Working with clients to assist in bringing innovative, safe new medicines and medical devices to patients in need.
GET A QUOTESAMPLE TRANSMITTAL FORMINTRODUCTION
In the present article we summarize what we’ve learned in the period since our earlier article appeared in Journal of GXP Compliance some years ago, entitled “Implementing Quality Agreements at the Contract Laboratory”1. Specifically, here we focus on how the lab gets the Quality Agreement “right” and then follows it throughout its lifecycle by identifying what we call key “check points”. Our experience has shown that there are points in the Quality Agreement process, by which we mean the process of drafting, approving, implementing and revising the Quality Agreement, that are particularly problematic because they require greater attention and/or pose greater risk and it is key to manage these.
Check Point ONE: Prepare to adopt agreements
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”.
Check Point TWO: Check that your quality house is in order
There should already be in place a strong quality program at the CRO and the QAG must make a good fit to this program. Those concerns (terms) that are rather common to Quality Agreements should already be addressed in the existing quality program at the CRO or else consideration must be given to incorporating them into the existing program. This approach is distinctly different and better than leaving them only in the QAG. To leave important concerns only in the QAG is tantamount to instructing the lab to only consider doing such quality work when working on this particular client’s project.
Check Point THREE: Understand the high-risk terms of the specific agreement
Understand and have input into specific terms of the agreement. It is important to make a careful review of the draft. Don’t agree to terms that you cannot meet. Take a team approach in order to foresee any/all difficulties in implementing the terms by involving lab Management, Principal Investigators and Quality Assurance in the draft review. Many of the high-risk terms are those that are specific to the client. By high-risk, we mean at risk of being ignored during the conduct of the study. For example, how and when the client is to be notified of an out-of-specification result is typically client-specific and terms involving notifications to be made to the client are generally high risk. Other terms that are often high risk center around how changes to laboratory procedures, testing protocols and written instructions in general are to be handled and how vendors and other 3rd parties providing materials and/or services in support of the client’s experimental study are to be qualified2.
Check Point FOUR: Train users on Quality Agreements in general and on each specific agreement they will be using
Quality Agreements are to be used. They are not to be put on the shelf and then forgotten. Keep track of who has been trained on each. This can be done with a training matrix. It is a good idea to keep an updated list of the agreements that are active (e.g. the training matrix) in a place highly-visible to all employees. To make it more useful, consider cross-referencing each QAG against the client and perhaps also to the Principal Investigators assigned to the client’s various experimental studies. To protect client-confidentiality, consider using a client code in place of the client name. A mechanism must be in place to ensure that the training is performed on a timely basis. Couple this training to some other required client-based activity that occurs at an appropriate time in the process of taking a client project from start to finish. For example, we start each experimental study using a kick-off meeting facilitated by the Principal Investigator (PI), who is guided by a kick-off meeting checklist3. Listed on the checklist is the requirement to review any applicable QAG. All scientists participating in the study are required to attend and to sign the attendance sheet. The attendance sheet is forwarded to the Training Coordinator, who uses it to update the Training Matrix. Any person missing the training must make it up and this will involve among other things, reading the QAG and passing a quiz on it. Attendance sheets are filed in the employees training binder.
Create an SOP on Quality Agreements. In it, provide background and details on the QAG system in place in general. All employees supporting client studies must be trained on this SOP.
Quality Assurance should keep the original QAG. QAGs are often 20 to 30 pages in length. It is a good idea for the QAU to also keep a one-to-two-page summary of key points appearing in the QAG and to note those points that the QAU anticipates causing more trouble to the lab or that may present a greater risk. The effective start and expiration dates along with client contact information should be included. The summary does not take the place of the QAG.
Check Point FIVE: Identify failures over time and feedback into the CAPA program
Any deviation from a QAG should be considered “major”, requiring a deviation investigation in order to identify root cause and corrective and preventative actions for Management and Quality Assurance review and approval4. The specific QAG will state whether or not the deviation is to be reported to the client and if so, how it is to be addressed with the client. Quality Assurance must query the CAPA database to determine if there is a pattern of failures to follow a specific QAG or QAGs, in general. Such feedback is critical to improving the QAG system and may help to improve specific Quality Agreements. Of course, the more effort put in at the front end to establish a good agreement and to train everyone on it, the less effort needed on the back end. Perhaps more importantly, like any quality system, a good QAG system will scale up with no difficulty as more and more clients desire to put QAGs into place with the lab, whereas a bad system simply will not.
SUMMARY
The effective implementation of a single Quality Agreement and the management of a quality agreement program at the Contract Research Laboratory, requires getting good QAGs into place and monitoring the lab’s performance in implementing them over time. Key to the management process is the identification and understanding of points in the workflow at which to make critical checks.
REFERENCES
Laboratory”, Journal of GXP Compliance, Vol. 16, Issue 4, 2012.
Contract Laboratory”, Journal of GXP Compliance, Vol. 17, Issue 3, 2013.
Laboratory”, Journal of GXP Compliance, Vol. 19, Issue 2, 2015.
Laboratory”, Journal of GXP Compliance, Vol. 21, Issue 2, Mar 2017.
Introduction
The demand for sophisticated drug delivery has led to the development of many innovative combinations of pharmaceuticals, biologics and medical devices across a broad range of therapeutic areas. While these combination products have the potential to fulfill major unmet medical needs, they also present unique challenges in development and approval. One of these challenges is the development and validation of analytical methods to assess the quality, safety and stability of combination products.
Types of Combination Products
The FDA Guidance for Industry Titled, “Current Good Manufacturing Practice Requirements for Combination Products (January 2017)” defines a combination product as “a product composed of two or more types of medical products (i.e. a combination of a drug, device, and/or biological product with another).” There are three types of combination products: single entity, co-packaged and cross labeled.
A single entity combination product has two or more related components (i.e. drug/device, biologic/device, drug/biologic or all three) that are physically, chemically, or otherwise combined to produce a single entity. Examples of single entity combination products would be a prefilled syringes or a drug eluting stent.
A co-packaged combination product has two or more components packaged together in a single package. Examples of a co-packaged combination product are surgical and first aid kits.
With a cross labeled combination product, each component is packaged separately and is intended only for use with an approved second product. An example of a cross labeled combination product would be a light activated drug requiring a light emitting device.
Single entity combination products (SECP) present unique analytical challenges compared to co-package and cross labeled combination devices. Most notably, a SECP runs the risk of impurities and degradants from the combining process and sterilization. In addition, an SECP may have specific design features that need to measure to ensure proper functioning of the product.
As a result of these unique analytical challenges for SECPs, this white paper will focus only on the analytical methods needed to support the development of an SECP. For brevity, this white paper will only discuss SECPs formed from a drug and a medical device. Similar approaches with the appropriate methodologies could be adapted for the other types of SECPs.
Analytical Methods for Assay, Related Substances and Degradation Products
There are three goals for these methods: to assay the drug content, to measure for known related substances and to detect degradation products of the drug. Like the analytical methods for more traditional drug products, the predominant instrumentation for these methods is high-performance liquid chromatography (HPLC)- UV, with one method usually able to accomplish all three tasks.
The main difference between analytical methods for SECPs and traditional drug product formulations like oral or parenteral dosage forms is in the sample preparation. The sample preparation is unique and specific to the SECP but usually involves complicated multi-step procedures. Based upon the SECP, sample preparation of SECPs often involves extensive extraction for complete recovery of the drugs. In addition, depending upon the size of the SECP and the location of the drug, the SECP may need to be disassembled or reproducibly cut, which can be a significant challenge when the SECP is made from hard plastics or metals.
Since the method is intended to be used as to evaluate stability of the SECP, the method will need to be proven to be stability indicating. If the SECP contains a new drug, a forced degradation study will be needed. If the SECP contains a generic drug, literature references to known degradation pathways and impurities can be used to evaluate if the method is stability indicating. However, if the combining process is significantly different from the literature references, additional degradation pathways may need to be explored experimentally.
When designing a forced degradation study for a SECP, the potential for the device to contribute to the drug degradation or the degrade itself, leading to detectable degradation products needs to be considered. The type of SECP will determine if the forced degradation study is done on just the drug, on the fully assembled SECP or on a combination of the drug and the parts of the device with direct drug contact. It is recommended to include the components of the SECP that have direct drug contact in the forced degradation study.
The conditions of the forced degradation study may also need to be adapted for an SECP. Stress conditions can be replaced with more relevant experimental conditions with proper scientific justification.
Once developed, the analytical methods need to be validated according to the ICH Harmonized Tripartite Guideline , Validation of Analytical Procedures: Text and Methodology (ICH Q2(R1)). One important additional consideration for validation of methods for a SECP is during ruggedness. In the ruggedness validation experiments, additional challenges should be added to evaluate ruggedness of the sample preparation. These experiments will vary depending upon the SECP and need to appropriately challenge each critical sample preparation step.
Residual Solvents Methods for a SECP
If organic solvents are used in the assembly of the SECP, the finished SECP will need to be tested for these residual solvents. A common example would be if the drug is sprayed onto part of a medical device during the assembly of the SECP. In this case, the solvents used in the spraying process would be considered residual solvents.
The residual solvents methods need to be developed and validated in accordance with USP <467>. One important additional consideration for residual solvents for a SECP may be the need for additional sensitivity. Because these residual solvents may be important parameters in developing the process to manufacture the SECP, the methods may be needed to reach sensitivities as much as 10 fold lower than the required specification to assist in development of the manufacturing process. Once the methods have been validated, residual solvents testing can be done as part of release testing or as part of the process validation.
Analytical Method to Evaluate the Drug Release/Elution from a SECP
If the SECP is not intended to mechanically infuse the drug into the patient but instead the drug is intended to passively diffuse into the patient, then analytical methods designed to measure the rate of drug release or elution are needed. To perform this experiment, the SECP 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 date.
Analytical Method to Evaluate Uniformity
For SECPs where the medical device is coated with a drug or drugs, the uniformity of this coating will need to be evaluated. Similar to the drug release methods, the instrument conditions used are usually those used for the assay; however, more sensitive methods may need to be developed depending upon the intended level of drug in the coating. In addition, the sample preparation will need to be adjusted so that samples are taken from all areas of the device to ensure uniformity of the coating. In some SECPs where the device is not a simple geometric shape, the surface area coated from the different sections of the device will need to be included in the determination of the level of the drug in the coating.
Analytical Methods for the Analysis of Leachables from the SECP
Leachables (a.k.a. migrants) from the medical device need to be considered when the drug is in direct contact with the medical device during the intended shelf storage, when the SECP is intended to be surgically implanted or when the SECP will have direct patient contact for an extended period of time.
For these types of SECPs, two types of studies are performed. The first study is a forced extraction study on just the medical device and the second study a migration study on the entire SECP.
In a forced extraction study, the medical device is extracted with two solvents at an elevated temperature. Usually the drug is not included in the forced extraction. The extraction solvents are selected so that one mimics either the drug formulation or the intended patient tissue that the SECP will contact and the second solvent is selected to represent a “worse case scenario” condition based upon either the drug formulation or the intended patient tissue. The sample extracts are analyzed by mass spectrometry (gas chromatography, liquid chromatography, or inductively coupled plasma (GC-MS, LC-MS, and ICP-MS respectively)) to attempt to identify all possible organic and inorganic extractables.
Analytical methods are then developed that can detect the extractables observed in the forced extraction studies as leachables in either the drug product or a model solvent that mimics the intended patient tissue. Analytical methods for leachables need to be extremely sensitive and usually require MS detection. An additional challenge commonly arises when the drug product is present at concentrations significantly higher than the levels required for detection of the leachables. In this case sample preparation steps and method adaption need to minimize the interferences from the drug. Once developed, the analytical methods need to be validated before proceeding to the migration study. The validation of the methods should be similar to validation of a method for related substances but allowances may be needed to reach the required level of sensitivity.
The migration study is the second study where the leachables (a.k.a migrants) are monitored. When the risk of leachables is deemed to be highest from the drug being in direct contact with the medical device during the intended shelf storage, the leachables should be evaluated as part of the stability study to determine the shelf life. When the risk of leachables is due to the SECP being surgically implanted or from direct patient contact, a simulated migration study is performed. In this case the SECP is exposed to a model solvent that mimics the intended patient tissue at 37°C for an appropriate length of time determine by the intended use. In both studies, previously validated analytical methods are used to evaluate the leachables entering the drug product or the model solvent.
Other Analytical Methods
Since many types of SECPs are in development with many unique critical features, analytical methods may be required to measure qualities specific to a given SECP that are expected to be critical to function properly. In this case the analytical method will need to be developed to address a specific attribute of the SECP.
These methods will still require validation even when few of the standard validation parameters apply. In this scenario, the validation should address at a minimum reproducibility and ruggedness. Ruggedness testing should include all method parameters that could impact the reported results.
Conclusion
As diverse and ingenious SECPs continue to be developed, analytical methods that can be used to support the development and ensure the quality of these products are needed. A thorough understanding of the SECP is critical to ensure that the analytical methods are monitoring the proper attributes of the SECP and the analytical chemist may need to be creative in developing methods and preparing samples to support these important therapeutic advances.
To support microbiome research, Pine Lake Laboratories has developed 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 drug treatments during pre-clinical and clinical trials. The method involves extracting the short chain fatty acids form feces then analysis by direct injection GC-MS.
This white paper will describe the method and highlight the performance of the method.
A. Sample Preparation
B. Standard and QA Preparation
Standards and QCs are prepared by spiking known levels of each short chain fatty acid into feces. The endogenous level of each short chain acid in the feces used to prepare the standards and QCs had previously been determined by standard addition. The final concentration is then the total of the amount added and the endogenous level.
Standards and QCs are then prepared for analysis same as the samples.
C. Instrumental Analysis
Samples are analyzed by GC-MS using a Nukol capillary column (15 m x 0.23 mm, 0.25 µm). Details available upon request.
A. Linearity and Range
For all three short chain fatty acids, the range of the method was 5 to 150 μg/mL with R2 > 0.990. Representative calibration curves can be found in Figures 1-3.
B. Precision in Matrix
A representative lot of feces was assayed in triplicate for all three short chain fatty acids. Results are in Table 1. Precision on all matrix samples (n=6) was excellent with %RSD values less than 5% for each SCFA within each of the three analyses. The total precision of the grand average was less than 10% for acetic and propionic acid. Butyric acid was higher at 26.6% but was still acceptable at less than 30%.
C. Accuracy and Precision
A representative lot of feces was spiked at three separate levels with known amounts of each of the three short chain fatty acids. This was repeated in three separate analyses. Results are in Table 2-4. Accuracy was acceptable for all three short chain fatty acids with average recovery within ± 15.0% of target for each level in each analysis and in the grand average. Similarly precision was acceptable at ≤15.0% for each level in each analysis and in the grand average.
This simple yet effective GC/MS analysis can be used to accurately determine the concentration of acetic, butyric, and propionic acid in feces. The method is specific, accurate, and precise with a large quantifiable range for short chain fatty acids. Additional short chain fatty acids not included in the method evaluation reported in this white paper can also be detected and quantitated using this method.
There should already be in place a strong quality program at the CRO and the quality agreement must make a good fit to this program. Those concerns (terms) that are rather common to Quality Agreements should already be addressed in the existing quality program at the CRO or else consideration must be given to incorporating them into the existing program. This approach is distinctly different and better than leaving them only in the quality agreement. To leave important concerns only in the quality agreement is tantamount to instructing the lab to only consider doing such quality work when working on this particular client’s project.
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.
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”
In addition to bioanalysis of small molecule drugs in plasma, at Pine Lake Laboratories we also have extensive experience developing bioanalytical methods by LC-MS/MS for drugs in tissue and other biological matrices. In general, tissues present a more complicated matrix than biological fluids but a well-designed mixed mode solid phase extraction method can usually handle the additional challenges of biological tissues. Below are examples of biological fluid and tissue matrices in which we have successfully developed methods:
Pine Lake Laboratories is a state of the art facility fully equipped to identify unknown degradation products and process impurities. We have successfully used our GC-MS, UPLC-QToF and ICP-MS to aid our clients in identifying unknown compounds. In the past we have adapted client’s methods to be MS compatible to aid in identification. If needed, we can even perform fraction collection to collect enough of the unknown to confirm the identity by NMR.
Peptides present some unique challenges for the bioanalysis by LC-MS/MS of these compounds in plasma and other biological matrices. One main challenge for LC-MS/MS bioanalytical methods for peptides is to get similar sensitivity and specificity compared to methods for small molecules. The challenge presented by peptide for sensitivity and specificity are due to the following properties common to most peptides:
Despite these inherent challenges from peptides, at Pine Lake Laboratories we have developed and validated LC-MS/MS bioanalytical methods for peptides that met the required sensitivity and specificity. We have used mixed mode solid phase extraction to help reduce background from similar peptides while still maintaining good recovery of the target peptide. When properly tuned and with optimized chromatographic conditions, our Waters Xevo TSQ UPLC-MS/MS instruments can achieve the needed sensitivity.
“We recognize this purchase is a significant milestone and growth opportunity for our staff and the Bristol community,” said Pine Lake Laboratories President, Kurt Moyer, Ph.D. “We plan to continue NSF International’s legacy of scientific and technical expertise and to serve our global clients. Moving forward as Pine Lake Laboratories, with our group of scientists and professionals, we will continue to grow our operations, enhance technologies and help our clients produce safer pharmaceuticals and medical devices here in Bristol.”
Along with Dr. Moyer, the Pine Lake Laboratories’ core leadership team includes group leaders Ulyana Matyugicheva and Cassandra Tellarini.
Dr. Moyer has more than 25 years of experience in pharmaceutical development. In his previous role as Director of Research at NSF International, he led all analytical, bioanalytical, and extractables and leachables studies. Prior to joining NSF International, he worked with Sanofi-Aventis as a senior research scientist and DuPont Pharmaceuticals as a senior research investigator. Dr. Moyer earned a Ph.D. in biochemistry from Villanova University and a bachelor of science in chemistry from Millersville University of Pennsylvania.
Pine Lake Laboratories is a GLP/GMP compliant analytical and bioanalytical contract testing laboratory. Our lab, located in Bristol, CT, is FDA registered and inspected and DEA licensed for Schedules 1 through V.
