As the dietary supplements industry grows and matures, there are inherent challenges we face. Most criticisms leveled at the industry by the media, State Attorneys General, the FDA and others have to do with quality and how that’s both defined and ensured. In our 20 years in business, one thing has never changed: a good testing lab is a gatekeeper for quality. What, then, do you need to know to help your lab help you prove your quality?
Identity & GMP Compliance
At the heart of regulatory compliance and providing safe and effective products to consumers is a basic question: What is it?
Knowing what is expected is key to cGMP compliance. The rules state that a scientifically valid method must be used to assess the identity of a product and be sure label claim is met, but they don’t state exactly what methods are to be used.
With identity testing being the first step of GMP compliance for botanicals, the identification aspect has to be very well thought out from beginning to end, from plant identification to record keeping. Traditionally, experienced herbalists relied on organoleptic identification of herbs: looking at it, smelling it and tasting a plant to confirm identify. Regarding the validity of organoleptic identification, the FDA says only when performed in a scientifically valid manner alongside legitimate training records proving the expertise employed, can this age-old technique be utilized. Obviously, this isn’t an option for powdered herbs, which is why a lab needs to be brought in here. And depending upon the herb, if it’s a likely subject for adulteration, identity testing is imperative.
While there has always been a testing component to cGMP compliance for dietary supplement manufacturers, New York Attorney General Eric Schneiderman brought it to the attention of people outside the GMP compliance department. His criticisms aside, we’ve seen a definite correlation over the last 20 years between our year after year growth and adherence to GMP compliance. Schneiderman’s attention created interest in understanding the nuts and bolts of testing, particularly test methods and technologies. That has led to conversations about which test methods to use, and our answer, every time, is that it always comes down to what is fit for purpose.
Fit For Purpose Test Methods
We all experienced the uproar based upon testing methods neither scientifically valid nor yet fit for purpose - the two cardinal rules upon which accurate testing is founded. While DNA barcode testing has a place in the pantheon of testing methods when performed by trained experts who understand the kinds of substances they are working with, it just can’t give accurate results on extracts because the commercial extraction process leaves little or no actual plant tissue, degrading DNA while leaving the actives present. It also doesn’t quantify, so the traces of substances not on the label - like rice via excipients - are identified but not quantified. DNA is also not capable of plant part differentiation, which is critical to an accurate ID. Obviously DNA testing alone will not provide the vital information you are required to obtain.
The World Health Organization and other government and non-profit organizations, including the U.S. Pharmacopeia and American Herbal Pharmacopeia, have published extensively on which methods are best for ensuring the quality of botanical materials which includes identity testing.
Examples of critical tools for botanical identity verification are Botanical Microscopy, HPTLC/ MS and HPLC fingerprinting and botanical reference standards. Chromatography – both TLC and HPLC – have been proven to be effective tools for helping the industry to unravel the complex chemistry of botanical products, thus proving quality and identity. Chromatography is very often appropriate for many of the samples that cross our lab benches, and as we continue to guide clients towards the right testing, HPLC and TLC are frequently part of the equation.
The usual tests a competent lab would perform for identity testing for finished products would include a purposeful combination of high performance liquid chromatography (HPLC) fingerprint analysis or high performance thin layer chromatography (HPTLC). While they do have some limitations, these methods have stood the test of time and are accepted by government agencies like the FDA, Australia’s TGA and Health Canada, among many others. In my experience, these tests work extremely well in the hands of specifically trained technicians who are committed to accurate results.
Extracts and Testing 101
For botanical extracts, it is critical to know what you are measuring. Botanical extracts contain a chemical profile or fingerprint, and depending on what solvents are used, that profile or fingerprint can change significantly.
Each morning I make an extract using hot water and ground Coffea Arabica seeds, otherwise known as espresso. If I were making a botanical extract for a dietary supplement, I'd then dry this liquid extract into a powder. Because water was used as the solvent, in science terms, it pulls out polar phytochemicals from the seeds and leaves behind others. That’s what we drink in our coffee, a solution mostly of water, pigments and polar compounds.
To test botanical extracts, you need to know what to look for. One easy and time-tested way is to compare the test sample to a reference sample of verified authenticity. That reference sample must be treated and prepared in the same way as the test sample in order to accurately compare the two. This is best done via high performance, thin layer chromatography (HPTLC) because it does comparative chemical fingerprinting.
While HPTLC is more than adequate, the other technique commonly used to ID a botanical extract is HPLC. HPLC is far more complicated and expensive to run and requires use of phytochemical standards or reference materials to compare and contrast a phytochemical fingerprint. An important point to note is that HPLC is designed to quantify and not to qualify. Methods are readily available to quantify the caffeine in your samples of Green Tea, however, the presence of caffeine does not mean you have Green Tea leaf, rather, you have something that has caffeine in it. It is not uncommon that HPLC is improperly used to assess the ID of a test sample. Saw dust spiked while caffeine might pass. Sometimes, those who use HPLC fingerprinting to ID a botanical do so because they have access to HPLCs and not HPTLC equipment. It’s a good question to ask those testing your botanical ingredients.
While HPTLC can successfully confirm the presence of specific botanical ingredients in a finished product, there are limitations with all methods, and one that confounds HPTLC is the practice 'fairy dusting.' Throwing in a smidgen of these and those ingredients just so they can be put on the label without offering therapeutic doses is not only unethical, it complicates testing.
Phytochemical reference standards are highly purified substances used in natural product testing. Their proper selection and use directly impacts the ability to assess the safety, quality and potency of both raw materials and finished products. Botanical reference materials are plant biomass materials that have been authenticated through a variety of analytical techniques such as macroscopic verification, microscopic identification and HPTLC. These materials are critical tools that help the industry meet cGMP ingredient identification requirements for botanical raw materials.
Finished Product Formula Testing
You can’t test quality into a product, you have to build it into the formula, which takes more than quality ingredients in therapeutic amounts. Establishing how it will be tested should be part of the process, and choosing time-tested ingredients will simplify the process.
Methods do not exist for complicated finished products until they are developed. Testing the caffeine content in Green Tea is fairly simple. Testing the caffeine in a product mixed with several other ingredients requires custom method development for a custom formula.
While the average FDA auditor is not generally trained in either Botany or Phytochemistry, they are skilled at finding loose threads. We have had several clients who chose not to invest in custom testing for custom products receive 483 notices from FDA.
If you are using a contract manufacturer, the good ones have in-house labs and should be able to provide you with methodology to prove each and every claim on that label. If it costs more, remind yourself that the extra investment is a trifle compared to the legal bills a 483 would require.
What happens when a material fails testing?
There is a learning curve when ramping up a robust testing program. For those in the early stages of doing so, an important thing to learn is how to handle a failed sample: the first priority is to identify why.
No one wants a failed sample, but the fact of the matter is, it happens. A positive result of all the negative publicity that the NY Attorney General sparked several years ago was that the number of companies finally starting to test their incoming ingredients and finished products more accurately, or at all, has been increasing. Understandably, this growth in the number of products we’re testing has increased the rate of sample failures we have been finding, giving rise to a lot of perplexed companies trying to figure out what the problem is and if they are doing things according to the cGMP’s, 21 CFR part 111 for dietary supplements.
A sample can fail to meet spec for botanical ID analysis or quantitative analysis. Whenever we fail a sample, it is frustrating and time consuming for all parties involved while we get to the bottom of the failure.
As an Analytical Testing Lab that has dealt with these problems for many years, we have a fairly rigorous method of investigating failures to rule out operator error. This may involve repeating the analysis with the same or a different method, etc. But, more importantly, we always perform a very thorough investigation of the submitted test sample by confirming that we have all the necessary information on our Sample Submission Form so that we can appropriately prepare the reference samples in the same or similar way to that used when the test sample was prepared. If this information was not available or was inaccurate, the samples are more likely to fail.
Because there are many reasons a sample can fail and in order for the testing lab to be confident in their results, it is essential that the Sample Submission Form be filled out accurately, so we compare the right sample and plant parts to the test sample.
Novel ingredients require special attention. Sometimes, the test sample was extracted in a very unique way that has removed unwanted phytochemical constituents, leaving typically only one component of interest that is not unique to the plant from which it was said to be derived. We have to fail the sample as not being chromatographically equivalent to the reference samples it was compared to. This is why it’s so important that suppliers of novel ingredients work with testing labs so they understand how the ingredients were processed and how they need to test it. It saves a lot of time, money, uncertainty and sleepless nights for all parties involved if suppliers of such novel ingredients interact with testing labs so they know how to test those ingredients.
Another obvious cause for failure is the test sample is simply not what it is supposed to be, or even not the same genus and species as the reference samples compared with. Since ingredient suppliers and/or manufacturers do not have to prove by third party testing that they sell what they say they sell, the buyer must perform testing to document identity.
Identifying a Competent Lab
How do you know if you are working with a lab that knows their stuff? One way is credentials.
An important lab certifying organization is the International Organization for Standardization (ISO) accreditation which offers a variety of certifications. 9001 is the most recognized certification as it applies to many industries. Just about every large manufacturer of just about any product is ISO 9001 or some variation thereof certified. The most meaningful accreditation to the dietary supplement industry is ISO 17025 which addresses how you manage a quality system, and confirms compliance with the AOAC Guidelines for Laboratories Performing Microbiological and Chemical Analyses of Food and Pharmaceuticals.
It’s important not to simply accept an ISO but to ask, “What is the scope of the accreditation?” It is possible for a lab to be ISO 17025 and not reveal what specifically they are accredited for, so it’s important not to assume across the board applicability. Perhaps it’s for a narrow scope like pH testing only, which may not be relevant to your needs. Ask what a company’s ISO 17025 testing covers, specifically.
Who certifies the certifiers? Any good accreditation body will be frequently under review by the International Laboratory Accreditation Cooperation (ILAC) Mutual Recognition Arrangement (MRA), International Accreditation Forum (IAF) and the Multilateral Recognition Arrangement (MLA). Alkemist Lab’s ISO accreditation is with A2LA. Without such, global acceptance of your ISO accreditation may be in question. Not all accrediting bodies are created the same. There are many to choose from, but the ones that offer the strictest of audits and requirements are the Laboratory Accreditation Bureau, ACLASS and Perry Johnson Laboratory Accreditation Inc.
A final note on what you should expect from your testing lab. Unfortunately, most labs don't reveal the entire method used to arrive at the information on the C of A that the client receives or they charge extra to provide it. That information should be included in information the lab provides to you, because if the test isn’t duplicable by another competent testing expert using the same methods, you’re vulnerable.
As the industry works to address the challenges that come with a maturing industry and global supply chain, a robust testing program can strengthen many of the attendant vulnerabilities. It’s my personal and professional mission to share information that helps that happen.