Practical Aspects of Transitioning to UPLC in the QC Laboratory

Senior Marketing Manager, Waters Corp

Douglas McCabe,

Project Leader (Quality & Compliance Lab Strategy), Johnson & Johnson

Rosalinda Devos,

2011 Waters Corporation

Understanding Some Proposed Changes to USP General Chapter Chromatography <621>

Doug McCabe
Senior Marketing Manager

2011 Waters Corporation

What is the USP?

The United States Pharmacopeia (USP) is an independent, official public standards-setting authority
Prescription and OTC medicines Healthcare products Food ingredients Dietary supplements Products marketed and sold in the United States must meet USP standards

USP standards recognized and used in >130 countries HQ in US with offices in India, China, Brazil and Switzerland

2011 Waters Corporation

What is the USP-NF?

The United States Pharmacopeia National Formulary (USPNF) is a book of pharmacopeial standards
Drugs substances & preparations monographs: USP Dietary supplements & ingredients monographs: USP Excipient monographs: NF More than 4500 monographs

The USP-NF is the official authority FDA-enforceable standards

Enforcement of USP standards is the responsibility of FDA and other government authorities in the U.S. and elsewhere

USP has no role in enforcement

The U.S. Federal Food, Drug, and Cosmetics Act designates the USPNF as the official compendia for drugs marketed in the United States
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Pharmacopeial Forum (PF)

USP standards are established and maintained through a public process
1. Sponsors provide draft standards and supporting data 2. USP scientific staff and volunteer experts review, test and forward new/revised monograph to PF 3. Monograph is refined, revised and finalized through a unique public-private collaborative process

PF is FREE bimonthly online journal where public review and comment takes place

You probably know this.so why am I telling you this?

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The Beginning of A Significant Change to Chromatography <621>

In early 2009, Waters was approached to write a Stimuli Article to modernize/add more flexibility to selecting LC columns in Chromatography <621> Chromatography <621> describes the adjustments allowed in the chromatography system when system suitability test fails In the summer of 2009 Dr. Uwe Neue et. al. wrote a paper (article) that basically describes UPLC technology and method transfer
Dr. Neues calculations are also the basis of the ACQUITY UPLC Columns Calculator

2011 Waters Corporation

Why are Such Changes to Chromatography <621> Important?

Question From CURRENT USP-NF Online FAQs1
Q. How much can I modify a chromatographic procedure and still be in compliance? Can column length, internal diameter, mobile phase composition be modified? A. Chromatography <621> contains a list of allowed adjustments to chromatographic systems. However, the user should verify the suitability of the method under the new conditions by assessing the relevant analytical performance characteristics potentially affected by the change (see section System Suitability under Chromatography <621>).

1http://www.uspnf.com/uspnf/scienceFAQ.html#q4
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2011 Waters Corporation

The Beginning of A Significant Change to Chromatography <621>

Stimuli Article PF 35(6) [Nov-Dec 2009]

Are you aware of this Article? What does the Stimuli Article propose?
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What is the PF 35(6) [Nov-Dec 2009] Stimuli Article?

Stimuli Article PF 35(6) [Nov-Dec 2009] allows the flexibility to change the column dimensions and/or particle size as long as an equivalent (or better) separation is achieved The ability to change LC columns in a method was restrictive and outdated What if the prescribed column is no longer available? What if a faster (more modern) separation can be obtained with another column (e.g., UPLC)? o In either case revalidation is required

2011 Waters Corporation

Stimuli Article PF 35(6) [Nov-Dec 2009]

Change the length to particle size ratio (l/dp) simultaneously Different lengths, IDs and particle sizes can be used Length to particle size (l/dp) must be 25% of original column Stationary phase characteristics must not change Flow rate scaled to particle size and column ID

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Current State of Chromatography <621> - A Quick Glance

Source: http://www.usppf.com/pf/pub/data/v373/CHA_IPR_373_c621.html#CHA_IPR_373_c621
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Allowable Adjustments in Chromatography <621>

Variable
Particle Size Column Length Flow Rate Column ID Injection Volume Column Temperature Mobile Phase pH

Allowable Changes Before Stimuli Article PF 35(6) After Stimuli Article PF 35(6)
-50% 70% 50% Any allowed Any reduction 10% 0.2 unit
Any allowed as long as column length to particle size (l/dp) is maintained (25%) Adjust for length, particle size and column ID Any allowed Any allowed 10% 0.2 unit

When Stimuli Article PF 35(6) [Nov-Dec 2009] officially becomes part of Chromatography <621>, analysts will have more Flexibility to utilize modern chromatographic techniques such as UPLC Technology

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Effect of Stimuli Article PF 35(6) [NovDec 2009] Change

OVERVIEW: Adjustments in column length, internal diameter, particle size, and flow rate can be used in combination to give equivalent conditions (same N), but with differences in pressure and run time. The table below lists some of the more popular column configurations to give equivalent efficiency (N), by adjusting these variables.1
Column Length Column ID Particle Size (mm) (mm) (dp, m) Relative Values l/dp F N Pressure Run Time 3.3 1.0 1.0 0.5 0.5 0.3 0.3 0.1 0.1

250 150 150 100 100 75 75 50 50

4.6 4.6 2.1 4.6 2.1 4.6 2.1 4.6 2.1

10 5 5 3.5 3.5 2.5 2.5 1.7 1.7

25,000 30,000 30,000 28,600 28,600 30,000 30,000 29,400 29,400

0.5 1.0 0.2 1.4 0.3 2.0 0.4 2.9 0.6

0.8 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0

0.2 1.0 1.0 1.9 1.9 4.0 4.0 8.5 8.5

EXAMPLE: If a monograph specifies a 4.6 x 150 mm, 5-m column operated at 1.5 mL/min, the same separation may be expected with a 2.1 x 75 mm, 2.5-m column operated at 1.5 mL/min x 0.4 = 0.6 mL/min, along with a pressure increase of about 4 times and a reduction in run time to about 30% of the original.1
1http://www.usppf.com/pf/pub/data/v373/CHA_IPR_373_c621.html#CHA_IPR_373_c621

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Example: USP HPLC Method for Budesonide Standard Separation

System Suitability N > 5500 plates Rs > 1.5 RTepimer A = 1.1 x RTepimer B

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Geometrically Scaled Transfer Using ACQUITY UPLC Columns Calculator

HPLC Method Conditions Calculated UPLC Method Conditions

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Example: Method Transferred to UPLC USP Budesonide Sample

System Suitability N > 5500 plates Rs > 1.5 RTepimer A = 1.1 x RTepimer B

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Example: Budesonide Method Transfer Summary All System Suitability requirements were met in UPLC method

Performed routine use study for more than 2850 injections

System suitability requirements were still being met

Benefits of transferring HPLC method to UPLC include:

1. 87% reduction in run time 2. 83% reduction in solvent consumption 3. 93% reduction in sample consumption

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Current Status
In PF 37(3) [May-Jun 2011], these changes are being proposed to <621> Chromatography1:
<621> Chromatography, USP 34 page 243. The Chemical Analysis Expert Committee is proposing to revise general test chapter Chromatography 621, to incorporate some of the ideas and approaches previously presented for public comments in the Stimuli article titled Transfer of HPLC Procedures to Suitable Columns of Reduced Dimensions and Particle Sizes by Neue et al., published in PF 35(6) [Nov.Dec. 2009], pages 16221626. The Stimuli article promoted the adoption of a more flexible approach to select HPLC column dimensions allowing the analyst to choose a combination of column length and particle size to achieve separation power equivalent to that obtained using the prescribed column. This approach markedly increases the range of options currently allowed in the chapter, and helps reducing solvent consumption and the time of the analysis, which is consistent with the current laboratory practices. Several examples are included to illustrate the proposed approach.

There was a 90-day comment period

Comment deadline ended: 31-Jul-2011
1http://www.usppf.com/pf/pub/data/v373/CHA_IPR_373_c621.xml
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USP-NF Development Process

Current Status: Comment Review

2011 Waters Corporation

Source: http://www.usp.org/USPNF/devProcess/

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Current Status of Stimuli Article PF 35(6) [Nov-Dec 2009]

Revised <621> Chromatography available for public comment beginning May 2011
Pharmacopeial Forum Pharmacopeial Forum Posting Date 2-May-11 Interim Revision Comment Announcement Due Date Posting Date 31-Jul-11 30-Sep-11 Interim Revision Announcement Official Date 1-Nov-11 Publication in Which Interim Revision Announcements are Incorporated First Supplement to USP 35-NF 30 Target Official Date

37(3)

1-Aug-12

Public comment period ended: 31-Jul-2011

Comments and changes incorporated: 01-Nov-2011 Target Official Publication that will incorporate 37(3) Interim Revisions: First Supplement to USP 35-NF 30 Target Date for First Supplement to USP 35-NF 30:

01-Aug-2012
2011 Waters Corporation

Source: http://www.usp.org/USPNF/publication.html

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Summary Upcoming changes to USP Chromatography <621> SHOULD remove a MAJOR objection to UPLC implementation Assuming no objections, USP Chromatography <621> will be updated on 01-Aug-2012 Proposed changes will allow regulated laboratories to realize the benefits of UPLC Technology

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Practical Aspects of Transitioning to UPLC in the QC Laboratory

Senior Marketing Manager, Waters Corp

Douglas McCabe,

Project Leader (Quality & Compliance Lab Strategy), Johnson & Johnson

Rosalinda Devos,

2011 Waters Corporation

23

Practical Aspects of Transitioning to UPLC in the QC Laboratory

QUESTIONS ?

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