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Influence of Postharvest Drying Temperatures on Alkaloid Levels In HORTSCIENCE 56(2):242–243. 2021. https://doi.org/10.21273/HORTSCI15500-20 alkaloids found in the plant. The goal of this study was to determine the influence of post- harvest drying temperature on alkaloid levels Influence of Postharvest Drying in goldenseal roots and rhizomes. Temperatures on Alkaloid Levels in Materials and Methods Goldenseal (Hydrastis canadensis L.) Plant material. In this study, goldenseal samples were removed from three spatially Grady H. Zuiderveen distinct (e.g., at least 20 m apart) colonies Department of Ecosystem Science and Management, Pennsylvania State within a wild population located in central University, University Park, PA 16802 Pennsylvania. Fourteen ramets were harvested from each plot in early April while plants were Eric P. Burkhart dormant. Voucher specimens were collected Department of Ecosystem Science and Management, Pennsylvania State and deposited at the Pennsylvania State Uni- versity Herbarium (PAC). University, University Park, PA 16802; and Shavers Creek Environmental Processing conditions. The underground Center, 3400 Discovery Road, Petersburg, PA 16669 portion of the plant consists of a horizontal rhizome roughly 1–1.5 cm in diameter, with Joshua D. Lambert multiple fibrous rootlets extending from the Department of Food Science, Pennsylvania State University, University rhizome. For this study, the roots and rhi- Park, PA 16802 zomes (subsequently referred to simply as rhizomes) were processed and analyzed to- Additional index words. alkaloid, medicinal plant chemistry, drying, quality control gether. This was done because industry typ- Abstract. Goldenseal (Hydrastis canadensis L.) is a medicinal forest herb native to ically does not differentiate between these Appalachia. Its roots and rhizomes are used as an antimicrobial and for the treatment of parts. Following harvest, rhizomes were intestinal ailments. Three alkaloids–berberine, hydrastine, and canadine–are recognized cleaned on a screen by hand under running as the major bioactive constituents in goldenseal. One important postharvest processing water to remove any soil. step for goldenseal is drying; however, it is not known how drying temperature influences Rhizome samples were dried using seven the concentrations of these alkaloids. In this study, pre-emergent (dormant) goldenseal different drying conditions (6 samples/drying samples were freeze-dried or air-dried at six different temperatures (26.7 to 54.4 8C) to temperature). Samples were dried until the determine the relationship between drying temperature and alkaloid content in the dried mass was 30% of the fresh weight, and rhizome and roots. High performance liquid chromatography analysis showed that the rhizomes could be broken cleanly (Davis berberine and hydrastine levels were unaffected by drying temperature, while canadine and Persons, 2014). Supplemental Table 1 levels decreased as temperature increased (0.55% w/w on average when samples were shows the final mass of the dried samples and freeze-dried, down to 0.27% w/w on average when dried at 54.4 8C). While canadine is the the mass of moisture lost. Air-dried samples least abundant alkaloid of the three, it is known to have key antibacterial properties. were dried in an adjustable Lindberg/Blue M Developing a more standardized drying protocol for goldenseal could lead to a more 260 Mechanical Oven (Model number predictable phytochemical profile. MO1490C-1; Thermo Scientific, Asheville, NC), with an air flow rate of 25.5 L/min and specific humidity of 8.9 g H2O/kg air at 26.7, Goldenseal (Hydrastis canadensis L., et al., 2008; Mahady and Chadwick, 2001; 32.2, 37.8, 43.3, 48.9, or 54.4 °C (nominal Ranunculaceae) is a medicinal herb indige- Scazzocchio et al., 2001). uniformity of 3.5% of the setpoint). The nous to eastern North American forestlands. Most of the commercially traded golden- drying temperatures selected cover the range It is included in many formulations used to seal is still wild harvested (Oliver and Lea- of current recommendations and reach the treat intestinal infections and digestive ail- man, 2018). While drying is an important lower limits that can be obtained while still ments, and it is known to have antimicrobial, postharvesting step for goldenseal (Lloyd and using a drying oven. One set of samples was anticancer, and immunostimulatory proper- Lloyd, 1884), no standardized protocol ex- freeze-dried using a VirTis Genesis Freeze ties (Predny and Chamberlain, 2005). The ists, and various drying methods are used Dryer (SP Industries, Warminster, PA) to medicinal properties of goldenseal are widely (Personal observation; Upton, 2001). Current provide a baseline of alkaloid concentrations attributed to three major isoquinoline alkaloids ‘‘folk’’ drying methods include drying the for comparison. After being dried, samples (Supplemental Fig. 1A): berberine, hydrastine, rhizomes in the sun and/or shade, in small were stored in air-tight containers at 4 °C and canadine (Abidi et al., 2006; Brown drying sheds, and in forced air dryers (Davis until analysis. and Persons, 2014; Personal observation). Chemical calibration standards. Berber- The most recent recommendations for gold- ine hydrochloride (purity > 98%) and cana- enseal rhizome suggest drying at 35 to dine (tetrahydroberberine) (purity > 98%) 37.7 °C to prevent mold growth and the were purchased from Quality Phytochemicals Received for publication 5 Oct. 2020. Accepted for b publication 4 Dec. 2020. decay of the rhizome (Davis and McCoy, LLC (East Brunswick, NJ); (-)- -Hydrastine Published online 12 January 2021. 2000). Davis and Persons (2014) state that (purity > 99%) was purchased from Chroma- This research was supported by the Pennsylvania rhizomes will lose 70% of their weight as Dex (Irvine, CA). State University College of Agriculture Graduate water evaporation during drying, and tem- High-performance liquid chromatography Student Competitive Grants Program (to G.H.Z.) peratures should be kept as low as possible analysis. Dried rhizomes were ground using a and was supported in part by USDA Hatch Project (ideally between 29.4 and 37.8 °C). The mortar and pestle. Fifty mg (±10%) of ground PEN 4565 and the Azzara Family Food Bioactives American Herbal Pharmacopoeia has identi- material was combined with 4 mL of extrac- Research Endowment (to J.D.L.). fied that determining the optimal drying tion solvent (70% water, 29.9% acetonitrile, Current address for G.H.Z.: USDA Forest Service, 0.1% phosphoric acid). Tubes were vortexed 201 14th Street SW, Washington, D.C. 20250. conditions for goldenseal is an area that needs E.P.B. is the corresponding author. E-mail: epb6@ further research (Upton, 2001). Developing a to mix, sonicated for 10 min at room temper- psu.edu. more standardized drying protocol for gold- ature, and centrifuged for 8 min at 3220 gn. This is an open access article distributed under the enseal could lead to more predictable health The supernatant was diluted 1:4 with 10% CC BY-NC-ND license (https://creativecommons. applications and outcomes by preserving the aqueous acetonitrile and filtered through org/licenses/by-nc-nd/4.0/). 0.2-mm nylon filters. 242 HORTSCIENCE VOL. 56(2) FEBRUARY 2021 High-performance liquid chromatogra- One explanation for the difference in find- method for the detection and/or quantifica- phy (HPLC) analysis was performed using a ings between the alkaloids is that berberine and tion of select alkaloids in goldenseal supple- Shimadzu HPLC system (Shimadzu Co., Co- hydrastine are both end products of biosynthetic ments and raw materials by reversed-phase lumbia, MD) equipped with two LC-20AD pathways, while canadine is an intermediate high-performance liquid chromatography. pumps, a SIL-20AC HT refrigerated auto step in berberine biosynthesis by the action of Pharm. Biol. 46(1–2):135–144, doi: 10.1080/ 13880200701735171. injector, a column oven maintained at the enzyme tetrahydro-protoberberine oxidase 24 °C, and an SPD-20AV ultraviolet/Vis (Mander and Liu, 2010). As such, an increase Correche, E.R., S.A. Andujar, R.R. Kurdelas, M.J.G. Lechon, M.L. Freile, and R.D. Enriz. detector. A binary gradient of water contain- in the enzymatic activity of the oxidase would 2008. Antioxidant and cytotoxic activities of ing 0.1% formic acid (solvent A) and meth- result in a decrease in canadine levels; and it is canadine: Biological effects and structural as- anol containing 0.1% formic acid (solvent B) known that as temperature increases, enzyme pects. Bioorg. Med. Chem. 16(7):3641–3651, with a flow rate of 0.75 mL/min was used. activity increases up to the point of enzyme doi: 10.1016/j.bmc.2008.02.015. The initial mobile phase was 20% B. The denaturation (Daniel and Danson, 2013; Elias Daniel, R.M. and M.J. Danson. 2013. Temperature concentration of B increased linearly for et al., 2014). The results for canadine are and the catalytic activity of enzymes: A fresh 15 min to 45%, and this was held at the consistent with this hypothesis. understanding. FEBS Lett. 587(17):2738–2743, concentration for 5 min. The mobile phase While canadine is the least abundant of doi: 10.1016/j.febslet.2013.06.027. wasthenreturnedto20%,andtheHPLC the three alkaloids, it is still potentially an Davis, J. and J.-A. McCoy. 2000. Commercial was re-equilibrated for 7 min. Analytes important contributor to the overall efficacy goldenseal cultivation. North Carolina State Univer- were separated using a Zorbax Eclipse of goldenseal. When isolated, canadine has sity, Raleigh. 8 Dec. 2020. <https://content.ces.nc- XDB-C18 column (4.6 · 150 mm, 3.5 mm been found to have significant activity su.edu/commercial-goldenseal-cultivation>. particle size, 80 A˚ pore size). Eluent was against numerous strains of bacteria, and it Davis, J. and W.S. Persons. 2014. Growing and monitored at 280 nm. is the only one of the three major alkaloids marketing ginseng, goldenseal and other wood- Statistical analysis. All statistical analysis found to be active against Pseudomonas aeru- land medicinals. New Society Publishers, was completed in R-studio version 3.5.1 ginosa and Staphylococcus aureus (Abbasoglu Gabriola Island, British Columbia.
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