• Application Note

Future–proofing the QC Laboratory for UPLC While Enabling the Faithful Analysis of Legacy HPLC Methods

Future–proofing the QC Laboratory for UPLC While Enabling the Faithful Analysis of Legacy HPLC Methods

  • Chris Henry
  • Mark Wrona
  • Richard Ladd
  • Andy Boughey
  • Waters Corporation

Abstract

This application note describes how the AstraZeneca QC department in Macclesfield has successfully transferred and run all registered QC methods on the Waters ACQUITY UPLC H-Class System with three high throughput products that were successfully developed and validated for UPLC. 

Benefits

  • The capability to faithfully and robustly run legacy HPLC methods while providing access to true UPLC technology when desired
  • No modifications required to provide seamless development, transfer and implementation of UPLC methods
  • Unprecedented efficiency savings along with solvent usage costs and corresponding solvent disposal costs

Introduction

In this application note, we describe how the AstraZeneca QC department in Macclesfield (a global center for developing new technologies for QC) has successfully transferred and run all registered QC methods on the Waters ACQUITY UPLC H-Class System with three high throughput products that were successfully developed and validated for UPLC. In efforts to update and modernize their labs, it was critical for Astra Zeneca to ensure new technology would be efficient, easy to adopt, and cost effective. AstraZeneca updated their LC platforms by implementing Waters UPLC in their pharmaceutical development department with the intention of developing  all new products on this platform. While future-proofing the QC department to receive newer UPLC methods, it was critical to retain the ability to faithfully and robustly run legacy chromatography methods. The technology of choice was the ACQUITY UPLC H-Class System, which has now been deployed throughout the AstraZeneca QC department based at Macclesfield, UK.

Within this body of work, we will give an example of a high profile compound ‘B’ legacy HPLC method transferred from an Agilent 1100 to a Waters ACQUITY UPLC H-Class System along with the newly developed UPLC method validated on the same instrument. 

Experimental

The new UPLC method for compound B degradant products was created using the ACQUITY UPLC Columns Calculator, to  simplify transfer and scale HPLC methodology quickly to UPLC conditions with equivalent performance (with significantly reduced runtimes and solvent savings) ensuring it satisfied the system suitability criteria stated  in the legacy HPLC method.

Impurities 1 and 2 of compound ‘B’ were validated over a range of 50% to 200% of their respective specification limits in the presence of the main compound ‘B’.

HPLC conditions (Agilent 1100 or Waters ACQUITY UPLC H-Class)

Column:

C8 4.6 mm × 250 mm, 5 μm

Flow rate:

1.3 mL/min

Injection volume:

50 μL

Run time:

30 min

Detection:

UV

UPLC conditions (ACQUITY UPLC H-Class equipped with ACQUITY TUV Detector)

Column:

Waters ACQUITY UPLC BEH 2.1 mm × 100 mm, 1.7 μm Column

Flow rate:

0.3 mL/min

Injection volume:

4.2 μL

Run time:

6.86 min

Data management

Empower 2 CDS (Chromatography Data System) Software

Results and Discussion

For the ACQUITY UPLC H-Class to be a successful forward facing platform for the quality control environment, it must first be able to faithfully and robustly reproduce the chromatography generated on the laboratory’s existing HPLC platform. Figure 1 shows the comparison of Compound B’s system suitability sample (SST) run on the Agilent 1100 (top), the Waters ACQUITY UPLC H-Class System in HPLC mode (middle), and the  ACQUITY UPLC H-Class System again using the newly developed UPLC method (bottom).

Figure 1. Comparison of Compound ‘B’ SST sample run on Agilent 1100 HPLC (top), Waters ACQUITY UPLC H-Class System in HPLC mode (middle), and Waters ACQUITY UPLC H-Class in UPLC mode (bottom). 

The ACQUITY UPLC H-Class System has reliably replicated the chromatography from the Agilent 1100 and reproduced  the relative retention times (RRT’s) of impurities 1 and 2 with respect to the main peak, as shown in Table 1. In HPLC mode, the ACQUITY UPLC H-Class System also consistently reproduced the peak areas of Compound B and its related impurities compared to those obtained on the legacy LC system, as shown in Table 2.

Table 1. Retention times / relative retention times of compound ‘B’ and impurities 1 and 2 generated using the legacy method on the Agilent 1100  and the Waters ACQUITY UPLC H-Class System, along with the newly developed UPLC method results obtained from Waters ACQUITY UPLC H-Class System. 
Table 2. Relative peak areas with respect to the main peak of impurities 1 and 2. Results show consistent relative areas between the Agilent 1100 and the Waters ACQUITY UPLC H-Class System in HPLC mode.

The UPLC method had a runtime of under seven minutes compared to the legacy method runtime of 30 minutes. There is also a marked improvement in peak symmetry. Impurities 1 and 2 have switched elution order, although this has not compromised system suitability criteria as shown in Figure 1. 

Validation

Once the newly developed UPLC method for Compound B degradants had satisfied system suitability criteria, the method was subject to a partial validation based on ICH Guidelines Q21 covering linearity, recovery, repeatability, and limits of detection  and quantitation (LOD and LOQ respectively).

The range of the the validation covered 50% to 200% of the impurities respective specification limits (this exceeds the ICH Guideline’s suggestion of 70% to 130% for added assurance of method robustness).2

Table 3 summarizes the validation data obtained. 

Raw linearity data is presented in Figure 2 and Table 3, method precision data in Table 4, and impurity 1 and 2 recovery raw data is presented in Tables 5 and 6 respectively.

Table 3. Linearity raw data.
Figure 2. The UPLC method for Compound ‘B’ degradants comfortably satisfied the acceptance criteria with linearity. Linearity was performed over the range of 50% to 200% of respective specification limits of impurities 1 and 2.
Table 4. Performed using six separate preparations of Compound ‘B’ standard spiked with impurities 1 and 2 at their respective impurity limits (figures adjusted for background impurity content in standard).

Recovery

Impurity 1 recovery data

Table 5. Recovery data for impurity 1 covering 50% to 200% of the range of specification limit. The 50% and 200% levels were prepared in  triplicate and the 100% level n=6 (100% data also used for precision).

Impurity 2 recovery data

Table 6. Recovery data for impurity 2 covering 50% to 200% of the range of specification limit. The 50% and 200% levels were prepared in  triplicate and the 100% level n=6 (100% data also used for precision).
Table 7. Estimated workflow efficiency and solvent cost savings with the implementation of UPLC Technology for all Compound ‘B’ methods based on AstraZeneca batch throughput.

Conclusion

The UPLC data detailed for Compound ‘B’ shows a time savings of between 77.1% to 81.8% equating to over 267 hours per month with solvent savings between 88.7% to 94.8% per month. This not only impacts solvents costs associated with purchase and disposal, but also reduces the need for large storage volume impacting space savings and health and safety.

The Waters ACQUITY UPLC H-Class System’s success in transitioning legacy methods within the Quality Control environment of AstraZeneca exemplifies the instrument’s ability to offer a seamless alternative to existing HPLC platforms while uniquely offering the option of true UPLC Technology when desired.

AstraZeneca have successfully run all registered QC methods on the ACQUITY UPLC H-Class System with three high throughput products transferred and validated successfully using UPLC Technology. 

The success of the Waters ACQUITY UPLC H-Class System in the Quality Control department of AstraZeneca Macclesfield has led to a wider adoption globally of the ACQUITY UPLC H-Class System by AstraZeneca.

References

  1. ICH Guidelines: Validation of analytical procedures: Text and methodology Q2(R1).
  2. USP General Chapter, <621> Chromatography, USP36-NF31, The United States Pharmacopeia Convention, official December 1, 2013.

720006615, July 2019

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