Pharmaceuticals

Bioanalytical Methods

• Method Development (non-GLP) – In guiding your product to market, ensuring its contents are accurately measured is of utmost importance. Avazyme experts take pride in developing analytical methods for acquiring the following product data and more:
  • Potency – Your product’s ability to elicit a specific response is inseparably tied to its value. We determine assays that will characterize your product, demonstrate its intended effects (e.g., antimicrobials), and be manufacturing-ready. We routinely produce the following potency data: the amount of active ingredient in intermediate manufacturing products, and formulated end-products, as well as criteria for identifying sub-potent batches and ensuring drug product consistency and that the product meets the required specifications (IS/OOS)..
  • Purity and Impurities – Manufacturing processes can naturally result in the production of substances other than the intended product. Such impurities can come from process contaminants, API and non-active component degradation, leachables, unreacted monomers/oligomers, and oxidized additives. We typically measure less volatile impurities with LC-MS and more volatile impurities with GC-MS, as the absence of a proper mobile phase can allow for a more accurate structural evaluation in some cases. Upon isolating your impurity, we have the capacity to perform further MS, NMR, and ICP or other specialized investigations.
  • Degradants – Various stress conditions are deliberately applied to your product for the purpose of estimating the degradants that will occur during stability studies. HPLC is then used to separate product peaks from degradant peaks for quantification. These studies are essential in proposing a shelf life to the FDA.
  • Moisture – In pharmaceutical product development, it is important to know the maximum allowable moisture content for each step in the process, as this information has implications for manufacturing especially, but also for shelf life and consumer safety. Compounds with functional groups such as amides and esters can be hydrolytically degraded, so moisture content control is quite important. Avazyme determines moisture content primarily using the following three methods: halogen heating moisture analysis, chemical titration, and moisture loss upon freeze-drying.
  • Dissolution (coming soon) – All solid oral dosage forms require dissolution testing, and product release and stability testing also benefit from dissolution testing. Dissolution testing can also be used to optimize drug release. Avazyme employs United States Pharmacopoeia (USP) dissolution apparatuses 1-7 for technical dissolutions, with 1 and 2 being most commonly used. The USP 1 basket is used with a dissolution bath to test products that would otherwise float, while the Paddle apparatus (USP 2) is used to test extended releases along with immediate and modified releases. USP 3-7 are used in other, more nuanced circumstances. In any event it is crucial that the dissolution methods are accompanied by the appropriate analytical determinations.
  • Residual Solvents – Toxicity is the primary basis for measuring the residual solvents present and produced in the manufacture of pharmaceutical products. A GC Limit Test is used to detect classes 1 and 2 of residual solvents, while class 3 residual solvents are detected using a separate LOD test. Quantitative data is acquired through further chromatographic analysis.
  • Stability-Indicating Assays – Forced degradation of your pharmaceutical product is used to generate samples representing the effect of time and other factors on its many properties. Avazyme achieves various levels of degradation by exposing your product to UV light, pH, temperature and certain oxidative/reductive reagents. Stability is indicated by the amount of active ingredient and degradation products as measured by GCMS or LCMS or other suitable methods (API specific, customized approach).
• Method Validation (GLP/non-GLP) – Our expert staff routinely uses GCMS and LCMS to identify new chromatographic peaks across a full range of reference standards. To validate these methods and prove that the analytical test system is capable of providing legitimate analytical data, we carefully document numerous assessments and associated data in accordance with GLP. These are a few common acceptance criteria we provide for method validation:
  • Limit of Detection
  • Limit of Quantitation
  • Linearity and Range
  • Precision and Accuracy
  • Robustness
  • Specificity
  • System Suitability

Bioanalysis (GLP/non-GLP) – A comprehensive biological regimen for drug testing ensures your product is in line with ICH and FDA standards and includes the following studies, among others:

• Bioanalysis (DMPK) – Screening to reduce drug-drug interactions and to understand drug toxicity levels and PK/PD relationships are some of the main reasons why DMPK studies are typically performed. These studies produce data on inhibition and induction, bioavailability, half-life, distribution volume, metabolics, and transporters. Avazyme has the capability to measure blood and urine samples with ELISA and LCMS (or other suitable methods).
• Large Molecule Analytical Chemistry – Avazyme determines the concentration of peptides, proteins, antibodies, and other large molecules using ligand binding assays via ELISA.
• Drug Metabolism (PK/PD) – Effect intensity, administration time, and dosage are combined to model your product’s biological effects. We partner with other laboratories to conduct the in-vitro or in-vivo in-life phase and then perform the bioanalytical determination of the analytes of interest in the biological samples, including tissue samples.
• Impurity Testing – Manufacturing processes can naturally result in the production of substances other than the intended product. Such impurities can come from process contaminants, API and non-active component degradation, leachables, unreacted monomers/oligomers, and oxidized additives. We typically measure less volatile impurities with LC-MS and more volatile impurities with GC-MS, as the lack of a proper mobile phase allows for a more accurate structural evaluation in some cases. Upon isolating your impurity, we have the capability to perform further MS, NMR, and ICP investigations.
• Dissolution (Coming Soon) – All solid oral dosage forms require dissolution testing, and product release and stability testing also benefit from dissolution testing. Dissolution testing can also be used to optimize drug release. Avazyme employs United States Pharmacopoeia (USP) dissolution apparatuses 1-7 for technical dissolutions, with 1 and 2 being the most common. The USP 1 basket is used with a dissolution bath to test products that would otherwise float, while the Paddle apparatus (USP 2) is used to test extended releases along with immediate and modified releases. USP 3-7 are used in other, more specific circumstances.
• Metals Testing – Avazyme performs the inorganic mass spectrometry necessary to detect heavy metals using an inductively coupled plasma mass spectrometer. We can detect the following elements in concentrations as small as parts per billion (ppb):
  • Aluminum
  • Arsenic
  • Antimony
  • Barium
  • Bismuth
  • Cadmium
  • Cobalt
  • Calcium
  • Copper
  • Germanium
  • Gold
  • Indium
  • Iron
  • Lead
  • Lithium
  • Magnesium
  • Manganese
  • Mercury
  • Molybdenum
  • Nickel
  • Palladium
  • Platinum
  • Potassium
  • Selenium
  • Silver
  • Sodium
  • Tin
  • Titanium
  • Thallium
  • Yttrium
  • Zinc
  • Zirconium
• Mass Spectrometry for Unknown Identification – Our GCMS, LCMS, and ICP-MS capabilities as well as access to NMR allow us to identify and quantify unknown components and degradants in your product. High peak resolution and separation ensure the unparalleled accuracy of chromatographic structural data related to your product, lending to stronger R&D turnouts and better understanding of your product’s specifications.
• NMR Analysis – Extremely useful in its power for quality control, NMR is utilized by Avazyme to confirm the structural identity of your product during R&D and product development. In instances where reference samples are not available, NMR is particularly useful in determining an unknown substance’s identity.
• Extractables & Leachables – Packaging system components intended to prevent contamination of a drug product sometimes themselves contaminate the product; these are called extractables and leachables. Avazyme identifies these contaminants by performing multiple extractions using solvents with a wide range of polarities and then referencing the samples against known compounds. Common quantitative methods for extractables and leachables include the following, among others:
  • GCMS, LCMS (Organics)
  • ICP (Metals)
  • UV/Vis Spectroscopy
  • pH, Total Organic Carbon
  • Karl Fischer Analysis (coming soon)
  • Particle Sizing Analysis (currently through partners, internal capacity coming soon)
• Release Testing – Avazyme tests your product for batch releases according to your specified release testing protocol, or we create a batch release protocol that ensures your product meets federal and state acceptance criteria.
• Stability Storage (All Zones)
  • Storage stability testing serves your product by gauging its chemical integrity or potency over time. Our stability chambers are validated per FDA and ICH guidelines (Q1A, Q1B, Q1C, Q5C) and we employ custom set points and chamber configurations to fit your needs. GLP documentation is used to track your product’s data throughout the entirety of testing and can be used for NDA filings and FDA registration.
• ADME (Nonradioactive) – As your drug gets closer to market, ADME studies are used to provide data regarding its potential effectiveness and safety. Avazyme identifies absorption, distribution, metabolism, and excretion properties using the following methods:
  • Chromatographic Identification of Metabolites
  • Protein Binding Assays
  • Metabolic Stability Assays
  • Customized Physiochemical Screens
• Identification of Degradation Products (Post-Stress Test) – Various stress conditions are deliberately applied to your product for the purpose of estimating the degradants that will occur during stability studies. HPLC is then used to separate product peaks from degradant peaks for quantification. These studies are essential in proposing a shelf life to the FDA.