Herbal Science Research - analytical chemistry

Phytochemical and analytical studies of extracts from Rhodiola rosea and Rhodiola quadrifida.

Site Editor — Mon, 11 Jun 2007 05:46:48 -0700

Phytochemical and analytical studies of extracts from Rhodiola rosea and Rhodiola quadrifida.: Pharmazie. 2007 Apr;62(4):308-11 Authors: Wiedenfeld H, Dumaa M, Malinowski M, Furmanowa M, Narantuya S

Column chromatography of hydrophilic extracts from Rhodiola rosea and Rodiola quadrifida led to the isolation of cinnamic alcohol, chlorogenic acid, rhodiooctanoside, rosiridin, rosavin and the phenolic compounds salidroside, rhodiolin and a novel compound consisting of viridoside with an attached arabinose unit (mongrhoside). HPLC analysis of plant material from different sources and from different collection periods showed a great variability in the composition and in the amount of pharmacologically active compounds contained.

Chasing molecules that were never there: misassigned natural products and the role of chemical synthesis...

Site Editor — Fri, 05 Jan 2007 19:47:05 -0800

Chasing molecules that were never there: misassigned natural products and the role of chemical synthesis in modern structure elucidation.: Angew Chem Int Ed Engl. 2005 Feb 4;44(7):1012-44 Authors: Nicolaou KC, Snyder SA

Over the course of the past half century, the structural elucidation of unknown natural products has undergone a tremendous revolution. Before World War II, a chemist would have relied almost exclusively on the art of chemical synthesis, primarily in the form of degradation and derivatization reactions, to develop and test structural hypotheses in a process that often took years to complete when grams of material were available. Today, a battery of advanced spectroscopic methods, such as multidimensional NMR spectroscopy and high-resolution mass spectrometry, not to mention X-ray crystallography, exist for the expeditious assignment of structures to highly complex molecules isolated from nature in milligram or sub-milligram quantities. In fact, it could be argued that the characterization of natural products has become a routine task, one which no longer even requires a reaction flask! This Review makes the case that imaginative detective work and chemical synthesis still have important roles to play in the process of solving nature's most intriguing molecular puzzles.

Steamed American Ginseng Berry: Ginsenoside Analyses and Anticancer Activities.

Site Editor — Wed, 03 Jan 2007 19:06:18 -0800

Steamed American Ginseng Berry: Ginsenoside Analyses and Anticancer Activities.: J Agric Food Chem. 2006 Dec 27;54(26):9936-9942 Authors: Wang CZ, Zhang B, Song WX, Wang A, Ni M, Luo X, Aung HH, Xie JT, Tong R, He TC, Yuan CS

This study was designed to determine the changes in saponin content in American ginseng berries after treatment by heating and to assess the anticancer effects of the extracts. After steaming treatment (100-120 degrees C for 1 h, and 120 degrees C for 0.5-4 h), the content of seven ginsenosides, Rg1, Re, Rb1, Rc, Rb2, Rb3, and Rd, decreased; the content of five ginsenosides, Rh1, Rg2, 20R-Rg2, Rg3, and Rh2, increased. Rg3, a previously identified anticancer ginsenoside, increased significantly. Two hours of steaming at 120 degrees C increased the content of ginsenoside Rg3 to a greater degree than other tested ginsenosides. When human colorectal cancer cells were treated with 0.5 mg/mL steamed berry extract (120 degrees C 2 h), the antiproliferation effects were 97.8% for HCT-116 and 99.6% for SW-480 cells. At the same treatment concentration, the effects of unsteamed berry extract were 34.1% for HCT-116 and 4.9% for SW-480 cells. After staining with Hoechst 33258, apoptotic cells increased significantly by treatment with steamed berry extract compared with unheated extracts. Induction of apoptosis activity was confirmed by flow cytometry after staining with annexin V/PI. The steaming of American ginseng berries augments ginsenoside Rg3 content and increases the antiproliferative effects on two human colorectal cancer cell lines. Keywords: Panax quinquefolius L.; berry; heating treatment; HPLC analysis; ginsenoside Rg3; antiproliferation; human colorectal cancer cells; apoptosis.

Isolation and characterization of yuremamine, a new phytoindole.

Site Editor — Fri, 09 Jun 2006 07:15:47 -0700

Isolation and characterization of yuremamine, a new phytoindole.: Planta Med. 2005 Nov;71(11):1053-7 Authors: Vepsäläinen JJ, Auriola S, Tukiainen M, Ropponen N, Callaway JC

Yuremamine was isolated and characterized from the stem bark of Mimosa tenuiflora. This plant is still used by indigenous peoples in North-eastern Brazil to make yurema, a psychoactive beverage that is used for medico-religious purpose ( jurema preta or vinho da jurema, in Portuguese). The characterization of this novel compound by NMR and mass spectrometry introduces a new class of phytoindoles.

Multiple aspects of hair analysis for opiates: methodology, clinical and workplace populations...

Site Editor — Fri, 09 Jun 2006 07:15:23 -0700

Multiple aspects of hair analysis for opiates: methodology, clinical and workplace populations, codeine, and poppy seed ingestion.: J Anal Toxicol. 2005 Oct;29(7):696-703

Authors: Hill V, Cairns T, Cheng CC, Schaffer M

Levels of morphine, 6-monoacetylmorphine (MAM) and codeine in hair in both clinical and workplace subjects are presented. Aggressive wash procedures, consisting of 1 isopropanol wash, three 30-min, and two 1-h buffer washes, followed by digestion, extraction and confirmation of digested samples, resulted in values from the cutoff of 2 ng morphine/10 mg hair to greater than 200 ng/10 mg hair. Both morphine and MAM were present above the cutoff in all hair samples from 69 clinical subjects. Only 39 of the 69 heroin-using subjects had urine tests positive for 6-MAM. In a study of morphine in hair following poppy seed consumption, ten subjects ingested 150 g of poppy seed over 3 weeks. Urine samples were collected on the days of poppy seed ingestion and hair samples were taken in the 5th week of the study. The range among the 10 subjects of the highest urine value for each subject was 2929 to 13,827 ng morphine/mL. Hair morphine levels were 0.05-0.48 ng/10 mg hair (average 0.17 ng/10 mg hair). Hair opiate levels of workplace subjects ranged somewhat lower than those of clinical subjects. While all clinical hair samples contained MAM, many workplace samples did not. From workplace samples, a maximum amount of morphine likely to be present from codeine use was 0-3.7% of the codeine in the hair.

Quantitative analysis of flavonol glycosides in Ginkgo biloba: a comparison of two analytical methods.

Site Editor — Fri, 09 Jun 2006 07:15:12 -0700

Quantitative analysis of flavonol glycosides in Ginkgo biloba: a comparison of two analytical methods.: Phytochem Anal. 2006 Jan-Feb;17(1):56-62 Authors: Gray DE, Upton R, Chandra A, Porter A, Harris RK

A comparison of testing methods for constituents of Ginkgo biloba was performed as part of the process for determining which of the many available analytical methods was most appropriate for inclusion in the American Herbal Pharmacopoeia (AHP). Three primary methods were reviewed: those adopted by the European Pharmacopoeia, the National Science Foundation-Institute for Nutraceutical Advancement (NSF-INA) and the United States Pharmacopeia. Methods were compared by evaluating sample preparation and hydrolysis, precision and methods of total flavonol glycoside calculation by two independent laboratories. The AHP adopted the method of NSF-INA for inclusion in the AHP monograph owing to observed superiority of this method in terms of precision, glycoside calculation, ease of sample preparation and chromatographic parameters.

Geographic information system method for assessing chemo-diversity in medicinal plants.

Site Editor — Fri, 09 Jun 2006 04:02:13 -0700

Geographic information system method for assessing chemo-diversity in medicinal plants.: Planta Med. 2005 Dec;71(12):1157-64 Authors: Moraes RM, Momm HG, Silva B, Maddox V, Easson GL, Lata H, Ferreira D

The spatial distribution of wild germplasm of Podophyllum peltatum L. (American mayapple) has been analyzed using the Geographic Information System (GIS) with the objective to develop a method and a database for evaluation of biotic and abiotic factors influencing drug yield, and to map elite genotypes for propagation and improvement. The field assessment followed a standard procedure including geographical coordinates of each accession, leaf biomass randomly harvested, identification of associate species, collection of herbarium specimen, soil sample and digital pictures of the site. By overlaying morphological and chemical data with geomorphic information, a thematic map was created locating the podophyllotoxin-rich accessions and the uniqueness of each site was recorded for post-collection analysis. This work has enabled the establishment of a database of P. peltatum germplasm in Mississippi with drug yield linked to spatial locations for rational utilization of our natural resources. While this method integrates information of well-characterized diverse in situ P. peltatum germplasm, it might become a strategy for curators to reduce cost for establishing and maintaining ex situ collections since the genetic material is geo-referenced. Abbreviations. GIS:geographical information system GPS:global positioning system DGPS:differential global positioning system DEM:digital elevation model.

Micellar electrokinetic chromatography for separation of a mixture of coptis alkaloids, scute flavonoids...

Site Editor — Fri, 09 Jun 2006 03:27:14 -0700

Micellar electrokinetic chromatography for separation of a mixture of coptis alkaloids, scute flavonoids, and rhubarb anthraquinones and bianthrones.: J Pharm Biomed Anal. 2005 Nov 15; Authors: Chang LC, Sun SW

Coptidis Rhizoma, Scutellariae Radix, and Rhei Rhizoma are three common Chinese herbs. There are many herbal formulas which contain either two or all three of the herbs mentioned above. Their bioactive components have already been identified, respectively. However, there is no report about separation of the 13 bioactive constituents of the three herbs at the same time. In order to assess these constituents of related Chinese herbal preparations, a micellar electrokinetic chromatography method was developed. While buffer pH and surfactant concentration affected the resolution of separation, acetonitrile percentage was found to significantly influence the resolution, peak shape, and elution window. Optimum separation of 13 compounds was achieved at pH 7.3 using a buffer mixture of 70% (v/v) 3mM di-sodium tetraborate, 10mM sodium dihydrogen phosphate, and 50mM sodium deoxycholate with 30% (v/v) acetonitrile. When applying the developed method to analyze a model preparation, San-huang-xie-xin-tang, which contains all three herbs, 8 of the 13 bioactive constituents, could be determined. The present study proposed a method to assess San-huang-xie-xin-tang within short analysis time and also provided a possible starting point to evaluate related herbal preparations containing Coptidis Rhizoma, Scutellariae Radix, and Rhei Rhizoma.

Determination of salvinorins and divinatorins in Salvia divinorum leaves by liquid chromatography/multistage mass spectrometry.

Site Editor — Fri, 09 Jun 2006 03:21:16 -0700

Determination of salvinorins and divinatorins in Salvia divinorum leaves by liquid chromatography/multistage mass spectrometry.: Rapid Commun Mass Spectrom. 2005 Dec 6;20(2):131-136 Authors: Medana C, Massolino C, Pazzi M, Baiocchi C

Salvinorin A is the most potent naturally occurring hallucinogen known and rivals synthetic LSD in potency. Structurally it belongs to the neoclerodane diterpenoids, and it is the only known non-nitrogenous kappa-opioid-selective agonist. Salvia divinorum (Diviner's sage) is a member of the mint family that was used in ancient Mexican traditional practices. Today it is widely cultivated in Europe as a recreational marijuana substitute; it is illegal to buy, sell or possess the plant or the active principle in some countries. Six different salvinorins and three divinatorins have been isolated from Salvia divinorum leaves. The ion fragmentation, separation and quantitation of these diterpenes by liquid chromatography/electrospray ionization multistage mass spectrometry (LC/ESI-MS(n)) are described. The importance of LC in herbal extract determination and the chemical diagnostic power of MS(n) in the analysis of classes of natural organic products are discussed. Copyright (c) 2005 John Wiley & Sons, Ltd.

HPLC-MS/MS analysis of willow bark extracts contained in pharmaceutical preparations.

Site Editor — Fri, 09 Jun 2006 03:12:31 -0700

HPLC-MS/MS analysis of willow bark extracts contained in pharmaceutical preparations. - Phytochem Anal. 2005 Nov-Dec;16(6):470-8 Authors: Kammerer B, Kahlich R, Biegert C, Gleiter CH, Heide L

Preparations containing willow bark extract are popular herbal remedies, but they are mostly standardised with respect to only one compound (usually salicin). RP-HPLC using a C18-column eluted with water:methanol:tetrahydrofuran and coupled to electrospray triple-quadrupole MS and MS/MS was used for the characterisation of dried extracts of Salix spp. and for the identification of their constituents. Comparison with reference substances led to the identification of 13 compounds (saligenin, salicylic acid, salicin, isosalicin, picein, salidroside, triandrin, salicoylsalicin, salicortin, isosalipurposide, salipurposide, naringenin-7-O-glucoside and tremulacin). Two pharmaceutical preparations containing willow bark extract, used in clinical trials and labelled Salix daphnoides and S. purpurea x daphnoides extracts, were compared using the described method and exhibited several clear differences, the most prominent of which was the possible presence of picein in the former preparation. The described method may be utilised for the characterisation of herbal medicines in order to ensure comparability of medication in further clinical trials.

Determination of DPPH free radical scavenging activity by reversed-phase HPLC

Site Editor — Fri, 09 Jun 2006 03:05:15 -0700

Determination of DPPH free radical scavenging activity by reversed-phase HPLC: A sensitive screening method for polyherbal formulations. - J Pharm Biomed Anal. 2005 Nov 15; Authors: Chandrasekar D, Madhusudhana K, Ramakrishna S, Diwan PV

The colorimetric method of evaluation of antioxidant activity by scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical is with certain shortcomings like failure to indicate antioxidant activity of certain drugs and interference from color pigments of natural products. A specific HPLC method was developed for evaluating the DPPH free radical scavenging activity of commercial polyherbal formulations using a LiChrospher((R)) 100 RP-18e column (250mmx4mm, 5muM). The mobile phase was a mixture of methanol and water (80:20, v/v) pumped at a flow rate of 1mL/min. The DPPH peaks were monitored at 517nm. The method was standardized using known antioxidants such as ascorbic acid, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (trolox), probucol and alpha-tocopherol. The 50% radical scavenging activity (IC(50)) determined by the HPLC method correlated well with that of colorimetry. This HPLC method was applied for the estimation of free radical scavenging activity of Silymarin and a few commercial hepatoprotective polyherbal formulations. While the colorimetric method failed to estimate the free radical scavenging activity of polyherbal formulations, HPLC method was free from interferences and was specific. The HPLC method is sensitive and can be used as a quality control tool for the rapid determination of free radical scavenging activity of variety of products including plant extracts, foods, drugs and polyherbal formulations.