Variation In Alkaloid Content Research Articles

Seasonal and Geographic Variation in Alkaloid Content of Kratom (Mitragyna speciosa (Korth.) Havil.) from Thailand

The objective of this study was to obtain data on the distribution of alkaloids in kratom plants grown in Thailand. Two collections were performed, covering the southern, central, and northern regions of Thailand and different seasons. The contents of alkaloids, including mitragynine (MG), paynantheine (PAY), and speciogynine (SG), were determined using the validated HPLC method. The 134 samples in the first collection were collected from Nam Phu subdistrict, Ban Na San, Surat Thani, Thailand, during June and October 2019 and January 2020. The maximum mitragynine content was 4.94% w/w in June (late summer), and the minimum content was 0.74% w/w in October (rainy season). To expand the study area after kratom decriminalization, 611 samples were collected in June-August 2021, October-December 2021, and January-April 2022. The accumulation of MG ranged from 0.35 to 3.46% w/w, 0.31 to 2.54% w/w, and 0.48 to 2.81% w/w, respectively. The meteorological data supported the climate's effect on alkaloid production. Soil analysis revealed the importance of Ca and Mg in promoting alkaloid production. Geographical locations played a role in the variation of MG in kratom leaves, but did not affect the color of leaf veins. In conclusion, the present study suggested that the alkaloid content in kratom diverges based on seasonal and geographical origin.

Seasonal variation in alkaloid content, phenolic constituent and biological activities of some Leucojum aestivum L. populations in Turkey

Summer snowflake (Leucojum aestivum L.), a bulbous plant in the Amaryllidaceae family, is a well-known source of two pharmaceutically significant alkaloids: galanthamine that is used to treat Alzheimer's disease and lycorine that has potent antiviral properties, especially against SARS-CoV-2 (COVID-19). Environmental factors such as temperature, light, soil, and so on, as well as harvest time and plant parameters such as genotype and plant parts, all have a quantitative and qualitative impact on secondary metabolite production such as alkaloids and phenols. This study hypothesized that different populations (genotype), growing seasons, environmental temperature and soil type can change the alkaloid (galanthamine and lycorine) and phenolic content, and biological activities (antioxidant and antibacterial) in the bulbs and leaves of this plant. Summer snowflakes were collected from 6 different locations in Turkey (Gölcük-Bolu, Yeniçağa-Bolu, Kaynarca-Sakarya, Delmece-Yalova, Uluabat-Bursa and Terkos-İstanbul) at 3 different growing periods (vegetative, reproductive and ripening). Methanol extract of the plant parts were quantitatively analyzed for their alkaloid content with HPLC-DAD, and for their antioxidant capacity and total phenol-flavonoid content with spectrophotometric determination. Besides, antibacterial activity was investigated with disc diffusion assay. The locations of Delmece-Yalova and Gölcük-Bolu provided galanthamine-rich population and optimal growing conditions for the cultivation of this species. Generally, vegetative and ripening periods were the most effective in alkaloid production. But, phenolic content accumulation was unrelated to the growing seasons. Higher temperatures in the bulbs and lower temperatures in the leaves supported the galanthamine and lycorine production. It was noticeable that alkaloid and total phenol accumulation in the bulbs showed similar tendency in that the highest temperature caused the highest alkaloid levels and phenolic content. Also, neutral to slightly alkaline soils with high organic matter content were effective in enhancing the alkaloid content and phenolic constituents. Moderate antioxidant activity and no antibacterial potential were observed with L. aestivum. The results found can be a guide for revealing the best population and environmental conditions for the cultivation of this plant, as well as showing the correct harvest time for bulbs and leaves to get the best galantamine and lycorine content.

Isolation of the antibacterial alkaloid distichamine from Crossyne Salisb. (Amaryllidaceae: Amaryllideae: Strumariinae)

Isoquinoline alkaloids of the Amaryllidaceae have proven to be of significant value in drug discovery and development. Crossyne guttata is frequently used in traditional medicine as an antiseptic and wound cleaner as well as to treat alcoholism. The aim of the study was to identify the variation in alkaloid content of the species in relation to published works on the genus Boophone. We isolated Distichamine 1 from an ethanolic fraction of Crossyne guttata using column chromatography. Spectroscopic analysis yielded three alkaloids, distichamine 1, crinamine 2 and epibuphanisine 3. Distichamine 1 is a rare β-crinane alkaloid previously isolated only from the genus Boophone. The presence of distichamine 1 in the genus Crossyne suggests that it may be more widespread in tribe Amaryllideae than originally thought.

Variation and plasticity in Epichloë alkaloid content of Festuca rubra across Europe

Abstract Many symbiotic microbes can aid their host plant's defense against herbivores and pathogens either directly or indirectly through plant-mediated mechanisms. Thus, microbes are of interest to agricultural research. In this study, we used vertically transmitted Epichloe species because these fungal endophytes of grasses produce alkaloids that have been shown to protect the host from herbivory. Depending on the amount, type, and profile of alkaloids, Epichloe species can promote the host's defense against invertebrate or vertebrate herbivores. Here, we used Festuca rubra colonized by Epichloe festucae from wild populations of different geographic origins in a reciprocal transplant experiment at three locations across Europe. We sought to determine whether genetic differences among Epichloe-colonized plants from different origins or phenotypic plasticity explained variations in alkaloid content and whether the alkaloid content was correlated with the growth and reproduction of the plants colonized by Epichloe. We found that the geographic origin of the Epichloe-colonized genotypes explained differences in ergovaline and peramine production. The ergovaline content also indicated chemotypic plasticity, as profiles changed when transplanted to a new environment, while the concentration of peramine was constant among the transplantation sites. The lack of a strong correlation between alkaloid content and plant performance (number of reproductive stems and total biomass) indicated that the costs of alkaloid production were too low to have consequences for fitness in terms of those parameters. Our findings emphasize the importance of estimating the fungal alkaloid content in multiple field environments in order to determine its potential toxicity and value for breeding programs that are aimed at the improvement of turfgrass and forage cultivars.

Biogeographical Patterns and Phenological Changes in Lapiedra martineziiLag. Related to Its Alkaloid Diversity

The aim of this work was to investigate the alkaloid patterns of Lapiedra martinezii and their relation to biogeography and phenology focused in a phylogenetic comparison. Plants from 14 populations of L. martinezii, covering almost its entire distribution area, were subjected to morphological, ecological, and phytochemical analysis. Experiments for different alkaloid-type content are proposed as a new tool for analysis of plant distribution. Several plants were transplanted for weekly observation of their phenological changes, and alkaloids from different plant organs were extracted, listed, and compared. The alkaloid pattern of L. martinezii comprises 49 compounds of homolycorine, lycorine, tazettine, haemantamine, and narciclasine types. The populations located in the north and south margins of the distribution area displayed alkaloid patterns different from those of the central area. Changes in these patterns during their phenological cycle may be related to a better defence for plant reproduction. L. martinezii is an old relict plant, and it has maintained some of the more primitive morphological features and alkaloid profiles of the Mediterranean Amaryllidaceae. The variations in alkaloid content observed could be interpreted in a phylogenetic sense, and those found in their phenological changes, in an adaptive one.

Alkaloid profiling of the Chinese herbal medicine Fuzi by combination of matrix-assisted laser desorption ionization mass spectrometry with liquid chromatography–mass spectrometry

A matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) method was developed for the high throughput and robust qualitative profiling of alkaloids in Fuzi—the processed lateral roots of the Chinese herbal medicine Aconitum carmichaeli Debx ( A. carmichaeli). After optimization, powdered roots – without any further sample preparation – could be used to screen for the presence of Aconitum alkaloids. Furthermore, the semi-quantitative potential of MALDI-MS was confirmed using liquid chromatography–mass spectrometry (LC–MS) as reference. In total over sixty alkaloids were detected by LC–MS and fifteen of them were tentatively identified. Both MALDI-MS and LC–MS analysis revealed significant variation in alkaloid content in different (commercial) samples. LC–MS analysis of three toxic alkaloids in 14 batches of Fuzi resulted in a variation of their concentrations expressed as RSDs of 138%, 99% and 221% for aconitine, hypaconitine and mesaconitine, respectively. The variation in concentrations (expressed as RSD) of about the ninety constituents detected were classified as follows: 13 constituents showed an RSD of 77–100%, 46 with an RSD of 100–150%, 21 with an RSD of 150–200% and 9 constituents with an RSD in concentration of 200–235%. These results demonstrate a strong difference in chemical composition of the various Fuzi and illustrate the necessity of adequate QA/QC procedures for both safety and efficiency of herbal medicine. The described analytical procedures for alkaloid profiling could play a role in these procedures.

Individual Variation in Alkaloid Content of Poison Frogs of Madagascar (Mantella; Mantellidae)

Brightly colored Malagasy poison frogs, Mantella spp., sequester lipophilic, basic alkaloids from arthropod prey for their own chemical defense. Consequently, microsympatric prey diversity is expected to influence alkaloid diversity observed in poison frogs. Twenty-two specimens of three Mantella species from four localities in moist forests of southeastern Madagascar were analyzed individually via gas chromatography-mass spectrometry, revealing that they contain over 80 known alkaloids. Frogs within a locality possessed significantly similar alkaloid content and diversity, while frogs from areas that varied in disturbance, elevation, and/or species showed greater differences. Based on dietary data, the larger frog species Mantella baroni consumed more and larger prey, and showed greater diversity in skin alkaloids than significantly smaller Mantella bernhardi. Additionally, frogs from the most pristine locality had the greatest number of alkaloids, whereas individuals from the most disturbed localities had the least. In a comparison of frog alkaloid profiles over a 10- to 14-yr period, alkaloid turnover, and thus presumably alkaloid-source arthropod turnover, was high in a disturbed locality and low in the pristine primary forest locality. We demonstrate that the nonlethal transcutaneous amphibian stimulator (TAS) is effective for harvesting alkaloids from poison frogs; future studies using this device could obtain larger sample sizes without harming local frog populations.

Variations in Alkaloid Content of Herbal Products Containing Goldenseal

To determine the concentration of hydrastine and berberine, the primary alkaloids in herbal products containing goldenseal. Descriptive comparison of 20 products purchased at local pharmacies or health food stores; 17 products were labeled as containing goldenseal root and 3 products contained goldenseal herb as the sole active herbal ingredient. Laboratory assessment of alkaloid content using high-performance liquid chromatography. Comparisons of hydrastine and berberine concentration, alkaloid ratio, and total alkaloid content among products. Hydrastine concentration in products labeled as containing goldenseal root ranged from 0 to 2.93%, whereas berberine concentration varied from 0.82% to 5.86%. Median total alkaloid concentration was 5.30%. Only 10 of 17 products met proposed United States Pharmacopeia (USP) standards for the hydrastine and berberine content of goldenseal root. Five products contained little or no hydrastine, unusual berberine:hydrastine ratios, and additional peaks not observed with other products. Alkaloid content in goldenseal root products was about fourfold higher than in products labeled as containing goldenseal herb. Alkaloid content of goldenseal products varies widely, with many products failing to meet proposed USP standards. In addition, the chromatographic pattern and alkaloid ratio for several products were atypical for goldenseal. Given the current absence of regulation of the quality of herbal products, pharmacists should take extra care to ensure that substandard goldenseal products are not sold in their pharmacies.

Geographic distribution of three alkaloid chemotypes of Croton lechleri.

Three known alkaloids, isoboldine (2), norisoboldine (1), and magnoflorine (8), have been isolated for the first time from Croton lechleri, a source of the wound healing latex "sangre de grado". An HPLC system was developed, and a large number of latex and leaf samples of C. lechleri from 22 sites in northern Peru and Ecuador were analyzed to gain an understanding of the natural variation in alkaloid content for the species. Up to six alkaloids were found to occur in the leaves including, in addition to those listed above, thaliporphine (3), glaucine (4), and taspine (9), whereas the latex contained only 9. Taspine (9) is the component that has been previously found to be responsible for the wound healing activity of C. lechleri latex, and its mean concentration throughout the range examined was found to be 9% of the latex by dry weight. In addition, three chemotypes are defined based on the alkaloid content of the leaves, and the geographic distribution of these chemotypes is discussed along with a quantitative analysis of the alkaloid content as a function of chemotype.

Content versus label claims in ephedra-containing dietary supplements.

The content of ephedra alkaloids in herbal dietary supplements containing ephedra (ma huang) was studied. The ephedra alkaloid content of 20 ephedra-containing supplements was determined by high-performance liquid chromatography. Contents of (-)-ephedrine, (+)-pseudoephedrine, (-)-methylephedrine, (-)-norephedrine, and (+)-norpseudoephedrine were measured. Ephedra alkaloid content varied considerably among products. Total alkaloid content ranged from 0.0 to 18.5 mg per dosage unit. Ranges for (-)-ephedrine and (+)-pseudoephedrine were 1.1-15.3 mg and 0.2-9.5 mg, respectively. (+)-Norpseudoephedrine, a Schedule IV controlled substance, was often present. Significant lot-to-lot variations in alkaloid content were observed for four products. For one product, lot-to-lot variations in the content of (-)-ephedrine, (+)-pseudoephedrine, and (-)-methylephedrine exceeded 180%, 250%, and 1000%, respectively. Half of the products exhibited discrepancies between the label claim for ephedra alkaloid content and actual alkaloid content in excess of 20%. One product was devoid of ephedra alkaloids. Assay of 20 ephedra-containing dietary supplements showed that alkaloid content often differed markedly from label claims and was inconsistent between two lots of some products.

Foliar storage and extraction methods for quantitative analysis of piperidine alkaloids from ponderosa pine (Pinus ponderosa)

Yields of 2,6-disubstituted piperidine alkaloids from ponderosa pine (Pinus ponderosa) foliage were compared for frozen, air- and oven-dried samples stored for 3 days and for 5 weeks. Oven-drying provided the most stable alkaloid yields, but biological variation was greater than the variation between the storage methods. Alkaloid yields were also compared for the classical liquid–liquid partitioning method and a solid-phase partitioning (SPP) method using Extrelut™ columns. Under proper conditions, both methods were reasonably accurate, but biological variation in alkaloid content was much greater than experimental variation from either extraction method. In order to optimize the SPP method, it was necessary to use chloroform rather than dichloromethane as the eluting solvent, and to rinse the column more than the manufacturer directs. The SPP method requires less solvent and time per sample, thus it may be preferable for ecological studies where numerous samples are required to detect differences against high background variation. Zinc dust treatment, to reduce possible alkaloid N-oxides in SPP extracts, failed to increase yields, suggesting the absence of such compounds. Copyright © 1999 John Wiley & Sons, Ltd.

Variation of Alkaloid Content in Erythroxylum coca Leaves from Leaf Bud to Leaf Drop

A sequential study describing the content (%) of alkaloids in leaves of Erythroxylum coca var. coca Lam. from leaf bud development to leaf drop has not previously been conducted. The length of time the leaf resides on the plant and its concurrent metabolic activity also has not been defined. In the present study, cocaine, methyl ecgonine, hygrine, tropinone, trans -cinnamoylcocaine, cis-cinnamoylcocaine, tropacocaine and cuscohygrine were monitored to determine: (1) their content and patterns of accumulation from leaf bud to leaf drop; (2) the time leaves resided on the plant; and (3) whether leaves were metabolically active until leaf drop. E. coca plants were grown in a controlled environment for 37 months. Leaves similar in chronological age were extracted and analysed for alkaloid content by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). Cocaine was highest in 7 d old rolled leaves (0·75%) and declined to 0·39% at leaf drop. Cocaine, methyl ecgonine, hygrine, tropinone, trans -cinnamoylcocaine, cis-cinnamoylcocaine, cuscohygrine and tropacocaine content in 35 d old (mature) leaves was 0·61, 0·59, 0·68, 0·08, 0·31, 0·55, 0·52, and 0·05%. respectively. Cocaine, methyl ecgonine, hygrine, cis -cinnamoylcocaine, and cuscohygrine displayed a gradual decline from week 2 to week 36 of leaf duration. Tropinone and tropacocaine were the least abundant of the alkaloids monitored. Cis-cinnamoylcocaine content exceeded cocaine at week 12, 16, and weeks 19 to 23 of leaf duration. Trans -cinnamoylcocaine was highest in rolled leaves (week 1) and in expanded leaves after week 30. The monitored alkaloids appeared to be part of the metabolically active pool of the leaf. Leaves remained on the E. coca plants for 36 weeks.

Assessment of inter- and intra-population variation in quinoline alkaloid profile of juvenile shoot cultures of Cinchona ledgeriana: effect of method of nutrient delivery on alkaloid profile

Intra-population quinoline alkaloid profiles surveying quinine, quinidine, cinchonine and cinchonidine were determined for each of five populations of Cinchona ledgeriana grown as shoot-culture for 125 days. No significant difference in respect of mean alkaloid content between populations was detected. In contrast, there was considerable between-seedling variation in alkaloid content within each population. When nutrients were delivered to shoot-cultures in droplet form by means of an aerosol spray (as compared to the supply of nutrients direct from agar-or liquid-based reservoirs) alkaloid profile was greatly perturbed; most notable in this respect was a four-fold increase in the production of cinchonidine concomitant with a four-fold decrease in the production of cinchonine. These data are discussed with reference to the optimisation of quinoline alkaloid production by juvenile shoot-cultures of Cinchona ledgeriana.

Lupin alkaloids from flowers of Echinosophora koreensis

(−)-Methyl 12-cytisineacetate (2) was isolated from methanol extracts of fresh flowers of Echinosophora koreensis together with seven known lupin alkaloids. Ethyl 12-cytisineacetate (3) was also isolated from ethanol extracts of the same flowers. Compounds 2 and 3 were artifacts and (−)-12-cytisineacetic acid (4) is assumed to be the principal source of 2 and 3. The variations in alkaloid content during growth of the flowers and the seedlings were also examined.

Monoamine oxidase inhibitors in South American hallucinogenic plants: Tryptamine and β-carboline constituents of Ayahuasca

Ayahuasca is a hallucinogenic beverage derived by boiling the bark of the Malpighiaceous liana Banisteriopsis caapi together with the leaves of various admixture plants, viz. Psychotria viridis, Psychotria carthagenensis, or Diplopterys cabrerana. B. caapi contains harmine, harmaline, and tetrahydroharmine while the admixtures contain N, N-dimethyltryptamine (DMT). DMT, a potent hallucinogen, is inactive orally due to degradation by visceral monoamine oxidase (MAO). The β-carbolines, however, are highly active reversible inhibitors of MAO and may protect the DMT from deamination by MAO and render it orally active. This mechanism has been proposed to underlie the oral activity of ayahuasca but has not been experimentally confirmed. In the present study the constituents of the admixture plants and the alkaloids of eight ayahuasca samples from Peru were qualitatively and quantitatively analyzed using two-dimensional thin-layer chromatography (TLC), high pressure liquid chromatography (HPLC) and gas chromatography/mass spectrometry (GC/MS). Several B. caapi cultivars were quantitatively compared for variations in alkaloid content. Three admixture plants used rarely in the manufacture of ayahuasca were also screened for alkaloids. A selected sample of β-carbolines were screened for activity as MAO inhibitors using an in vitro assay system, and structure/activity relationships were compared. Inhibition observed with single compounds was compared with the activity of selected samples of ayahuasca which were screened in the system and also with the activity of mixtures of β-carbolines. The levels of DMT and β-carbolines found in the ayahuasca samples examined in the present study were an order of magnitude greater than the levels reported in a previous study. Ayahuasca was found to be an extremely effective inhibitor of MAO in vitro and the degree of inhibition was directly correlated with the concentration of MAO-inhibiting β-carbolines. Inhibition experiments using mixtures of β-carbolines indicated that their effects in combination are additive, rather than synergistic or antagonistic. Implications of the results in understanding the pharmacology of ayahuasca are discussed.

Survey of the Genus Lycopersicon for Variability in α-Tomatine Content1

Abstract Twenty-seven accessions from 6 species of the genus Lycopersicon and 1 accession of Solanum pennellii were surveyed for content of the glycoalkaloid, α-to-matine. Significant variation in alkaloid content was found to exist among the accessions. L. esculentum cultivars were found to contain much lower concentrations of α-tomatine than accessions of L. esculentum var. cerasiforme, L. pimpinellifolium, and the L. peruvianum accession, LA 462.

Thebaine Content of Selections of Papaver bracteatum Lindl. at Different Ages

Approximately 200 accessions of Papaver species were evaluated for identification as P. bracteatum Lindl. and for thebaine content. Fifteen authentic P. bracteatum accessions were selected on the basis of chromosome count. Statistical analyses of variation in alkaloid content were made for these accessions. Four of the 15 warranted further study based on the vigor of the plant and the total thebaine present in the tissue. Significant variability in thebaine content was found in wild strains of P. bracteatum, thus requiring genetic selection studies. The data showed that total yield potential of thebaine should be considered in selecting P. bracteatum strains for commercial growth rather than concentrating on strains that produce the highest thebaine concentrations. A method for the quantitative estimation of thebaine from P. bracteatum is presented.

Studies on parasitic cultivation of ergot [Claviceps purpurea (Fr.) Tul.] in India

Experiments were undertaken to standardise the techniques involved in large-scale cultivation of ergot [Claviceps purpurea (Fr.) Tul.]. It was found that the rye crop planted from the last week of September up-to the middle of October to be more suitable for obtaining a good yield of ergot. The spore suspension of the ergot fungus raised on wheat or rye grain medium retained its infective ability even after storing for about 3 days at room temperature (20±2° C.) and for about 5 days when stored under refrigerated conditions (5°C.). Among the four synthetic liquid media tested the medium described by Amiciet al. (1967) proved to be more suitable for raising the ergot inoculum on a large scale and it also retained its infectivity even after diluting twenty times of its volume. Rye plots, if inoculated two times by the needle board puncture method with the inoculum containing about 41,000 spores/mm.3 was found to be sufficient to yield maximum quantity of ergot sclerotia. The yield of the ergot sclerotia obtained from the plots inoculated by the needle board puncture method was found to be almost two times the yield obtained from those plots which were inoculated by spray method. Variation in alkaloid content has been observed in the ergot sclerotia obtained from the same strain and also within the same sclerotium. Experiments carried out on the probability of contamination of the ergot infection to the other cereal crops indicated that there are very little chances of ergot infection spreading to the other cereal crops and also the over-summering of the ergot sclerotia in the field under the natural conditions of the Jammu region.