Another mass spectrometry technique that has been usedfor the analysis of pharmaceutical drug formulations isdesorption electrospray ionisation mass spectrometry (DESIMS).InDESIMSchargeddropletscreatedbyanelectrospraysource are concentrated onto a solid sample, resulting insurface interaction creating secondary ions that can bedetected using MS. DESI MS has been used to studypharmaceutical drug formulations
and for the screeningof illicit Ecstasy tablets.
Whilst DESI requires no samplepreparation, is relatively fast and is a soft ionisationtechnique, the spatial resolution available for imagingexperiments is only of the order of 0.25mm
and selectiveextraction based on the desorption solvent used wouldappear to be a signiﬁcant issue in DESI experiments.Matrix-assistedlaser desorption/ionisation(MALDI) MS, asoft ionisation technique, utilises a chemical matrix coatedonto the sample of interest to act as an energy transferintermediatebetweenthelaserandthesample.
Theroleof the matrix is primarily to absorb at the laser wavelength andtransfer energy to the analyte in order to promote efﬁcientionisation. The most commonly used lasers in MALDI MSexperiments are UV lasers – nitrogen (
337nm) andNd:YAG (
266nm) and, since the majorityof MALDI MS experiments that have been published to datehave been conducted in positive ion mode, the most commonmatrices employed in suchstudiesarestrongly UV-absorbingorganicacids.Themostcommonlyusedorganicacidmatricesinclude
-cyano-4-hydroxycinnamic acid (
-CHCA), dihy-droxybenzoic acid (DHB) and sinapinic acid (3, 5-dimethoxy-4-hydroxycinnamic acid).Matrix-assisted laser desorption/ionisation mass spec-trometry imaging (MALDI MSI)
is a relatively newtechnique that offers all of the beneﬁts of conventional massspectrometry such as rapid analysis time, speciﬁcity andsensitivity. It can be used for the analysis of a range of samples. Data is obtained by recording mass spectra as asample is moved by set increments under a stationary laser.By plotting the spatial dimensions of the x and y axes againstthe abundance/intensity of selected ion/ions, images can begenerated. Mass spectrometry permits the parallel detectionof multiple analytes thus providing a wealth of informationabout the sample of interest and this is why MALDI MSI has been used in a wide range of biomedical applications.MALDI MS has been applied previously to the analysis of ‘‘street’’ tablets containing prohibited substances.
Thetablets were ground into a ﬁne powder and dissolved inmethanol in order to permit conventional MALDIanalysis to be performed using a combination of
-CHCA andcetyltrimethylammonium bromide (CTAB) in a water/acetonitrile solution (50:50 v/v) as matrix.The work presented herein demonstrates the use of MALDI MSI to study whole tablets for the ﬁrst time. Theaim of this work was to map the distribution of the activedrug throughout all the excipients contained within tablets.MALDI MSI has been used to directly image the distributionof the active drug throughout the tablet, and also to providesome quantitative information in relation to the manufac-tured dose of the tablet.The following tablets were obtained for MALDI MSIanalysis: an unknown tablet formulation, tablet X (placebo,1mg, 3mg and 6mg dosage forms); sildenaﬁl citrate (Viagra25mg); paracetamol (500mg); aspirin (75mg); Anadin Extra(a mixture of aspirin (300mg), paracetamol (200mg) andcaffeine (45mg) and Solpadeine (a mixture of paracetamol(500mg) and caffeine (65mg)).Principal component analysis (PCA) is a form of multi-variate analysis that can be used to simplify multidimen-sional datasets. In previous published work, PCA has beenincorporated into the interpretation of MALDI MSI data todiscriminate between different regions in mammalian tissuesections
andtoevaluatedatafromtheanalysisofporcineskin that had been treated with hydrocortisone.
PCA iscommonly used to reduce the dimensionality of multivariatedata sets while retaining most of the original informationcontent. Discriminant analysis (DA) is often used to deﬁne aclassiﬁcation from an observation of pre-deﬁned groups.PCA-DA is a two-stage supervised statistical techniquewhere the number of variables in the data set is ﬁrst reduced by the use of PCA before a second discriminant analysisis performed. PCA-DA takes into account user-suppliedinformation about external variables (group information)when reducing the dimensionality of the datasets in the ﬁrststage. Therefore, it is better suited than PCA for clusteringanalysis.
Here we have demonstrated how PCA-DA can beusedtodifferentiatebetweentheactivedrugcomponent andtheexcipients containedwithinthetabletandalsohowPCA-DA can be used to provide semi-quantitative information.
-Cyano-4-hydroxycinnamic acid (
-CHCA), ethanol andtriﬂuoroacetic acid (TFA) were purchased from SigmaAldrich (Poole, UK). The following tablets were purchasedcommercially: paracetamol 500mg (Wallis Laboratory Ltd.,Luton, UK), aspirin 75mg (M & A Pharmachem Ltd., Bolton,UK), Anadin Extra (Wyeth Consumer Healthcare, SouthTaplow, UK), and Solpadeine (GlaxoSmithKline Ltd.,Dungarvan,Ireland). Sildenaﬁl citrate (Viagra)25mg dosageform and tablet X in placebo, 1mg, 3mg and 6mg dosageformsweresuppliedbyPﬁzer(Sandwich,UK).Thechemical
Photograph of tabletX showing the curved surfaceof the tablet and the position at which the horizontal sectionwas obtained using a commercially available tablet cutter.
2010 John Wiley & Sons, Ltd.
Rapid Commun. Mass Spectrom.
: 1665–1672DOI: 10.1002/rcm
1666 C. J. Earnshaw