Working document QAS/17.699/Rev.

2
July 2018
Draft document for comments

1 PROTOCOL TO CONDUCT EQUILIBRIUM SOLUBILITY

2 EXPERIMENTS FOR THE PURPOSE OF
3 BIOPHARMACEUTICS CLASSIFICATION SYSTEM-BASED
4 CLASSIFICATION OF ACTIVE PHARMACEUTICAL
5 INGREDIENTS FOR BIOWAIVER

6 (July 2018)
7 TAKEN FROM DRAFT NOTES ON THE CONDUCT OF SOLUBILITY STUDIES (AUGUST 2017)

8 REVISED DRAFT FOR DISCUSSION

Should you have any comments on the attached text, please send these to Dr Valeria Gigante, Technical
Officer, Medicines Quality Assurance, Technologies Standards and Norms (gigantev@who.int) with a
copy to Mrs Xenia Finnerty (finnertyk@who.int ) by 30 September 2018.

Working documents are sent out electronically and they will also be placed on the Medicines website
(http://www.who.int/medicines/areas/quality_safety/quality_assurance/guidelines/en/) for comments
under the “Current projects” link If you have not already received our draft guidelines, please send
your e-mail address to jonessi@who.int and we will add you to our electronic mailing list.

10 © World Health Organization 2018

11 All rights reserved.
12 This draft is intended for a restricted audience only, i.e. the individuals and organizations having received this draft. The
13 draft may not be reviewed, abstracted, quoted, reproduced, transmitted, distributed, translated or adapted, in part or in whole,
14 in any form or by any means outside these individuals and organizations (including the organizations' concerned staff and
15 member organizations) without the permission of the World Health Organization. The draft should not be displayed on any
16 .
website.
17 Please send any request for permission to:
18 Dr Sabine Kopp, Group Lead, Medicines Quality Assurance, Technologies Standards and Norms, Department of Essential
19 Medicines and Health Products, World Health Organization, CH-1211 Geneva 27, Switzerland, fax: (41-22) 791 4856,
20 e-mail: kopps@who.int.
21 The designations employed and the presentation of the material in this draft do not imply the expression of any opinion
22 whatsoever on the part of the World Health Organization concerning the legal status of any country, territory, city or area or
23 of its authorities, or concerning the delimitation of its frontiers or boundaries. Dotted lines on maps represent approximate
24 border lines for which there may not yet be full agreement.
25 The mention of specific companies or of certain manufacturers’ products does not imply that they are endorsed or
26 recommended by the World Health Organization in preference to others of a similar nature that are not mentioned. Errors
27 and omissions excepted, the names of proprietary products are distinguished by initial capital letters.
28 All reasonable precautions have been taken by the World Health Organization to verify the information contained in this
29 draft. However, the printed material is being distributed without warranty of any kind, either expressed or implied. The
30 responsibility for the interpretation and use of the material lies with the reader. In no event shall the World Health
31 Organization be liable for damages arising from its use.
32 This draft does not necessarily represent the decisions or the stated policy of the World Health Organization.
 Working document QAS/17.699/Rev.2
page 2
33 SCHEDULE FOR THE PROPOSED USE OF DOCUMENT QAS/17.699/Rev.2:

34 PROTOCOL TO CONDUCT EQUILIBRIUM SOLUBILITY EXPERIMENTS FOR THE

35 PURPOSE OF BIOPHARMACEUTICS CLASSIFICATION SYSTEM-BASED
36 CLASSIFICATION OF ACTIVE PHARMACEUTICAL INGREDIENTS FOR
37 BIOWAIVER

Presentation to the forty-seventh meeting of the WHO Expert

Committee on Specifications for Pharmaceutical Preparations (ECSPP). October 2012
ECSPP recommended to update the WHO Biowaiver List.

Presentation to the forty-eighth meeting of the WHO ECSPP. ECSPP

recommended to continue revisions and to align the WHO Biowaiver October 2013
List with the 18th Model List of Essential Medicines (EML).

Discussion during an informal consultation held together with

regulatory experts from national regulatory authorities, the WHO
Prequalification Team: medicines (PQTm) and the WHO Collaborating 5–6 July 2014
Centre for Research on Bioequivalence Testing of Medicines,
Frankfurt/Main, Germany.

Presentation to the forty-ninth meeting of the ECSPP. ECSPP

recommended to continue revisions of the WHO Biowaiver List in a October 2014
format that facilitates updating in line with the EML.

Presentation to the fiftieth meeting of the ECSPP and discussion of the

remaining revisions before public consultation. Adoption of the revised
Multisource (Generic) Pharmaceutical Products: Guidelines on
Registration Requirements to Establish Interchangeability (WHO
October 2015
Technical Report Series, No. 992, 2015) which includes a chapter on in
vitro equivalence testing in the context of the Biopharmaceutics
Classification System (BCS) and qualification for a biowaiver based on
the BCS.

Visit of the WHO Collaborating Centre for Research on Bioequivalence February 2016
 Working document QAS/17.699/Rev.2
page 3

Testing of Medicines, from Frankfurt/Main, Germany, to review and

discuss the progress made.

Presentation of the WHO Biowaiver List and revisions at the Biowaiver

Working Group of the International Generic Drug Regulators May 2016
Programme (IGDRP).

Discussion during an informal consultation held together with

regulatory experts from national regulatory authorities, PQTm and the 8–9 July 2016
WHO Regulatory Systems Strengthening team (RSS).

Presentation to the 51st meeting of the ECSPP. October 2016

 Development of the proposal for conduct of solubility studies by Dr

John Gordon in connection with the Proposal to Waive In Vivo
Bioequivalence Requirements for WHO Model List of Essential March 2017
Medicines, Immediate-Release, Solid Oral Dosage Forms (WHO
Technical Report Series, No. 937, 2006, Annex 8).

Proposal discussed with experts from national quality control

April–June 2017
laboratories and specialists in assessing bioequivalence data.

Proposal discussed during the Joint Meeting on Regulatory Guidance

for Multisource Products with Regulators with the WHO Medicines
Quality Assurance group, the Prequalification of Medicines team 7–8 July 2017
(PQTm), and the Regulatory Systems Strengthening team held in
Copenhagen, Denmark.

Revision of text including feedback received. August 2017

Mailing and posting of the working document on the WHO website for
September 2017
public consultation.

Compilation of comments received. October 2017

Presentation to the 52nd ECSPP .* 16–20 October 2017

Preparation of draft Protocol to conduct equilibrium solubility

November 2017
experiments for the purpose of BCS-based classification of APIs for
Working document QAS/17.699/Rev.2
page 4
biowaiver and the Model Certificate of equilibrium solubility
experiments for BCS-based classification of APIs for biowaiver (Annex
1) by Dr Valeria Gigante, WHO Medicine Quality Assurance group
(MQA), Dr John Gordon from PQTm, Professor Giovanni Pauletti,
Professor Marival Bermejo, Professor Virginia Merino Sanjuán,
Professor Carla M. Caramella, Mr Xu Mingzhe and Professor Jin
Shaohong.

Circulation for comments to experts and compilation of comments

received for the Protocol and the Model Certificate of equilibrium
December 2017
solubility experiments for BCS-based classification of APIs for
biowaiver.
Start of APIs equilibrium solubility studies for BCS-based classification
by Professor Giovanni Pauletti, Professor Marival Bermejo, Professor
Virginia Merino Sanjuán, Mr Xu Mingzhe and Professor Jin Shaohong.
February 2018
Collation of regulatory information from the European Medicines
Agency (EMA), Agência Nacional de Vigilância Sanitária (ANVISA)
and PQTm.

Protocol and Model Certificate (Annex 1) optimization during the WHO

March–April 2018
pilot project for BCS-based classification of API for biowaiver.
Presentation of regulatory documents and tests results during the Joint
Meeting on Regulatory Guidance for Multisource Products 18–19 May 2018
with MQA and PQTm in Copenhagen, Denmark.

Updated Protocol and Model Certificate of equilibrium solubility

studies for BCS-based classification of APIs mailed and posted for June–September 2018
public consultation.

Consolidation of comments received. September 2018

Presentation of updated Protocol and Model Certificate of equilibrium
solubility experiments for BCS-based classification of APIs for 22–26 October 2018
biowaiver to the 53rd meeting of the ECSPP.

Any other follow-up action as required.

 Working document QAS/17.699/Rev.2
page 5

38 *Background Information

39 Extract from the 52nd ECSPP. WHO Technical Report Series, No. 1010, 2018:

40 8.2.1. Revision of the biowaiver list

41 As part of its 2006 guidance on waiving of bioequivalence requirements for immediate-release oral solid dosage
42 forms on the WHO EML, WHO had provided a list of APIs that are eligible for biowaivers. The intention is for this
43 list to be updated and maintained as a living document on the WHO website. In 2016 it was agreed that the list
44 should be based on verified laboratory data instead of a literature-based approach. The WHO Secretariat intends
45 to coordinate a new multicentre project to determine the solubility profiles of APIs contained in medicines on the
46 WHO EML to enable an informed decision on whether a biowaiver could safely be granted. The WHO Secretariat
47 proposed that the Expert Committee contribute to updating the biowaiver list by proposing appropriate
48 laboratories to perform the tests, review experimental protocol templates, review laboratory results, determine the
49 APIs’Biopharmaceutics Classification System (BCS) class, and/or participate in the publication of results. A
50 number of members responded positively to the WHO Secretariat’s call for support for the envisaged studies.

51 The Committee noted the report on the update of the WHO biowaiver list. WHO gratefully acknowledged the
52 support offered by the members of the Expert Committee.

53 8.2.2 Conduct of solubility studies

54 During the design of studies to support the revision of the WHO biowaiver list, it became apparent that more
55 guidance was needed on how to design and conduct solubility studies for the purpose of classifying APIs within
56 the BCS. A guidance text was drafted in March 2017, building on recently adopted WHO guidance on
57 equilibrium solubility experiments and on the general chapter on solubility measurements included in the
58 Brazilian Pharmacopoeia in 2016. The proposed guidance was discussed with relevant specialists and at a
59 joint meeting on regulatory guidance held in July 2017. The proposal was further revised and circulated for
60 public consultation in September 2017. It was presented to the Committee at its fifty-second meeting together
61 with comments received.

62 The Committee noted the update on this draft guidance and endorsed the proposed approach to conducting the
63 solubility studies, using this guidance as part of the protocol. Equilibrium solubility experiments for the
64 purpose of classification of active pharmaceutical ingredients according to the biopharmaceutics classification
65 system, Appendix 2 to Multisource (generic) pharmaceutical products: Guidelines on registration requirements
66 to establish interchangeability. In: WHO Expert Committee on Pharmaceutical Preparations. Fifty-first report.
67 Geneva: World Health Organization; 2017: Annex 6 (WHO Technical Report Series, No. 1003).

68 The Committee endorsed the proposed approach to conducting solubility studies for the purpose of revising the
69 WHO biowaiver list.”
 Working document QAS/17.699/Rev.2
July 2018
Draft document for comments

70 PROTOCOL TO CONDUCT EQUILIBRIUM SOLUBILITY EXPERIMENTS FOR

71 THE PURPOSE OF BIOPHARMACEUTICS CLASSICIATION SYSTEM-BASED
72 CLASSIFICATION OF ACTIVE PHARMACEUTICAL INGREDIENTS FOR
73 BIOWAIVER

74

75 1. OBJECTIVE OF THE DOCUMENT

76 The objective of this document is to provide guidance on the design and conduct of equilibrium
77 solubility studies undertaken for the purpose of active pharmaceutical ingredient (API)
78 classification within the Biopharmaceutics Classification System (BCS) (1,2). Notably, the
79 definition and guidance given in this document to perform solubility studies apply to APIs and
80 there might be differences in requirement from the conditions for dissolution studies applicable
81 to finished solid pharmaceutical products (FPP).

82 A study protocol has been developed to provide a harmonized approach when performing
83 solubility studies.

84 2. EXPERIMENTAL CONSIDERATIONS

85 Overall, the API sample should be dissolved/suspended in the buffer, then separated by
86 appropriate methods and, in the end, its concentration should be determined in the liquid
87 phase/supernatant.

88 According to the World Health Organization (WHO) definition given in the guidance document
89 Multisource (Generic) Pharmaceutical Products: Guidelines on Registration Requirements to
90 Establish Interchangeability (3), an API is considered highly soluble when the highest single
91 therapeutic dose as determined by the relevant regulatory authority, typically defined by the
92 labelling for the innovator product, is soluble in 250 mL or less of aqueous media over the pH
93 range of 1.2–6.8. The pH solubility profile of the API should be determined at 37 ± 1 °C in
94 aqueous media. A minimum of three replicate determinations of solubility at each pH condition
95 is recommended (3).
 Working document QAS/17.699/Rev.2
page 7

96 In general, equilibrium solubility experiments should be employed. However, in exceptional

97 cases, where the API is not available in sufficient quantities, and is prohibitively expensive, or
98 when it is not possible to maintain the pH of the medium with pharmacopoeial buffers,
99 experiments where the highest therapeutic single dose as recommended by the approved
100 label/summary of product characteristics (SmPC) is examined in a 250 mL volume, or a
101 proportionally smaller amount examined in a proportionally smaller volume of buffer, can be
102 considered. As these are equilibrium solubility experiments, small volumes of solubility media
103 (e.g. 50 mL) may be employed without issue if the available experimental apparatus will permit
104 it.

105 The source and purity of the API should be reported according to the Model Certificate of
106 equilibrium solubility experiments for BCS-based classification of APIs for biowaiver (Annex
107 1). Additional characterization of the solid API used in the solubility experiments may be
108 necessary. The depth of the characterization will depend on the existing knowledge of the solid-
109 state properties of the API in question. For example, if it has been established that the API
110 exists as a single polymorphic form, then less solid-state characterization is necessary.
111
112 The “shake flask” method for solubility determination is recommended; a mechanical agitation
113 device should be used (e.g. orbital shaker) and an appropriate validation method should also be
114 employed. The device used should be capable of maintaining a temperature of 37 ± 1 °C and an
115 appropriate agitation speed that ensures particle contact with the diluent. The agitation speed
116 should be optimized based on the shape of the flask or tube and volume of the liquid in order to
117 prevent particle agglomeration and ensure particle contact with the diluent. Vortex formation
118 should be avoided. With an optimized agitation rate, it is expected that samples will generally
119 reach equilibrium within 24 hours (4). Samples should be taken at several time points to verify
120 that equilibrium has been reached.
121
122 To address issues such as poor wettability and the tendency of the API to float on the surface of
123 the solubility medium, it may be necessary to include tools such as glass microspheres to aid in
124 the de-aggregation of the particles with agitation or sonication (4). Once wetting is successfully
125 achieved, the solubility experiment should proceed toward equilibrium.
126

127
 Working document QAS/17.699/Rev.2
page 8
128 3. BUFFERS FOR EQUILIBRIUM SOLUBILITY DETERMINATION

129 The pH-solubility profile of the API should be determined over the pH range of 1.2–6.8, with
130 the API’s solubility classification being based on the lowest solubility measured over this pH
131 range. Measurements should be made in triplicate or more, according to the study variability,
132 under at least three pH conditions, pH 1.2, 4.5 and 6.8 using, for example, 0.1 M HCl solution or
133 simulated gastric fluid without enzymes pH 1.2, acetate buffer pH 4.5, and phosphate buffer pH
134 6.8 solution. If there are any known solubility minima for the API in aqueous media within that
135 pH range (for example the pKa of the API when within this range), data should also be collected
136 at that pH. Pharmacopoeial buffer solutions are recommended for use in solubility experiments
137 as reported below. Adjust the pH of the buffers at the same temperature as the equilibrium
138 solubility experiments are performed, i.e. at 37 °C +1 °C. The pH should be verified after
139 addition of the API and at the end of the experiment with a calibrated pH meter. If the pH of the
140 buffer changes upon combination with the solute, adjustment of the pH with an appropriate acid
141 or base solution may be sufficient to address the issue, or a buffer with a stronger buffering
142 capacity may be employed. After adjustment of the pH, the solution should be allowed to re-
143 equilibrate for at least an hour before a sample is taken.

144 3.1 Buffers composition

145  Buffer pH 1.2, TS (test solution)

146 Dissolve 2.52 g of sodium chloride R (Reagent) in 900 mL of water R, adjust

147 the pH to 1.2 with hydrochloric acid (~70 g/L) TS and dilute to 1000 mL with
148 water R.

149  Buffer pH 4.5, TS

150 Dissolve 2.99 g of sodium acetate R in 900 mL of water R, adjust the pH to

151 4.5 by adding about 14 mL of acetic acid (~120 g/L) TS and dilute to 1000 mL
152 with water R.

153  Buffer pH 6.8, TS

154 Dissolve 6.9 g of sodium dihydrogen phosphate R and 0.9 g of sodium

155 hydroxide R in 800 mL of water R, adjust the pH to 6.8 with sodium
156 hydroxide (~80g/L) TS and dilute to 1000 mL with water R.
 Working document QAS/17.699/Rev.2
page 9

157 Information on the reagents to be used can be found by consulting the International
158 Pharmacopoeia, Reagents, Test Solutions and Volumetric Solutions Section,
159 http://apps.who.int/phint (5).
160
161 4. EXPERIMENTAL DESIGN
162
163 The details of the solubility experiment’s design should be based on the characteristics of the
164 API under investigation. It is recommended that preliminary testing be conducted to assess the
165 amount of API required and the length of time required for the pivotal solubility experiment.
166
167 5. PRELIMINARY ASSESSMENT OF TIME TO EQUILIBRIUM AND
168 EXPECTED SOLUBILITY
169
170 Preliminary estimation of solubility from chemical structure is suggested as a starting point,
171 using an open source tool (e.g. ChemSpider www.chemspider.com; Virtual Computational
172 Chemistry Laboratory http://www.vcclab.org; Swiss ADME http://www.swissadme.ch ) or
173 by estimating this data.
174
175 From this calculation, determine the amount of solid needed to have approximately 30–40%
176 excess of undissolved solid in 5 mL (or the selected working volume) of buffer solutions at
177 pH 1.2, 4.5, and 6.8. Weigh this amount of solid and put in, for instance a 10 mL tube, if 5
178 mL of the buffer will be used (corresponding to the expected minimum solubility condition).
179 If the solubility is greater than expected, reduce volume to 3 mL while if the expected
180 solubility is low and the API is available in sufficient quantity, use higher volumes.
181
182 Alternatively, the volume can be kept at 5 mL and the mass can be increased or reduced as
183 appropriate.
184
185 Check for the presence of undissolved solid; if the entire solid dissolves when adding the
186 buffer, additional solid should be added until such time that some solid remains undissolved
187 in the tube. To solve any potential issues related to solid wettability or agglomeration, put the
188 tubes in the agitation system such as shaker or magnetic stirrer adding glass beads.
 Working document QAS/17.699/Rev.2
page 10
189 When the amount of solute and volume of buffer has been determined to obtain a saturated
190 solution, minimum three replicates samples for each pH should be prepared to allow
191 measurements at 2, 4, 8, 24, 48, and 72 hours to identify the equilibration time.
192
193 Filtration is normally recommended to remove undissolved API from collected samples,
194 although centrifugation is a valid alternative method, particularly if the medium volume is small.
195 Filter the samples (using, for example, a filter pore size of 0.45 μ) immediately after withdrawal
196 or separate dissolved from undissolved API by centrifugation as appropriate.
197
198 Solubility experiments are performed at 37 +1 ºC; therefore, if samples are to be left at room
199 temperature until analysis, samples should be diluted immediately after centrifugation or
200 filtration in order to avoid precipitation of the solute. This should be taken into account for
201 back calculations.
202
203 To determine equilibrium solubility, the concentration of the solution should be measured at
204 different time points (approximately six; i.e. 2, 4, 8, 24, 48, and 72 hours) until it does not
205 deviate significantly between each measurement. The shortest time needed for reaching the
206 plateau of drug concentration against time could be considered a suitable equilibration time.
207 Collect samples over time to establish a plateau for the amount of solute dissolved and also to
208 monitor stability of the API at each pH (see below).
209
210 Measure the pH value of the buffer solutions after establishing the time to obtain equilibrium.
211
212 6. STABILITY
213
214 The API’s stability across the pH range should be monitored in order to measure true solubility
215 (6,7).
216
217 To distinguish the drug substance from its degradation products, a validated, stability-indicating
218 analytical method should be employed for solubility determination of APIs such as those
219 indicated in the Ph. Int. or other Pharmacopoeias adapted as appropriate, if available, e.g. high-
220 performance liquid chromatographic (HPLC) analysis (see chapter 1.14.4 High-performance
221 liquid chromatography in The International Pharmacopoeia (5)) or an alternative, validated
222 stability-indicating assay. An advantage of an HPLC method over a spectrophotometric method
 Working document QAS/17.699/Rev.2
page 11

223 is that the HPLC method can also be employed to detect impurities and instability (6,7). If
224 degradation of the drug substance is observed as a function of buffer composition and/or pH, it
225 should be reported. If a stability-indicating analytical method is not available, separate stability
226 experiments will be necessary to demonstrate that the API is stable in the buffer media
227 employed.
228
229 7. RECOMMENDATIONS FOR THE ANALYTICAL METHOD
230
231 Construct calibration curves, ideally with 5–6 standards for regression and estimation of
232 intercept, slope, and correlation coefficient and three additional control standards
233 independently prepared for precision and accuracy estimation.
234
235 If necessary, dilute samples to be on the range of the calibration curve, recording the dilution
236 factor for back calculations.
237
238 Run each sample in duplicate and establish a calibration curve. It is anticipated that at least 3
239 to 4 analytical runs are expected (e.g., the first for the samples of 2, 4, 8 hours and then
240 possibly another three for 24, 48 and 72 hours samples), depending on the stability of the
241 samples. In the end, data for intra, inter-day accuracy and precision estimation should be
242 available. In general, the control standards are intercalated with the samples.
243
244 To check filter influence, control standards could be injected without and after filtration.
245 Recovery should be between 98–102% (if less than this value there is some adsorption
246 happening; if more some filter component is affecting the analysis). If necessary, change the
247 filter type.
248
249 Estimate specificity, linearity, range, accuracy, repeatability, and intermediate precision (7),
250 which should meet the minimum acceptance limits.
251
252 8. PIVOTAL EXPERIMENT

253 Pivotal experiments should be designed considering the results of preliminary experiments. The
254 following steps are presented as a general example of a pivotal solubility experiment:
 Working document QAS/17.699/Rev.2
page 12
255 1. in triplicate, for each pH condition to be evaluated, weigh approximately a 10%
256 excess amount of API (as determined during the preliminary test) and combine
257 with an appropriate volume of the necessary buffer solutions (at least pH 1.2, 4.5
258 and 6.8 buffers) in a flask;
259 2. mix and measure the pH value;
260 3. stop and secure the flask to the orbital shaker with controlled temperature and
261 shaker speed:
262 4. collect an aliquot of the supernatant solution at the time equilibrium was
263 established in the preliminary experiment;
264 5. immediately separate dissolved from undissolved API by filtration or
265 centrifugation;
266 6. record the PH value of the solution at the end of the experiment;
267 7. dilute the sample to avoid precipitation before quantifying; and
268 8. determine the concentration of the API.
269
270 9. REPORTING OF RESULTS
271
272 Test results should be reported in the Model Certificate of Equilibrium Solubility Experiments
273 for Biopharmceutics Classification System-based classification of Active Pharmaceutical
274 Ingredients for Biowaiver appended to this Protocol as Annex 1. The report should include
275 information on the API (chemical structure, molecular weight, known dissociation constants,
276 etc.), the actual experimental conditions, including information on buffer composition and the
277 analytical method, results (raw data plus mean values with standard deviations), and any
278 observations such as, for example, the degradation of an API due to pH or buffer composition.
279 The section describing the experimental conditions should include initial and equilibrium pH of
280 solutions and de facto buffer concentrations. If samples are filtered, the type and pore size of
281 the filter should be recorded, along with data from filter adsorption studies. If samples are
282 centrifuged the conditions of centrifugation should be recorded. A graphic representation of
283 mean pH-solubility profile should be provided.
284
285 Any deviations from the protocol should be noted and duly justified.
286
 Working document QAS/17.699/Rev.2
page 13

287 The solubility should be reported in mg/mL. The relative standard deviation (RSD) between the
288 obtained solubility results should not be more than 10% between the replicates of each test
289 condition.

290 The dose: solubility volume (DSV) represents the volume of liquid necessary to completely
291 dissolve the highest single therapeutic dose of the API (as recommended by the approved label/
292 SmPC) at the pH where lowest solubility was observed. Based on the lowest solubility
293 calculated in mg/mL, the DSV can be calculated by dividing the highest therapeutic dose (in
294 milligrams) by the solubility (in milligrams per milliliter) obtained in the study. An API is
295 considered highly soluble when the DSV is < than 250 mL over the entire pH range 1.2-6.8.
296
297 REFERENCES

298 1. Amidon GL, Lennemas H, Shah VP, Crison JR. A Theoretical Basis for a
299 Biopharmaceutic Drug Classification: The Correlation of In Vitro Drug Product
300 Dissolution and In Vivo Bioavailability. Pharmaceutics Research, 1995, 12:413–
301 420.
302
303 2. Proposal to Waive In Vivo Bioequivalence Requirements for WHO model list of
304 essential medicines, Immediate-Release, Solid Oral Dosage Forms. WHO
305 Technical Report Series, No. 937, 2006, Annex 8.
306
307 3. Multisource (Generic) Pharmaceutical Products: Guidelines on Registration
308 Requirements to Establish Interchangeability. WHO Technical Report Series, No.
309 1003, 2017, Annex 6.
310
311 4. Apley M. et al., 2015. Determination of Thermodynamic Solubility of Active
312 Pharmaceutical Ingredients for Veterinary Species: A New USP General Chapter.
313
314 5. The International Pharmacopoeia, http://www.who.int/medicines/
315 publications/pharmacopoeia/en/.
316
317 6. Stability Testing of Active Pharmaceutical Ingredients and Finished
318 Pharmaceutical Products. WHO Technical Report Series, No. 1010, 2018, Annex
319 10.
 Working document QAS/17.699/Rev.2
page 14
320 7. Analytical Method Validation, Appendix 4 in Supplementary Guidelines on Good
321 Manufacturing Practices: Validation. WHO Technical Report Series, No. 937, 2006,
322 Appendix 4.
323
324 FURTHER READING
325
326 1. Guidance for Organizations Performing In Vivo Bioequivalence Studies. WHO
327 Technical Report Series, No. 996, 2016, Annex 9.
328
329 2. Model Certificate of Analysis. WHO Technical Report Series, No. 1010, 2018,
330 Annex 4.
331
332 3. Good Practices for Pharmaceutical Quality Control Laboratories. WHO
333 Technical Report Series, No. 957, 2010, Annex 1.
334
335 4. WHO Good Manufacturing Practices for Pharmaceutical Products. WHO
336 Technical Report Series 986, 2014, Annex 2.
337
338 5. Determinação da Solubilidade Aplicada à Bioisenção de acordo com o Sistema de
339 Classificação Biofarmacêutica. Edition 5, Brazilian Pharmacopoeia, II supplementary,
340 2017.
341
 Working document QAS/17.699/Rev.2
page 15

342 ANNEX 1

343

344 Header:

345 Logo of the laboratory/company issuing the certificate (if applicable)

346 Identification No. of the study: page X of Y

347

348

349 MODEL CERTIFICATE OF EQUILIBRIUM SOLUBILITY

350 EXPERIMENTS FOR BIOPHARMCEUTICS CLASSIFICATION
351 SYSTEM-BASED CLASSIFICATION OF ACTIVE
352 PHARMACEUTICAL INGREDIENTS FOR BIOWAIVER

353

354 Name and address of the laboratory issuing the study report:
355
356
357 Name of the API (international nonproprietary name (INN), brand name, etc.):
358
359
360 Certificates of analysis (CoAs) from manufactures provided: assay within specifications
361 <Yes/No>
362
363
364 Batch number:
365
366 Date received: Quantity received:
367

368 Date of manufacture (if available):

369
370 Expiry date/retest date:
371
 Working document QAS/17.699/Rev.2
page 16
372 Name and address of the original manufacturer:
373
374
375
376 Telephone:
377
378 E-mail:
379
380 Information about the API
381
382 Chemical structure (please report here):
383

384

385
386
387 Nature of the drug (i.e. acid, basic, neutral, amphoteric):
388
389 Dissociation constants [i.e. pKa(s)]:
390
391 Molecular weight (g/mol):
392
393 Equilibrium solubility experiment
394
395 Apparatus:
396
397
398 Highest therapeutic dose:
399
400 Recorded temperature (target 37 °+ 1 °C):
401

402 Volume of the buffer:

403
 Working document QAS/17.699/Rev.2
page 17

404 Sampling times:

405
406 Time to equilibrium:
407
408 Buffer composition (please indicate if different buffers from those recommended in the
409 Protocol to conduct equilibrium solubility experiments for the purpose of BCS-based
410 classification of APIs for Biowaiver where used):
411
412
413 Separations of samples (please indicate how and when samples were filtered, filter type and
414 pores size. If samples are centrifuged, the conditions of centrifugation should be recorded. If
415 separated through different methods, this should be justified):
416
417
418
419
420 Stability (report and discuss any problems with pH-related stability of samples):
421
422
423
424 Solubility method and conditions:
425
426
427
428
429
430 Brief summary of analytical methods including validation:
431
432
433
434
435
436
437
 Working document QAS/17.699/Rev.2
July 2018
Draft document for comments

438 Result of the preliminary solubility experiment

Theoretical Individual Final pH Adjusted Final pH API Concentrati CV % Conc. Conc. Conc. Conc. Conc. Conc. Conc.
d
pH pH before with (ml of corrected equilibrium on mean (mg/ml) (mg/ml) (mg/ml) (mg/ml) (mg/ml) (mg/ml) (mg/ml)
measurement correction 0.1 N HCL concentrati (mg/ml) c 2h 4h 6h 12 h 24 h 48 h 72 h
or NaoH)a on (mg/ml)
b

pH 1.2 1
2
3
pH 4.5 1
2
3
pH 6.8 1
2
3
Potential 1
additional 2
pH 3
439
440
441 a
Amount of acid or base needed to adjust the measured pH. The measurement should be conducted in triplicates and, per each measurement, the corresponding pH values should be reported.
442 b
Report here the three measurements registered at each Ph.
443 c
Report here only the mean of the individual values reported in the previous column.
444 d
Coefficient of variation.
445

446

447

448
 Working document QAS/17.699/Rev.2
page 20
449 Result of the pivotal solubility experiment

450

Theoretical Individual pH Final pH Adjusted Final pH Time to APIs Weight Buffer API API CV % d
pH measurement before with (ml of corrected equilibrium volume equilibrium equilibrium
correction 0.1 N HCL concentratio concentration
or NaoH)a n (mg/ml) b mean (mg/ml)c
pH 1.2 1
2
3
pH 4.5 1
2
3
pH 6.8 1
2
3
Potential 1
additional 2
pH 3
451
452
453 a
Amount of acid or base needed to adjust the measured pH. The measurement should be conducted in triplicates and, per each measurement, the corresponding pH values should be reported.
454 b
Report here the three measurements registered at each pH.
455 c
Report here only the mean of the individual values reported in the previous column. It is the mean solubility value for each pH.
456 d
Coefficient of variation.
457
458
459
460
461
462
463
464
465
466
 Working document QAS/17.699/Rev.2
July 2018
Draft document for comments

467 Plot of solubility

468
469 To identify the pH of minimum solubility: plot concentration at saturation versus pH and provide a
470 graphical representation of the results. Include error bars on mean.
471
472 Example chart

473

<API> solubility profile

5
Equilibrium concentration

3
(mg/ml)
mean

0
pH 1.2 pH 4.5 pH 6.8
474
475
Please report here the intermediate calculation for the DSV at (add additional pHs tested as
appropriate):
 pH 1.2 Highest therapeutic dose (mg)/Solubility (mg/ml)] [Concentration mean] =
 pH 4.5 Highest therapeutic dose(mg)/Solubility (mg/ml) [Concentration mean]=
 pH 6.8 Highest therapeutic dose(mg)/Solubility (mg/ml)] [Concentration mean] =
476

477 Conclusion: is the highest single therapeutic dose (according to the approved originator
478 product labelling) soluble in 250 mL of buffer over the pH range of 1.2 - 6.8 at 37 ± 1 °C i.e.,
479 in all buffers tested including buffers at pH 1.2, 4.5, and 6.8?

480 <Yes>/<No>

481
482
 Working document QAS/17.699/Rev.2
page 22
483 Solubility classification (please refer to Multisource (generic) pharmaceutical products:
484 guidelines on registration requirements to establish interchangeability. WHO Technical
485 Report Series, No. 1003, 2017, Annex 6):
486
487 <High> / <Low>
488
489 Name of the head of laboratory or person authorized to approve the certificate:

490 ________________________________________________________________________________

491 Telephone: E-mail:

492 Signature: Date:

493

494

495 ***