Norman J. Doorenbos

CULTIVATION, EXTRACTION, AND ANALYSIS OF CANNABIS SATIVA L.*

Cannabis sativa L. (marijuana) has been cultivated at the University of Mississippi since 1968 under contract support from the National Institute of Mental Health. The primary objective is to provide fully defined standardized marijuana for research. Confiscated marijuana of unknown history is often adulterated. C. sutiva was grown on three sites in Mississippi in 1968 and on two sites in 1969 and 1970. The soil types were loess (clay) on campus (site A), sandy loam some 300 miles south of campus (site B), and clay loam in the delta region of the state (site C) . Site C was used only in 1968. Land was prepared by conventional agricultural methods and treated with slag at the rate of two tonslacre. Seeds were planted mechanically or by hand at depths of about one-half inch in 40-inch-wide rows with a side dressing of 13-13-13 fertilizer at the rate of 300 pounds/acre. Plantings were made in May, June, and July. Seeds germinated in less than one week if the soil was moist. Weeds were controlled by cultivators and hand hoes. No herbicides or insecticides were used. Weed control was especially important to the young plants, since their growth was greatly inhibited by competition for light, water, and soil nutrients. Plants were watered at least once a week when it did not rain. Plants were usually thinned to 12-18 inches apart when 12-24 inches high. This was best accomplished by clipping an inch above the ground, since uprooting frequently set back or killed neighboring plants. This method allowed greater branching and leaf and flower development. Plants were also side-dressed one or two additional times during the growing season. Many seed types have been planted in this study. One cannot help being amazed at the many morphological variations observed among plants produced by these seed types, and it must be concluded that Cannabis is either a genus composed of more than 100 species or a single species that has not stabilized and that has many variations. We prefer to look upon these plants as a single species, Cannabis sutiva, as first named by Linnaeus. Plants within a given seed type tended to resemble each other and grow at similar rates, although some marked differences were occasionally observed. Plants grew as rapidly as two feet a week during peak growth and were three to eighteen feet in height at maturity. Branches measured from a few inches to eight feet in length and leaves from two to twelve inches across, and the number of leaflets per leaf ranged from one to eleven, almost always odd in number (FIGURES 1, 2, and 3). Plants of several variants were observed to contain many

A simple gas liquid chromatography procedure for determination of cannabinoidic acids inCannabis sativa L.

La chromatographie gazeuse des extraits de la marihuana donne la quantité complète de chaque composant cannabinoïque; tandis que la silylation des extraits précédant la chromatographie gazeuse, fournit des informations indépendentes sur les formes acides et non acides de chaque composant cannabinoïque.

RUSCODIBENZORURAN, A NEW DIBENZOFURAN FROM RUSCUS ACULEATUS L. (LILIACEAE)

The structure of ruscodibenzofuran, a new naturally occurring dibenzofuran from the roots of Ruscus aculeatus L. (Farn. Liliaceae), was determined by spectral means, the synthesis of model compounds and was confirmed by X-ray diffraction analysis.

Structure-activity relationship of grayanotoxin derivatives using a tetrodotoxin-antagonized spasmodic response of brine shrimp larvae (Artemia salina)

Abstract Grayanotoxins and related diterpenoids with an andromedane skeleton are common in several genera of the plant family Ericaceae. Grayanotoxin I caused the nauplii of Artemia salina to exhibit an unusual spasmodic response, affecting motility, which was antagonized in trace amounts of tetrodotoxin, resulting in the restoration of normal swimming behavior. This inhibited motility of brine shrimp has been used to assay 15 andromedane compounds. Activity was diminished when the 6β-hydroxyl group was substituted, but only slightly modified by the acetylation of the 14β-hydroxyl group. A complete abrogation of toxicity resulted from the introduction of aromaticity at the 6-position, and of 3,6-diesterification in the molecule of grayanotoxin III.

Thin-layer chromatographic and spectroscopic characterization of some diterpenes of the grayanotoxin type

Abstract In a study of toxic polyhydroxylated andromedane diterpenoids, several novel derivatives of grayanotoxin III are described. Their spectral and physical characteristics are compared to ericaceous natural products. Separations on aluminium oxide and silica gel indicated that the absorption affinity of hydroxy groups on the perhydroazulene carbon skeleton was in teh order of 14γ-OH > 3β-OH > 6β-OH > 2α-OH. Partition chromatography using ethylene glycol as stationary phase exhibited migration largely in accordance with Reversed Traubes rule. This chromtographic-spectroscopic combination procedure is suitable for the micro-screening of these toxins.

The Next Green Revolution. Essential Steps to a Healthy, Sustainable Agriculture

‘‘Gentlemen don’t need mistletoe’’ said the Christmas whiskey billboard I saw recently in Florida, a garish reminder of how much mistletoe lore is embedded in western society. Mistletoe The Genus Viscum, one in the series of medicinal and aromatic plants for industry and academic researchers, centers on the mistletoe in this advertisement, the common European mistletoe,Viscum album. There are helpful discussions on African, Asian, and Argentine mistletoes and their uses but the corpus of the book deals with V. album. Viscum album, considered sacred by the Druids, is no doubt the most widely used parasitic angiosperm for various health concoctions. As a result, much has been learned about its biochemistry and pharmaceutical potential. A great deal of the work on the culture and utilization ofV. album has been done by investigators at Institute Hiscia Center for Cancer Research in Switzerland where votaries of Rudolf Steiner’s distinct form of homeopathy have used mistletoe extracts (‘‘Iscador’’) for many years in cancer treatment. Mistletoe collected from less common hosts are considered to have greater efficacy in preparation of Iscador. As a result, there is a helpful chapter on culturing V. album which is not as difficult as might be imagined for an obligate parasite. Most of the chapters deal with the medicinal aspects of mistletoe including toxicology, clinical aspects, chemistry, and biochemistry emphasizing the lectins which are one of the more desirable compounds produced by the parasite. I found the treatment by Bu ̈ssing (Biological and pharmacological properties of Viscum album L.) helpful because it reviews the link between folk and modern medicine. However, several relevant papers were omitted which are included in a recent excellent review of mistletoes (Watson 2001). Like many books with a diversity of authors, the quality of chapters varies. Obviously produced as a reference volume, Mistletoe lacks an extensive index. Especially aggravating is the lack of an author’s index, limiting the value of the book for anyone looking for specific papers. These days,

Mississippi-grown Cannabis sativa L.: preliminary observation on chemical definition of phenotype and variations in tetrahydrocannabinol content versus age, sex, and plant part

70 for a hardbound monograph is reasonable. Color images are well-produced but some of the black and white figures are blurry in my copy. This volume will be of value to ethnobotanists, anyone interested in alternative medicines, and students of mistletoes and parasitic plants. LITERATURE CITED