Veratrum spp. grow throughout the world and are especially prevalent in high mountain meadows of North America. All parts of Veratrum plants have been used for the treatment of ailments including injuries, hypertension, and rheumatic pain since as far back as the 1600s. Of the 17–45 Veratrum spp., Veratrum californicum alkaloids have been proven to possess favorable medicinal properties associated with inhibition of hedgehog (Hh) pathway signaling. Aberrant Hh signaling leads to proliferation of over 20 cancers, including basal cell carcinoma, prostate and colon among others. Six of the most well-studied V. californicum alkaloids are cyclopamine (1), veratramine (2), isorubijervine (3), muldamine (4), cycloposine (5), and veratrosine (6). Recent inspection of the ethanolic extract from V. californicum root and rhizome via liquid chromatography–mass spectrometry has detected up to five additional alkaloids that are proposed to be verazine (7), etioline (8), tetrahydrojervine (9), dihydrojervine (10), 22-keto-26-aminocholesterol (11). For each alkaloid identified or proposed in V. californicum, this review surveys literature precedents for extraction methods, isolation, identification, characterization and bioactivity to guide natural product drug discovery associated with this medicinal plant.

Veratrum californicum contains steroidal alkaloids that function as inhibitors of hedgehog (Hh) signaling, a pathway involved in the growth and differentiation of cells and normal tissue development. This same Hh pathway is abnormally active for cell proliferation in more than 20 types of cancer. In this current study, alkaloids have been extracted from the root and rhizome of V. californicum, followed by their separation into five fractions using high performance liquid chromatography. Mass spectrometry was used to identify the presence of twenty-five alkaloids, nine more than are commonly cited in literature reports, and the Bruker Compass Data Analysis software was used to predict the molecular formula for every detected alkaloid. The Gli activity of the raw extract and each fraction were compared to 0.1 µM cyclopamine, and fractions 1, 2, and 4 showed increased bioactivity through suppression of the Hh signaling pathway. Fractions 2 and 4 had enhanced bioactivity, but fraction 1 was most effective in inhibiting Hh signaling. The composition of fraction 1 consisted of veratrosine, cycloposine, and potential isomers of each.

Glycomacropeptide (GMP) is isolated from whey and used as an ingredient in phenylketonuria-safe foods because it does not contain phenylalanine. GMP is highly glycosylated and has several sites where N-acetylneuraminic acid (NANA) is bound. In the dairy industry, quantification of NANA from dairy proteins is accomplished by colorimetric, fluorometric, enzymatic, and chromatographic procedures; there is no uniformly accepted industry-wide standard method. In this investigation, NANA quantification methods were evaluated using GMP, and a comparison was made based on the length of time to complete the assay, protein-specificity, linearity, precision, and accuracy. From the methods evaluated, the chromatography protocol was determined to have the greatest benefit for use as a dairy industry standard to measure NANA on GMP. The average mass percent of NANA in 10 statistically independent replicates from a commercial GMP product was measured to be 6.18% ± 0.12%, with a relative standard deviation of 1.94%, which was the lowest of all the methods tested. The accuracy of the chromatographic approach was validated using spike and recovery experiments that provided an average recovery of 90.25%.