The Use of Biorelevant Dissolution Media

to Forecast the In Vivo Performance of a Drug

AAPS Workshop on the Role of Dissolution

in QbD and Drug Product Life Cycle
April 28-30, 2008, Arlington, VA

Dr. Sandra Klein

Institute of Pharmaceutical Technology
Johann Wolfgang Goethe University Frankfurt
„Classical“ applications of
dissolution testing

• to evaluate product quality immediately

after manufacture and during the shelf-life

• to evaluate product quality after changes

in composition or manufacturing method
(SUPAC)

• to compare formulations during

development

Dr. Sandra Klein, 04/2008

„Modern“ applications of
dissolution testing

• examining bioequivalence of different drug products

• predict the in vivo release in terms of IVIVC

Is it possible to predict the in vivo performance of a drug and the

release of a dosage form using simple dissolution tests ?

Æ biorelevant dissolution tests

Æ simulation of conditions in the human gastrointestinal tract

that may affect the dissolution process

Dr. Sandra Klein, 04/2008

Human GI tract

Stomach
ƒ pH = 1-3 (fasted)
ƒ pH = 4-7 (fed)

Dr. Sandra Klein, 04/2008

Human GI tract

Duodenum
ƒ pH = 4-6
ƒ bile and pancreatic secretion

Dr. Sandra Klein, 04/2008

Human GI tract

Jejunum
ƒ pH = 6-7, slightly lower in the fed state

Dr. Sandra Klein, 04/2008

Human GI tract

Ileum
ƒ pH = 7-7.5

Dr. Sandra Klein, 04/2008

Human GI tract

Colon
ƒ pH = 5-7

Dr. Sandra Klein, 04/2008

Selection of test conditions

• classify the drug substance according to the BCS

Æ characterize the drug solubility

over the pH range 1.2 to 6.8
(BCS classification)

Æ run dissolution tests with the pure drug

to determine whether
there are any wetting problems

• choose appropriate media (pH, composition) and volumes

Dr. Sandra Klein, 04/2008

Biopharmaceutics Classification
Scheme
Classification of drug substances according to the BCS

CLASS 1 CLASS 2
highly soluble poorly soluble
highly permeable highly permeable

CLASS 3 CLASS 4
highly soluble poorly soluble
poorly permeable poorly permeable

Guidance for Industry: Immediate release solid oral dosage forms, FDA, 1995

Dr. Sandra Klein, 04/2008

Dissolution tests for highly
soluble drugs (Class I and Class III)
Class I:
Acetaminophen: 14.5 mg/ml → D/S ca. 22.4 ml
neutral
85-90% bioavailability
Metoprolol: > 1 g/ml (-tartrate) → D/S < 1 ml
pKa 9.7 (weak base)
~ 100% bioavailability

Class III:
Aciclovir: 1.3 mg/ml → D/S ca. 150 ml
neutral
ca. 20% bioavailability
Dr. Sandra Klein, 04/2008
Dissolution media for highly
soluble drugs (Class I and Class III)
• a simple medium is sufficient!

• USP media or aqueous buffers, e.g.:

→ SGFsp pH 1.2

→ Acetate buffer pH 4.5

→ SIFsp pH 6.8

• avoid water: no buffer capacity → variable pH

Dr. Sandra Klein, 04/2008

Proposed simplified test method for IR
dosage forms with highly soluble drugs
Test setup
• Paddle apparatus
• 500 ml medium
• SIFsp or IP Phosphate buffer pH 6.8
• 75 rpm
• 37°C
• sampling after 30 min

Specification
• > 85 % release within 30 min

http://whqlibdoc.who.int/trs/WHO_TRS_937_eng.pdf

Dr. Sandra Klein, 04/2008

Developing dissolution tests for poorly
soluble drugs (Class II and Class IV)
Weak acids D/S LITERS !
Troglitazone 2 µg/ml pKa 6.1 100
Glyburide < 0.1 µg/ml pKa 5.3 >> 36
Phenytoin 27 µg/ml pKa 8.3 3.7

Neutral substances
Danazol 0.5 µg/ml 200
Atovaquone 0.1 µg/ml (pKa 9) 1,785
Felodipine 1 µg/ml 10
Griseofulvin 15 µg/ml 16.6

Weak bases
Ketoconazole 4.7 µg/ml pKa 2.9; 6.5 44.4
Itraconazole < 0.001 µg/ml pKa 3.7 >> 100,000
Tamoxifen 3.3 µg/ml pKa 8.8 12

Dr. Sandra Klein, 04/2008

Solubility in the GI tract compared
to aqueous solubility
• solubility of lipophilic compounds (typically logP > 2) is better in
presence of bile salt / lecithin micelles

• solubility of weak bases is often best in the stomach, that of

weak acids in the small intestine

• solubilites in the GI tract may far exceed the aqueous solubility

→ dissolution is often better in vivo !!!

Dr. Sandra Klein, 04/2008

Dissolution media for Class II and
Class IV substances
Location pre-/postprandial Medium

Stomach preprandial SGFsp (USP)

plus surfactant
z.B. Triton X 100, SDS
FaSSGF

Stomach postprandial Ensure® Plus,

Milk

Small intestine preprandial FaSSIF

Small intestine postprandial FeSSIF

Dr. Sandra Klein, 04/2008

In vitro simulation of the preprandial
Stomach, „SGFplus“

SGF plus pH 1.8

Sodium chloride 2g
Hydrochloric acid conc. 3g
Triton X 100 1g
Deionized water qs ad 1 liter

pH 1.8
Osmolality [mOsmol/kg] 120
Surface tension [mN/m] 31

Dr. Sandra Klein, 04/2008

Fasted State Simulated Gastric
Fluid
„Biorelevant“ Simulation of conditions in the fasted stomach

Dr. Sandra Klein, 04/2008

In vitro simulation of the
postprandial stomach

• whole milk or a standardized nutritional 3,3 % proteins

fluid e.g. Ensure® Plus to simulate initial 4,7 % carbohydrates

composition after a standardized breakfast 4% fat

Macheras et al. Int. J. Pharm. 33: 125-136 (1986)

Klein et al. J. Pharm. Pharmacol. 56: 605-610 (2004)

Dr. Sandra Klein, 04/2008

Postprandial Stomach

„Biorelevant“ Simulation of the physicochemical properties in the

stomach immedaiately after administration of a standard breakfast

Standard Breakfast FDA Division of Biopharmaceutics

2 slices of toasted white bread with butter

2 eggs fried in butter
2 slices of bacon
2 ounces of hash-browned ( fried shradded ) potatoes = 56,7 g
8 ounces of whole milk

Carbohydrate: 58g, 232 kcal, 972 kJ, 24 %

Protein: 33g, 132 kcal, 552 kJ, 14 %
Fat: 67 g, 603 kcal, 2533 kJ, 62 %

Æ ENSURE PLUS®
Dr. Sandra Klein, 04/2008
In vitro simulation of the fasted SI
Fasted State Simulated Intestinal Fluid

FaSSIF
Sodium taurocholate 3 mM
Lecithin 0.75 mM
NaH2PO4 3.438 g
NaCl 6.186 g
NaOH qs ad pH 6.5
Deionized water qs ad 1 liter
pH 6.5
Osmolality [mOsmol/kg] ~ 270 *
Buffer capacity [mEq/pH/L] ~ 12
Surface tension [mN/m] 54

Dr. Sandra Klein, 04/2008

In vitro simulation of the fed SI
Fed State Simulated Intestinal Fluid

FeSSIF
Sodium taurocholate 15 mM
Lecithin 3 mM
Acetic acid 8.65 g
NaCl 11.874 g
NaOH pellets 4.04 g
Deionized water qs ad 1 liter
pH 5.0
Osmolality [mOsmol/kg] ~ 670*
Buffer capacity [mEq/pH/L] 72
Surface tension [mN/m] 48

Dr. Sandra Klein, 04/2008

In vitro simulation of the colon
Simulated Colonic Fluid

SCoF pH 5.8

1M Acetic acid 170 ml

1M NaOH 157 ml
Deionized water qs ad 1 liter
pH 5.8
Osmolality [mOsmol/kg] 295
Buffer capacity [mEq/pH/L] 29
Surface tension [mN/m] 70

Dr. Sandra Klein, 04/2008

Does a meal increase the bio-
availability of a drug substance ?
Case example: Danazol (neutral compound, BCS Class II)

Aqueous solubility: 1µg/ml D:S = 200 litres H20

Dose: 200 mg 20 litres FaSSIF
pKa: neutral 6 litres FeSSIF
logP: 4.53
Dr. Sandra Klein, 04/2008
Danazol Dissolution profiles in
various media at 100 rpm

postprandial

preprandial

compendial

Dr. Sandra Klein, 04/2008

Food effects on bioavailability of
Danazol

z preprandial
{ postprandial

Charman et al., J. Clin. Pharm. 33: 1207-1213 (1993)

Dr. Sandra Klein, 04/2008

 Official Test Method

Danazol Capsules (USP)

Medium: 0.75 % sodium lauryl sulfate solution, 900 mL

Apparatus 2: 75 rpm

Time: 30 minutes

Procedure: measure UV absorbance at 286 nm in comparison

with a standard solution having a known concentration
of USP danazol

Tolerance: n.l.t. 75% (Q) of the labeled amount of danazol is

dissolved in 30 minutes

Dr. Sandra Klein, 04/2008

pH/Solubility profile of weak
acids/bases
100

10

pH
Solubility [µg/ml]

1
1 2 3 4 5 6 7 8 9 10

0,1

0,01

0,001

Free Acid Free Base

Dr. Sandra Klein, 04/2008

Dissolution properties of poorly
soluble weak acids
• dissolution in the stomach is likely to be poor due to the low pH
and likely to begin in the more neutral small intestine

• the pH of the medium should be pH >> pKa where possible

ªIf applicable, conditions of the upper or mid small intestine
should be used (pH 5-7)
ªa typical pH would be pH 6.8 (e.g. SIF pH 6.8)

• if the compound is lipophilic, bile components may boost the

solubility in vivo

Dr. Sandra Klein, 04/2008

Is it possible to predict the plasma
profile of weak acids?
Case example: Troglitazone (BCS Class II)

Aqueous solubility: 2 µg/ml

Dose: 200-400 mg (antidiabetic)
pKa: 6.1; 12

Dr. Sandra Klein, 04/2008

Dissolution profiles of Troglitazone
in the fasted and the fed state
Romozin® tablets

Nicolaides et al. Pharm. Res. 16: 1876-1882 (1999)

Dr. Sandra Klein, 04/2008

Bioavailability of Troglitazone

Nicolaides et al. Pharm. Res. 16: 1876-1882 (1999)

Dr. Sandra Klein, 04/2008

Is it possible to predict the plasma
profile of weak acids?
Case example: Phenytoin (BCS Class II)

Aqueous solubility: 27 µg/ml

Dose: 100 mg (antiepileptic)
pKa: 8.3

Dr. Sandra Klein, 04/2008

 drug in solution (ug/ml)

0
10
20
30
40
50
60
W
at
SG er
Fs
p
U
SP
test media

28
SG
Fs
p
m
od
Ac .
et
at
e
bu
ffe
Bl r
an
k
Fa
SS
IF
Bl
an
k
Fe
SS
IF

Fa

Dr. Sandra Klein, 04/2008

SS
IF

Fe
SS
IF
Ac
et
at
e
Solubility of Phenytoin in different

bu
ffe
SI r
Fs
p
U
SP
2 8
Dissolution profiles of Phenytoin
in the fasted and the fed state
Phenytoin AWD 100 mg tablets

25 FeSSIF pH 5.0

20
% Release

FaSSIF pH 6.5
15

10

Blank FeSSIF pH 5.0

5

0
0 30 60 90 120 Blank FaSSIF pH 6.5
Time (min)

Dr. Sandra Klein, 04/2008

Bioavailability of Phenytoin

A Melander et al. Europ. J. Clinical Pharmacol. 15 269-274 (1979)

Dr. Sandra Klein, 04/2008

Dissolution properties of poorly
soluble weak bases
• to characterize the compound, perform dissolution in all
biorelevant media

ª the pH will be most favourable in the fasting stomach, but (if

lipophilic) the bile salts may also contribute to solubilization

• in the fasted state the primary site of dissolution is usually the

stomach, but , caution: many patients are also receiving proton
pump inhibitors or H2 receptor antagonists

• in the postprandial state, the greater concentration of bile in the

GI tract can compensate for the initial higher gastric pH

Dr. Sandra Klein, 04/2008

Is it possible to predict the plasma
profile of weak bases?
Case example: Ketoconazole (BCS Class II)

Aqueous Solubility: 4.7 µg/ml

pKa: 2.9 / 6.5

Dr. Sandra Klein, 04/2008

Ketoconazole Dissolution profiles
In various media at 100 rpm
100

80

60
Release [%]

300 ml SGFsp
40 1000 ml FeSSIF
500 ml FaSSIF
900 ml SIFsp
20

0
0 20 40 60 80 100 120
Time [min]

Dr. Sandra Klein, 04/2008

Bioavailability of Ketoconazole

Dr. Sandra Klein, 04/2008

Summary

• screen the physicochemical properties of the drug

• start with examining the drug under different pH conditions

• characterize the drug solubility over the pH range 1.2 to 6.8

• run dissolution tests with the pure drug to determine whether

there are any wetting problems

• after characterization of the drug select an adequate biorelevant

medium to predict the in vivo performance of your formulation

Dr. Sandra Klein, 04/2008

Literature
z FIP Guidelines for dissolution testing of solid oral products. In e.g. Die Pharmazeutische Industrie 59:760-766
(1997), or Dissolution Technologies 4:5-14 (1997).

z JB Dressman, GL Amidon, C Reppas, VP Shah. Dissolution testing as a prognostic tool for drug absorption:
immediate release dosage forms. Pharm. Res. 15: 11-22 (1998)

z E. Galia, E Nicolaides, D Hörter, R Löbenberg, C Reppas, JB Dressman. Evaluation of various dissolution

media for predicting in vivo performance of Class I and II drugs. Pharm. Res. 15: 698-705 (1998)

z E. Galia, J Horton, JB Dressman. Albendazole Generics – A comparative in vitro study. Pharm. Res.
16:1872-1876 (1999)

z R. Löbenberg, J Krämer, V Shah, GL Amidon, JB Dressman. Dissolution testing as a prognostic tool for drug
absorption: Dissolution behaviour of glibenclamide, a case II compound. Pharm. Res. 17: 439-444 (2000)

z JB Dressman, H Lennernäs. Oral Drug Absorption, Drugs and the pharmaceutical sciences Volume 106.
Marcel Dekker (www.dekker.com) ISBN 0-8247-0272-7. (2000)

z JB Dressman, J Butler, J Hempenstall, C Reppas. The BCS: Where do we go from here? Pharmacetutical
Technology 25: 68-76 (2001)

z B Rohrs. Dissolution method development for poorly soluble compounds.

Dissolution Technologies 8:6-12 (2001)

Dr. Sandra Klein, 04/2008

Literature
z E Stippler Biorelevant Dissolution Test Methods to Assess Bioequivalence of Drug Products. Doctoral Thesis.
Institut für Pharmazeutische Technologie. Frankfurt, Johann Wolfgang Goethe-Universität.
Shaker Verlag: ISBN 3-8322-3218-4 (2004).

z S Klein. Biorelevant Dissolution Test Methods for Modified Release Dosage Forms. Doctoral Thesis. Institut
für Pharmazeutische Technologie. Frankfurt, Johann Wolfgang Goethe-Universität:
Shaker Verlag: ISBN 3-8322-4276-7 (2005).

z S Klein. J Butler, J Hempenstall, C Reppas, JB Dressman. Media to simulate the postprandial stomach I.
Matching the physicochemical characteristics of standard breakfasts. J Pharm Pharmacol 56(5): 605-10
(2004).

z S Klein, JB Dressman. Simplification of biorelevant media to prescreen solubility and dissolution performance
of poorly soluble drugs. Abstract, AAPS Annual meeting, Baltimore (2004)

Dr. Sandra Klein, 04/2008