Assurance and Quality Control in Manufacturing Process Validation of powder for Extemporaneous Oral Suspension
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1 Assurance and Quality Control in Manufacturing Process Validation of powder for Extemporaneous Oral Suspension Miusa C. Coelho ab, Ricardo J. Grilo a, Maria D. Marques b a Laboratórios Atral SA, Rua da Estação 42, Vala do Carregado, P Castanheira do Ribatejo, Portugal b BSEL - BioSystems Engineering Lab, Instituto Superior Técnico, University of Lisbon, Portugal Abstract This study aimed to provide necessary competence and autonomy for the development and theoretical analysis / practice Manufacturing Process Validation documents in the pharmaceutical industry. The validation process was conducted in Cephalosporin Production Unit Labs Atral SA, as per the requirements of current Good Manufacturing Practices. The proposal is a prospective study of manufacturing process validation where analytical tests are described in all stages of the process, results and conclusion. The analytical results of these tests were positive and were within specification limits demonstrating that this manufacturing process, validated, is robust and practicable, being able to produce quality medicines. Thus, after the results can be concluded that the process in question is in a position to be transport to an industrial scale with the aim of marketing the resulting product. Keywords: Manufacturing Process Validation, extemporaneous oral suspension, Quality Assurance, Pharmaceutical Ingredient, Pharmaceutical Industry. 2. Material and methods 1. Introdution This dissertation on the topic "assurance and quality control in the powder manufacturing process validation for Extemporaneous Oral Suspension (EOS)", is designed to obtain a master's degree within the Master in Pharmaceutical Engineering (MEFARM), where for six months I carried out a theoretical and practical training in laboratories Atral, SA, belonging to the Portuguese pharmaceutical group AtralCipan. Three consecutive batches (X001, X002 and X003) of Extemporaneous Oral Suspension, each with dimensions of 6000 units were produced. Raw materials In Table I are presented the raw materials that make up the extemporaneous oral suspension [1]. Formula for calculation the actual amount of API Equation 1
2 Table I - Raw materials required for the production of Extemporaneous Oral Suspension Raw materials Function Weight (g) Percentage (%) Trihydrate SA A API Xanthan Gum Suspending agent Hipromelose 15 (HPMC) Stabilizing agent Sodium Benzoate Antimicrobial preservative Sodium lauryl sulfate Wetting agent Aspartame Sweetening agent Orange Essence Flavoring agente Tutti Frutti Essence Flavoring agente Ground sugar (Sacarose) Viscosity-increasing agent, sweetening agent Control tests performed during production Bulk Density: The tests carried out during the development of the new formulation were: API identification; bulk and tapped density test; water content; API content; torque; uniformity of weights. Tapped Density: Equation 2 Bulk and tapped densities Equation 3 Equipment and materials: compression apparatus, capable of causing 250 ± 15 strokes per minute at 3 ± 0.2 mm high; beakers of 250 ml; Support cylinder with its clamping device [2]. Sample size: 30 g powder. Technique: introducing the dry cylinder without compacting, 30 g of sample (volume corresponding register) or a test portion to an apparent volume of 50 ml (corresponding register mass). Fix the beaker on support, read the uncompressed apparent volume with an accuracy of 1 ml. Submit to 10, 500 and 1250 strokes and read the corresponding apparent volumes V10, V500 and V1250 with an accuracy of 1 ml. If the difference between V500 and V1250 is greater than 2 ml, making other strokes in 1250 [2]. 2.2 Analytical validation tests for the evaluation of the quality of the product during the blending stage (VP01) Organoleptic characteristics: This test is done visually, the acceptance criterion is white or almost white powder. API and Sodium Benzoate contents Assay Method API Sample size: 1.5 g. Equipment: HPLC (Merck), analytical balances (Mettler Toledo) with integrated printer, centrifuges, ultrasound, ph meter (Metrohm), water purification system Milli-Q [3]. Reagents: purified water, tetrabutylammonium hydroxide solution 40% 2
3 HPLC grade acetonitrile, methanol for HPLC, API reference and phosphoric acid 85% [6]. Acceptance criteria for the results: 90.0 to 110.0% Analytical conditions: column: Merck Purospher Star RP 18e, 125 x 4 mm, 5 um; column oven temperature: 40 C; Solution A: 20.5 ml of tetrabutylammonium hydroxide solution 40% in 1000 ml of purified water; adjusted ph to 6.5 with H 3 PO 4 (phosphoric acid) 85%; Mobile phase: Solution A: Acetonitrile (75:25, v /v); Flow rate: 1.0ml /min; Detection: UV at λ = 290 nm; Injection volume: 20 µl; Run time: 15 min [3]. Preparation of solutions: 749 mg to a volumetric flask of 100 ml; add 10 ml of methanol and submit to ultrasound for 5 min at room temperature and complete the volume with mobile phase; submit to ultrasound for more 5 min and centrifuge an aliquot at 4000 rpm for 5 min; dilute an aliquot of 2 ml of the supernatant in a 25 ml volumetric flask and complete with mobile phase. Preparation of reference solution: in a volumetric flask of 50 ml dissolve 28 mg of API reference in 10 ml of methanol and dilute to 100ml with mobile phase; from this solution dilute an aliquot of 2 ml to 25 ml with mobile phase. HPLC procedure: inject six standard solutions and two solutions for each of the ten samples of the mixture [3]. Assay Method preservative Sodium Benzoate HPLC grade acetonitrile, methanol for HPLC, reference sodium benzoate and phosphoric acid 85% [4]. Acceptance criteria for the results: 90.0 to 110.0% Analytical conditions: Column: Merck Purospher Star RP 18e, 125 x 4 mm, 5 μm; Autosampler temperature: 5 C. Solution A: 20.5 ml of tetrabutylammonium hydroxide solution 40% in 1000 ml of purified water, adjusted the ph to 6.5 with H 3 PO 4 (phosphoric acid) 85%; Mobile phase: Solution A: Acetonitrile (75:25, v/v); Flow rate: 1.0ml/min; Detection: UV at λ = 254 nm; Injection volume: 20 µl; Run time: 20 min [4]. Preparation of solutions: transfer 1.873g to a 250 ml volumetric flask; complete the volume with mobile phase and submit to ultrasound for 5 min at room temperature; centrifuge an aliquot of 10 ml at 4000 rpm for 5min; inject the supernatant of the solution [4]. Preparation of reference solution: dissolve 12.5 mg of sodium benzoate reference substance with the mobile phase and dilute to 100 ml volumetric flask, with mobile phase; take an aliquot of 2 ml and dilute to 10 ml with mobile. HPLC procedure: inject six reference solutions and two solutions for each of the samples. Calculations for determining the API content and Sodium Benzoate Concordance of the calculation formula of Sodium Benzoate Reference Standard Sample size: 3.75g. Equipment: HPLC (Merck), analytical balance (Mettler Toledo) with integrated printer, centrifuge, ultrasound, ph meter (Metrohm), the water purification system Milli-Q [4]. Reagents: purified water, tetrabutylammonium hydroxide solution 40% Average weight calculation formula Equation 4 3
4 Equation 5 Calculation formula of the API content and sodium benzoate as a percentage Equation 6 Dose - Standard sample size; A - Average area; A - Área. Calculation formula Control reference Sample size: 20 flasks, (beginning, middle and end of the filling). Sample size for the finished product: 10 flasks [5]. Equipment and materials: analytical balance (Mettler Toledo) with integrated printer, spatulas and cups for weighing [5]. Acceptance Criteria: ± 5% 36 g. Technique: Individually weigh the content of the units 20 flasks beginning, middle and end of the filling; determine the average weight; no more than 2 of the individual weights may deviate from the average weight found in higher proportion than the 10% and in no case can the difference exceeds 20% [5]. Calculation formula for determining the maximum deviation in uniformity of weights. - Área Média; standard - Average area reference. Equation 7 Equation 10 Calculation formula for determining the minimum deviation in uniformity of weights API content calculation formula in mg / 5ml Equation 8 Calculation formula of Sodium Benzoate content in mg / 5ml is the average. Average weight Equation 11 Equation Analytical test to assess the quality of the filling operation (VP02) The calculation of the average weight of the stage from the beginning, middle and end of the filling process is performed for each batch on an analytical balance. The average weight results are presented in section, and represent the average of samples from each phase (beginning, middle and end) of the filling process [5]. Acceptance Criteria: g. Uniformity of weights 2.4 Analytical tests for the evaluation of the final product (VP03) 4
5 Measurement of the ph of the reconstituted suspension Sample size: three bottles per batch. Equipment and materials: ph meter (Metrohm), volumetric glassware. Technique: move the vial until the powder is loose; add a little water, close and shake; complete with water to the existing brand on the label for the suspension after reconstitution transfer it into a glass and then introduce it in her bosom an electrode to determine the ph [6]. The acceptance criteria for the ph of the reconstituted suspension are Determination of water content Technique: reconstruct each of the ten flasks with 120 ml of water and stir individually; take care not to form air bubbles; placing the contents of each flask in individual beakers, in which the capacity not exceeding two times and a half the volume to be measured; in this case 200 ml beakers were used; during the transfer of the contents of each vial even allow to drain for a period not exceeding 30 minutes until no observe air bubbles; read the volumes. Microbial contamination tests and related substances were performed by employees in the microbiology laboratory and in the HPLC laboratory respectively; the methods are not described in this work. Size of sample: 300 mg of each flask, two vials per batch. Equipment and materials: Karl Fischer titrator, analytical balance (Mettler Toledo) with printer. Technique: weigh individually in a glass boat 300 mg of the suspension and record its weight; register the Karl Fischer titrator product data, enter 300 mg sample of the titration container; after titration it prints the value of the existing moisture in the sample; the acceptance criterion is 2.0% water content. Reconstitution Reagent: purified water. Technique: move the Flask until powder is loose; add a little water, close and shake; complete with water to the existing brand on the label for the suspension; after reconstitution should get a stable homogeneous suspension with smell of fruit. Deliverable volume after reconstitution Sample size: ten flasks per batch. Materials: 200 ml beakers. Reagent: purified water. 3. Results and discussion This section will present the results of analytical tests described in Materials and Methods. The data presented below are for the three batches studied. 3.1 Results of Process Control In Table II, it can be seen that the three batches test results are according to the specifications described in Materials and Methods. Table II - Results of control tests during the batches production Density: - Bulk (g.ml -1 ) - Tapped (g.ml -1 ) Identification API 0,60 0,88 0,58 0,86 0,58 0,85 Positive Positive Positive Moisture (%) 1,3 0,8 1,1 Asaay API (%) 101,1 98,6 99,8 Table III verification and weights in the three batches are conform to the acceptance criteria. These results demonstrate that the filling 5
6 process present robustness to produce EOS to be placed on the market. Tabela III - Control of the average weights during the filling operation Verification C C C Weight (g): Average Minimum (%) Maximum (%) 36,14 35,18 36,40 36,16 35,08 36,55 36,07 35,03 36,30 Figure 1 shows the control chart for variables performed with the results of weighing of EOS samples (during the filling step) using Microsoft Excel. It can be seen that from the all batches (X001, X002 e X003) representing 100% of the samples that are within the specification limits ( g). This filling method can be considered robust to the manufacturing process. I Figure - Control charts the weights of Oral Suspension Extemporaneous during filling of the three batches 3.2 Mixture results of quality assessment API contents In Table IV are present the results of the API contents of the batches X003 and X002. All samples were within specification limits. The X001 batch was the only one with two non-compliant results more with little variation over the limits specifications ( %). The standard deviation for the three batches was 5% as specification limits. As for visual description all samples were compliant. 6
7 IIV Table - Assay API and Aspect Samples Aspect - C C C Assay API 1 97,8 98,7 95,0 2 98,9 96,7 97, ,4 100,4 97, ,0 96,9 103,1 5 97,7 96,9 97, ,4 100,3 99, ,2 99,4 103, ,5 94,9 99,5 9 97,7 98,1 99, ,2 99,6 100,1 Average (%) 100,5 98,2 99,3 Minimum (%) 97,7 94,9 95,0 Maximum (%) 107,0 100,4 103,7 rsd (%) 3,3 1,8 2,7 Control charts of assay API Figure 2 shows the control charts for variables of API assays of the three batches studied. All the batches presented conforming results, with slight deviation in relation to the specification limits (95-105%), thereby confirming the homogeneity of the powder batch analysis. II Figure - Control charts of assay API for the three batches Results of the Sodium Benzoate Assay X001 and X002 batches showed the best results, with all values within specification limits (90-110%). The X003 batch only one sample showed a result (89.4%) six tenths below the minimum specification 7
8 limit (90-110%). The rsd for all batches was below the specification limit ( 5%). As for the aspect of the samples are comply. V Table - Assay Sodium Benzoate and Description in mixing Samples Aspect - C C C Assay Sodium Benzoate 1 98,8 102,5 99,1 2 94,1 100,1 97,2 3 97,8 94,9 89,4 4 95,2 101,4 96,4 5 98,8 98,2 98,3 6 91,7 97,4 97,1 7 99,9 96,3 96,8 8 96,9 97,5 96,2 9 98,5 101,2 97, ,4 102,1 98,3 Average (%) 96,6 99,2 96,7 Minimum (%) 91,7 94,9 89,4 Maximum (%) 99,9 102,5 99,1 rsd (%) 2,7 2,7 2,8 Control charts of sodium benzoate Figure 3 shows the control chart for variables in the determination of sodium benzoate for the three batches. In these batches, only sample 3 showed a batch X003 result (89.4%) outside the range of specification limits ( %), and the difference 6 tenths, but the rsd for all batches was below the specification limit ( 5%), these results demonstrate the robustness of the method used. Figura III - Control charts of determination assay of sodium benzoate for the three batches 8
9 3.3 Results evaluation the filling operation quality In Tables VI, VII and VIII presents the results of analytical tests carried out at the beginning, middle and end of the filling. The dosages of the API and sodium benzoate for the three batches are within the interval limits of the specifications for API (95-105%) and benzoate (90-110%), respectively. The results of the average weights for the three batches are within the specification limits ( g) with a uniform weight having a rsd below the limits ( 5%). These results show the homogeneity of the EOS. VI Table - Results of begin filling Aspect C C C Assay (%): API Sodium benzoate Average weight (g) Mass uniformity (%): Min. deviation Max. deviation 103,4 101,2 100,3 96,0 97,0 99,6 35,9 36,3 36,2-0,7 +1,4-1,3 +1,6-1,2 +1,0 IIIII Table - Results of Middle filling Aspect C C C Assay (%): API A Sodium benzoate Average weight (g) Mass uniformity (%): Min. deviation Max. deviation 96,2 102,2 100,4 96,5 IVIII Table - Results of end filling 98,8 94,8 36,1 36,2 35,9-1,4 +1,5-1,4 +1,2-1,3 +1,2 Aspect C C C Assay (%): API A Sodium benzoate Average weight (g) Mass uniformity (%): Min. deviation Max. deviation 99,1 93,5 97,7 96,4 96,0 97,6 36,8 36,04 36,2-0,9 +1,2-1,2 +1,4-1,6 +1,1 3.4 Results of the finished product All analytical tests, carried out on the finished product had results as the specification limits described in materials and methods. IX Table - Results of analytical tests performed to evaluate the finished product Aspect C C C Identification API Positive Positive Positive ph 3,9 3,9 3,8 Moisture content (%) 0,9 1,1 0,9 Reconstitution Satisfies Satisfies Satisfies Deliverable Volume: average (ml) individual (ml) , 122, 122, 122, 122, 122, 122, 120, 122, , 120, 120, 120, 120, 120, 122, 118, 122, , 120, 122, 118, 122, 122, 120, 122, 122, 120 9
10 IX Table - Results of analytical tests performed to evaluate the finished product Weight Uniformity (%): desviation minumum desviation maximum Assay (%): API Sodium Benzoate Related substances (%): Impurities A Impuities D Impuities E Impuities unknown maior Total Impurities Microbial control:* bacterium fungus Escherichia coli -0,8 +1,4 98,3 99,0 0,15 0,15 Absent -0,7 +0,8 99,3 96,3 0,15 0,052 0,2 Absent -0,7 +0,5 100,0 97,3 0,15 0,052 0,2 Absent 4. Conclusion The results showed to be within the specification limits, except for three of the assays obtained in the mixing step with little significant variation. Based on all the results obtained during the development of this new formulation is possible to prove that the manufacturing process of the EOS (Generic Drug) meets the requirements of quality, safety and efficacy to be validated and could be part of the production lines ATRAL laboratories, SA, to commercialize in the Pharmaceutical Market. 5. References [1] Raymond C Rowe, Paul J Sheskey and Marian E Quinn (2009), Handbook of Pharmaceutical Excipients, 6th Edition, Pharmaceutical Press, UK and the American Pharmacists Association, USA, pp.326, 627, 651, 703, 782. [2] Volume Aparente Comissão da Farmacopeia Europeia (2010), Farmacopeia Europeia 7.0, vol. I, Council of Europe, Strasbourg, France. [3] Laboratório de Estudos Farmacêuticos, (2009), Analytical Validation Report, Analytical Methodology of HPLC for the determination of Pharmaceutical Active in the Drug Product, Powder for oral suspension, Atral Cipan, S.A, Carregado. [4] Laboratório de Estudos Farmacêuticos (2009), Analytical Validation Report, Analytical Methodology of HPLC for the determination of Sodium Benzoate in the Drug Product, Powder for oral suspension, Atral Cipan, S.A, Carregado. [5] Uniformidade de massa da dose dispensada pelos recipientes multidose, Comissão da Farmacopeia Europeia (2010), Farmacopeia Europeia 7.0; vol I, Council of Europe, Strasbourg, France. [6] Determinação potenciométrica do ph, Comissão da Farmacopeia Europeia (2010), Farmacopeia Europeia 7.0, vol I, Council of Europe, Strasbourg, France. 10
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