Rapid Thermal Profiling of Pharmaceutialsby Direct Analysis in Real Time for

Detection of Ingredients, DegredationProducts and Other Trace Ingredients in

Pharmaceutical Products

Brian Musselman, Elizabeth CrawfordJoseph Tice and Jordan Krechmer

IonSense, Inc. Saugus, MA USA

Introduction to DART®

• Direct Analysis in Real Time enables rapid analysis of gas,liquids or solids by using metastable atoms and molecules

• Steps necessary for DART– Electrical Discharge into a gas to generate metastable atoms and

ions– Heating of the gas to increase its temperature for more efficient

desorption– Stripping ions from the desorbing gas– Directing the flowing gas at the sample– Collection of the gas phase ions by an API MS for mass analysis

Solid Tablet AnalysisThermal Profiling

DART

VAPUR

Position Sample on movablesurface

Set DART gas Temperature to initialvalue of 150

oC, 250

oC, 350

oC, 450

oC

Push samples through thedesorption ionization region

Repeat at successively highertemperatures

Direct Analysis of LiquidDip tubes intoliquid

Position tip indesorptionionization region

Vacuum drawsgas containingions towards theMS API inlet Single liquid coated tip sampling

150 ºC

250 ºC

350 ºC

450 ºC

Temperature Profile Phase of Method DevelopmentDaytime Caplet – 4 DART-ET Temperatures - Search for 3 Active Ingredients

A

A

A

A

P

P

D

D

D

Deploying DART

• Everyone knows what’sin their products

• But then, there are allthose suppliers…

• And shippers…

• And formulators…

• So how do you reallyknow?

Shapes and Sizes

• For tablets you cansubsequently load up to 10tablets then analyze forcontent by DART-MS inunder 2 minutes

• Small tablets are held itplace by a spring loadedplate pushing the tabletsagainst a template

• Large tablets can simply beplaced in the tablet tray

DART-SVP with Angle

• Exposed Surface ofTablet Limited

• Vary the carrier gastemperature whilescanning across thesurface to thermallydesorb layer afterlayer of product

• Operation at up to45 degree angle forefficient desorption

150 ºC

250 ºC

350 ºC

450 ºC

Temperature Profile Phase of Method DevelopmentDaytime Caplet – 4 DART-ET Temperatures - Search for 3 Active Ingredients

A

A

A

A

P

P

D

D

D

Chlorpheniramine Maleate 0.4%

Dextromethorphan HBr

Nighttime CapletDART-ET 350 ºCLinear Rail Speed 0.5 mm/s

1 Chlorine Isotope Pattern

2mg/450mg

Detection of API in Tablets at Low %DL

Purpose: Evaluate ability to use DART to analyze API in tablets at low %DL

200 400 600 800 1000 1200 1400 1600 1800 2000

m/z

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

95

100

Re

lative

Ab

un

da

nce

241.40

624.07

863.96

342.37523.40385.34

703.99421.39820.28 879.87 1103.98 1484.311247.76 1724.081565.001364.21 1986.891868.37

Gas Temp

(ºC)

API PeakIntensity(n x 1000)

200 5

250 42

300 680

350 750

Results: DART able to detect low level API in RC platformformulation. Adjusting temperature plays significant rolein signal intensity.

DL = Drug Load

Organic Process Research and Development , in press 2010,

Rapid Prep-HPLC Fraction Analysis

Which fraction contains the bromo imp ?

It was never separated on theprep-column

R1

Organic Process Research and Development , in press 2010,

Utilization of DART for Automated Force DegradationWorkflow

Oxidative Stress

Degradation solutions sampleddirectly

R2 N

O O

OH

Exact Mass: 351.052 Exact Mass: 323.057

R1

R3

R2 N

O

OH

R1

R3

H2O2

Mk 1918 Hundreth N HCL_080131185320#3 RT: 0.01 AV:1 NL: 3.69E7T: ITMS + c NSI Full ms [100.0-1200.00]

100 200 300 400 500 600 700 800 900 1000 1100 1200

m/z

0

2000000

4000000

6000000

8000000

10000000

12000000

14000000

16000000

18000000

20000000

22000000

24000000

26000000

28000000

30000000

32000000

34000000

36000000

Inte

nsity

AcidBaseThermalAIBNControl

352352.14

334.20127.12 380.10

324.24 394.19168.11 214.36 480.90 537.13

Mk1918h2O2_080131185320#2 RT:0.01 AV:1 NL:6.41E6

T:ITMS+cNSIFullms[100.00-1200.00]

100 200 300 400 500 600 700 800 900 1000 1100 1200

m/z

0

200000

400000

600000

800000

1000000

1200000

1400000

1600000

1800000

2000000

2200000

2400000

2600000

2800000

3000000

3200000

3400000

3600000

3800000

4000000

4200000

4400000

4600000

4800000

5000000

5200000

5400000

5600000

5800000

6000000

6200000

6400000

Inte

nsity

352.10

324.15

127.09

310.09 674.00518.78370.97184.12 592.62442.26 733.42 936.43804.06 873.88 1151.431014.34

Peroxide

352

324

701.68656.84 1051.68757.87 838.27 981.99 1107.29

Organic Process Research and Development , in press 2010,

What aboutPowders?

Ambient pressure desorptionionization methods use

pressurized gas

Displaced chemicals can clog or otherwisecontaminate the API inlet of the MS

Simple Sample Preparation

Position inDesorption

Ionization Region

Mix Metal withPowder

Tylenol Cold Tablet

• Nighttime Caplet– Acetaminophen

• 325 mg/caplet

– Phenylephrine HCl

• 5 mg/caplet

– ChlorpheniramineMaleate

• 2 mg/caplet

Investigation of Off-axis Sampling

Glass slide with magnets affixed to bottomApply sample by transfer from weaker magnetic field

Or use a powder pipettor

Sample Analysis Movie

Gas temperature and speed ofsample movement

Methodology

•10 mg tablets of Clairitin-brand loratadine (Schering-Plough) were crushedinto powder in a mortar and pestle and evenly distributed into 1.5 dramVWR vials

• The caps were removed and the open vials were placed in an oven andheated to 35 degrees C. Each vial was removed at a specific time invervalfor analysis.

• The powders were analyzed at time intervals of 24 hours, 48h, 72h, 7days, and

• Once vials were removed. .75 dram of magnetite (VWR #100202-020)was added to each vial and shaken until the mixture color was uniform. Acustom magnetic DIP-it (IonSense) was used to pick up the powder andinsert it into the ionization region.

• The spectrum at each time interval was compared to data taken with acontrol vial, which was kept sealed and not inserted into the oven.

Loratadine (Clairitin Brand) MS/MS of M+ ion

M+H+

- H

Loratadine (Clairitin Brand) Control VialPositive-Ion Spectrum

(M+H)+

Loratadine Degradation after 72-hours in OvenPositive-Ion Spectrum

(M+H)+

Loratadine Degradation after 7 Days in OvenPositive-Ion Spectrum

(M+H)+

Conclusions• Tablet Carriers designed to permit multiple

tablet analysis with various sizes

• Automation of DART-based ionizationmethod improving quantitative analysiscapability

• New methodology for rapid analysis ofpowders without requirement of dissolution

• Utilization of magnetic fields to manipulatesample position and control sample quantityinvestigated