P0125

ABSTRACT

The objective of this research study is to compare

the release properties of poorly water-soluble

progesterone from gel formulation using USP

<1724> specified enhancer immersion cells and a

modified USP apparatus IV incorporated with

Float-A-Lyzer dialysis cells. The membrane,

receiving medium, and hydrodynamic speed were

evaluated for both enhancer immersion cells and

Float-A-Lyzer cells, respectively. Compared to

traditional enhancer immersion cells, Float-A-

Lyzer dialysis cells provide consistent release

profile, better reproducibility and sufficient

discrimination power for three product strengths

(1.1, 2.2, and 3.3% of Progesterone). In addition,

the sample preparation procedures are simplified

for Float-A-Lyzer cells and a faster diffusion rate

was achieved due to larger surface area.

INTRODUCTION

METHODS

METHODS (CONTINUED)

RESULTS

In-vitro release testing (IVRT) in the

pharmaceutical industry is a powerful tool for

formulation design and quality control of

semisolid dosage form for non-oral products. In

this study, since the drug is in a dissolved state in

the vehicle, it has to be diffused and released

prior to becoming available to penetrate into skin.

Thus, the diffusion rate of drug from carry vehicle

becomes a critical parameter to reflect quality,

efficacy, and product consistency with scale-up

and post-approval changes. Although the FDA

SUPAC-SS guidance includes general

methodology descriptions of diffusion systems, it

does not specify a particular test methodology

due to the absence of compendial in-vitro release

test methodology for semisolid dosage forms.

In our laboratory, several shortcomings with

current IVRT methods were discovered during

studies with a topical gel. For instance,

evaporation of the receiving medium may

potentially impact test results. In addition, the

rational for selecting membrane filters is poorly

understood. For IVRT studies of semi-solid

dosage forms, enhancer immersion cell

technologies have been the standard system

used.

The application of a Float-A-Lyzer cell offers

distinct advantages compared with the enhancer

immersion cell because it is easier to use and

readily adaptable for use with the flow-through

apparatus. Moreover, it does not suffer from the

problem of having to remove air bubbles at the

membrane–sample interface, which commonly

occurs when using enhancer immersion cells.

Historically, dialysis cells were developed as a

tool to study tissue biochemistry in animals and it

has been available as a preclinical and clinical

tool for human drug pharmacokinetic studies. For

instance, Float-A-Lyzer cells were used for

release testing of suspensions proves the

feasibility of utilizing dialysis membranes with

controlled surface area and MWCO by providing

consistency in sample introduction and accurate

results that would be needed for a product

containing micro and nano particles for product

development and routine QC performance

testing. These cells have potential to be in semi-

solid dosage forms as well.

Table 2. IVRT and UV/VIS Conditions for

Progesterone Gel using Float-A-Lyzer Cells. REFERENCES

CONCLUSIONS

ACKNOWLEDGEMENTS

DISCLOSURES

Both Enhancer Immersion Cell and Float-A-

Lyzer Cell are capable of accessing the

“sameness” of products during quality

testing, formulation development, and

SUPAC testing if appropriate receiving

medium and membrane are used.

The advantages of Enhancer Immersion Cell

are that the commercial availability of basic

USP dissolution apparatus and the large

volume range makes it easier to adapt this

system to study the release of products

with low drug loads where active

ingredients are difficult to analyze. The

disadvantage of enhancer cells would be

their tedious equipment and cell assembly

procedures. In addition, Teflon cells require

time consuming temperature equilibrium

between the formulation and the receptor

medium.

The main advantage of flow-through cells in

corporate with Float-A-Lyzer cells is the

ease of operation in terms of sample

loading and cell assembling. The problem

of having to remove air bubbles at the

membrane will not be a concern for this

type of cell configuration. In addition, Float-

A-Lyzer cells could offer even larger sample

sizes compared to Enhancer Immersion Cell

to ensuring more consistent results.

An Exploratory Study of In-vitro release testing (IVRT) for Semisolid Dosage Form Products using USP

<1724> and differing sample cell Technologies Xi Shao1,2 , Jian-Hwa Han1, Gregory K. Webster1

1 Global Research & Development, AbbVie, North Chicago, IL 60064;

2 Corresponding Author (xi.shao@abbvie.com)

Presented at the USP Workshop on Quality Attributes of Drug Products Applied to the Skin, September 21–22, 2015, Rockville, MD

Materials

Three research formulations containing

progesterone at strength of 1.1%, 2.2%, and

3.3% were prepared in our laboratory. The

excipients used in preparing the gels were:

Propylene glycol, Oleyl alcohol, Ethyl alcohol,

Carbopol 980, purified water, and sodium

hydroxide.

IVRT Methods using Enhancer Immersion

Cells

An USP Apparatus II (VK7000, Vankel

Industries, NJ, USA) with modified 200 mL flat

bottom flask was used for IVRT of Progesterone

gel using Enhancer Immersion Cells. Enhancer

Immersion Cells were purchased from Hanson

Research Corp. (p/n 65-190-043). The Sampler

preparation steps are shown in Figure 1.

Samples were collected by a fraction collector

(VK8000, Vankel Industries, NJ, USA) and

assayed using a HPLC system (Agilent 1100

series, Agilent Technology, Santa Clara, CA,

USA). The results were reported as mean ± SD

(n = 3). The detailed IVRT and Chromatography

conditions are listed in Table 1.

Figure 1. IVRT of Progesterone Gel on USP

App II with Enhancer Immersion Cells.

Table 1. IVRT and Chromatography Conditions

for Progesterone Gel using Enhancer Immersion

Cells.

Apparatus Modified USP App II (mini paddles, 10 mm from top of the ointment cell to the bottom of the paddle)

Versapor® - 3000 Membrane Filter, 3.0 µm, 25 mm diameter, P/N 66386 (PALL Life Sciences)

MFTM-Membrane Filter, 3.0 3.0 µm SSWP, 25 mm diameter, REF SSWP02500 (Merck Millipore)

Agitation 50 rpm

Medium 0.25% SDS, 0.5% SDS, 20% EtOH, and 40% EtOH

Medium Volume 150 mL

Temp 32 ± 2.0 ºC

Sampling Time Points 0.5, 1, 1.5, 2, 3, 4, 6, 8 hrs

Sampling Volume 2.0 mL

Instrument Agilent 1100 Series HPLC system

Colmun Kinetex C8 100A, 2.6 µ, 100 × 3 mm

Colmun Temp 40 °C

Mobile Phase ACN : Water = 60 : 40

Injection Volume 10 µL

Flow Rate 0.5 mL/min

Detection 254 nm

IVRT Conditions

Chromatography Conditions

Membrane Filter

IVRT Methods using Float-A-Lyzer Cells

An USP apparatus IV in conjunction with

22.6mm sample cells (SOTAX CE70, Sotax,

Westborough, MA) were employed to examine

the feasibility of Float-A-Lyzer cells in the IVRT

of Progesterone gels. Float-A-Lyzer cells were

purchased from Spectrum Labs. An Agilent 8453

UV-Visible spectrophotometer was integrated

into this closed-loop system to monitor the

release rate (µg/cm2) of Progesterone from gel

formulation through mixed cellulose esters

membrane. The steps of sample preparation

and assembling Float-A-Lyzer cells in 22.6mm

sample cells are demonstrated in Figure 2.

Detailed IVRT and UV/VIS conditions are listed

in Table 2.

Figure 2. IVRT of Progesterone Gel on USP

App IV with Float-A-Lyzer Cells.

Apparatus USP App IV (SOTAX CE70, Sotax, Basel, Switzerland)

Float-A-Lyzer CellsBiotech grade Cellulose Ester (CE), MWCO's: 100-500 Da, 500-1000 Da, 3.5-5 KDa, 1000 Kda

(Spectrum Lab, Inc)

Flow Rate 5, 10, and 15 mL/min

Medium 40% EtOH

Medium Volume 200 mL

Temp 32 ± 2.0 ºC

Sampling Time Points 0.5, 1, 1.5, 2, 3, 4, 6, 8 hrs

Instrument Agilent 8453 UV-Visible spectrophotometer

Passlength 10 mm

Detection 254 nm

IVRT Conditions

UV/VIS Conditions

To minimize potential impact for evaporation of

the receiving medium and still provide sufficient

partitioning from the semisolid, aqueous solution

containing 0.25% and 0.5% SDS, 20% and 40%

ethanol solutions were screened using PVDF

membranes, 3.0 µm. The release rates of

Progesterone gel, 3.3% in four receiving media

are shown in Figure 3. As expected, the release

of hydrophobic progesterone was faster in

ethanol solutions than aqueous solutions

containing surfactant. The release rates listed in

Figure 4 showed that hydrophilic mixed

cellulose esters membrane, 3.0 µm had

significantly different permeability characteristic

compared to hydrophobic PVDF membranes.

However, the release rate of active component

was still able to reach ~ 2000 µg/cm2 within 8

hours, which represents approximately 30% of

the investigated molecule has been released.

With the same receiving medium, both PVDF

and mixed cellulose esters membranes

demonstrated sufficient discriminatory power for

progesterone gels with different strength in

Figure 5 & 6.

Figure 3. Release profiles of Progesterone Gel,

3.3% in SDS and EtOH solutions, USP App II,

PVDF membrane.

Figure 4. Release profiles of Progesterone Gel,

3.3% in 0.5% SDS & 40% EtOH solutions, USP

App II, Mixed Cellulose Membrane, 3 µm vs.

PVDF membrane, 3 µm.

RESULTS (CONTINUED)

Figure 5. Release profiles of Progesterone Gel,

1.1, 2.2, and 3.3% in 40% EtOH solutions, USP

App II, PVDF membrane.

Figure 6. Release profiles of Progesterone Gel,

1.1, 2.2, and 3.3% in 40% EtOH solutions,

Mixed Cellulose Ester membrane.

Figure 9 shows the release rates of

Progesterone gel with three dosage strengths

when Float-A-Lyzer cells (1000 KDa) were

used. From the release profiles, it is clear that

these innovative dialysis cells were able to

provide consistent release profile reproducibility,

discrimination for different dosage strengths,

and robustness, as traditional Enhancer

Immersion Cells could offer. However, the

deviation of release behavior from linearity was

observed after 6 hours in high dosage strength

(3.3%). The Progesterone gel, 1.1% and 2.2%

maintained the linear release behavior well

within 8 hours and about 7 hours, respectively.

RESULTS (CONTINUED)

Figure 7. Release profiles of Progesterone Gel,

3.3% in 40% EtOH solutions, USP App IV

(Float-A-Lyzer cells: 0.1-0.5 KDa, 0.5-1 KDa,

3.5-5 KDa, and 1000 KDa).

Figure 8. Release profiles of Progesterone Gel,

3.3% in 40% EtOH solutions, USP App IV

(Float-A-Lyzer cells, 1000 KDa), Flow rate: 5,

10, and 15 mL/min.

Figure 9. Release profiles of Progesterone Gel,

1.1, 2.2 & 3.3% in 40% EtOH solutions, USP

App IV (Float-A-Lyzer cells, 1000 KDa).

This study was funded by AbbVie. AbbVie

participated in the study design, research, data

collection, analysis and interpretation of data, as

well as writing, reviewing, and approving the

publication. Xi Shao, Jian-hwa Han, Gregory K.

Webster, Paul Curry, Jr. and Bryan Erickson are

AbbVie employees and may own AbbVie

stock/options.

Since the pump on USP apparatus IV is used to

force receiving medium flow through acceptor

compartment and provide homogeneity force for

the closed-loop system, the flow rate did not

make a significant impact on the release rate of

progesterone. Three different flow rates: 5, 10,

and 15 mL/min were screened and results are

shown in Figure 8.

INTRODUCTION (CONTINUED

An evaluation of USP <1724> apparatuses

(enhancer immersion cell and flow-through cell),

as well as a dialysis cell will be conducted to

address the concerns and improve on current

IVRT methodology. Test parameters employed

in the IVRT methods will be compared to ensure

the accuracy in providing consistent release

profile, robustness, and discrimination capability

for different dosage strengths as well as

differences in excipients. The evaluation on

flow-through and dialysis cells will expand our

knowledge and capability for IVRT method and

semi-solid dosage form development moving

forward.

The 40% Ethanol solution was directly adopted

as the receiving medium used for USP

apparatus IV configurations. Float-A-Lyzer cells

with four MWCO ranges were screened and

results are shown in Figure 7. All four MWCO’s

membranes offered least-possible resistance to

the diffusion of active compounds. However,

membranes with a larger MWCO range always

provide faster release rate as well as better

reproducibility.

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http://www.spectrumlabs.com/dialysis/FloatALyzer.html

The authors would like to thank Paul Curry, Jr. and

Bryan Erickson for their technical assistance.