LPN 2226

ABSTRACTThe current Good Manufacturing Practice (cGMP) guidelines require appropriate laboratory testing of in-process and bulk materials to ensure specification conformity during the production of pharmaceutical products. Depending on the complexity of the manufacturing process, the number of in-process samples and the cycle time of the related analytical methods can delay successive production steps. Therefore, it is important to minimize the time between obtaining the sample and obtaining the final analytical result to minimize production life cycle times.

This poster demonstrates a method to substantially decrease the cycle time of an existing in- process control (IPC) HPLC method by using an optimized LC system. In addition, a state of the art chromatography data system is used to instantly calculate the results and compare them against the specification, allowing the production time to be shortened significantly.

INTRODUCTIONNovosis AG is a pharmaceutical company focusing on the development and production of innovative biodegradable implants and transdermal therapeutical systems (TDS, Figure 1). These pharmaceuticals are used for the treatment of medical complaints such as depression, blood hypertension, hormone replacement therapy, and moderate to severe pain. According to the cGMP guidelines, every interstage product has to undergo reasonable in-process controls with suitable analytical methods (e.g., HPLC), prior to a release of materials to the successive manufacturing step. IPC is a valuable means of steering a manufacturing process. To increase production throughput and to get fast product quality feedback, it is beneficial to receive analytical results of IPC samples as quickly as possible. Two factors to accelerate the analytical process are using a fast LC method and a fast chromatography management system (CMS). In this poster we discuss the speed-up of an existing analytical HPLC IPC method for the analysis of dissolution samples of a TDS in combination with fast data processing.

Figure 4. Method speed-up calculator tool for fast, easy, and convenient transfer of fast LC method parameters.

Chromeleon and UltiMatre are registered trademarks of Dionex Corporation.

Hypercarb is a trademark of Thermo Fisher Scientific, Inc.

RESULTSThe major restriction in accelerating this customer method was the maximum 400 bar backpressure capacity of the separation column. To accelerate the existing gradient separation on a column with defined dimensions, the flow rate, together with the gradient profile, was adapted accordingly. Unfortunately, an increase of flow rate equals an increase of system backpressure which could have been partially compensated with a higher column temperature and hence lower eluent viscosity. In this case, however, a change of the column temperature decreased the resolution between the critical peak pair at 3.9 min at 305 nm (Figure 6). Therefore, it was necessary to maintain the initial column temperature at 30 °C and increase the flow rate by a factor of 2.2. As a result, the overall IPC analysis time needed for the entire manufacturing process was cut down to 45% of the initital HPLC method (see Table 3).

Figure 7. Chromeleon report for dissolution testing replaces any third-party spread sheets and reduces data transcription errors and data evaluation times to a minimum. A customer can immediately see whether the results pass the specification.

SUMMARYThis poster demonstrates how to save valuable analytical response time during the manufacturing process of a transdermal CombiPatch containing two active pharmaceutical ingredients. The HPLC method cycle time of a dissolution method was reduced by 55%, and the data and result evaluation time was reduced by 92.5% of the initial value (see Table 3). Using optimal HPLC instrumentation and software, the described method speed-up concept is available for transfer to other HPLC methods such as assay, related substances, and identity, provided individual application requirements allow for such a transfer.

Fast in-process control analysis helps accelerate pharmaceutical manufacturing processes and improve product quality by providing prompt delivery of analytical results.

ACKNOWLEDGEMENTSThe authors would like to thank Novosis AG for providing samples, and for their assistance.

METHOD SPEED-UP CONCEPTA typical approach to speeding up an existing method is to either shorten the column or increase the flow, and to decrease the particle size used to compensate for the decrease in separation efficiency. For this application, it was not possible to shorten the column or decrease the particle size as the existing stationary phase (Hypercarb 3 µm, Thermo Scientific) was not available in a smaller size. However, the excess resolution between API B and Peak 2 at 305 nm (Figure 3) left enough room for acceleration by increasing the flow rate as the predicted drop in efficiency still provided baseline separation.

Thomas Piecha1, Fraser McLeod1, Dirk Schenk2

1Dionex Corporation, Germering, Germany; 2Novosis AG, Miesbach, Germany

Fast Release of In-Process Materials with Short LC Run Times and High-Speed Data ProcessingFast Release of In-Process Materials with Short LC Run Times and High-Speed Data ProcessingThomas Piecha1, Fraser McLeod1, Dirk Schenk2

1Dionex Corporation, Germering, Germany; 2Novosis AG, Miesbach, Germany

Figure 1. CombiPatch containing two active pharmaceutical ingredients (API) in significantly different con-centrations of 0.4% API A and 11% API B.

CUSTOMER WORKFLOWFigure 2 illustrates the manufacturing process of a TDS. Several steps, including coating, punching, packaging, and labelling, require repeated IPC analysis. Analytical testing for intermediate materi-als comprises rheology, optical inspection, adhesive strength, cold-flow behavior, matrix weight, concentration of active pharmaceutical ingredients (API), identity of API, assay, related substances, residual solvents, and size.

Figure 2. Customer workflow for in-process control sampling during an exemplary transdermal patch manufacturing process.

Table 1.Time Consumption for IPC Testing Before Method Speed-UpProduct No of

SamplesNo of Injections Incl. SST* and

Validation Samples

Analysis Time [h] (Initial Method)

Data Evaluation Time [min]

(3rd Party Spreadsheet)Coated Laminate 12 122 27.8 80Punched Patches 12 122 27.8 80Final Product 6 70 15.9 40

— 314 71.5 200

Table 1. Number of drawn samples and related time effort for HPLC run times and data evaluation with a method cycle time of 13.67 min/sample.

*SST= System Suitability Test

EXISTING ANALYTICAL METHODTable 2 lists the chromatographic parameters of the dissolution method for HPLC analysis before and after speed-up. In the early stage of pharmaceutical product development, one has to take into account that additional peaks may arise during long-term stability studies and hence provide sufficient peak spacing. At a later stage a method may be optimized and revalidated to cut down analysis time to an optimum. The initial method was developed with a strong focus on robustness and resolution. Note the changes in flow rate and gradient profiles before and after speed-up.

Table 2. Method Parameters Before and After OptimizationMethod Parameter Setting Before Speed-up Setting After Speed-upColumn Hypercarb 3 × 50 mm, 3 µm

(Thermo Scientific)Hypercarb 3 × 50 mm, 3 µm (Thermo Scientific)

Guard column Hypercarb (Thermo Scientific) Hypercarb (Thermo Scientific)Flow rate 0.65 mL/min 1.430 mL/minEluent A HPLC grade water + 0.15% TFA HPLC grade water + 0.15% TFAEluent B CH3CN / water, 1:1 (v/v)

+ 0.15% TFACH3CN / water, 1:1 (v/v) + 0.15% TFA

Gradient profile -00.1 min %B = 31 00.0 min %B = 31 03.5 min %B = 31 08.5 min %B = 50 08.6 min %B = 31 12.0 min %B = 31

0.000 min %B = 31 1.591 min %B = 31 3.864 min %B = 50 3.909 min %B = 31 5.455 min %B = 31

Sample volume 60 µL 60 µLColumn temperature 30 °C 30 °CDetection API A 205 nm 205 nmDetection API B 305 nm 305 nm

Figure 3. Customer‘s initial HPLC method for the quantification of API A and API B in dissolution samples at a UV wavelength of 205 nm (A) and 305 nm (B). Peak resolution between API B and Peak 2 at 305 nm is 2.9.

EXPERIMENTAL SYSTEM AND SOFTWARE SETUPFor the analytical experiments the following UltiMate® 3000 system was used:

•AnalyticalquaternarylowpressuregradientpumpLPG-3400A with built-in online vacuum degasser and additional low-volume mixing device

•ThermostattedanalyticalautosamplerWPS-3000TSL•ColumnCompartmentTCC-3000•DiodearraydetectorPDA-3000with13µLanalytical

flow cell•ThermoFisherScientificHypercarb™ column,

3 × 50 mm, 3 µm•Chromeleon® Chromatography Management System,

Version 6.80, SP4, Build 2361•DionexMethodSpeed-upCalculator,Version1.11i

Whenincreasingflow,itisnecessarytoadaptthegradientprofileaccordingtothe“Gradient- Volume” principle. Instead of manual calculation, the Dionex Speed-Up calculator was used (Figure4).Withthiscalculatoritispossibletoinputtheoriginalmethodconditions(columnused,flow rate, injection volume, and gradient table) and the column to be used for a new method. In addition, a boost factor can be used to define the flow speed-up. In our example, the original and new columns remain the same, but a boost factor of 2.2 was used to calculate the new flow rate and gradient profile.

Figure 6. The accelerated customer method, with cycle time reduced to 45% of the initial value. Resolution of API B and Peak 2 at 305 nm is 1.7.

Table 3. Comparison of Time Consumption to Run the IPC Samples and Evaluate the Data with the Initial Method and Optimized Method

ProductNo of

Samples

Analysis Time [h]

(Initial Method)

Analysis Time [h]

(New Method)

Data Evaluation Time [min]

(Initial Method)

Data Evaluation Time [min]

(New Method)Coated Laminate 12 27.8 12.5 80 5Punched Patches 12 27.8 12.5 80 5Final Product 6 15.9 7.2 40 5

— 71.5 32.2 200 15

Table 3. Comparison of IPC times required to run the samples and evaluate the raw data using both initial and optimized methods. The initial process required 74.8 h, while the optimized process required only 32.5 h. This represents a total time savings of 55% for the HPLC analysis, and 92.5% for data evaluation.

In addition, the Chromeleon Data Management software instantly calculated the dissolution profile and visually showed whether or not results passed specification (Figure 7). This eliminates the need to transfer the raw data into a third-party application for calculation purposes. Using this powerful tool, the data and result evaluation time was reduced from 200 min to 15 min—a reduction of 92.5% compared to the previous method (see Table 3).

–100

0

200

mAU

0 1 2 3 4 5 5.5–10

0

50

mAU

Minutes

B

A

1. A

PI A

– 1

.563

3. A

PI B

– 3

.613

Peak

4 –

3.9

48

1. A

PI B

– 3

.614

Peak

2 –

3.971

Optimized Method WVL: 205 nm

WVL: 305 nm

25369

–250

0

500

mAU

0 12–10

50

mAU

Minutes

0

1. A

PI A

–3.

272

1. A

PI B

–7.

843

2. AP

I B –

7.841

Peak

3 –

8.558

Peak

2 –

8.435

A Initial Method

B

WVL: 205 nm

WVL: 305 nm

25368

Coating SolutionSamples

Assay(HPLC)

Not OK

InvestigateReasons

Release & StartCoating Process

Coated LaminateSamples

Punched PatchSamples

Final ProductSamples

Release FinalProduct

Not OK

Not OK

Not OK

OK

OK

OK

OK

InvestigateReasons

InvestigateReasons

InvestigateReasons

Release & StartPunching Process

Dissolution(HPLC)

Dissolution(HPLC)

Release & StartLabeling Process

Dissolution(HPLC)

25367

Figure 5. HPLC system setup for method optimization. Backpressure capability up to 50 MPa and temperature range up to 85 °C allow for use of fast-LC- compatible methods.