As reported before, the employed dose was 10 mg/ml is in according with commonly used dose.

Transcription

1 REPORT N. BF/STAB/DF/15/13 Object Stability Study of Flucloxacillin in DOSI-FUSER Elastomeric Infusion Pump Manufactured by Leventon S.A.U. Authors Biofordrug s.r.l. - Spin-off Università degli Studi di Bari INTRODUCTION The purpose of this report is to study the stability of Flucloxacillin into the elastomeric infusion pump DOSI-FUSER in Normal Saline (NS) and Water for Injectables (WFI), at storage periods and temperatures similar to those the drug is usually prepared and stored in Pharmacies and Hospitals for administration to patients. In particular, Flucloxacillin 10 mg/ml, in NS and WFI, has been studied in the elastomeric pump DOSI-FUSER and the study was performed at two different temperatures: 25 C and 4 C. The obtained data have been compared with data reported in following literatures: 1.Sarif Niroush Konari, Jane T. Jacob. Stability-indicating LC-analytical method development and validation for the simultaneous estimation of flucloxacillin and amoxicillin in pharmaceutical dosage form. Journal of Taibah University for Science, 2015, 9, Dhiraj S. Nikam, Chandrakant G. Bonde, S.J. Surana, G. Venkateshwarlu, P.G. Dekate. Development and Validation of RP-HPLC Method for Simultaneous Estimation of Amoxicillin trihydrate and Flucloxacillin sodium in capsule dosage form International. Journal of PharmTech Research 2009, 1, The part of device that is on contact to the drug is the elastomeric membrane and the infusion plastic line at the moment of the sampling. As reported before, the employed dose was 10 mg/ml is in according with commonly used dose. In all performed studies, the stability-indicating method involves the use of a combined HPLC/LC-MS approach allowing an unambiguous assessment of drug purity, stability and

2 compatibility with medical devices. The development and validation of the stabilityindicating HPLC/LC-MS method have been established matching the recommendations of official guidelines of the International Conference on Harmonization (ICH) and European Pharmacopeia (Ph. Eur.). The stability-indicating protocol was done according to the internal protocol POQ 10 Rev.1-01/10/2015. The details are reported in the document Doc. n. BF/STAB/DF Biofordrug s.r.l. is certified ISO Reg. N A for the following products-services: Pharmaceutical Products, in-vitro Diagnostic Kits, QC analysis and Release Certification for the Market.

3 I. PROTOCOL 1. Test product SUMMARY 2. Filling procedure 3. Medical device 4. Storage temperature 5. Preparation of test samples and sampling 6. Sample Collection 7. HPLC Apparatus 8. LC/MS Apparatus 9. Robustness 10. Stability Criterion II. RESULTS III. CONCLUSIONS IV. EXPERIMENTAL SECTION 1. Chemicals and Reagents 2. HPLC apparatus 3. HPLC Operating Conditions 4. Test sample preparation and testing frequency 5. Chromatograms 6. LC-MS analyses

4 I. PROTOCOL N A 1. Test product. The employed drug is described in the Table 1 Drug Vol Filled Concentration Diluent Brand Manufacturer Batch Expiry date Flucloxacillin 250 ml 10 mg/ml NS, WFI Flucacid1g Euro- Pharma S.r.l /2017 Table 1 2. Filling Procedure The filling procedure of the elastomeric pumps was executed according with protocols of the Unit of Antineoplastic Drug Handling (U.Ma.CA.) at the Cancer Institute "Giovanni Paolo II", Bari. The manipulation was performed in accordance with international standard guidelines on Injectables. The details of the procedure are reported in Doc. n. BF/STAB/DF In each pump the final concentration is C= 10 mg/ml and the total volume is V= 250 ml. The number of filled elastomers is reported in the Table 2: Concentration 10 mg/ml Diluent NS WFI N of Devices 4 4 Table 2

5 3. Medical device. DOSI-FUSER Model: 250D1; LOT L; Dispensing speed: 10.4 ml/h; Expiry date Storage temperature. Elastomeric pumps have been stored at two different temperatures as reported in Table 3. Concentration 10 mg/ml Diluent NS WFI Storage Condition 4 C* 25 C** 4 C* 25 C** N of Devices Table 3 **Oven: Binder Drying and Heating chamber with natural convection MODEL ED 115 *Refrigerator: Bosch Energy A+ 5. Preparation of test samples and sampling. Samples were taken as reported Doc. n. BF/STAB/DF The determinations are indicated in the Table 4.

6 Concentration 10 mg/ml Diluent NS, WFI Storage Condition 4 C 25 C Determinations* 10 d 24 h Table 4 (* h= hours, d=days) 6. Sample Collection. The collection was carried out directly by the terminal part of the plastic infusion line in a glass tube and then the solution was transferred into vials for HPLC and LC-MS analysis. The samples were used as such in HPLC and LC-MS Analysis. 7. HPLC Apparatus. HPLC Analyses were performed on Agilent 1260 Infinity instrument equipped with 1260 DAD VL + detector controlled by OpenLAB CDS ChemStation Edition Software. The employed column was a C18 reversed phase column Vydac 218TP 5 µm. HPLC operating conditions (Wavelength, Flow, Mobile phase, Injection volume, Retention time, Column temperature) are reported in Experimental Section. 8. LC/MS Apparatus. LC-MS analyses were performed on an Agilent 1100 LC/MSD trap system V spectrometer equipped with an electrospray ionization (ESI-MS n ) system. The samples (2 ml) were analysed after Lyophilization and subsequent extraction with an appropriate solvent (AcOEt).

7 9. Robustness. N A The precision of the method was verified by repeatability in a single day and the intermediate precision by different analysts on different days. Repeatability and intermediate precision were maintained by analysing the samples in triplicate. The results are summarized for each drug and the corresponding typical chromatograms have been reported. The critical parameters of flow rate and temperature were changed to assess the method performance. Moreover the Method Validation was performed as reported in Doc. n. BF/STAB/DF Stability Criterion For Flucloxacillin the qualitative and quantitative results have been compared with the data reported in literature about substance identification, drug shelf life, safe dose and degradation products. In general the maximum accepted variability, in terms of concentration, is ± 10% respect to the expected concentration.

8 II. RESULTS N A In this study the employed method involved the use of a combined HPLC-LCMS approach. The samples were taken over the studied period and submitted to HPLC analysis and LC- MS evaluation allowing an unambiguous assessment of drug purity, stability and compatibility. The used procedure for establishing purity and stability of the tested drug was to compare the HPLC and LC-MS results with that of a reference solution. For the quantitative evaluation, the areas of peak under interest have been compared with that of the reference solution to appreciate the stability of drug concentration during the period of analysis. Final quantitative results are reported in term of average % change in areas and average concentration of samples (two different determinations related to two different elastomers for each experimental condition) in Tables 5 and 6. The details about the protocol are reported in Doc. n. BF/STAB/DF For the qualitative evaluation the HPLC and LC-MS analyses of all samples have been compared with the results obtained with a standard solution of the drug. Moreover, particular attention was given to the monitoring of colour and clarity of the solution into the elastomer and into the taken samples. In this regard, there are no findings to communicate.

9 Flucloxacillin in NS T h Area 1 (%) Area 2 (%) Average concentration (mg/ml) Average Concentration Change (%) C 24 h C 10 d Table 5 Flucloxacillin in WFI T h Area 1 (%) Area 2 (%) Average concentration (mg/ml) Average Concentration Change (%) C 24 h C 10 d Table 6

10 I. CONCLUSIONS Qualitative The obtained HPLC-chromatograms and LC-MS analyses of reference solution and of all samples displayed that Flucloxacillin is stable in the Dosifuser device for all experimental conditions reported in Table 4. In fact, as reported in HPLC and LC-MS studies other peaks due to degradation of drug and/or to medical device-drug interaction are not detected. Quantitative The study of % variation of areas has been employed to appreciate the stability of drug concentration during the requested period of observation. As reported in Table 5, very low variability at the all studied period and at all experimental conditions has been observed. This finding allows to confirm that for each determination, the drug concentration is constant and superimposed with respect to reference solution and the sample at T=0.

11 IV. EXPERIMENTAL SECTION 1. CHEMICALS AND REAGENTS. All chemicals and reagents were of the highest purity. All solvents were HPLC grade quality and all chemicals were purchased from Sigma- Aldrich Co. (St. Louis, USA). 2. HPLC APPARATUS HPLC: Agilent 1260 Infinity OpenLAB CDS Column: Vydac 218 TP C18 5µm Detector: DAD 2. HPLC Operating Conditions Wavelength: 280 nm Flow: 1 ml/min Mobile phase: (CH 3 CN: Phosphate Buffer 10 mm) = (80:20) ph= 3.5 Injection volume: 2 µl Retention time: about 2 min Column temperature: 25 C 3. Test sample preparation and testing frequency The samples, taken on the indicated days, were used as such for HPLC Analyses.

12 4. Chromatograms N A The chromatograms obtained by HPLC analysis of taken samples are reported below. a) T = 0, NS b) T = 24 h, NS at 25 C

13 c) T = 24 h, WFI at 25 C d) T = 10 d, NS at 4 C

14 e) T = 10 d, WFI at 4 C 5. LC-MS Analyses An example of ESI-positive analyses of samples is reported below. Flucloxacillin (MW = 453 g/mol): ESI + /MS m/z: 476 [M+Na] +. ESI+/MS/MS m/z: 317(100), 182(70). Bari, September 29, 2015