WO 2016/0S8985 PCT/EP2015/073852 Teach unknown Impurity | = 0. 201% 201% = 01% Total related substances | = 1.0% < 1.0% < 1.0% < 1.0% 7 Microbiology Sterile serie fo serie & Endotoxins = 30 EU ym | complies | == 9. Assay LC (melatonin) | 95.0 105.0% | 100.2% | 99.8% | 99.3% | 99.7% | 99.1% Example 8 Preparation of 2000 ml of oral solution of melatonin (25 mg MLT/ml).. Materials: MLT (assay 99.1%) commercial batch n. 194343002; Polyoxyl 40 hydrogenated castor oil commercial batch n. 23660956P0; Ethanol 96% commercial batch n. 14073-02. Auxiliary ingredients: Glycerol (£422) commercial product: 10.0 %; Xylitol (E967) commercial product: 7.0 %; Sodium saccharinate (E054) commercial product: 0.15 %; Cherry essence commercial product: 4.0 %; Purified water commercial product. Glassware as necessary; ingredients, container (38 ml amber glass bottle) and stopper are from qualified manufacturers and additionally tested by the quality control. All quantities are reported to 100% assay. Cleanroom B equipped with dosing machine for bottles (temperature 21°C; lightening system with UV filters). Preparation of 500 mi of bulk solution (Solution A) Step 1 - 50.45 g MLT (assay 99.1%) and 50.0 g polyoxyl 40 hydrogenated castor oil, both accurately weighed with a precision balance, were placed in a laboratory scale stainless steel mixer and thoroughly kneaded to yleld an homogeneous pasty mass (about 15 minutes); Step 2 - 500.0 ml of ethanol 96 per cent were added to pasty mass inside the mixer (before under vacuum and then using nitrogen as a purge gas) and stirred slowly to complete dissolution while a clear and transparent solution is obtained (about 15 minutes). The resulting bulk solution is 62WO 2016/0S8985 PCT/EP2015/073852 placed into a 3000 ml amber glass bottle that is well closed with a suitable stopper, protected from light (about 465 ml; step yield 93.0%). Step 3 - 3.0 mi sample of the above bulk solution withdrawn from the bottle was sent to the laboratory for analysis (MLT assay: 98.8 mg/ml) Step 4 - Preparation of 1500 ml of auxiliary ingredients (Solution B) In a 5 litres laboratory stirring tank (stainless steel) the following auxiliary ingredients (calculated to yield 2000 mi final oral concentrated solution) glycerol 200 g, xylitol 140 g, sodium saccharinate 3.0 g were thoroughly mixed in 1500 ml purified water until complete dissolution. Step 5 - Preparation oral concentrated solution (theoretical 2000 ml). 465 ml bulk solution (melatonin assay correction factor of 98.8/100=459 ml of 100% melatonin) was placed into a laboratory stirring tank from stainless steel and the cherry essence (8.0 g) was firstly dissolved into bulk solution A and stirred to complete dissolution. Thereafter 1395 ml of bulk solution B (by weight), representing the 75% of the final solution, were added and admixed to yield the final volume of 1854.0 mi of oral concentrated solution. Step 6 - 3.0 mi sample of the diluted solution withdrawn from the bottle was sent to the laboratory for analysis (MLT assay : 24.95 mg/ml). Step 7 - Once the oral concentrated solution received approval, it was passed through a membrane filter and the filtrate was collected directly into a suitable amber glass bottle. The bottle containing the diluted solution was passed to the Cleanroom B where the solution was apportioned by means of a dosing machine into amber glass bottles (38 ml capacity) from glass Type I (Ph. Eur.). A total 51 glass bottles filled with 30.0 mi diluted solution dosed at 25 mg MLT/mi were obtained; bottles were automatically closed by a suitable child proof anti-pilfer cap. The bottles were then labelled and transferred into a dark box protected from light. Two complete bottles were sent to quality control; solution passed physico- chemical tests (clear and transparent), melatonin assay (24.90 mg/ml), complied with pH range (6.5-7.5), filling volume and with microbiological enumeration test (TAMC max 107 CFU/g; TYMC max 10 CFU/g). The 63WO 2016/0S8985 PCT/EP2015/073852 product was packed in a folding box along with a dosing plastic device to adapt the solution dosage according to its use. The released product (n. 49 bottles; cumulative yield 74%) were partially used for the programmed stability tests. Example 9 Stability studies of pharmacy and industrial bulk solution package of melatonin for oral use of Example 8 of the invention - 30,0 ml multidose bottle. Study protocol Pharmaceutical form : 30.0 ml (25 mg MLT/ml) amber glass bottle (multidose) Batch n.° : bottle from Example 8 (A) Type of stability : accelerated Conditions : 40°C + 2°C / RH. 75% + 5% (climatic monitored storage) Duration : 6 months Intervals : 0 (initial), 3 and 6 months. Tests Specifications | TimeO | Smonths | 6 months T. Appearance Free of particles | conform | conform [conform 2. Clarity Clear solution cear_| dear clear 3. Colour Corourless colourless | colourless | colourless % Mean volume (vial) | 30.0- 31.5 ml sizmt | = 5. pH 60-75 69 70 65 @ Related Substances =5-methoxytryptamine | < 0.5% <05% | <05% | 0.3% = each unknown impurity | s 0.1% <0.1% | <0.1% | 0.2% Total related substances | = 1.0% <1o0% | <10% | 05% 7. Microbiology (CFU) Tame $10? 10? = 107 TMC 510 10 10 &, Assay LG (melatonin) | 95.0 - 105.0% 3.7% | 993% | 99.2% (B) Type of stability : long term 64