OUR SERVICES FOR YOU

Transcription

1 OUR SERVICES FOR YOU Valid from May 2014

2 40 YEARS EXPERIENCE YOUR ADVANTAGE Founded in 1973, BioChem GmbH is an independent family-owned analytical contract laboratory, located in Karlsruhe, Germany, offering high quality laboratory services and competence in all areas of physico-chemical microbiological molecular biological testing for the pharmaceutical, chemical, medical and cosmetics industry. Our scientific and practical experience, combined with the expertise of more than 60 highly trained specialists at our side, guarantees highest quality, safety, and efficiency. Our expertise includes analytical testing of raw materials, intermediate or finished products in any dosage form as part of quality control or pharmaceutical development, EU re-analysis or ICH stability studies, and we are approved for handling cytostatic drugs and narcotics. We offer analytical testing in accordance with current guidelines (e.g., Ph. Eur., USP, JP, BP, HAB, DAB, AMG, MPG, Chemicals Act, OECD or EC) and we also comply with other national or international and customer specific requirements. As your trusted business partner and internationally-oriented modern service provider, BioChem GmbH always finds solutions to technical problems for your projects and requests. You will find an overview of our research methods and general cost guidelines in the following. We will be happy to provide you with product and project-specific service offers, which of course take into consideration quantity-specific synergies in the context of combinatory or graduated pricing. Give us a try. OUR SERVICE - YOUR ADVANTAGE 2

3 CONTENTS 1. QUALITY MANAGEMENT / ASSURANCE Page 5 2. ANALYTICAL TESTING Page 7-13 Analysis of Chemical and Physical Properties Sample Preparation Chromatography Spectroscopy / Spectrometry Electrochemical Analysis and Titration Analysis of Physical Properties Total Organic Hydrocarbon (TOC) Visual Inspections Testing for Particles Further Investigations on Request Special Tests for the Pharmaceutical Industry Release and Stability Testing for in-process Control Pharmaceutical Quality Control and Development Stability Studies (Storage and Testing) Pharmaceutical Development Photostability Testing Pharmaceutical Approval, Expert Opinions Testing in Accordance with GLP Guidelines 3. MICROBIOLOGICAL ANALYSIS Page Microbiology for Almost any Question Testing for Microbial Contamination of Non-sterile Products Identification of Micro-organisms Sterility Testing Testing of Biological Indicators Bacterial Endotoxin Testing using the LAL-Test Microbiological Assay of Antibiotics Efficacy of Antimicrobial Preservation On Site Environmental Monitoring Other Microbiological Tests 4. MOLECULAR BIOLOGY TESTING Page PCR Analysis Real-time PCR Analysis Fluorometric / Luminometric Detection Enzymatic Detection Protein Analysis / Immunochemistry Bioanalytics Validation / Testing under GLP Conditions Detection of Plants and GMOs by PCR Animal Species Differentiation 5. GENERAL ADDendum Page ORDER CONTRACT Page 33 3

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5 1. QUALITY MANAGEMENT / ASSURANCE BioChem GmbH's Quality Management (QM) and Quality Assurance (QA) is based on the extensive experience of our motivated staff, and our desire to continuously improve our procedures. The implementation of current pharmacopoeia specifications, relevant regulations and guidelines, and the safe handling of your products and data are a given for us. The analyses conducted by the company BioChem GmbH within the meaning of 14 para 4 no. 3 of the German Medicines Act (Arzneimittel Gesetzes, AMG) are registered with the responsible supervisory authority in accordance with 67 para 1 AMG. Pursuant to 64 para 1 AMG, BioChem GmbH is subject to regular monitoring for GMP and GLP compliance by the Control Centre for Drug Monitoring, Baden-Württemberg, and by the Chemicals Act or Directive 2004 / 9 / EG by the State Office for Environment, Measurements and Environmental Protection of Baden-Württemberg. In addition, BioChem GmbH is registered under the Facility Establishment Identifier (FEI) with the U.S. Food and Drug Administration (FDA) and has received unrestricted authorization as an independent, internationally operating contract laboratory in two pre-approval inspections (2009 and 2013). BioChem GmbH performs contract work according to the current state of science and technology and complies with recognized regulations and relevant statutory provisions, in particular in accordance with 14 and 23 of the German Ordinance on the Manufacture of Medicinal Products and Active Ingredients (Arzneimittel- und Wirkstoffherstellungsverordnung, AMWHV), and in accordance with the GMP principles of the EU GMP guidelines of good manufacturing practice for medicinal products and substances, the U.S. Code of Federal Regulations (21 CFR Part 210 / 211) and the Medical Devices Act (MPG). Regular customer-based audits (> 30 per year) ensure the maintenance of the high GMP and GLP standards that apply to the pharmaceutical environment. 5

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7 2. ANALYTICAL TESTING 2.1 ANALYSIS OF CHEMICAL AND PHYSICAL PROPERTIES Page Sample Preparation Chromatography Thin layer chromatography (TLC) High Performance Liquid Chromatography (HPLC) Ion Chromatography (IC) Capillary Gas Chromatography (GC) Special Chromatography Methods Spectroscopy / Spectrometry UV / VIS Spectroscopy / Photometry Infrared Spectroscopy (FT-IR) Atomic Absorption Spectroscopy Inductive-coupled Plasma Mass Spectrometry (ICP-MS) Electrochemical Tests and Titration Analysis of Physical Properties Total Organic Hydrocarbon (TOC) Visual inspections Testing for Particles Additional Tests on Request 2.2 SPECIAL PHARMACEUTICAL TESTS Page Pharmaceutical Quality Control and Development as part of In-Process Control, Release and Stability Testing Tests according to a Monograph Method Development, Validation, Method Transfer Technological Parameters of Drugs in Solid Forms Uniformity of Content / Mass Release of Active Ingredients Water for Pharmaceutical Purposes Stability Studies (Storage and Testing) Pharmaceutical Development Photostability Testing Pharmaceutical Registration, Expert Opinions 2.3 TESTING PER GLP GUIDLINES Page 13 7

8 2.1 ANALYSIS OF CHEMICAL AND PHYSICAL PROPERTIES SAMPLE PREPARATION The following established sample preparation techniques are standard at BioChem: Preparation of sample, standard and reference solutions Mechanical sample preparation, e.g., crushing, washing, homogenization, recrystallization Concentration of solutions by rotary evaporator Distillation Extraction Solid phase extraction Filtration Ashing Drying Digestion, e.g., closed digestion, open digestion, microwave digestion Sample analysis by column chromatography Sample preparation for chromatography Derivatization Preparation of mixed samples, etc. The costs are calculated on a time and material basis CHROMATOGRAPHY Thin Layer Chromatography (TLC) (Includes photographic documentation) TLC identity testing... TLC purity testing, (semi-quantitative), limit test... Application of specific spray reagents... TLC content determination* High performance liquid chromatography (HPLC) (Detection: UV-VIS, diode array detector DAD, refractive index RI, fluorescence detector and light scattering detector ELSD) HPLC identity testing... HPLC identity / content testing... HPLC purity testing / related substances Ion chromatography (IC) Conductivity detection; anions / cations with / without suppressor technology IC - identity testing... IC - identity / content testing... IC - purity testing Capillary gas chromatography (GC (Detection: flame ionization detection FID, electron capture detection ECD, phosphorus nitrogen detection PND, thermal conductivity detection WLD or TCD // carrier gases: helium, hydrogen, make up gases: helium, nitrogen // sampling systems: liquid: autosampler or manual, gaseous: headspace) GC content determination... GC limit test... * Tests are conducted at a partner laboratory. 8

9 GC testing for related substances... GC testing for solvent residues... GC fatty acid composition... GC ethylene oxide EO / dioxane... GC residual oxygen determination... GC ethylene oxide EO / ethylene chlorohydrin ECH / ethylene glycol EC... (detection: mass spectrometry MS)* Special Chromatography Methods Chromatographic enantiomer separation... Amino acid analysis SPECTROSCOPY / SPECTROMETRY UV / VIS Spectroscopy / Photometry UV / VIS identity testing... UV / VIS content determination Infrared Spectroscopy (FT-IR) (Measurement liquid or solid / KBr pellet or by ATR) FT-IR identity: KBr-pellet... Film... Measurement by ATR... FT-IR quantitative Atomic Absorption Spectroscopy AAS flame, graphite furnace or cold vapour / hydride technique... Additional special techniques and work up steps in case of interference by the sample matrix Inductively coupled plasma mass spectrometry (ICP-MS) Qualitative and quantitative determination / identity, content, purity and residue analysis... Additional special work up steps in case of interferences by the sample matrix ELECTROCHEMICAL ANALYSIS AND TITRATION ph (potentiometric)... Ions with selective electrodes... Conductometry (conductance)... Titration (visual end point determination)... Titration (potentiometric)... Karl Fischer water content (volumetric) MEASUREMENT OF PHYSICAL PROPERTIES Polarimetry, spec. rotation at 589 nm... Refractometry, refractive index at 20 C... 9

10 Gravimetry to mass constancy (e.g. drying loss)... Density by densimeter (oscillation measurement)... Density by pycnometer - liquids... Melting point / crystallisation range... Boiling point... Ubbelohde viscosity... Viscosity by rotation viscometry (spindle)... Surface tension... Osmolality (by freezing point depression)... Osmolality (by vapour pressure osmometry)... Consistency (micro-penetrometer) TOTAL ORGANIC HYDROCARBON (TOC) Total organic carbon (TOC)... Additional preparation steps VISUAL TESTS Clarity and opalescence of solutions... Coloration of solutions TESTING FOR PARTICLES Testing for visible particles... Testing for non-visible particles Measurement by light blockage (Method 1)... Microscopic determination (Method 2)... additional methods on request ADDITIONAL ANALYSES ON REQUEST (e.g.)* Elemental analysis, each element... NMR spectroscopy... Mass spectrometry MS SPECIAL PHARMACEUTICAL ANALYSES IN-PROCESS CONTROL, RELEASE AND STABILITY TESTNG FOR PHARMACEUTICAL QUALITY CONTROL AND DEVELOPMENT We accompany you in all stages of the analytical life cycle management of your raw materials and pharmaceutical product. From the first pharmaceutical development within Phase I, II, III and IV clinical trials, or for comparative studies with generic products development, through registration and routine quality control for launch and market release. The prices for individual testing of identity, purity, and content correspond to the basic prices of the chemical, physical and instrumental analyses listed above. We create project-based individual quotations under consideration of the desired methodology, matrix, and specified completion date. Please contact us. * Tests are conducted at a partner laboratory. 10

11 Testing According to a Monograph Complete analysis in accordance with specifications of the respective current Pharmacopoeia or client test procedure Method Development, Validation, Method Transfer Method development / optimization... Validation / verification of procedure / product-specific validation in accordance with ICH guidelines including test plan and report Preparation in consultation with client (test scope)... Methods transfer including test plan and report preparation Technology Parameters of Solid Drug Forms Dimensions n = Abrasion / friability... Breaking strength n = Disintegration time n = Uniformity of Content / Mass Uniformity of mass (n = 20) Tablets, film-coated tablets... Divided tablets... Hard gelatine capsules... Uniformity of content depending on the analytical procedures (Photometry, HPLC, titration, etc.), n = Divisibility studies per Ph. Eur Active Substance Dissolution Paddle or rotary basket (n = 6) depending on the analytical method (HPLC, photometry, etc.)... Comparative dissolution profiles (f 2 test) Water for pharmaceutical purposes Chemical / physical tests in accordance with current Ph. Eur. Aluminium... Ammonium... Calcium, magnesium... Chloride... Conductance... Nitrate... Oxidizable substances... Acidic or alkaline reacting substances... Sulphate... Heavy metals... TOC... Non-visible particles... 11

12 2.2.2 STABILITY STUDIES (STORAGE AND TESTING) BioChem is your experienced partner available for all questions regarding stability of pharmaceutical products (stress and photostability, short-term and long-term stability studies, on-going studies under ICH conditions). Computer-controlled and alarmed environmental chambers are available for storage of test samples. In addition to the usual ICH conditions (5 C, 25 C / 60% r.h., 30 C / 65% r.h., 40 C / 75% r.h.) other environmental settings can be applied upon request. Narcotics can be stored. The cost for storage, logistics and analysis depends on the specifications of the test material, the quantities to be stored, and the deposited study protocols. Please specify the number of batches, the volume of the test samples, the storage conditions, and, if already determined, the test parameters and testing schedule so that we can provide you with an accurate quote. We will also gladly advise you. Basic Charges for Storage and Testing at BioChem: Initial storage (receipt, etc.) of samples and preparation of stability testing schedules... Storage per batch, storage condition and month... Preparation of stability reports per test time point... Return shipment of test samples... Stability testing dependent on extent of testing PHARMACEUTICAL DEVELOPMENT We are at your side for all analytical questions relating to pharmaceutical development: starting from selecting appropriate raw material batches, to the analysis of the first product formulations and testing of their suitability, first short-term stability testing and analysis of pilot batches based on your provisional release and shelf life specifications, and to the testing of first approval batches, which includes preparation of analytical documentation for registration. Naturally, this includes screening of raw materials for contaminations, creating comparative dissolution profiles, and analysis by f 2 -test, or performing packaging trials to optimize the shelf life and other tests according to your specifications. For such complex and long-term pharmaceutical development, a BioChem project member is designated to you, who is at your side with competency and advice in all phases of the project to lead your project to ultimate success: approval PHOTOSTABILITY TESTING Irradiation in accordance with ICH Q 1B PHARMACEUTICAL APPROVAL, EXPERT ASSESSMENT We will be happy to assist you with the planning and implementation of projects for approval. We support you with preparation of study protocols, and, together with our partners, also address questions around regulatory approval, ranging from defect notice to preparing the complete approval documentation package. Please ask for a specific quotation relating to your current project. 12

13 2.3 TESTING IN ACCORDANCE WITH GLP GUIDELINES BioChem offers the following tests in accordance with GLP guidelines: Method test parameters OECD 101 UV-VIS absorptions spectrum... EU A.1 / OECD 102 Melting point / range... EU A.2 / OECD 103 Boiling point... EU A.3 / OECD 109 Relative density (solid / liquid)... EU A.5 / OECD 115 Surface tension... EU A.6 / OECD 105 Water solubility*) Flask method... Column method... EU A.8 / OECD 107, 117 Distribution coefficient*)...oecd OECD EU A.10 Flammability (solid)...screening......complete... OECD 112 Dissociation constant in water... OECD 114 Viscosity... OPPTS Aggregation state... Other test parameters in collaboration with our qualified partner laboratories. *) Price when validated methods are available. We gladly offer method development / validation (on time and material basis). 13

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15 3. MICROBIOLOGY TESTING 3.1 MICROBIOLOGY FOR ALMOST ANY QUESTION Page TESTING FOR MICROBIAL CONTAMINATION OF NON-STERILE PRODUCTS Page Microbial Enumeration Test Testing for specified micro-organisms with enrichment methods Test according to acceptance criteria of Ph. Eur., chapter Testing of herbal medicinal products for oral use and extracts used in their preparations according to Ph. Eur, Chapter and and per acceptance criteria section Addendum to testing microbial contamination Product-specific test method validation 3.3 IDENTIFICATION OF MICRO-ORGANISMS Page STERILITY TESTING Page Membrane filtration method Direct inoculation method Addendum to sterility testing Product-specific test method validation 3.5 TESTING OF BIOINDICATORS Page BACTERIAL ENDOTOXIN TESTING USING LAL-TEST Page Gel clot and kinetic-turbidimetric methods Kinetic-chromogenic method Addendum to the LAL-Test Product-specific test method validation 3.7 MICROBIOLOGICAL ASSAY OF ANTIBIOTICS Page EFFICACY OF ANTIMICROBIAL PRESERVATION Page USP Chapter <51> of current version Ph. Eur Chapter of current version Addendum to preservative challenge test Product-specific test method validation 3.9 ON SITE ENVIRONMENTAL MONITORING Page OTHER MICROBIOLOGICAL TESTS Page 25 15

16 3.1 MICROBIOLOGY FOR ALMOST ANY QUESTION We offer method development, model testing, concept validation, and complete expert opinions for microbiology-related questions in addition to the following tests. 3.2 TESTING FOR MICROBIAL CONTAMINATION OF NON-STERILE PRODUCTS MICROBIAL ENUMERATION TEST Performed according to Ph. Eur., Chapter , and USP Chapter <61> of the respective current version TAMC (Total Aerobic Microbial Count)... TYMC (Total Yeast and Moulds Count)... Combination TAMC and TYMC... Required sample volume: at least 10 g or ml Depending on the microbiological purity requirements for the product to be tested and its properties, either the pour plate method, surface spread method or spiral plate method are used. Bioburden determination by membrane filtration without additional rinsing of the membrane filter with one culture medium... with two culture media... MPN-Method... Membrane filtration method including rinsing of the membrane filter with 100 ml rinsing volume per membrane... Membrane filtration method including rinsing of the membrane filter with 3 x 100 ml rinsing volume per membrane... Qualitative detection of anaerobic micro-organisms... Quantitative detection of anaerobic micro-organisms... Testing of the sample homogenates for fastidious anaerobes is performed before and after heat treatment (80 C, 10 min) TESTING FOR SPECIFIED MICRO-ORGANISMS WITH ENRICHMENT METHODS Performed according to Ph. Eur., Chapter , and USP Chapter <62> of the respective current version. Qualitative detection of Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella species, Escherichia coli and other bile tolerant gram-negative bacteria in 1 g / ml or 10 g / ml... Microbial enumeration test (TAMC and TYMC) and qualitative detection of specified micro-organisms (according to and 3.2.2)... required sample size: at least 30 g or ml 16

17 Microbial enumeration test (TAMC and TYMC) and quantitative determination of bile tolerant gram-negative bacteria and qualitative tests for other specific microorganisms... required sample size: at least 30 g or ml TESTING ACCORDING TO ACCEPTANCE CRITERIA TEST OF THE PH. EUR., CHAPTER Preparations for rectal use Microbial enumeration test (TAMC and TYMC)... required sample size: at least 10 g or ml Preparations for oromucosal, gingival, cutaneous, nasal and auricular use Microbial enumeration test (TAMC and TYMC) and qualitative detection of P. aeruginosa and S. aureus in 1 g / ml... required sample size: at least 10 g or ml Aqueous and non-aqueous preparations for oral use Microbial enumeration test (TAMC and TYMC) and qualitative detection of E. coli in 1 g / ml... required sample size: at least 10 g or ml Special Ph. Eur. Provision for oral dosage forms Microbial enumeration test (TAMC and TYMC) and qualitative detection of Salmonella in 10 g / ml, E. coli, S. aureus and the quantitative determination of bile tolerant gram-negative bacteria in 1 g / ml... required sample size: at least 30 g or ml Preparations for vaginal use Microbial enumeration test (TAMC and TYMC) and qualitative determination of P. aeruginosa, S. aureus and C. albicans at 1 g / ml... required sample size: at least 10 g or ml Inhalables Microbial enumeration test (TAMC and TYMC) and qualitative determination of P. aeruginosa, S. aureus and bile tolerant gram-negative, bacteria in 1 g / ml... required sample size: at least 20 g or ml Transdermal patches Microbial enumeration test (TAMC and TYMC) by membrane filtration and qualitative detection of P. aeruginosa and S. aureus in 1 g / ml... required sample size: at least 10 patches TESTING OF HERBAL MEDICINAL PRODUCTS FOR ORAL USE AND EXTRACTS USED IN THEIR PREPARATIONS ACCORDING TO PH. EUR., CHAPTER AND AND ACCEPTANCE CRITERIA PER CHAPTER Category A products Microbial enumeration test (TAMC and TYMC) and quantitative detection of E. coli in 1 g / ml and qualitative determination of Salmonella in 25 g / ml... required sample size: at least 35 g or ml 17

18 Category B and C Products Microbial enumeration test (TAMC and TYMC) and qualitative determination of E. coli in 1 g / ml and Salmonella in 25 g / ml and quantitative determination of bile tolerant gram-negative bacteria in 1 g / ml... required sample size: at least 45 g or ml All micro-organisms detected within the scope of the enrichment procedures are identified according to the state of the art methods (see 3.3 Identification of micro-organisms) to provide the client with a comprehensive GMP-compliant view of the microbial status. In addition, per client request, identification of the micro-organisms may be performed. Verification of the accuracy and / or suitability of the test method is required per Ph. Eur. and USP for every formulation of different composition Addendum to TESTING FOR MICROBIAL CONTAMINATION Special sample preparation (e.g., emulsification in Tween 80, sonication, Ultra Thurrax, Stomacher)... ph adjustment of the stock solutions... Buffer and nutrient media volumes larger than those specified by pharmacopoeia (e.g., 500 ml up to 1000 ml) after method modification following validation... A maximum of two larger volumes are charged for each test. Larger volumes required beyond are not charged additionally. Additional weighing... Work with highly potent and toxic substances (e.g., cytostatics, etc.)... Tests according to validated methods with increased effort on request PRODUCT-SPECIFIC TEST METHOD VALIDATION Effort depend on the dosage form, test method and number of simultaneously ordered validations... including a validation report with detailed description of the method, presentation and evaluation of results in German or English. We will gladly advise you and, on request, generate an individual quotation for each specific question. Ask our experts. 3.3 IDENTIFICATION OF MICRO-ORGANISMS Basic macroscopic identification to family or genus level in the context of testing for microbial contamination as per Ph. Eur Identification to the species level in the context of testing for microbial contamination as per Ph. Eur with biochemical reactions (API)... Identification to species level with molecular biology methods (PCR / sequencing) 3... Identification of colonies on submitted on agar media by biochemical reactions (API)... 18

19 Identification of micro-organisms submitted in nutrient solutions with biochemical reactions (API)... Increased effort for preparation of subcultures for purity control and / or identification of morphologically identical colonies prior to biochemical identification STERILITY TESTING Performed according to Ph. Eur., USP Chapter and Chapter <71> of the respective current version. Testing is performed in an Isolator MEMBRANE FILTER METHOD Testing of 1 to maximally 20 individual containers without rinsing the membrane filter two culture media, incubation time 14 days DIRECT INOCULATION METHOD Growth media volumes 200 ml, ratio sample / growth medium 1:10, for liquid formulations, 1:100 for solid, insoluble preparations Incubation time 14 days, two growth media... Replicate tests for compliance with the minimum sample volumes per culture medium for every additional culture with 2 growth media within the same test ADDendum to STERILTY TESTING Rinsing both membrane filter for inhibitor removal with 3 x 50 ml each (3 x 100 ml per test)... with 6 x 50 ml each (6 x 100 ml per test)... with 9 x 50 ml each or to a maximum of 5 x 100 ml each (9 x 100 ml or 5 x 200 ml per test)... Dissolving lyophilized products, powders and the like (up to 20 sample vials) Depending on volume... Penicillinase treatment... Preparation of sterile-filtered isopropyl-myristate up to 100 ml including sterility testing... Preparation of mixed samples with creams and eye ointments from a maximum of 10 tubes... Larger (>200 ml) volumes of growth media (must be individually prepared and tested for growth properties and sterility prior to release) up to 2000 ml for 2 culture media... Detection of microbial growth with substantial turbidity at the conclusion of sterility testing by transfer of aliquots from the nutrient broth into fresh broth... 3 Tests are performed at a partner laboratory. 19

20 With suspected microbrial growth, subcultures from the growth media at the end of sterility testing by streaking out with inoculation loops... Colony identification with positive sterile test. Working with highly potent and toxic substances (e.g., hormones, cytostatic drugs, narcotics, etc.)... Tests according to validated methods with increased effort... Testing for air tightness of primary vessels during hydrogen peroxide sterilization in the Isolator... Notes on sterility testing The analysis include batch-specific testing and documentation of sterility and growth properties of culture media and solutions used; in addition, microbiological environmental monitoring using settle plates, contact agar plates and active air sampling is conducted for every test. Verification of suitability of the test method is required per Ph. Eur and USP for every formulation of different composition PRODUCT-SPECIFIC TEST METHOD VALIDATION Effort depending on test, product and simultaneously ordered validations... including a validation report with detailed description of the method, presentation and evaluation of results in German or English language. We will gladly advise you and, on request, will generate an individual quotation for any question. Ask our experts. 3.5 TESTING OF BIOINDICATORS Sterility testing (e.g., for checking the efficacy of sterilization processes) one bioindicator + positive control... each additional bioindicator at the time of result presentation in one combined report... Quantitation of spore population on spore strips, discs and in spore suspensions. Individual testing of 3 units. In reference to USP Preparation and testing of one 10-fold dilution series... According to USP Preparation and testing of two 10-fold dilution series... 20

21 3.6 BACTERIAL ENDOTOXIN TESTING WITH LAL-TEST Performed following the procedure per Eur. Chapter and USP Chapter <85> in the respective current version GELCLOT AND KINETIC-TURBIDIMETRIC METHODS One sample or one prepared test solution of the sample to be tested up to the maximum valid dilution (MVD) (MVD) in duplicate... One sample or one prepared test solution of the preparations to be tested as a mixed sample of one container each from the beginning / middle / end to the maximum valid dilution (MVD) (MVD) in duplicate... 3 samples or 3 test solutions of the sample to be tested from one vessel each at the beginning / middle / end to the maximum valid dilution (MVD), each in separate duplicate together KINETIC-CHROMOGENIC METHOD One sample or prepared test solution of the sample to be tested up to the maximum valid dilution (MVD) in duplicate... One sample or a prepared test solution of the sample to be tested as a mixed sample from one vessel each of the beginning / middle / end to the maximum valid dilution (MVD), in duplicate... 3 samples or 3 solutions of the prepared test solution to be tested from one vessel each of the beginning / middle / end to the maximum valid dilution (MVD), in separate duplicates... All tests according to the requirements of the current pharmacopoeia including standard series, negative controls with water-lal / buffer in duplicate, positive controls with water-lal / buffer, each in duplicates, positive controls for each measured test solution in duplicates Addendum to LAL-TEST Semi-quantitative determination by serial dilution with a positive result at MVD by gel clot method and with results greater than the highest concentration of the standard curve in the kinetic-turbidimetric-kinetic method, or with invalid positive controls... Use of endotoxin-free Tris buffer to adjust the ph or dilution medium for sample solution PRODUCT-SPECIFIC TEST METHOD VALIDATION Effort dependent on method, product, and number of batches per product ordered together... including a validation report with detailed description of the method, presentation and evaluation of results in German or English. We will gladly advise you and on request, will generate an individual quotation for any question. Ask our experts. 21

22 3.7 MICROBIOLOGICAL ASSAY OF ANTIBIOTICS Performance per Eur. Chapter of the respective current version, agar diffusion method. Testing against official Pharmacopoeia reference or laboratory standards, including variance analysis of antibiotic content / activity as active substance or in the finished product... Simultaneous testing of an antibiotic from at least two preparations... Combination preparations against mixing standards per antibiotic... Antibiotics containing cytostatics including surcharge for cytostatics... Testing of combination preparations... Extraction of antibiotics from ointment bases and other complex sample preparations... Non-aliquotable reference standard substances (declared in IU / container or µg / vessel) per standard substance and container at cost. Product-specific test method validation following ICH guidelines Performance per Ph. Eur by agar diffusion method against official standards by multiple 3-dose assay with randomized block design. Determination of reproducibility (same analyst, same instrument, same laboratory, and short time frame) with at least 6 independent assays performed with individual sample preparation for every product or antibiotic as described in Q2B-Note for Guidance on Validation of Analytical Procedures: Methodology CPMP / ICH / 281 / 95; determining the accuracy of the method, intermediate precision and linearity of the reproducibility within a measurement interval of e.g., 70% to 130% relative to the 100% nominal concentration of the antibiotic to be determined. Within this measurement interval, at least 3 spiking concentrations in triplicate each are required. To this end, we request the corresponding spiked samples as experimental starting base. Effort depending on the antibiotic and the formulation on a time and material basis between... including a validation report in German or English with a detailed description of methods, presentation of results with calculation of the coefficient of variation for each individual result, including 95% confidence limits and test for validity. We will gladly advise you and on request, will generate an individual quotation for any question. Ask our experts. Other validation parameters (e.g., robustness, selectivity, testing by a second employee)... 22

23 3.8 EFFICACY OF ANTIMICROBIAL PRESERVATION USP CHAPTER <51> IN THE CURRENT VERSION 5 test organisms for category 1 Products (parenterals, sterile nasal and ophthalmic products)... for category 2 products (non-sterile topical and nasal products)... for category 3 products (oral products)... for category 4 products (antacids) PH. EUR. CHAPTER IN THE CURRENT VERSION At the relevant re-sampling time points (see below) depending on product type parenteral and ophthalmic preparations (4 test organisms)... Oral preparations (5 test organisms)... Topical preparations (4 test organisms)... resampling time points: Parenterals and ophthalmic preparations Bacteria: t 0, 6 hours, 24 hours, 7 and 28 days Fungi: t 0, 7, 14 and 28 days (corresponding to the requirements per Criteria A and B) Topical preparations Bacteria: t 0, 48 hours, 7, 14, 28 days Fungi: t 0, 14 and 28 days (corresponding to the requirements per Criteria A and B) Oral preparations Bacteria and fungi: t 0, 14 and 28 days Addendum to THE PRESERVATIVE CHALLENGE TEST Each additional test depending on the number of resampling time points and on test organism... Each additional resampling time point in addition to the required resampling time points... Complex sample preparation such as emulsifying ointment with Tween 80 per reference micro-organism and sampling time... modified tests... 23

24 3.8.4 PRODUCT-SPECIFIC TEST METHOD VALIDATION Product-specific validation of test methods, depending on the test method, product, product category and simultaneously ordered validations... including a validation report with detailed description of the method, presentation and evaluation of results in German or English. We will gladly advise you, and on request generate an individual quotation for each specific question. Ask our experts. Special note The effort for microbiological tests as specified in 3.2 to 3.8 is based on the parameters specified in the Pharmacopoeia test procedure for which costs of testing can be reasonably estimated. Depending on the particular test samples, an increased effort may be incurred due to sample preparation and test performance. Should these costs be known in advance or become apparent on the basis of validation studies, we will notify the client in advance of potential additional costs incurred. This applies equally to testing according to additional specifications that are not reflected in this price list. Please ask our professionals for information; they will gladly advise you. On request, we would be happy to send you an individually quotation for each question. Naturally, depending on the contracted quantity, we also offer discounts in the form of an annual rebate. Just ask us. 3.9 ON SITE ENVIRONMENTAL MONITORING Active air sampling is performed with the RCS Plus air sampler collector, settle plates and surface monitoring with Rodac plates as well as swab tests including media supplied by BioChem. Trip to and from site... On-site sampling... Incubation and analysis of active air sampling strips, settle plates, and Rodac plates... On request, upon growth, identification of micro-organism type genus by colony morphological and microscopic characteristics... With suspicion of pathogenic relevant micro-organisms, identification of micro-organism to species level on request Rental of active air sampling device including days of dispatch and return... Supply of culture media for environmental monitoring... 24

25 3.10 OTHER MICROBIOLOGICAL TESTS Drinking water testing per German Drinking Water Regulation (Trinkwasser Verordnung, TrinkwVO) Determination of the minimum inhibitory concentration Testing for bactericides and fungicides Microbiological process controls, e.g., in disinfection, sterilization, endotoxin decrease Growth promotion tests of culture media Simulated microbiological in-use stability tests Container Closure Integrity Tests Testing of medical products of all kinds for Bioburden Sterility Bacterial endotoxins and a variety of other microbiology-related questions The microbiological tests shown in this list essentially represent the tests required as part of the pharmaceutical quality control procedures according to current Pharmacopoeias. A variety of other available tests performed in our laboratory for different questions are not listed here. We will gladly provide you with information on other tests for microbiology-related questions on request. Talk to our specialists. 25

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27 4. MOLECULAR BIOLOGY TESTS 4.1 PCR ANALYSIS Page REAL TIME PCR ANALYSIS Page FLUOROMETRIC / LUMINOMETRIC DETECTION Page ENZYMATIC DETECTION Page PROTEIN ANALYSIS / IMMUNOCHEMISTRY Page BIOANALYSIS Page VALIDATION / TESTING UNDER GLP CONDITIONS Page IDENTIFICATION OF PLANT AND GMO'S BY PCR Page DIFFERENTIATION OF ANIMAL SPECIES Page 29 27

28 4.1 PCR ANALYSIS Mycoplasma detection... RT-PCR... Further analysis per customer request (e.g., detection of pathogens or determination of identity); cost on a time and material basis. 4.2 REAL-TIME-PCR ANALYSIS DNase detection in pipette tips, microtiter plates, test tubes, PCR tubes, solutions, etc... Detection of human DNA in pipette tips, microtiter plates, test tubes, PCR tubes, solutions, etc... Detection of bacterial DNA in pipette tips, microtiter plates, test tubes, PCR tubes, solutions, etc... RNase detection in pipette tips, microtiter plates, test tubes, PCR tubes, etc... RNA detection in pipette tips, microtiter plates, test tubes, PCR tubes, solutions, etc... Detection of PCR inhibitors in pipette tips, microtiter plates, test tubes, PCR tubes, solutions, etc... Detection of mycoplasma by SYBR-Green... Detection of mycoplasma using probes... More real-time PCR and real-time RT-PCR detection by SYBR Green and probes on request, on a time and material basis. 4.3 FLUOROMETRIC / LUMINOMETRIC DETECTION ATP detection... Fluorometric DNA determination ENZYMATIC DETECTION E.g., Chromogenic tests Streptokinase activity... Anti-factor Xa or anti-iia activity... Determination of anti-factor Xa-activity / anti-factor IIa-activity factor add... 28

29 4.5 PROTEIN ANALYSIS / IMMUNOCHEMISTRY Molecular Weight Determination by SDS-PAGE (reducing and non-reducing) In single determination... In duplicate... ELISA... Isoelectric focusing (IEF) In single determination... In duplicate... Protein Quantification According to Bradford (eight replicates)... Photometric (single determination)... Other methods on request (Kjeldahl, Lowry, BCA, Biuret, fluorometric) Radial immunodiffusion (RID)...price on request 4.6 BIOANALYSIS Capillary electrophoresis E.g., Purity verification of aprotinin or ropivacaine according to monograph... Enantiomeric purity (e.g., phenylephrine) VALIDATION / TESTING UNDER GLP CONDITIONS Validation of above tests... Performance of above tests under GLP-conditions IDENTIFICATION OF PLANTS AND GMO'S BY PCR Species detection (sugar beet, potato, tomato, canola, corn, soy, wine, etc.), screening of transgenic plants (35S promoter, Nos terminator and nptii gene from Tn5 transposon) and specific detection of GMOs ANIMAL SPECIES DIFFERENTIATION Species differentiation by PCR of chicken, turkey, beef, pork, goat, ostrich, horse, donkey, etc... 29

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31 5. GENERAL Addendum Project meeting First project meeting at no charge... Materials Special materials (chromatography columns, reference substances, special reagents) will be charged at cost, including shipping, unless they are provided by the client... Test performance Generation of a validation plan (if not included) in German or English... Working with highly potent substances (e.g., cytostatics) or substances with high or unknown risk potential Inactivation and disposal... Handling of narcotics... Reporting Interim reports and reissue of audit reports 1-page... multiple pages... Copy of raw data (including control and authorization) per page... Translation of reports in German or English... OOS procedures Administrative expenses for the processing of OOS procedures in the event of non-obvious laboratory error... Failure Investigation Performance of failure Investigation and preparation of a report for Sterility testing and non-sterile results or OOE / OOT results... Client-specific work procedures Preparation of testing specifications / work instructions... Test release on behalf of pharmacies by cross sample expert according to 65 para 4 AMG. Miscellaneous Notarization of documents without Apostille... with Apostille... Translation of raw data and other documentation... 31

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33 BioChem GmbH Contact Daimlerstraße 5b Phone +49 (0) Karlsruhe Fax +49 (0) biochem@biochem.de Test order to BioChem Offer no. (if applicable): Client Company Address Zip code Contact Phone City Fax Date Signature Item Material Batch Quantity BioChem receipt no. (internal use) 1. to be completed by BioChem 2. to be completed by BioChem 3. to be completed by BioChem Tests to be performed Specification Testing procedure (Ph. Eur., USP-NF, other...) Specific risks none Cytostatics* toxic Narcotics* very toxic potential allergen irritant to mucous membranes Other precautions Comments: Desired project start date: Storage Conditions room temperature 2-8 C -18 C protect from light protect from moisture *requires additional charge; see price list 33

34 A5 Siemensallee BioChem Daimlerstraße B36 Siemensallee allee Exit Nr. 7 B10 B36 B36 Direction Mannheim Hertzstraße Moltkestraße Kaiserallee Karlsruhe centre B36 Siemensallee B36 B10 (K9657) B10 Kriegsstraße B10 Exit Karlsruhe Mitte A5 Basel Frankfurt Direction Landau / KA-Center B10 (K9657) How to find us: On A5 exit Karlsruhe centre, then continue on B10 (K9657) toward Landau / KA-centre, exit 7 direction Mannheim / Knielingen / Rheinhafen, follow B36, turn left into Siemensallee and take the first right (Daimlerstr.) BioChem Labor für biologische und chemische Analytik GmbH Daimlerstraße 5b Karlsruhe Phone +49 (0) Fax +49 (0) biochem@biochem.de Conception / Design FORMAT WERBEAGENTUR GmbH Ettlingen