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At the end of the stability study, results from the leachables analyses will be reported. There are two basic outcomes to a leachables study:
Pine Lake Laboratories is fortunate to have Dr. Michael Ruberto as a retained consult available to support all extractable and leachable studies.
Dr. Ruberto is the President of Material Needs Consulting, LLC which provides consulting services to manage the development and commercialization of medical devices and packaging, with a special emphasis on material selection, extractables and leachables, and supply chain management. He has been an active member of various pharmaceutical working groups that have developed “best practices” for characterizing and evaluating the safety of container closure systems and packaging for several different drug dosage forms. Some of these teams include:
Dr. Ruberto was formerly the Head of Regulatory Services for the NAFTA region at Ciba Specialty Chemicals where he was responsible for worldwide notifications of new products, food contact notifications, and regulatory compliance of Ciba chemicals. At Ciba Dr. Ruberto also served as the Director of Analytical Research. Dr. Ruberto was employed by Ciba for fifteen years.
Dr. Ruberto received a B.S. with thesis from Stevens Institute of Technology and a Ph.D. in Analytical Chemistry from Seton Hall University.
To learn more about Dr. Ruberto, please visit www.materialneedsconsulting.com
For bioanalytical methods for small molecule drugs in biological matrices, sample preparation is a critical step. A balance must be achieved between a sample preparation method that reduces interferences while still being affordable and fast. At Pine Lake Laboratories, we have successfully developed and validated a wide variety of bioanalytical methods that achieved this balance. We have experience with mixed mode extraction, liquid-liquid extraction, solid phase extraction, protein precipitation and enzymatic digestion sample preparation methods. We have the experience and expertise to develop the bioanalytical method needed to help advance your drug to the patients who need it.
A major challenge at the contract lab (CRO) is to handle Quality Agreements (QAGs) through some standardized procedure, while at the same time allowing for the diverse needs of a wide client base. This becomes more evident when one considers that the QAG typically originates with the client, in their format, and therefore contains many client-specific idiosyncrasies. Furthermore, the contract lab may be performing laboratory testing under GLP, GMP and/or ISO laboratory testing protocols, which, although greatly similar to one another, will each have its own unique requirements. Procedures, to be useful, must be detailed enough for personnel to be trained on and to follow. However, to be standardized across the wide client base, they must not be too detailed. One cannot have a procedure for each client. Management must therefore “see the forest for the trees”.
At Pine Lake Laboratories, we have developed a standard methodology using Ion-Exchange HPLC-UV to quantitate therapeutic oligonucleotides and pegylated oligonucleotides in plasma. This methodology has been adapted and optimized for multiple compounds across a wide variety of therapeutic areas. For most compounds chromatographic resolution can be achieved between the parent compound and the N-1 to N-X metabolites. Both gradient and column temperature are important in achieving good separation. The sample preparation before HPLC analysis includes an overnight enzymatic digestion. Carryover is a common problem but a strategy of including wash injections minimizes the impact of carryover. All other validation parameters will meet the acceptance criteria. One common area for instability is for the drug in plasma at room temperature. Stability usually can be improved by keeping samples on ice during preparation or using higher concentrations of EDTA. Validated methods have been used to support GLP and human clinical studies without any method related issues. Most methods have column lives that exceed 1000 injections.
Leachables are compounds that migrate into a drug product from the sample container closure system (CCS) under normal storage condition. Leachables can enter any type of drug product including solid dosage forms. Generally, orally inhaled and nasal drug products (OINDP) and parenteral and ophthalmic drug products (PODP) are the most common drug products at high risk of leachables.
Both the primary CCS in direct contact with the drug product (metered dose inhaler, prefilled syringe, eye dropper, IV bag, HDPE bottle, LDPE ampoule, etc.) and the secondary CCS which does not contact the drug product (printed label, cardboard box, foil pouch, environmental exposure, etc.) can be sources of leachables. Leachables present a potential risk to the patient both from the toxicity of the leachable and from the possible negative impact upon stability and efficacy of the drug product. Examples of common leachables can be seen in the table below.
An exaggerated extraction study on a medical device is a forced extraction study to generate a complete extractable profile for hazard identification and is required by ISO 10993-12 to be exhaustive. An overview of the exaggerated extraction study can be found below. The key decision in study design is solvent selection. For an exaggerated extraction study, the extraction solvents are selected based upon the anticipated tissues the device will encounter. The extraction type is based on the solvent type and the analytical methods for analysis of extractables are the same for all extractions. For exaggerated extractions, the extraction must be proven to be exhaustive, therefore extraction time is established experimentally. Extractables are identified by MS and quantitated against structurally similar standards.
Overview of Exaggerated Extraction Study for Medical Devices
Solvent Extraction Type Analytical Methods 1. Polar – water, phosphate buffered saline, culture media without serum
2. Non-polar – ethanol/water, ethanol/saline, dimethyl-sulfoxide.
1. Low boiling neat solvents : Soxhlett
2. Mixed solvents, buffers and high boiling neat solvents: Batch extraction with agitation or circulation
1. Volatile organic extractables by GC-MS
2. Non-volatile organic extractables by LC-MS
3. Inorganic extractables by ICP-MS (aqueous extract only)
To support microbiome research, Pine Lake Laboratories has available an assay for short chain fatty acids (acetic, butyric, and propionic acid) in feces. These targeted metabolites can be used as an indicator of microbiome activity in research subjects and as biomarkers to evaluate the effect of various pharmaceutical treatments during pre-clinical and clinical trials. The method involves extracting the short chain fatty acids from feces then analysis by direct injection GC-MS. This method has been used to support studies in both humans and multiple animal species. For more information, please see the white paper in our technical library titled “Short Chain Fatty Acid Analysis”
The FDA has finalized guidance on validation of bioanalytical methods. Differences between the draft and final guidance, which is 10 pages longer, include a re-working of the text, a new title for Section III, which was previously named “Chromatographic methods” but is now “Bioanalytical method development and validation,” and new sections on parameters of chromatographic assays (CCs) and ligand binding assays (LBAs).
“This final guidance incorporates public comments to the revised draft published in 2013 and provides recommendations for the development, validation, and in-study use of bioanalytical methods,” FDA said. “The recommendations can be modified with justification, depending on the specific type of bioanalytical method. This guidance reflects advances in science and technology related to validating bioanalytical methods.”
If the single entity combination product is not intended to mechanically infuse the drug into the patient but instead the drug is intended to passively diffuse into the patient, then the rate of drug release or elution needs to be determined. To perform this experiment, the single entirty combination product is placed in an appropriate media designed to model the target tissue. The analytical method is then used to measure the increase in drug concentration in the media over time. The instrument conditions used are usually similar to those used for the assay; however, more sensitive methods may need to be developed if the release of drug is expected to be slow. The method will need to be validated similar to an assay method with special consideration to sensitivity and additional ruggedness testing for the sample preparation factors that might impact the release rate.
Just like methods for traditional drug products, the assay and related substances/degradation products methods for single entity combination products need to be proven to be stability indicating. If the single entity combination product contains a new drug, a forced degradation study will be needed. If results of the stability of the drug product alone are available, these results should be used to evaluate if the methods are stability indicating. However, additional degradation experiment on the intact single entity combination product under appropriate conditions will likely be required. If the single entity combination product contains a generic drug, literature references on the stability of the drug product can similarly be used but degradation experiment on the intact single entity combination product under appropriate conditions will still likely be required.
During the PQRI PODP Workshop at USP Headquarters on April 18-19, the PQRI toxicology sub-team announced that they could not get the FDA to agree to the analytical evaluation threshold (AET) concept for ophthalmic drug products. FDA felt that the human eye was a unique organ that was especially susceptible to local topical effects and that ocular irritation was a key endpoint to consider. Therefore, they rejected the use of an AET for reporting leachables. PQRI ophthalmic drug sub-team stated in their presentation that the “unwritten FDA expectations” for ophthalmic drug packaging systems was that:
PQRI further stressed in this presentation that many pharmaceutical companies actually go above and beyond these FDA expectations and report, identify, and qualify all leachables > 1 ppm for ophthalmic drug products.