

many laboratories. With the newPPSQ-50 protein sequencer series,Shimadzu has now further devel-oped the successful PPSQ series.Popular features like robustness,reproducibility and low runningcosts have been retained whilesensitivity has been increased sig-nificantly by using a far more sen-sitive detector.

In addition to its customary simplicity of use, the newLabSolutions PPSQ software forinstrument operation and dataevaluation now also offers option-al compliance with the 21 CFRPart 11 guidelines of the US Foodand Drug Administration (FDA).Two versions of the sequencer areavailable:

that, little by little, all N-terminalamino acids are analyzed.

Meanwhile, the PTH amino acidsare injected into the HPLC unitwhere they are identified chroma -tographically via the specific re -tention times of each amino acidand comparison with a standard.Because the separation takes placeisocratically on a C18 column, themobile phase can be continuously

Sequencing according to Edman degradation

In the main unit of the PPSQ, theN-terminal amino acids of a pro-tein or a peptide which is immobi-lized on a membrane are deriva-tized and cleaved off via the Ed -man degradation reaction.

The remaining protein or peptideis available for the next cycle so

• the PPSQ-51A equipped withone reactor

• the PPSQ-53A with three reac-tors enabling simultaneousplacement of up to three sam-ples into the instrument withsubsequent sequential process-ing. This ensures the best possi-ble system utilization, for in -stance during the night or overthe weekend.


New generation of protein sequencerPPSQ-50 with increased sensitivity and FDA compliance

Minutes

PPSQ-51A/53ACycle 17 PTH-Val

Cycle 21 PTH-Ile

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Figure 1: Sample: Horse myoglobin 10 pmol; results show the subtraction chromatograms by PPSQ-50 (A, C) and PPSQ-30 (B, D) connected in tandem after Edman degradation

A B

C D


PRODUCTS

LATEST NEWS

recycled and reused. This not onlyreduces purchase and disposalcosts of the eluent, but also en -ables quick startup, even when thesystem has not been used for lon -ger periods since gradient optimi -zation is not necessary. Anotheradvantage of isocratic analysis isthe baseline and retention timestability.

Increased sensitivity – alsoupgradable

The design of the new PPSQ-50(figure 2) is more compact thanthe predecessor series and requiresless laboratory bench space. In addition, the new detectorachieves a significantly higherdetection sensitivity in compari-son with the PPSQ-30 series.

Figure 2 shows a tandem analysisin which the sample was analyzedusing the new detector as well asthat of the predecessor model. It is easy to see that detection viathe new unit is far more sensitive.In this way, it is possible to deter-mine long protein sequences, evenfor samples of which only smallamounts are available. As the sep-

classic version (LabSolutionsPPSQ) and a version with inte-grated database (LabSolutionsPPSQ DB), which are both suit-able for use in stand-alone instru-ments. A new feature is the possi-bility to work in accordance withthe 21 CFR Part 11 guidelines ofthe FDA. This includes securitysettings, user administration, his-tory documentation (audit trail),as well as data storage in a data-base.

The third version is a client/serversolution (LabSolutions CS) thatallows network integration of theinstrument so that data evaluationcan be carried out from any com-puter. Clients already using aPPSQ-30 system can easily switchto this new software, which makesworking in a regulated environ-ment much easier thanks to thecompliance achieved.

strument operation as well as dataevaluation. In addition to auto-mated sequence determination, theuser-friendly software also enablesreprocessing and overlaying ofchromatograms, as well as a sub-traction mode that shows differ-ential chromatograms of two se -quential ones for simple and fastsequence determination.

The LabSolutions PPSQ softwareis available in three versions: The

aration column, the eluent and theanalytical conditions are still thesame, it is possible to upgradepreviously acquired instrumentsof the PPSQ-30 series and therebyalso profit from the increased sen-sitivity.

Software solutions for regulated environments

The newly developed LabSolutionsPPSQ software incorporates in -

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the MOAH fraction. For this rea-son, an established evaluationmethod is at present not available.

Multidimensional GC increases sensitivity and selectivity

Gas chromatography with a massselective detector offers one possi-bility to obtain detailed informa-tion on the composition of theMOAH fraction. In particular, theso-called comprehensive GCxGCseparation is the technology ofchoice for increased sensitivityand selectivity.

Are newspapers the source of printing ink migration intopackaged products?

Newspapers and magazines arepart of the raw materials for theproduction of recycling productsfor many different applications.The mineral oil containing print-ing inks are considered to be themain sources of contamination inrecycling products.

LATEST NEWS

T he presence of saturated andaromatic mineral oil hydro-carbons (MOSH – Mineral

Oil Saturated Hydrocarbons andMOAH – Mineral Oil AromaticHydrocarbons) in paper and card-board packaging for food and hy -giene products is an issue that isbeing widely discussed at the mo -ment. The MOSH fraction con-sists of linear and branched alka-nes as well as alkyl-substitutedcycloalkanes, whereas the MOAHfraction consists of alkylated poly -aromatic hydrocarbons with up tofour aromatic rings. The mainfocus is on the aromatic fraction,as it may contain potential carci -nogenic and mutagenic compounds[1]. The proportion of the aromat-ic fraction is approximately 15 -30 % of the total mineral oil frac-tion. The main sources of mineraloil compounds in paper and card-board are printing inks, which areeither applied directly onto thepackaging or introduced via therecycling processes during whichthey are not fully removed.

The migration of mineral oil hy -drocarbons from packaging intopackaged products takes placethrough direct contact or via thegas phase, by evaporation fromthe packaging and re-condensa-tion in the packed product. Thesepro cesses can re sult in a relevantmi gration of mineral oil com-pounds with carbon chain lengthsof up to n-C28.

Mineral oil fraction migratesinto packed products

The concentration of mineral oilhydrocarbons ≤ n-C28 in paperproducts is up to 1000 mg/kg [2],of which up to 70 % can migrateinto the packed product. Based on the acceptable daily intake of0.01 mg/kg body weight, as speci-fied by the Joint FAO/WHO Ex -pert Committee on Food Addi -

Concentration of MOSH andMOAH is determined in this way.

Large-volume injection (LVI) withflame ionization detection (FID)is used to achieve sufficient sensi-tivity and to avoid detector dis-crimination. This, however, causesfractions to elute as non-separated‘mineral oil humps’ (broad peaks)and identification of individualcompounds is not possible (figure1). Because of this lack of detailedinformation, it is difficult to eval-uate the mutagenic potential of

tives ( JEFCA) in 2002, a specificmigration limit for MOSH of 0.6 mg/kg and for MOAH of 0.15 mg/kg packed product hasbeen defined under assumptions of standard conditions.

At present, analytical determina-tion of MOSH and MOAH ismainly carried out via online oroffline high-performance liquidchromatography coupled with gaschromatography-flame ionizationdetection and large-volume injec-tion (HPLC-GC-LVI-FID) [4].


Mineral oil compounds in paperand cardboard packagingInvestigating the composition of the aromatic mineral oil fraction

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Figure 1: GC-FID chromatogram of a MOSH and MOAH fraction [5]

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Figure 2: Phenylalkanes (Scan and m/z 92) in standard and sample (newspaper)

Figure 4: Detection of different branchings and substitutions exemplified by 1-phenylpentane


LATEST NEWS

As starting material, various dailynewspapers were analyzed. In ad -dition, different packaging papers(fresh fiber paper and recyclingcardboard) were analyzed.

Sample preparation was carriedout by extracting the paper sam-ples and separation of MOSH andMOAH via HPLC. Subsequently,GCxGC-MS analysis of theMOAH fraction was executedusing Shimadzu’s GCMS-QP2010Ultra with a Zoex cryogenic mo-dulator. Details of the analyticalparameters are shown in table 1.

For identification of individualcompounds, a standard mix con-sisting of the internal standards of the MOSH/MOAH separation[2, 5], alkylated aromates and poly -aromatic hydrocarbons (PAHs) asnon-alkylated marker substanc eswas used.

GCxGC-MS analysis identifiesphenylalkanes as dominatingaromatic substance group

Using multidimensional GCxGC-MS analysis, it was possible toidentify linear and branched phe -nylalkanes as the dominating aro-matic substance group. An unequi -vocal identification of the phenyl -alkanes was carried out using astandard mix containing linearphenylalkanes with a carbon chainlength of C5 to C18 (figure 2).

Through extraction of the massfragment m/z 92, characteristic forthe phenylalkanes, diagonal linesof unknown compounds becomeapparent that are also character-ized by the mass fragment m/z 92originating from the linear phenyl -alkanes. The exact analysis ofthese unknown compounds re -vealed that all compounds lyingon one diagonal originated fromthe same mass fragment as the lin-ear marker substance.

This is illustrated for phenylunde-cane isomers in figure 3. Throughextraction of the m/z 92 fraction,it is possible to recognize the dia -gonals originating from the linearphenylalkanes. By extracting themass fragment m/z 232 that is typical for phenylundecane, it be -comes ap parent that all compoundson a diagonal are characterized bythe same molecular mass.

An identical molecular masspoints towards differently substi-tuted and branched analogs. Thiscould also be shown by verifica-tion of the compounds 1-phenyl -pentane, isopentylbenzene and (1-methylbutyl)benzene containedin the standard mix (figure 4).Extraction of the characteristicmolecular mass fragment m/z 148revealed that these three com-pounds are on a straight line (dotted line). An unequivocalidentification of the unknown dif-ferently substituted and branchedanalogs of linear phenylalkaneswith longer carbon chains is,however, currently not possibledue to a lack of available stan-dards.

All samples were contaminatedwith mineral oil components

The results revealed that all sam-ples were contaminated with mi -neral oil components, whereupona comparison between fresh fiberand recycled samples showed thatin recycled samples there was aconsiderably higher proportion ofmineral oil compounds than infresh fiber samples (figure 5, page24).

Phenylalkanes could be detectedin all samples analyzed and, there-fore, represent a substantial pro-portion of the MOAH fraction.The highest concentration for thiscompound class was detected in

daily newspapers, followed byrecycled papers. The concentra-tion in fresh fiber samples was, asexpected, considerably loweralthough still clearly detectable,which raised the question con-cerning the source of phenylalka-nes in fresh fiber products. �

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Abbildung 3: Phenylalkane und unterschiedlich substituierte/verzweigte AnalogaFigure 3: Phenylalkanes and differently substituted/branched analogs

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24

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Figure 5: 2D-plot of a fresh fiber and a recycling sample

Column 1st dim.Column 2nd dim.InjectorModulator frequencyTemp. programCarrier gasIon source temp.Interface temp.Solvent cut timeData acquisition

Zebron HT 1 (30 m x 0.25 mm i.d. x 0.25 µm df)SGE BPX 50 (2.5 m x 0.15 mm i.d. x 0.15 µm df)OPTIC IV, 1 µl splitless, 270 °C5 s, 280 °C hot jet 350 ms50 °C (1 min) - 10 °C/min to 120 °C - 2 °C/min to 280 °C (5 min)Helium 155 kPa column head pressure, constant flow200 °C270 °C7 minScan 50 - 340 amu, 50 scans/s

Analytical parameters of the GCxGC analysis

Table 1: Analytical parameters of the GCxGC analysis

Non-aromatic compounds alsoin the aromatic MOAH fraction

In addition to the phenylalkanesand aromatic substance groupssuch as biphenyls and diisopro -pylnaphthalenes, various non-aromatic compounds such as sat-urated hydrocarbons and ter-penes were, however, also identi-fied in the aromatic MOAH frac-tion (figure 5). In particular, satu-rated hydrocarbons that shouldhave been constituents of theMOSH fraction could lead tofalse-positive values in the deter-mination of the MOSH andMOAH sum parameters.

In addition to this problem, thecomplexity of the MOAH frac-tion becomes clearly apparenteven in GCxGC separation,where chromatographically non-separated retention ranges occur.

Literature[1] R. Lorenzini, M. Biedermann, K. Grob,

D. Garbini, M. Barbanera, I. Braschi,

Food Addit Contam 30 (2013) 760 -770

[2] S. Moret, M. Scolaro, L. Barp, G. Purcaro,

M. Sander, L.S. Conte, Food Chem 157

(2014) 471- 475

[3] M. Biedermann, K. Grob, Eur Food Res

Technol 230 (2010) 785 -796

[4] M. Biedermann, K. Grob, J Chromatogr A

1255 (2012) 76 - 99

[5] K. Fislier, F. Grundböck, K. Schön, O. Kap -

penstein, K. Pfaff, C. Hutzler, A. Luch, K.

Grob, J Chromatogr A 1271 (2013) 192-200

This is especially true for sampleswith high mineral oil concentra-tions such as recycling and news-paper samples.

Conclusion

In summary it can be said thatmultidimensional GCxGC iswell able to obtain valuable in -formation on the chemical andstructural composition of mineraloil contaminations in paper andcardboard products. Neverthe -less, more progress still needs tobe made in the elucidation of thecomposition, which could even-tually enable correct assessmentof risk.

AuthorsDipl.-Ing. A. Jurek, Ao.Univ.-Prof. Dipl.-Ing.

Dr. techn. E. Leitner, Institute of Analytical

Chemistry and Food Chemistry, Technical

University Graz, Austria

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