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Saw Palmetto Extract Laboratory Guidance Document

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Saw Palmetto Extract Laboratory Guidance Document Saw Palmetto Extract Laboratory Guidance Document By Stefan Gafner, PhD* American Botanical Council, PO Box 144345, Austin, TX 78714 Serenoa repens *Correspondence email Photo ©2019 Steven Foster Keywords: Adulteration, animal fatty acids, canola oil, coconut oil, palm oil, saw palmetto, Serenoa repens, sunflower oil, vegetable oil Citation (JAMA) style: Gafner S. Saw palmetto extract laboratory guidance document. Austin, TX: ABC-AHP-NCNPR Botanical Adulterants Prevention Program. 2019. 1. Purpose There is documented evidence of the adulteration of saw palmetto fruit extracts with a number of vegetable oils, such as canola (Brassica napus ssp. napus, Brassicaceae), coconut (Cocos nucifera, Arecaceae), olive (Olea europaea, Oleaceae), palm (Elaeis guineensis, Arecaceae), peanut (Arachis hypogaea, Fabaceae), and sunflower (Helianthus annuus, Asteraceae) oils. The partial or complete substitution of saw palmetto fruit extracts with mixtures of fatty acids of animal origin was first documented in 2018,1 and seems particularly common in materials sold as saw palmetto originating from China. This Laboratory Guidance Document (LGD) presents a review of the various analytical technologies used to differentiate between authentic saw palmetto extracts and ingredients containing adulterating materials. This document can be used in conjunction with the Saw Palmetto Botanical Adulterants Bulletin, rev. 3, published by the ABC-AHP-NCNPR Botanical Adulterants Prevention Program in 2018.2 2. Scope does not reduce or remove the responsibility of laboratory Various analytical methods are reviewed here with the personnel to demonstrate adequate method performance specific purpose of identifying their strengths and limita- in their own laboratories using accepted protocols. Such tions in differentiating saw palmetto fruit extracts from protocols are outlined in the United States Food and Drug potentially adulterating materials. Less emphasis is given to Administration’s Good Manufacturing Practices (GMPs) the authentication of whole, cut, or powdered saw palmetto rule (21 CFR Part 111) and those published by AOAC Inter- fruit and distinguishing it from potential confounding national, International Organization for Standardization materials, e.g., the Everglades palm (Acoelorrhaphe wrightii, (ISO), World Health Organization (WHO), and Interna- Arecaceae), by macroscopic, microscopic or genetic analy- tional Conference on Harmonisation (ICH), and national sis. Analysts can use this review to guide their selection pharmacopeial bodies, as may be applicable, depending on of appropriate analytical authentication techniques. The the regulatory requirements of the country in which the suggestion of a specific analytical method for testing saw saw palmetto extract is being offered for sale, re-sale, and/ palmetto materials in their particular matrix in this LGD or processing into finished consumer products. Saw Palmetto Extract - Laboratory Guidance Document • 2019 • www.botanicaladulterants.org 1 3. Common and Scientific Names 4. Botanical Description and Geographical Range Saw palmetto grows as a small shrub, occasionally a small 3.1 Common name: Saw palmetto tree with creeping, horizontal, branched stems, usually to a height of 2-7 feet (0.6-2.1 m), although it may reach up to 3.2 Other common names for saw palmetto 25 feet (7.5 m). The stem systems run parallel to the soil English: Scrub-palmetto, sabal palm, saw palmetto berry surface, eventually branching beneath the substrate to form Chinese: Ju zonglu (锯棕榈) rhizomes. Saw palmetto leaves are fan-shaped, evergreen French: Sabal, palmier nain, palmier scie and about 3 feet (1 m) wide. The margins of the petioles German: Sabal, Sägepalme, Zwergpalme are lined with sharp spines that have given saw palmetto Italian: Palma nana, cavolo di palma its common name. The flowers are cream-colored and Russian: Сереноя ползучая (Serenoa repens), Сабаль fragrant, with three petals at the end of stalked panicles пильчатый (Sabal serrulata), карликовая пальма (karliko- that grow from the leaf axils. The fruit is a drupe, green or vaya palma, “dwarf palm”), пальма cереноа, co пальметто yellow at immature stages, and black when ripe (between Spanish: Sabal, palma enana americana3,4 August and October), resembling black olives in size and Swedish: sågpalmetto shape.6,7 The plant is endemic to the southeastern United States, growing from the coastal plains of Louisiana across 3.3 Latin binomial: Serenoa repens (W. Bartram) Small the Florida peninsula and up to South Carolina.6 3.4 Synonyms: Chamaerops serrulata Michx., Corypha 5. Adulterants and Confounding Materials obliqua W. Bartram, Corypha repens W. Bartram, Diglos- See Table 1 below. sophyllum serrulatum (Michx.) H. Wendl. ex Drude, Sabal serrulata (Michx.) Nutt. Ex Schult. & Schult. f., Serenoa 6. Identification and Distinction using serrulata (Michx.) G. Nicholson5 Macroanatomical Characteristics Botanical descriptions of saw palmetto fruit have been 3.5 Botanical family: Arecaceae published in a number of pharmacopeial monographs Table 1. Scientific Names, Family, and Common Names of Plants Used as Sources of Vegetable Oils Known as Saw Palmetto Fruit Extract Adulterants* Speciesa Synonym(s)a Family Common nameb Other common namesc Arachis hypogaea L. A. nambiquarae Hoehne Fabaceae Peanut Groundnut Brassica napus L. Brassica napus ssp. napus Brassicaceae Canolad Colza, rape, rapeseed Cocos nucifera L. Calappa nucifera (L.) Kuntze Arecaceae Coconut Coconut palm Cocos indica Royle C. nana Griff. Elaeis guineensis E. dybowskii Hua Arecaceae African oil palm Oil palm Jacq. E. madagascariensis (Jum. & H. Perrier) Becc. E. melanococca Gaertn. Helianthus annuus L. H. aridus Rydb. Asteraceae Sunflower H. jaegeri Heiser H. lenticularis Douglas H. macrocarpus DC H. ovatus Lehm. Olea europaea L. Oleaceae Olive aThe Plant List and the Kew Medicinal Plant Names Services database.8,9 A comprehensive list of synonyms can be accessed through both websites. bAmerican Herbal Products Association’s Herbs of Commerce, 2nd ed.3 cAmerican Herbal Products Association’s Herbs of Commerce, 2nd ed.,3 and the USDA GRIN database.10 dAccording to the Canadian Food Inspection Agency,11 canola oil may be obtained from Brassica rapa and B. juncea as well. *Note: Species other than those listed in Table 1 that are used for production of edible fatty oils are also known to be used as adulterants. In addition, admixture or substitution of saw palmetto extracts with “designer blends,” mimicking the saw palmetto fatty acid composition, which include fatty acids from animal sources have been described.1,12 The few reports of adulteration of saw palmetto berries with berries from related species, i.e., dwarf palmetto (Sabal minor),13,14 queen palm (Syagrus romanzoffiana),14 and Everglades palm (Acoelorrhaphe wrightii)15 of the palm family (Arecaceae) seem to suggest that such adultera- tion is rare. Fruits of dwarf palmetto (6-12 mm) and the Everglades palm (10-15 mm) are smaller and spherical compared to saw palmetto fruit, which is oval-shaped, of ca. 15 mm width and 12-25 mm length.4,13,14,16 Compared to saw palmetto, the fruit of queen palm is larger (20-25 mm) and heavier.14 2 Saw Palmetto Extract - Laboratory Guidance Document • 2019 • www.botanicaladulterants.org and books.4,17-19 Criteria to distin- guish saw palmetto fruits from other fruits in their whole form have been published by many authors.16,20,21 The macroscopic assessment is the method of choice to distinguish unripe (green) from semi-mature or mature (black) berries and is sufficient for identify- ing saw palmetto to species. For obvi- ous reasons, macroscopic identifica- tion is not applicable to saw palmetto extracts. 7. Identification and Distinction using Microanatomical Characteristics Microscopic descriptions of saw palmetto are found in the pharma- copeias of Europe and the United States, and the American Herbal Pharmacopoeia’s textbook on micro- scopic characterization of botanical medicines.17,18,22 Details of the fruit anatomy of saw palmetto, Everglades palm, and dwarf palmetto have been 21 published by Zona; however, no clear Figure 1: Color of authentic saw palmetto ethanol extracts (1,2) and CO2 differentiation criteria for the fruits of extracts (3,4); adulterated ingredients labeled as saw palmetto extract (5-8). these palms using botanical micros- Image provided by Euromed, SA (Mollet del Vallès, Spain). copy were provided. (18S, ITS, ms, prk, rpb2) were used to establish the rela- 8. Organoleptic Identification tionship among members of the palm family.25,26 Genome Saw palmetto berries are initially sweet, then pungent, skimming was applied to assemble the entire chloroplast acrid, and saponaceous. The aroma is strongly aromatic nucleotide sequence in leaf samples of 29 palm species, and foul, reminiscent of foul-smelling socks. Saw palmetto including saw palmetto and the Everglades palm.23 Genetic extracts also have a distinct aromatic and foul odor. Color data on commercial saw palmetto products are less abun- can provide an indication to detect adulterated ingredients dant. Nevertheless, Little and Jeanson15 investigated the authenticity of 37 commercial saw palmetto products (Figures 1 and 2). Ethanol, hexane, and high pressure CO2 extracts of saw palmetto from ripe berries typically have containing dried, cut and sifted plant material using mini- a dark green-brown color due to the extraction of chloro- barcodes from the matK and the rbcL regions. Amplifiable phyll with these solvents. Low pressure
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