Developing Drugs? Take a Powder!

Welcome Speakers Uwe Preckwinkel Marketing & Sales Manager, XRD Madison, WI Holger Cordes Senior Applications Scientist, XRD Madison, WI Today s Topics Use of powder X-ray Diffraction (XRD) in various stages of the drug development chain Differences between X-ray and other analytical methods to help you make informed decisions about the best approach for analysis and process control

Applications of XRD in the Pharmaceutical Industry Synthesis of new materials Solid-state characterisation Crystallinity Thermal behaviour Hygroscopical behaviour Grindability / compressability Polymorphism Bioavailability Scale-up Quality control assay Failure Analysis Patenting Counterfeiting

XRD Applications for Pharmaceutical Samples XRD & XRD 2 Single Crystal Several Grains Powder Finished Product Solutions Qualitative Phase ID Quantitative Rietveld analysis Quantitative analysis with standards X-ray movie, non-ambient Structure solution, indexing Microdiffraction, mapping Shape analysis High-throughput screening Grain size determination ( ) % Crystallinity ( ) ( )

Powder X-ray Diffraction Basics Diffraction of an ideal powder Diffraction of textured materials Diffraction of a small number of crystallites ("spotiness effect")

X-ray Diffraction Systems for Powders D8 ADVANCE: modular, expandable D4 ENDEAVOR: compact system, high throughput 6 Bruker Confidential 31.01.2008

Detector Options for XRD Systems Point detectors Scintillation detector Sol-x detector Commonly used for routine analysis with Bragg-Brentano geometry Position-sensitive detectors (PSD) VÅNTEC-1 LynxEye High speed analysis, quality control Area detectors GADDS-HISTAR VÅNTEC 2000 For microdiffraction, polymorph screening, non-ideal powders with poor statistics

Audience Poll Please use your mouse to answer the question on the right of your screen: What process analytical techniques are you currently using for the analysis of your materials? (Check all that apply): Chromatography (HPLC) Thermal Analysis (DSC) Infrared spectroscopy Raman Spectroscopy Mass Spectroscopy Powder X-ray Diffraction Single Crystal X-ray Diffraction Particle size analysis Other

Small Sample Analysis: Gabapentin in Bragg-Brentano Geometry Lin (Counts) 80000 60000 40000 Lin (Counts) 10000 8000 6000 4000 2000 0 10 20 Several mg of sample dusted on silicon lowbackground holder VÅNTEC-1 detector Measurement range: 3-50 20000 2-Theta - Scale Total measurement time: 1:43 min! 0 3 10 20 30 40 50 2-Theta - Scale

Small Sample Analysis: Low-angle Msmts with Position-sensitive Detectors 8000 Ibuprofen scan without air scatter screen Ibuprofen powder with air scatter screen 6000 4000 Lin (Counts) 2000 0 3 10 2-Theta - Scale

Small Sample Analysis: VÅNTEC-1 Performance VÅNTEC-1 at low angles with air scatter screen Lin (Counts) 600 400 200 0 3 10 20 2-Theta - Scale Ibuprofen 0.3 fixeddivergence slits, 2.5 soller slits on both sides 0.015 step size, 0.1 sec/step, 4- min scan from 3 to 40 deg Scan on lowbackground holder without sample

Small Sample Analysis: Ibuprofen Sample in Capillary Lin (Counts) 5000 4000 3000 2000 1000 0 Lin (Counts) 5000 4000 3000 2000 1000 0 13 20 5 10 20 30 40 50 2-Theta - Scale 2-Theta - Scale 32-1723 (*) - Ibuprofen - C13H18O2 60 mm Goebel mirror Capillary stage Radial Soller slit Step size 0.02 Time per step 0.1 sec 0.7 mm glass capillary Measurement time: less than 4 minutes

Small Sample Analysis: Microdiffraction with VÅNTEC-1 detector, Ibuprofen Lin (Counts) 30000 20000 10000 0 3 10 20 3 2-Theta - Scale All measurements with 0.1 sec/step and step size 0.023 Measurement time: 2.5 min 40 mm Goebel mirror, 1 mm exit slit, no collimator (unscaled) 40 mm Goebel mirror + 1 mm collimator (scaled with factor 15) 40 mm Goebel mirror + 0.5 mm collimator (scaled with factor 80)

Small Sample Analysis: Mapping

Small Sample Analysis: API Distribution in a Pill

Small Sample Analysis: Detection Limit ~3 ng * of silicon powder adhered to a 0.1 mm loop Lin (Cps) 0.003 0.002 6000 sec frame with background subtracted 0.001 0 21 30 40 50 60 7 * Estimated by quantitative analysis of x-ray diffraction patterns of silicon powder [~ 5.8 cps/μg (111) reflection] 2-Theta - Scale? Frame: c:\bhuv\si\si2\micro_frame: c:\bhuv\si\si2\micro_2_01.001 - File: si_micro_less.raw - Type: 2Th alone - Start: 7.000 - End: 70.000 - Step: 0.050 - Step time: 10035.8 s - Temp.: 25 Operations: Import 27-1402 (*) - Silicon, syn - Si - Y: 50.00 % - d x by: 1. - WL: 1.54056 - Cubic - I/Ic PDF 4.7 - Powder pattern of Si to 70 o 2θ

High-throughput Screening (HTS) Multiple Samples Library Analysis Screening Properties Screening Results

High-throughput Screening (HTS) D8 DISCOVER powder diffractometer with 2D detector and XYZ sample handling Reflection mode (CS) Transmission mode (CST)

D8 DISCOVER GADDS HTS: 1 + 1 = 3 + = CST CS HTS

Crystallization Glass reactor bottoms 1 mm above block Crystals in direct access to XRPD

PILOT Software for HTS

IμS Microfocus Source: Small Spot Analysis

IμS & VÅNTEC-2000 vs. Classic Setup: Corrundum Comparison IμS & VÅNTEC-2000 45 kv, 0.650 ma, 0.3 mm snout Total counts: 1235K Single 40 mm Goebel mirror, 45 kv, 40 ma, 0.3 mm snout Total counts: 78K

Comparison After Data Integration 21 20 19 18 17 Black: Sealed Tube Red: IμS & VÅNTEC-2000 16 Lin (Cps) 15 14 13 12 11 10 9 8 7 6 Observation - (104) reflection Black: Max Int: 1.25 cps FWHM : 0.189 Red: Max Int: 21.1 cps FWHM: 0.187 5 4 3 2 1 0 22 30 40 50 2-Theta - Scale corundum6282007 - File: corundum6282007_05.raw - Type: 2Th alone - Start: 22.000 - End: 55.200 - Step: 0.020 - Step time: 100. s - Temp.: 25 C (Room) - Time S 1) corundum6282007 - Obs. Max: 35.144 - Max Int.: 1.25 Cps - FWHM: 0.189 Operations: Import Corundum06192007_newsource - File: Corundum06192007_newT_01.raw - Type: 2Th alone - Start: 22.000 - End: 55.200 - Step: 0.020 - Step time: 100. s - Temp.: 1) Corundum06192007_newsource - Obs. Max: 35.150 - Max Int.: 21.1 Cps - FWHM: 0.187 Operations: Import

Comparison of Intensities Optics IμS UBC UBC UBC UBC Collimator 0.3mm 0.8mm 0.5mm 0.3mm 0.3mm front pinhole removed Frame Intensity (cps) Peak Intensity (cps) Corundum (104) FWHM 12.3K 10.2K 3K 0.78K 8.9K 21.1 15.2 4.6 1.25 10.4 0.187 0.212 0.202 0.189 0.265

IμS & VÅNTEC-2000 vs. Classic Setup: Millisecond Snapshot IμS & VÅNTEC-2000 45 kv, 0.650 ma, 0.3 mm snout Total counts: 1942 Single 40 mm Goebel mirror, 45 kv, 40 ma, 0.3 mm snout Total counts: 607

Quartz Powder Symmetrical reflection 600 seconds Sample to detector: 15 cm

The Five Fingers of Quartz 300 Lin (Counts) 200 100 0 66 67 68 69 70 D8 DISCOVER GADDS with IμS and VÅNTEC-2000

Transmission Beam Path

Ibuprofen, Measured in Transmission with Sealed Tube Sample to detector: 29 cm 0.3 mm collimator VÅNTEC-2000 detector Measurement time: 120 sec

Ibuprofen, Measured in Transmission with IμS and VÅNTEC-2000

Gabapentin, Measured in Transmission with Sealed Tube Sample to detector: 29 cm 0.3 mm collimator VÅNTEC-2000 detector 120 sec collection time

Gabapentin, Measured in Transmission with IμS and VÅNTEC-2000 15 sec collection time

High-resolution Screening System with Vαrio1 and VÅNTEC-1 detector D8 ADVANCE HTS High throughput and high resolution for transmission samples

Application: VÅNTEC-1 Detector with Vαrio1 in Transmission 18000 16000 14000 Lin (Counts) 8000 6000 4000 Sample: 3 mg Citric Acid Hydrate between Prolene foils Lin (Counts) 12000 10000 8000 2000 0 13.1 14 15 16 17 18 19 20 21 22 23 24 2-Theta - Scale File: 3mg Citric acid H2O transmission vario 6mm exit slit vantec slits 8-12 0.085_0.2sec.raw 00-016-1157 (*) - Citric acid - C6H8O7 00-015-0985 (*) - Citric acid hydrate - C6H8O7 H2O 6000 4000 2000 0 4 10 20 30 40 2-Theta - Scale File: 3mg Citric acid H2O transmission vario 6mm exit slit vantec slits 8-12 0.085_0.2sec.raw 00-016-1157 (*) - Citric acid - C6H8O7 00-015-0985 (*) - Citric acid hydrate - C6H8O7 H2O

Polymorph Screening Very powerful tool for QC: the ability to make sure that the drug substance you want to be in the final product is actually the one that is being produced in manufacturing Influencing factors Changes in temperature Changes pressure Changes in raw materials Stirring rate etc. Technique can also be used to verify that the correct excipients are in the final product, as well as the drug substance

Polymorph Screening Searching Against a Defined Database The manual method allows for direct comparison but it is subjective The user must use their objectivity to make the comparison The other drawback of the manual method is the fact that you do not have an unequivocal comparison of what the materials present are. In other words you say it is the same or not, but if it is not the same then you are left guessing as to the difference. This is the advantage of the screening method when comparing against a database Not only can you say it is the same.you can also identify materials that do not belong

Polymorph Screening Searching Against a Defined Database Lin (Counts) 40000 30000 20000 10000 0 Here is an XRD pattern from a drug sample. We have set up a database of all of the drug substances and excipients that could be present for this type of sample. This particular sample should be only the pure drug polymorph. There should be nothing else in the material. So we search the database to determine if this is the case. 7 10 20 30 40 2-Theta - Scale sugar [003] - File: Front loaded sugar 0.5 div 0.2 det slit sol-x [003].raw- Type: 2Th/Th locked - Start: 2.000 - End: 89.980 - Step: 0.017 - Step time: 6005. s - Temp.: 25 C ( Operations: Range Op. Merge Import [003]

Polymorph Screening Searching Against a Defined Database Lin (Counts) 40000 30000 20000 10000 0 7 10 20 30 40 2-Theta - Scale sugar [003] - File: Front loaded sugar 0.5 div 0.2 det slit sol-x [003].raw- Type: 2Th/Th locked - Start: 2.000 - End: 89.980 - Step: 0.017 - Step time: 6005. s - Temp.: 25 C ( Operations: Range Op. Merge Import [003] 00-039-1503 (*) - Acetaminophen paracetamol - C8H9NO2 - Y: 33.33 %- d x by: 1. - WL: 1.5406-0 - After searching the database, the software has found that indeed the polymorph drug substance is present and is Acetaminophen. However there are other peaks in the pattern that the database pattern of Acetaminophen does not account for. Conclusion: There are other materials in the pattern. At a cursory glance, this material can be rejected, but for the purpose of troubleshooting we need to know what the contamination is.

Polymorph Screening Searching Against a Defined Database Lin (Counts) 40000 30000 20000 We then search the database for the remaining peaks. The software now reports that the remaining peaks are from the material Sucrose. We now know the material is contaminated and what it is contaminated with. 10000 0 7 10 20 30 40 2-Theta - Scale sugar [003] - File: Front loaded sugar 0.5 div 0.2 det slit sol-x [003].raw - Type: 2Th/Th locked - Start: 2.000 - End: 89.980 - Step: 0.017 - Step time: 6005. s - Temp.: 25 C( Operations: Range Op. Merge Import [003] 00-024-1977 (*) - Sucrose - C12H22O11 - Y: 79.17 %- d x by: 1. - WL: 1.5406-0 - I/Ic PDF0.7 - S-Q 100.0 %-

Polymorph Screening Searching Against a Defined Database 40000 Acetaminophen Sucrose 30000 Lin (Counts) 20000 10000 0 7 10 20 30 40 2-Theta - Scale sugar [003] - File: Front loaded sugar 0.5 div 0.2 det slit sol-x [003].raw - Type: 2Th/Th locked - Start: 2.000 - End: 89.980 - Step: 0.017 - Step time: 6005. s - Temp.: 25 C ( Operations: Range Op. Merge Import [003] 00-039-1503 (*) - Acetaminophen paracetamol - C8H9NO2 - Y: 33.33 % - d x by: 1. - WL: 1.5406-0 - 00-024-1977 (*) - Sucrose - C12H22O11 - Y: 79.17 % - d x by: 1. - WL: 1.5406-0 - I/Ic PDF 0.7 -

Polymorph Screening Searching Against a Defined Database Searching against a defined database is a very powerful tool for polymorph screening. Not only can you tell if a sample meets specifications, you can also determine the materials, or lack of materials, that caused the sample to fail. However, the ideal polymorph screen would be to provide the information to a manual search and a database search all in one with no user interaction. This is called pattern recognition and is available in PolySNAP software.

PolySNAP Pattern Matching Full profile analysis: PolySNAP pattern matching is based on a statistical comparison of each measured data point in each pattern. This true, full pattern analysis approach takes advantage of all pattern information, including presence or absence of peaks, peak shoulders, and background regions. PolySNAP provides an easy-to-use interface to several powerful and novel statistical methods to rank patterns in order of their similarity to any selected sample, allowing known as well as unknowns to be identified quickly. Every data point in every pattern is used to compare samples!

PolySNAP Full Pattern Analysis

PolySNAP Auto-detection of Identical Samples

PolySNAP Auto-detection of Different Samples Automatic phase ID of knowns Identical colors = identical samples

PolySNAP Auto-detection of Unknown or Unexpected Phases or Samples Automatic phase ID of knowns Identical colors = identical samples Different colors = different samples

PolySNAP Auto-detection of Mixtures Automatic phase ID of knowns Identical colors: identical samples Different colors: different samples Automatic detection of unknown or unexpected phases or patterns

PolySNAP Auto-detection of Mixtures Form B+C Automatic phase ID of knowns Identical colors = identical samples Different colors = different samples Form B Form C Automatic detection of unknown or unexpected phases or patterns

PolySNAP Auto-detection of Amorphous Phases Automatic phase ID of knowns Identical colors = identical samples Different colors = different samples Automatic detection and quantification of mixtures Form B+C Automatic detection of unknown or unexpected phases or patterns Form B Form C

PolySNAP Full Pattern Analysis Automatic detection of amorphous phases Automatic phase ID of knowns Identical colors = identical samples Different colors = different samples Automatic detection and quantification of mixtures Form B+C Automatic detection of unknown or unexpected phases or patterns Form B Form C

PolySnap Visualization Options 6-dimensional Plots Selected sample pattern Sample crystallization conditions Display of crystallization parameters as - symbol size - symbol shape - symbol color Video image of sample well

PolySnap True Full Pattern Matching Pharma Approved

D8 SCREENLAB Combined XRD and Raman Spectroscopy

D8 SCREENLAB Combined XRD and Raman Spectroscopy No sample reloading between XRD and Raman measurements PILOT software that controls XRD and Raman measurements PolySNAP software for combined full pattern matching of XRD and Raman patterns Analysis of amorphous phases using Raman spectroscopy

Methodology PXRD + Raman n XRPD Patterns Full profile matching All patterns against all patterns nxn Correlation matrix nxn Distance matrix XRD results Combined results Combine nxn Distance matrix n Raman Patterns Full profile matching All patterns against all patterns nxn Correlation matrix nxn Distance matrix Raman results

Combined Datasets Example 1 48 patterns of 3 forms of Sulfathiazol (forms 2, 3 and 4) PXRD and Raman data collected PXRD Data only: splits Form 3 into two separate clusters Form 4 Form 3 Form 2 Form 3

Combined Datasets Example 1 48 patterns of 3 forms of Sulfathiazol (forms 2, 3 and 4) PXRD and Raman data collected Raman data only: doesn t distinguish between Form 3 and Form 4 Forms 3 & 4 Form 2

Combined Datasets Example 1 48 patterns of 3 forms of Sulfathiazol (forms 2, 3 and 4) PXRD and Raman data collected Combined PXRD + Raman using Automatic Weights: does much better than the individual methods alone Form 4 Form 3 Form 2

Combined Results Example 2 46 patterns of 2 anhydrous forms of Carbamazepeine (Forms 1 & 3) PXRD and Raman data collected PXRD data only: E3 and F7 in different clusters

Combined Results Example 2 46 patterns of 2 anhydrous forms of Carbamazepeine (Forms 1 & 3) PXRD and Raman data collected Raman data only: E3 and F7 in same cluster!

Combined Results Example 2 46 patterns of 2 anhydrous forms of Carbamazepeine (Forms 1 & 3) PXRD and Raman data collected PXRD & Raman data combined: F7 highlighted as an outlier due to this inconsistency Other outliers (yellow) are mixtures of the 2 forms

Combined Results Matching method does very well in distinguishing forms automatically using either Raman or PXRD data Combined results using Automatic Weights seem to be better than either PXRD or Raman alone Identification of pure phases / mixtures improved Use of combined data highlights any inconsistencies in separate analyses Such inconsistencies would not be obvious with only one data source User can then examine outliers manually in detail Seeing similar clustering from multiple original data sources increases confidence in the results

Quality Control Given a set of reference patterns, new patterns can be considered to be similar enough to the references to pass, or different enough to fail. Graphical representation: new samples within the green Pass surface are OK, samples falling outside the surface fail.

Quantitative Analysis We have now seen many different ways to screen samples to make sure we have made what we want to make. The next question that needs to be asked is Am I making these materials in the correct amounts? This can be answered with quite a few different methods Reference intensity ratio Full pattern scaling Standard-based quantification Quantitative Rietveld Analysis

Audience Poll Please use your mouse to answer the question on the right of your screen: Which method do you prefer for quantitative phase analysis? Conventional standard-based quantification Reference-intensity ratio Full pattern scaling based on reference scans of pure phases Standardless Rietveld Analysis (because of peak overlap or because standards are not available)

Quantitative Analysis Scaling Method Lin (Counts) 21000 20000 19000 18000 17000 16000 15000 14000 13000 12000 11000 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 Acetaminophen Sucrose 10.5 11 12 13 14 2-Theta - Scale sugar [003] - File: Front loaded sugar 0.5 div 0.2 det slit sol-x [003].raw- Type: 2Th/Th locked - Start: 2.000 - End: 89.980 - Step: 0.017 - Step time: 6005. s - Temp.: 25 C (Room) - Operations: Range Op. Merge Import [003] 00-039-1503 (*) - Acetaminophen paracetamol - C8H9NO2 - Y: 41.66 %- d x by: 1. - WL: 1.5406 - Monoclinic - I/Ic User 0.1 - S-Q97.2 %- 00-024-1977 (*) - Sucrose - C12H22O11 - Y: 8.40 %- d x by: 1. - WL: 1.5406 - Monoclinic - I/Ic PDF 0.7 - S-Q2.8 %- This method takes the information from the database and, using the scaling factors that can be applied to the patterns, quantitative information can be obtained. The user simply has to scale the intensity of the database pattern to the intensity of the unknown. This pattern is a zoom region of the Sucrose, Acetaminophen sample. The scale factors here are obviously wrong.

Quantitative Analysis Scaling Method Lin (Counts) Acetaminophen Sucrose 21000 20000 19000 18000 17000 16000 15000 14000 13000 12000 11000 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 10.5 11 12 13 14 2-Theta - Scale sugar [003] - File: Front loaded sugar 0.5 div 0.2 det slit sol-x [003].raw- Type: 2Th/Th locked - Start: 2.000 - End: 89.980 - Step: 0.017 - Step time: 6005. s - Temp.: 25 C(Room) - Operations: Range Op. Merge Import [003] 00-039-1503 (*) - Acetaminophen paracetamol - C8H9NO2 - Y: 20.83 %- d x by: 1. - WL: 1.5406 - Monoclinic - I/Ic User 0.1 - S-Q68.5 %- 00-024-1977 (*) - Sucrose - C12H22O11 - Y: 67.19 %- d x by: 1. - WL: 1.5406 - Monoclinic - I/Ic PDF0.7 - S-Q31.5 %- However the user can simply scale these patterns with the mouse and obtain the correct match to the unknown samples. Once the data are properly scaled, the software will automatically report the correct concentrations. Material % Acetaminophen 2.8% Sucrose 97.2%

Quantitative Analysis: Lower Detection Limits with Faster Detectors Sqrt (Counts) 2e5 1e5 1e4 1000 0 100% peak of impurity peak (0.08wt.%) 3 10 20 30 4 00-032-1723 (*) - Ibuprofen - C13H18O2 2-Theta - Scale Example: Ibuprofen with known amount of impurity Measurement circle: 500 mm VÅNTEC-1 detector 40 kv, 40 ma Step size: 0.015 Step time: 1 sec/step 0.08 wt% of known impurity was added Actual detection limits depend on: Crystallinity Peak overlap Crystal symmetry Preferred orientation Crystallite statistics

Standard-based Quantitative Analysis: Powder Samples, Comparison Lin (Counts) 30000 20000 10000 Polymorph A Polymorph B For quantification based on single peak areas, the peak at approx. 22.1 can be used to best distinguish Form B from Form A, because there is no overlap 0 19 20 21 22 23 24 25 26 2-Theta - Scale 90089 poly a ground 3 0.5div 500mm LynEye2.5 dg - Step: 0.019 Operations: Import PolyB ground lynxeye 0.5dg div 2.5 dg soller - Step: 0.020 Operations: Import

Standard-based Quantitative Analysis: Quantification of Polymorph B with DQuant The peak area (highlighted green) was used to quantify Polymorph B. The yellow areas are background areas. This also works for the degree of crystallinity quantification of partially amorphous samples.

Standard-based Quantitative Analysis: Calibration Curve Consistent sample preparation with identical sample volumes is necessary to get sufficient accuracy

Failure Analysis: Quality Control on Tablets Measurement on small tablet with 50 mg API without sample preparation Pure Polymorph A Pure Polymorph B Tablet with 50 mg API: Polymorph B present Tablet with 50 mg API: no Polymorph B Higher background of tablet scans caused by excipients. LynxEye detector: 0.5 divergence slit 4 soller slits 0.019 /step 1 sec/step Measurement time: 10 minutes from 18 to 25

Quantitative Analysis: Basic Principle of the Rietveld Method The Rietveld method is a full-profile approach to quantitative phase analysis using powder diffraction data. The Rietveld method generates a calculated diffraction pattern that is compared with the observed data. Least-squares procedures are used to minimize the difference between the complete observed and calculated diffraction patterns. The following parameters is simultaneously refined: the structural parameters of each phase (lattice parameters, atomic coordinates, site occupancies). These are normally obtained from a data base or the literature the various experimental parameters affecting the pattern (displacement correction, peak shape, background, etc.) The Rietveld method is standard-less. The Rietveld refinement method can be used to characterize several phases simultaneously. The relative masses of all phases contributing to the diffraction pattern can be derived from the refinement.

Quantitative Analysis of Test-Mixture: Rietveld Analysis using TOPAS 13.2 wt% beta-d- Mannitol was added to Ibuprofen as a test mixture Both crystal structures are known and available in databases There is considerable peak overlap between the two phases and preferred orientation for Ibuprofen

Quantitative Analysis of Test-Mixture: Rietveld Analysis using TOPAS Individual calculated curves are highlighted The full pattern can be used for quantitative analysis despite considerable peak overlap 14,000 12,000 beta d-mannitol 13.32 % Ibuprofen 86.68 % 10,000 8,000 Counts 6,000 4,000 2,000 0-2,000-4,000 12 14 16 18 2Th Degrees 20 22 24

Percent Crystallinity Lin (Cps) 400 300 200 100 0 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 2-Theta - Scale POLYMERDATA Converted fromuxdformat byconverted fromuxdformat by XCHVersion 1 FFT Smoothed - File: Poly.raw- Type: 2Th alone - Start: 8.600 - End: 58.360 Operations: Bezier Background 4.571,1.000 Import QUARTZ - File: Quartz.raw- Type: 2Th/Th locked - Start: 18.000 - End: 90.000 - Step: 0.020 - Step time: 10. s - Temp.: 25 C(Room) - Time Started: 0 s - 2-Theta: 18.000 - Theta: Operations: YScale Mul 0.083 Import Another critical piece of information that is important to the pharmaceutical community is how crystalline is a sample XRD is an excellent tool for determining this parameter Crystalline peaks are very sharp and defined Amorphous or noncrystalline peaks are very broad By simply dividing the areas under each of the peaks the percent crystallinity can be easily obtained

Percent Crystallinity Lin (Counts) 3000 2000 1000 0 When both amorphous and crystalline phases are present in the same material you will get a pattern that is a combination of both A large amorphous region with a crystalline region overlaid on top Again by obtaining the area under the curves and simply dividing the percent crystallinity can be obtained 9 10 20 30 40 50 2-Theta - Scale POLYMERDATA Converted fromuxdformat byconverted fromuxdformat by XCHVersion 1 FFT Smoothed - File: Poly.raw- Type: 2Th alone - Start: 8.600 - End: 58.360 Operations: Import POLYMERDATA Converted fromuxdformat byconverted fromuxdformat by XCHVersion 1 FFT Smoothed - File: Poly.raw- Type: 2Th alone - Start: 8.600 - End: 58.360 Operations: Bezier Background 5.623,1.000 Import

Percent Crystallinity Lin (Counts) 3000 2000 1000 0 9 10 20 30 40 50 2-Theta - Scale POLYMERDATA Converted fromuxdformat byconverted fromuxdformat by XCHVersion 1 FFT Smoothed - File: Poly.raw - Type: 2Th alone - Start: 8.600 - End: 58.360 Operations: Import POLYMERDATA Converted fromuxdformat byconverted fromuxdformat by XCHVersion 1 FFT Smoothed - File: Poly.raw - Type: 2Th alone - Start: 8.600 - End: 58.360 Operations: Background 5.623,0.000 Import The easiest way to accomplish this is to have the user subtract the amorphous portion of the pattern and have the software calculate the area under the curves The software then tells us that this sample is 25% crystalline and 75% amorphous The only issue with this technique is again the user intervention that needs to occur

Percent Crystallinity The best way to accomplish this is by mathematically fitting the unknown pattern and then allowing the software to automatically calculate the areas and thus the crystallinity This can also be completely automated to the point that all the user has to do is put the ample in the instrument and walk away

Structure Determination from Powder Data Peak Finding (FPA) Indexing, Space Group Determination Intensity Extraction LeBail, Pawley Structure Determination Structure Refinement "Profiling" LeBail, Pawley Structure Determination AND Refinement from y i (obs) TOPAS Approach Coelho (2000)

D8 ADVANCE Vαrio1 Monochromator for Transmission and Reflection Johansson-type monochromator for pure Kα 1 radiation Six pre-defined geometries for reflection and capillary transmission measurements Geometry change by moving the Vαrio1 along the track

Indexing of Ibuprofen with TOPAS Get the first 20 d-spacings by profile fitting for input file seed index_zero_error Bravais_Cubic_sgs Bravais_Trigonal_Hexagonal_sgs Bravais_Tetragonal_sgs Bravais_Orthorhombic_sgs Unique_Monoclinic_sgs Bravais_Triclinic_sgs load index_d { 14.51938 good 7.244643 6.922005 6.332719 6.003281 5.333514 5.29781 5.266562 5.009033 4.82278 4.72698 4.651116 4.542544 4.392206 4.111215 4.053623 3.974692 3.897449 3.806073 3.692002 3.666344 3.620241 }

Indexing of Ibuprofen Output after approx. 150 sec calculation time Space Group Un-indexed peaks GOF Zero error Lattice parameters Figure of merit versus cell volume

Indexing of Ibuprofen Run whole powder pattern fitting for best matching unit cells

Ibuprofen: Comparison with ICDD Database Lin (Counts) 20000 10000 Note the missing lines in ICDD file With those lines not being resolved or detected, indexing from powder data becomes more difficult Pure Kα 1 radiation really does help for indexing 0 11 12 13 14 15 16 17 18 19 20 21 22 2-Theta - Scale 00-032-1723 (*) - Ibuprofen - C13H18O2

Structure Determination TOPAS software for powder crystallography Indexing Structure solution Structure refinement Caffeine Anhydrous V = 4453.7 Å3 C8H10N4O2 5 molecules (rigid bodies) in the asymmetric unit 70 non-hydrogen atoms in asymmetric unit

Non-ambient Measurements: Stages for Use with Area Detector Anton Paar DHS 900 MRI BTS

Environmental Stages: Humidity Stage D8 ADVANCE powder diffractometer with integrated Hot-Humidity System

Lactose Monohydrate: Dehydration and Hydration in MRI Humidity Stage Lin (Counts) Room temperature sample as received Dehydrated at 160 C and cooled to 45 C Rehydrated at 40 C and 76% relative humidity 30000 20000 10000 LynxEye detector: 0.5 divergence slit 0.019 /step 0.1 sec/step 0 10 20 2-Theta - Scale File: lactose monohydrate at RT_0.5dg div Lynxeye1.5dg.raw Y + 20.0 mm - File: lactose monohydrate at 45C_after heating 0.5dg div Lynxeye1.5dg.raw Y + 40.0 mm - File: lactose monohydrate after rehydration at 40C 76pc hum.raw 00-027-1947 (I) - Lactose hydrate - C12H22O11 H2O

Temperature Study with VÅNTEC-1 Detector in Snapshot Mode Sample: beta d-mannitol 140 fixed scans with a 10 2theta angular coverage The sample was heated to 170 C and slow-cooled Measurement time for each snapshot: 2 sec The measurement was performed in air using a Pt strip heater Heating rate and cooling rate: 0.2 /sec Cursor at 169 C

Temperature Study with VÅNTEC-1 Detector in Snapshot Mode, Level Plot Sample: beta d-mannitol Alpha D-Mannitol + Beta D-Mannitol heating cooling Beta D-Mannitol

21CFR Part 11 is Good for You... Benefits of being compliant are numerous for areas where the FDA is currently not asking for records, or maybe never will: It is easier and cheaper to buy new equipment with Part 11 in mind now, than to deal with unnecessary risk assessments and future Part 11 remediation Exact records support any patent filing or later patent disputes Electronic records have less space requirements and can be more easily retrieved Dividing line between the discovery and development stages are not clear cut, and drug candidates often cycle between the two stages

Meeting Quality System Regulations at Bruker AXS Bruker AXS quality system: Hardware and software are being developed by applying a formal design process and product development life cycle according to Bruker AXS's ISO9001 certified product development procedures For software, additional written standards exist, such as coding standards, configuration management, programmer qualifications, software version control, maintenance, formal testing of software/hardware, incident reporting and tracking, and disaster recovery (Bruker AXS SW404)

Meeting Quality System Regulations at Bruker AXS External system testing (holistic testing at the customer site) Bruker AXS IQ/OQ/PQ Procedure for regulated industries Bruker AXS Instrument Verification Procedure for all other customers* IQ OQ Internal system testing (component-based testing in the test field) Test procedure for internal IQ/OQ/PQ before shipping including a Final holistic test using the Bruker AXS Instrument Verification Procedure * System acceptance test required by Bruker AXS (subset of the Bruker AXS IQ/OQ/PQ Procedure) PQ

Meeting Quality System Regulations at Bruker AXS 21 CFR Part 11 To integrate into an FDA 21 CFR Part 11 (or OECD) compliant laboratory, DIFFRAC plus BASIC offers several tools to provide and guarantee authenticity, integrity and confidentiality of electronic records and electronic signatures, including: Secure system log-ins Automatic audit trail generation Electronic signatures with reports and data Network security with Windows NT4 / 2000 Tamper-proof data files with the ability to discern invalid or altered records White Paper DIFFRAC plus BASIC: Meeting the Requirements of the FDA s 21CFR Part11 Regulation

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