Analytical Services

Analytical Services Sample Applications

Standardized Procedure for Determination of Total Phenolics Concentration in Natural Extracts

Standardization of natural extracts is a necessary step in the manufacture of commercial extracts to ensure product consistency and batch-to-batch equivalency. One test that has been used extensively to standardize natural extracts is the measurement of the total phenolics content. The total phenolic content of a sample is determined by derivatizing the polyphenols in the sample and comparing the UV-VIS absorbance to a standard curve of a representative polyphenol, typically gallic acid. The results are then reported as gallic acid equivalents (GAE). This method is applicable to a wide range of natural extracts.

Analytical Reference Standard Purification and Qualification

High purity, fully characterized reference standards are necessary for validation of analytical methods during API development and batch testing during manufacturing. Cayman Chemical offers a full suite of services to ensure our Analytical Reference Standards are of the highest quality available on the market. HPLC purity, mass spectral data, NMR, Karl Fischer water content, melting point, and residual solvent testing are performed routinely and additional tests such as IR, optical rotation, and heavy metals, among others are available upon request.

Automated Multi-Gram Chiral Purification of Nutlin-3

The protein p53, often called the ‘guardian of the genome,’ is a transcription factor that is activated in response to cellular stress (low oxygen levels, heat shock, DNA damage, etc.) and acts to prevent further proliferation of the stressed cell by promoting cell cycle arrest or apoptosis. Its role as a tumor suppressor is evident by the observation that approximately 50% of human tumors have mutated or non-functional p53. Mdm2, a key negative regulator of p53, which is over-expressed in many human tumors, functions by binding to and targeting p53 for proteasomal degradation. (-)-Nutlin-3 is a potent inhibitor of p53-Mdm2 interaction. It potently inhibits Mdm2-p53 binding with an IC₅₀ value of 0.09 µM, induces the expression of p53-regulated genes, and exhibits potent antiproliferative activity in cells with functional p53, but not in cells with mutated p53. (-)-Nutlin-3 inhibits the proliferation of exponentially growing human skin and murine fibroblasts with IC₅₀ values of 2.2 and 1.3 µM, respectively, and induces substantial tumor shrinkage in mice expressing LnCaP, 22Rv1, or MHM cancer cell lines when treated orally with a 200 mg/kg dose twice daily for two weeks. An automated, chiral HPLC method has been developed to purify racemic Nutlin-3 into its more active (-)-enantiomer and less active (+)-enantiomer. Purity and enantiomeric excess >98% is routinely achieved and the process can be scaled up to multi-gram scale.