Sterile Pharmaceutical Preparations PDF

Sterile pharmaceutical preparations Dr. Abdulkarim Alzomor 1 Dr. Alzomor Sterile preparations include: Parenteral preparations Ophthalmic preparations Dialysis solutions Irrigation solutions Radiopharmaceuticals Plasma expanders 2 Dr. Alzomor References 1.Aulton's Ph...

Sterile pharmaceutical preparations Dr. Abdulkarim Alzomor 1 Dr. Alzomor Sterile preparations include: Parenteral preparations Ophthalmic preparations Dialysis solutions Irrigation solutions Radiopharmaceuticals Plasma expanders 2 Dr. Alzomor References 1.Aulton's Pharmaceutics The Design and manufacture of Medicines 2.Pharmacy practice (Ballington) 3.Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems 4.Remington The science and Practice of Pharmacy 5.British Pharmacopeia (BP) 6.British National Formulary (BNF) 3 Dr. Alzomor Parenteral preparations Introduction and definitions Advantages and limitations Types of injections Classifications of injections Specifications and requirements Components of parenteral products Production process Compendial quality control testing 4 Dr. Alzomor Parenteral preparations Para = beyond; entrone = intestine Sterile dosage forms that are delivered to the patient by a way outside (bypassing) the alimentary canal B.P.: “Parenteral preparations are sterile preparations intended for administration by injection, infusion or implantation into human or animal bodies” 5 Dr. Alzomor What is the difference between the term “IV injection” and “IV infusion”? IV Injection involves administration of Small volume parentrals (SVP) medications by a syringe into the a vein directly to blood stream IV infusion involves administration of Large volume parentrals LVP via a catheter into the vein directly to blood stream 6 Dr. Alzomor IV infusions can provide the following for hospitalized patients: 1.Parenteral nutrition (PN) and total parenteral nutrition (TPN) 2.Correction of fluids or electrolyte imbalance 3.Large doses continuous administration of drugs (antibiotics, chemotherapy) 7 Dr. Alzomor General Advantages (Rational) 1.Drugs with instability in GIT (insulin, heparin) 2.Drugs with poor GIT absorption (streptomycin) 3.GIT irritating drugs 4.Unconscious or uncooperative patients 5.Rapid correction of fluid and electrolyte imbalance 6.Nauseated and vomiting patients 7.Shorter onset of action (emergency) 8.Lower dose and side effects (why?) 9.Drugs targeting specific organs 8 Dr. Alzomor General limitations 1.Invasive and painful (lower compliance) 2.Traumatic injury from insertion of needle or catheter into the body 3.Mistakes (toxic dose, incorrect drug or presence of contaminants) are difficult to be treated. 9 Dr. Alzomor  Types of injections 1.Intramuscular route (IM): injections made into muscles (5 ml) 2.Intravenous route (IV) Direct injection into the vein 3.Subcutaneous route (SC): injections made under the skin (1.5 ml)(insulin injection) 10 Dr. Alzomor 4. Intradermal injection (ID): injections made into the skin dermis (allergic testing; Penicillin ) 5.Intraspinal (Intrathecal) injection : Injection through interthecal spaces into the spinal fluid (spinal anesthesia) 6.Intraperitoneal injection (IP): injection into the peritoneal cavity (animal studies) 11 Dr. Alzomor  Common types of injections 1. Intravenous route Advantages 1. The IV route fastest method of systemic administration among others 2. The preferred administration route for emergency situations 3. IV infusions (LVP) can provide large doses of fluids, electrolytes , nutrition and drugs for hospitalized and unconscious patients For patients having serious GIT problems (ulcers, 12 Dr.intoxication) Alzomor  Administration and precautions 1. A danger for IV route is the risk for introduction of pyrogens, toxic agents or microorganisms directly into blood stream (irreversible way) 2. The solution must be Free of particles or air bubbles (Fatal blockade of blood vessels= embolism) 3. Not suitable for suspension-based injections (great restriction) or oily injection 4. Cannot achieve depot effect 5. Infiltration 13 Dr. Alzomor  Infiltration  Breakdown or collapse of veins that allows the drug to leak into sites surrounding site of needle causing edema and tissue damage.  IV administration needs an expert 14 Dr. Alzomor  2. IM injection  Slower onset of action compared to IV. (disadvantage)  Longer duration (depot effect)  Suspension and oil-based injections (slowly dissolved, slowly absorbed)  More practical for use outside the hospital compared to IV injections  Volume of injection up to 5 ml. 15 Dr. Alzomor  Classification of parenterals 1.Pharmaceutical classification 2. Volume-based classification 16 Dr. Alzomor  1. Pharmaceutical classification 1.Solutions 2. Suspensions 3. Emulsions 4. Dry powder 17 Dr. Alzomor  1. Solutions Water (grade?) Aqueous Water+ co-solvent (IM & IV) (Solubility and stability) EXAMPLE? Solutions Non-aqueous Oils (fixed) (IM) Refer to components of parenterals (solvents and additives) 18 Dr. Alzomor  2. Suspensions  For insoluble drugs in water  Given via IM  Prolonged (depot) effect  This will ↓ drug dissolution rate in tissue fluids and result in ↓ absorption rate and sustained effect  Common particle size range 5-10 µm  IV injection? 19 Dr. Alzomor  2. Suspension (cont.)  Drawbacks: 1. Caking 2. Syringeability  Characteristics of the suspension while drawing it into the syringe  Ease of withdrawal from the container into the syringe, clogging and foaming tendency and accuracy of dose measurement. 20 Dr. Alzomor  3. Emulsions  O/W emulsions for IM and IV injections  Parenteral IV emulsions are rare: comment  It is necessary to achieve a stable droplet size lower than 1 µ (emolism)  Ex. Vit K . Refer to BNF 21 Dr. Alzomor  4. Dry powder  For unstable drugs in solution (ex antibiotics)  Powder should be reconstituted immediately before injection  May be reconstituted into solution or suspension  The pharmacist should be totally aware with the final reconstitution volume , form and expiry date. 22 Dr. Alzomor  2. Volume based classification Large volume Small volume parenterals (LVP) parenterals (SVP) Volume 100-1000 ml < 100 ml Can be added Additives No additives (example?) Type of fluid Solutions and emulsions Solution, emulsion, suspension Dose (container) Single dosed containers Single (ampoules) or multiple dosed (vials) Route IV IV, IM 23 Dr. Alzomor How to achieve controlled drug delivery in parenteral preparations? 1. Increasing particle size (suspension) and viscosity (oil solutions and suspensions or viscosity imparting agents):↓ drug dissolution rate in tissue fluids and result in ↓ absorption rate and sustained effect 2. Use of implants (solid sterile DF) 3. Use of less soluble salts (Insulin-Zn) 4. Use of crystalline rather than amorphous form (Insulin Lente) 24 Dr. Alzomor  Insulin lente Insulin zinc suspension An intermediate acting porcine or human insulin with zinc salt added such that the solid phase of the suspension contains a ratio of 7:3 Crystalline to amorphous insulin 25 Dr. Alzomor  Assignment 1 Investigate different formulations of insulin in market with variable duration and onset of action. 26 Dr. Alzomor  Assignment 2 Collect some examples from market on the following pharmaceutical forms of parenteral products: Dry powder for reconstitution Oily injections Long acting injections Emulsion based parenterals Regarding: Active constituent, different doses; Label (excipients used); method of 27 reconstitution Dr. Alzomor ) Specifications of parenteral preparations 1.Sterility 2.Clarity (limit for particulate matter) 3.Absence of Pyrogens 4.Tonicity 5.pH 28 Dr. Alzomor  1. Sterility Sterile preparations are completely free from microorganisms and contaminants Presence of pathogen in parenteral preparations is very serious (fatal in IV) To keep product sterility, preservatives (antimicrobial agents) are used. 29 Dr. Alzomor  Methods of sterilization Moist heat sterilization Dry heat sterilization Ionizing radiation sterilization Gaseous sterilization (ethylene oxide) Filtration (bacterial filters ): cannot be used for suspensions? 30 Dr. Alzomor  Preservation Antimicrobial agents are added to multi-dose vials Protect the injection from contamination during production, use and storage. Preservatives are not added to LVP (single dose containers discarded after opening); why? Not used if the drug itself has antimicrobial effect (methohexital sod. Injection) 31 Dr. Alzomor  Preservation. cont Ideal preservative must be: 1.Effective against wide range of bacteria 2.Nontoxic in the concentration used 3.Stable 4.Compatible with injection components (No interaction) Preservative uptake is more significant with natural and neoprine rubber Much less uptake with butyl rubber closures. 32 Dr. Alzomor Commonly used preservative in multi- dose injections Preservative Concentration (% w/v) Benzyl alcohol 1-3 Chlorocresol 0.1- 0.3 Cresol 0.25 – 0.5 Methyl para hydroxybenzoate 0.1 (Methyl paraben) Propyl para hydroxybenzoate 0.1 – 0.2 (propyl paraben) Phenol 0. 25 – 0.6 Thiomersal 0.01 NB: Benzalkolium chloride (4ry ammonium salts) 33 Dr. Alzomor is not used for injection (toxic).  2. Clarity Parenteral solutions must be clear: Completely free of any foreign particles (dust, glass, fiber): WHY?: Foreign particles are non-biodegradable, if injected will accumulate: IM: muscle granuloma IV: Embolism (thrombosis): Fatal 34 Dr. Alzomor  3. Absence of pyrogens Parenteral solutions must be non-pyrogenic (pyrogen free) Pyrogens are fever producing endotoxins Found in the outer membrane of Gram- negative bacteria (most common) Pyrogens are composed of lipopolysaccharides (Lipid A linked to a central polysacharide core) 35 Dr. Alzomor  Sources of pyrogens: 1.Solvents (water) is the main source 2.Equipment 3.Packing materials 4.Raw materials used 36 Dr. Alzomor  Biological activity of pyrogens Injection of pyrogens can produce toxic effect Contamination of LVP can specifically cause serious problems: 1.Lipid A can affect thermoregulatory centre in the brain (fever) 2.High doses of pyrogens will activate the coagulation system and produce shock and ultimately death. 37 Dr. Alzomor  Characteristics of the pyrogens 1.Water soluble 2.Non-volatile 3.Pass through bacterial filters 4.Heat stable 38 Dr. Alzomor  Depyrogenation Depyrogenation is the elimination of all pyrogens from the solvents, equipment and raw materials Achieved either by removal or inactivation 39 Dr. Alzomor  1. Water depyrogenation A. Distillation (BP) is the principle method to avoid water contamination with pyrogens (non- volatile) B. Reverse osmosis (USP) Filtration of water under high pressure through semi-permeable membrane that separate all substances and most ions C. Ultrafiltration can remove pyrogens from 40 solutions Dr. Alzomor  2. Packing materials and equipment Removing pyrogens from surfaces by rinsing with non-pyrogenic water Inactivation of pyrogens by dry heat at 250 ◦C for 30 minutes (for hermostable glass ware and equipment; not for plastic) 41 Dr. Alzomor  4. pH (cont.) Ideal pH of the parenteral preparations is 7.4 HOWEVER, some injected drugs require different pH values than neutrality (WHY?) Some drugs are insoluble at pH 7.4 others are unstable at this pH. 42 Dr. Alzomor  What is the solution? 1. Non-neutral SVP (IM, SC, IV): Formulate at the suitable pH for the stability or solubility Add a buffer system of low buffering capacity (why?) Allows rapid neutralization of the SVP by the natural buffer systems Buffers used: acetate (1-2%), phosphate (0.8-2%) and citrate (1-5%) buffers 43 Dr. Alzomor  1. Non-neutral SVP (cont.) The acceptable pH range for non-neutral SVP is 4-9 for tissues (other routes than IV) Body tissues: Higher or lower values are very irritant and damaging to sensitive cells. N.B: Borate buffer use in parenteral 44 formulations is forbidden Dr. Alzomor  Non-neutral LVP IV infusions should not contain buffering system (comment?) 1. Large volume needs large amount of buffer (toxicity) 2. To allow neutralization (full neutralization of large amount of buffer is difficult even if low buffer capacity is used) 45 Dr. Alzomor  Non-neutral LVP (cont.) IV infusions of non-neutral pH values should be injected very slowly ; low infusion rate (justify) No change (acidosis or alkalosis) in blood pH by the large volume (allow rapid neutralization) Avoid precipitation of the drug in the circulation (No sudden change in formulation pH) 46 Dr. Alzomor  Non-neutral LVP (cont.) The accepted pH range of non- neutral LVP is 3-10.5 Any decrease or increase in blood pH can result in systemic acidosis or alkalosis (life threatening) 47 Dr. Alzomor  5. Tonicity (osmotic pressure) If a semi-permeable membrane separates water from electrolyte solution, water will pass from its compartment to the salt compartment The pressure required to overcome water pressure is the osmotic pressure 48 Dr. Alzomor  Tonicity Parenteral preparations should be isotonic Have the same osmotic pressureas blood plasma Solutions having OP plasma are hypertonic 49 Dr. Alzomor  Tonicity Injection of hypertonic solutions Painful to tissues with nerve supply (IM, SC) Shrinkage of RBCs (IV, reversible) Destructive to delicate nerve cells in the spinal cord (intrathecal injections) Intrathecal injections must be isotonic (avoid serious changes in the tonicity of cerebrospinal fluid) Injection of hypotonic solutions IV: Haemolysisof RBC (irreversible, FATAL) WHAT 50 Dr. IS Alzomor THE SOLUTION?  Hypertonic parenteral solutions Hypertonic solutions for injection can be made isotonic by…dilution…………. Hypertonic solutions for IM and SC injections can be injected (slightly painful) Hypertonic solutions of IV infusions can be injected slowly or via central catheter 51 Dr. Alzomor  Comment In some cases, hypertonic IV infusions are deliberately prescribed to the patient. In cases of TPN where high concentrations of nutrients are required (25% dextrose) 52 Dr. Alzomor  Hypotonic parenteral solutions Made isotonic by addition of electrolytes as sodium chloride or osmotic agents as mannitol or glucose. The latter two agents are incompatible with some drugs. Solutions are made isotonic with isotonic saline solution (0.9 % NaCl). 53 Dr. Alzomor  Tonicity adjustment The amount of solute required to be added or the dilution required can be calculated using different methods 1.The freezing point depression method 2.Sodium chloride equivalent 3.Molar concentrations 4.Serum osmolarity Refer to tutorial session 54 Dr. Alzomor  Lecture Quiz Compare between SVP and LVP solutions regarding the following points: SVP LVP Sterility Clarity Absence of pyrogens Use of buffers (type/ example) Injection of non-neutral solutions (4-9) Injection of Hypotonic solutions Injection of Hypertonic solutions 55 Dr. Alzomor  Components of parenteral products 1.Container 2. Active (packaging) constituent 3.Solvent 4. Additives 56 Dr. Alzomor  1. Packaging (containers) 1. Glass ampoules 2. Rubber Stoppard vials 3. Glass and plastic bottles 4. Glass and plastic syringes 57 Dr. Alzomor 5. Prefilled syringes  Packing materials Rubber Glass Plastic 58 Dr. Alzomor  Ampoules Single dose glass container for SVP The glass neck should be scratched (or self-breaking) NB: may release some glass particles into the solution (filled under vacuum) 59 Dr. Alzomor  Vials Multi-dose and single dose SVP(1-100 ml) Glass bottle with rubber cap and sealed with aluminum seal covered with plastic cover Rubber material may be natural or synthetic (butyl rubber) 60 Dr. Alzomor  Vials Limitations: 1.Incomplete sealing of multi-dose 2.Air contamination 3.Release of rubber particles 4.Adsorption of injection components (preservatives) 61 Dr. Alzomor  Pre-filled syringes  Advantage: overcome problems of particle release and air contamination  Limitations: Expensive, need special type of machines 62 Dr. Alzomor  Plastic containers LVP Single dose IV infusion bottles (100-1000 ml) Polymer (Polyethylene or polypropylene or PVC) +additives (plasticizer, opacifier) Advantages: Handling; transportation Disadvantages: 1.Leaching Some drugs (fat emulsions, Paclitaxel, Amiodarone hydrochloride (Cordarone Xc), blood) leach the plasticizer DEHP(Diethylhexyl phthalate) from PVC container 2.Adsorptionof some drugs on PVC (nitrglycerin) 3.Low 64 clarity Dr. Alzomor and inspction Flexible (PVC) Adsorb some drugs (nitroglycerin) No need for air tube WHY?(Solutions drains out by gravity due to bag flexibility. Semi-Rigid (PE) More compatible Needs aeration for solution drainage (contamination) 65 Dr. Alzomor  Glass bottles Advantages: ❑ Transparent; chemically inert ❑ Used with incompatible materials with plastic Disadvantage:  Breakage; transportation  Need for aeration tube or needle (contamination) 66 Dr. Alzomor  Glass bottles  Glass Chemically consists of silicone dioxide + other oxides; three types: 1. Type III glass ❖ Soda lime glass (SiO2+Na2O) ❖ Releases alkalinity with water (for dry powder, non- aqueous soln.) 2. Type II glass ❖ sulphated glass (common use) ❖ Internal surface treated with sulphur oxide to neutralize alkalinity 67 Dr. Alzomor  Glass (cont.)  Type II glass cannot be reused (repeated wash and autoclaving remove internal layer)  Type II glass cannot be used with high pH products 3. Type I glass (SiO2+ boron oxide)  No release of alkalinity  Multiple use  Expensive 68 Dr. Alzomor  Solvents for injection  Constitutes the highest proportion of the formulation  Should be non-toxic and inert Aqueous Non-aqueous Water Oil Co-solvents Oily material 69 Dr. Alzomor  Grades of water 1. Water for injection (WFI)  Potable water may be contaminated with organisms, particles, dissolved gases or minerals  WFI is completely free of pyrogens and of high chemical purity  Common use as vehicle for preparation of parenteral products 70 Dr. Alzomor  WFI (cont.)  WFI can be prepared by distillation (BP) Feed water for distillation should be pretreated by  Filtration  Chemical softening  Deionization  pH adjustment  Reverse osmosis 71 Dr. Alzomor  2. SWFI WFI is packed in sealed containers (single dose) followed by sterilization of whole container by moist heat Used to dissolve or dilute parenteral products before patient administration 3. Bacteriostatic water for injection: SWFI containing bacteriostatic agent (multidose) 72 Dr. Alzomor  Co-solvents  Water-miscible solvents are mixed with water in some parenteral preparations (why?) 1. Enhance solubility of some drugs 2. Act as stabilizers  The most common: glycerin, ethyl alcohol, propylene glycol and PEG 300  Higher concentrations may be toxic, irritant and painful (ethanol)  Can be given both IV and IM 73 Dr. Alzomor  Non-aqueous solvents (oils)  Used for solubilization (digoxin)  Stabilization of water hydrolysable drugs (barbiturates)  Sustained effect (steroids) Refer to some official injections in oil (USP) (assignment) 74 Dr. Alzomor   Major class of non-aqueous solvents is fixed vegetable oils  Example: Corn oil, cottonseed oil, peanut oil and seasame oil  The oil in the product must be stated on the label  Some oils may cause sensitivity reactions to some patients  Mineral , volatile and animal oils never been used in injections. 75 Dr. Alzomor  Additives  To provide effective, safe and elegant preparation 1. Antimicrobial agents 2. Buffers 3. Tonicity adjusting agents 4. Antioxidants 5. Surfactants 76 Dr. Alzomor  Antioxidants  Added for the purpose of stability (how?)  Preferentially oxidized and gradually consumed over the shelf life of the products  Examples:  Ascorbic acid (0.01-0.5%),  Cysteine (0.1-0.5%),  Sodium metabisulfite (0.1-1%)  Tocopherol (0.05-0.5%) 77 Dr. Alzomor  Surfactants  Utilized in parenteral suspensions for wetting, to prevent crystal growth and provide good syringeability  Used also in parenteral emulsions  Examples: Sorbitan monooleate (0.05-0.25%) and polyoxyethylene sorbitan monooleate (0.1-0.5%) 78 Dr. Alzomor  Production of parenteral solutions Parenteral preparations are produced and filled into containers in a high standard clean room environment Any mistake in the productions leads to serious contamination with microorganism, pyrogens or particulate matter : 1. Raw materials 2. Personnel 3. Production area 79 Dr. Alzomor  Production of parenteral solutions 1. Raw materials Special grade for injections (pyrogen free) 2. Personnel: High degree of training High personnel hygiene Special clothes (fiber free and sterile) to prevent foreign material contamination 80 Dr. Alzomor  3. Production area Sterile rooms fitted with Laminar flow hoods and HEPA filters Air is filtered through HEPA filters that remove 99.97% of all particles larger than 0.3 μm. and microbial contaminants as well. 81 Dr. Alzomor  NOTE THAT: Laminar flow is not a means of sterilization, it only maintains an area free from particles and microbial contaminants, specially in IV admixtures When one or more sterile products added to an IV fluid for administration, the resulting combination is known as IV admixture. Manipulation (Operation) of IV admixtures should be under aseptic environment (maintain sterility, clarity and pyrogen free). 82 Dr. Alzomor Quality control testing and evaluation (BP) I. Tests for container II. Tests for III. Tests for whole samples of batch the batch Stick to the compendial methodology 83 Dr. Alzomor  I. Tests for the container A. Tests on glass containers B. Tests on plastic containers C. Tests on rubber closures 84 Dr. Alzomor  A. Glass test (glass alkalinity) 1. Surface test (for surface alkalinity) Fill the container with water then autoclave and titrate with sulfuric acid solution The titer should not exceed certain value. 2. Powdered glass test (Total alkalinity ) Crush the container, add water then autoclave Proceed as above. 85 Dr. Alzomor  B. Test for plastic containers (leaching) Fill the container with water and autoclave (at lower temperature. Water is tested chemically for color, pH, turbidity, UV absorbance and reducing substances Water is tested biologically in some cases of new plastic substances Injected into mice and any toxic symptoms are observed. 86 Dr. Alzomor  C. Test for rubber closure 1. Test for releasing materials Soak closure in water then autoclave Test water chemically as in leaching test 2. Permeability test To test the force needed for piercing the closure The force should not exceed certain value 87 Dr. Alzomor  3. Resealing test To test efficiency of vial resealing as a multi- dose container 4. Fragmentation test To test the number of rubber fragments inside the vial Should not exceed certain value 88 Dr. Alzomor  II. Tests for the whole batch A. Leaker test Test for complete sealing (no leakage) of the ampoules Immerse the ampoules directly after autoclaving in water bath containing methylene blue Test the ampoules visually and reject any colored ampoules) 89 Dr. Alzomor  B. Clarity test Test the presence of visible particles › 50 μm in the injection The injections are tested visually by human inspection under good light against white and black backgrounds Any container having visible particles is rejected 90 Dr. Alzomor  III. Tests for samples of the batch A. Sterility tests (refer to microbiology) B. Pyrogen tests 1. Rabbit pyrogen test 2. Bacterial endotoxin test (LAL test) C. Extractable volume test D. Clarity test for sub-visible particles 1. Direct measurement 2. Microscopical measurement 91 Dr. Alzomor  Pyrogen testing 1. Biological test: Rabbit pyrogen test Qualitative test Inject the tested solution IV into 3 rabbits and measure the increase in rabbit body temperature Pyrogenic solutions would cause elevation in body temperature in 3 hours The increase in each rabbit body temp not more than 0.6◦C The increase in the all rabbits body temp. not more than 1.4 ◦C 92 Dr. Alzomor  1. Rabbit pyrogen test (cont.) Advantages Give reaction similar to that in man Detect all types of pyrogens Disadvantages Expensive Time consuming Low sensitivity Some components of the injection may interfere with the test 93 Dr. Alzomor  2. Bacterial endotoxin test LAL (Limulus Amebocyte Lysate ) test. An in-vitro test Depends on gel formation when pyrogens are added to LAL reagent The LAL agent contains clotting proteins while pyrogens help coagulation process 94 Dr. Alzomor  LAL test (cont.) Advantages Rapid , easier to perform Inexpensive Quantitative determination of endotoxins Disadvantages Many factors may affect the test results (pH, ions in solution) Detect pyrogens of Gram negative bacteria (more dangerous and common) but not that of Gram positive or pyrogenic material 95 Dr. Alzomor  C. Extractable volume test Measured the volume that can be extracted from the container It should not be lower than the labeled volume D. Clarity test for sub-visible particles 1. Direct measurement 2. Microscopical measurement 96 Dr. Alzomor  1. Direct measurement Measure the number and size of the particles present in the solution directly Using coulter counter or laser diffraction: LVP Max 25 particles ≥ 10 μm/ml Max 3 particles ≥ 25 μm/ml SVP Max 6000 particles ≥ 10 μm/ container Max 600 particles ≥ 25 μm/container 97 Dr. Alzomor  2. Microscopical measurement Filter a certain volume of injection through membrane filter Calculate the number and size of particles microscopically: LVP: Max 12 particles ≥ 10 μm/ml Max 2 particles ≥ 25 μm/ml SVP: Max 3000 particles ≥ 10 μm/container Max 300 particles ≥ 25 μm/container 98 Dr. Alzomor