Bioactive Marker Compounds Research Articles

Antidepressant-like effects of salidroside on olfactory bulbectomy-induced pro-inflammatory cytokine production and hyperactivity of HPA axis in rats

Salidroside (SA) is the primary bioactive marker compound in the standardized extracts from Rhodiola rosea. Although it has potential antidepressant activity in a rat behavioral despair model, the mechanisms of antidepressant effect for SA remain unclear. The objective of this study was to evaluate the antidepressant effects of SA and to discuss the potential mechanisms in olfactory bulbectomized (OBX) rats. SA of 20, 40mg/kg (p.o.) for 2weeks notably alleviated OBX-induced hyperactivity in open field test, decreased immobility time in TST and FST. Chronic treatment with SA could remarkably reduce TNF-α and IL-1β levels in hippocampus. Western blot showed that SA could markedly increase glucocorticoid receptor (GR) and brain-derived neurotrophic factor (BDNF) expression in the hippocampus. Besides, SA could also attenuate corticotropin-releasing hormone (CRH) expression in hypothalamus, as well as reducing significantly the levels of serum corticosterone. In conclusion, this study demonstrated that OBX rats treated with SA could significantly improve the depressive-like behaviors. The antidepressant mechanisms of SA might be associated with its anti-inflammatory effects and the regulation of HPA axis activity. Reversal of abnormalities of GR may be partly responsible for those effects. These findings suggested that SA might become a beneficial agent to prevent and treat the depression.

Optimization of spray drying conditions for production of Bidens pilosa L. dried extract

Spray drying is a technique commonly used to produce powdered phytopharmaceutical ingredients from medicinal plants. Product quality is related to operating conditions and the drying carriers used. Bidens pilosa L. is a South American plant with proved antiinflamatory, antimalaric, antitumoral and hepatoprotector activities. The aim of this work was to optimize the processing variables inlet drying temperature, plant extract feed flow rate and drying composition on the properties of the powdered B. pilosa compositions. Experimental planning, regression and multiple response analysis were used in data processing and to optimize the drying conditions. The responses evaluated were total flavonoid content, antioxidant activity, concentration of selected plant compounds, particle size, residual moisture content, solubility, flow properties, product recovery and drying efficiency. Results showed that inlet drying temperature was the factor that showed significant influence in most of the responses. The concentration of the bioactive marker compounds monitored decrease conversely with inlet spray drying temperature. Powders obtained at low inlet drying temperatures presented higher residual moisture and particle size, favouring flow properties. The optimized conditions based on the mathematical models fitted to experimental data were: Aerosil®:Cellulose 13:37%, extract feed rate 8.95mg/min and inlet gas temperature of 155.2°C.

Simultaneous determination of bioactive marker compounds from Gardeniae fructus by high performance liquid chromatography

Gardeniae fructus is one of the medicinal herbs that have been used in Far Eastern countries, such as Korea, China, and Japan. Gardeniae fructus is the dried ripe fruit of Gardenia jasminoides Ellis (Rubiaceae) and has been used as a yellow dye. It is widely used as traditional herbal medicine for reducing fever, cholagogue, diuretic and antiphlogistic effects. We established an analytical method that was useful to evaluate the quality control, and standardize quantification monitoring of 68 samples of Gardeniae fructus collected from Korea and China. While numerous previous studies have focused on the simultaneous analysis of geniposide, which constitutes the higher proportion of Gardeniae fructus, and crocin, which determines its color, no simultaneous analysis of gardenoside and geniposide, the major components of Gardeniae Fructus, has been performed. However, previously reported methods are not considered accurate enough because only geniposide or gardenoside was chosen to be the marker component for the quality control of Gardeniae fructus. Thus, we developed the method using simultaneous determination of four components including geniposide, gardenoside, geniposic acid and chlorogenic acid. Against this backdrop, this study aims to propose a new calculation for gardenoside and geniposide concentrations by analyzing their concentrations in Gardeniae fructus.

Quality changes during spouted bed drying of Pepper‐Rosmarin extract

AbstractThis study was undertaken in order to evaluate the effects of processing and feed composition variables on the retention of volatile compounds and product properties during spouted bed drying of pepper‐rosmarin extract (Lippia sidoides Cham.). The product loss on drying, LD, moisture content, Xp, water activity, Aw, concentration of bioactive markers (flavonoid, TF, polyphenol, TPF and thymol contents, TT), antioxidant activity, IC50, powder flow properties, powder morphology and crystalline state were the responses analysed. Processing conditions and feed composition significantly affected the product quality, leading to noticeable changes in the retention of bioactive substances. Results of this study show that the concentration of bioactive markers in the dried product tended to increase with the feed flow rate of the feed composition relative to the maximum dryer evaporation capacity, Ws/Wmax, and conversely with the concentration of drying carriers added to the feed composition. The dried extract showed significant antioxidant activity, although significant losses of bioactive marker compounds (TF, TPF and TT) were observed in some processing conditions.

Analysis of nine compounds from Alpinia oxyphyllafruit at different harvest time using UFLC-MS/MS and an extraction method optimized by orthogonal design

BackgroundThe dried fruits of Alpinia oxyphylla Miq have been widely used as an herbal medicine for the treatment of diarrhea and enuresis in China. Medicinal chemistry studies revealed that the tepenes, diphenylheptanes and flavones were the main components. Therefore, these three kinds of components should be chosen as the bioactive marker compounds for the quality control of A. oxyphylla fruits. Moreover, multiple active components has been widely recognized to be a more feasible method for the quality control of herbal medicines. This study firstly provided a better method for comprehensive component analysis of A. oxyphylla fruits. Meanwhile, the different harvest time was also evaluated.ResultsThe solvent-to-sample ratio was the most important factor comparing with solvent, extraction time and temperature. The highest yield of nine compounds was achieved with 70% ethanol-water and a solvent-to-sample ratio of 20:1 at 60°C for 30 min. The optimized analytical method for ultra fast high performance liquid chromatography (UFLC) was a gradient elution using water containing 0.04‰ formic acid (A) and methanol containing 0.04‰ formic acid (B), at a flow rate of 0.3 mL/min. Using this optimized method, nine compounds were simultaneously separated and quantified by UFLC coupled with tandem electro-spray ionized mass spectrometry (MS/MS).ConclusionsThe contents of the six bioactive compounds were reported in A. oxyphylla for the first time. The contents of nine compounds of different harvest time fruits of A. oxyphylla were assessed under the optimized extraction and UFLC-MS/MS analytical conditions, and the 45-day culture fruit had the highest content levels.

Quantitative determination and pattern recognition analyses of bioactive marker compounds from Dipsaci Radix by HPLC

In this study, quantitative and pattern recognition analyses were developed using HPLC/UV for the quality evaluation of Dipsaci Radix. For quantitative analysis, five major bioactive compounds were assessed. The separation conditions employed for HPLC/UV were optimized using ODS C18 column (250 × 4.6 mm, 5 μm) with a gradient of acetonitrile and water as the mobile phase at a flow rate of 1.0 mL/min and a detection wavelength of 212 nm. These methods were fully validated with respect to linearity, accuracy, precision, recovery, and robustness. The HPLC/UV method was applied successfully to the quantification of five major compounds in the extract of Dipsaci Radix. The HPLC analytical method for pattern recognition analysis was validated by repeated analysis of 17 Dipsaci Radix and four Phlomidis Radix samples. The results indicate that the established HPLC/UV method is suitable for quantitative analysis.

Stability testing and shelf live prediction of a spouted bed dried phytopharmaceutical preparation from Maytenus ilicifolia

AbstractIn this work the storage stability of a Maytenus ilicifolia phytopharmaceutical preparation produced by spouted bed drying was investigated. Powder samples of M. ilicifolia extracts were submitted to stress storage conditions at temperatures of 25, 34 and 45°C and relative humidities of 32.4% and 63.5%. The effect of the storage conditions on the physical and chemical properties of the product were evaluated through total polyphenols, tannin and catechin determination, moisture content, macroscopic changes in powder appearance and X‐ray difractograms. The results showed that high storage temperature and high relative humidity lead to increased degradation of the bioactive marker compounds. The shelf life of the spouted bed dried extracts was 276.2 days when the powder is stored at 25°C and 32.4% relative humidity.

A standardized phytopreparation from an Indian medicinal plant (Dalbergia sissoo) has antiresorptive and bone-forming effects on a postmenopausal osteoporosis model of rat

The aim of this study was to evaluate the skeletal effects of an extract made from the leaves and pods of Dalbergia sissoo (butanol-soluble standardized fraction [BSSF]) on ovariectomized rats, a model for postmenopausal osteopenia. Adult Sprague-Dawley rats were ovariectomized and administered BSSF (50 and 100 mg/kg per day) or 17β-estradiol orally for 12 weeks. The sham-operated group and the ovariectomy + vehicle group served as controls. Bone microarchitecture, bone turnover markers (serum osteocalcin and C-telopeptide fragment of collagen type I), biomechanical strength, new bone formation (based on mineral apposition rate and bone formation rate), and skeletal expressions of osteogenic and resorptive gene markers were studied. Uterine histomorphometry was used to assess estrogenicity. Bioactive marker compounds in BSSF were analyzed by high-performance liquid chromatography. One-way analysis of variance was used to test the significance of effects. In comparison with ovariectomized rats treated with vehicle, BSSF treatment in ovariectomized rats resulted in an improved trabecular microarchitecture of the long bones, increased biomechanical strength parameters of the vertebra and femur, decreased bone turnover markers (osteocalcin and type I collagen) and expression of skeletal osteoclastogenic genes, and increased new bone formation and expression of osteogenic genes in the femur. Overall, the osteoprotective effects of BSSF were comparable to those of 17β-estradiol. BSSF did not exhibit uterine estrogenicity. Analysis of marker compounds revealed the presence of osteogenic methoxyisoflavones, including caviunin 7-O-[β-D-apiofuranosyl-(1→6)-β-D-glucopyranoside] (a novel compound), biochanin A, and pratensin. Oral doses of BSSF in the preclinical setting are effective in preventing estrogen deficiency-induced bone loss by dual action: inhibition of bone resorption and stimulation of new bone formation.

Qualitative and quantitative determination of bioactive marker compounds from Gastrodia elata by HPLC-DAD-MS

Qualitative and quantitative determination of four bioactive compounds including gastrodin, gastrodigenin, p-hydroxybenzaldehyde, bis(4-hydroxybenzyl)ether isolated from Gastrodia elata has been developed using high performance liquid chromatography-diode array detector coupled with electrospray ionization/mass spectrometry (HPLC-DAD-MS). The analysis of four standard compounds showed good linearity, intra- and inter-day precisions, and accuracy of them. In addition, metabolite profiling of 23 different Gastrodia elata extracts was carried out to identify their origins. Total analyzed metabolites were applied to multivariate statistical analysis including principal component analysis and hierarchical clustering analysis for pattern analysis.

Cytochrome P450 Inhibition Assay for Standardized Extract of Terminalia chebula Retz.

The hydroalcoholic extract of fruit pulp of Terminalia chebula Retz. was standardized and evaluated for its safety through cytochrome P450 (CYP 450) inhibition assay. Standardization was performed through high performance thin layer chromatography (HPTLC) using gallic acid (GA) standard. Cytochrome P450-CO complex microplate assay was performed using rat liver microsomes. The effect of standardized extract, its fraction and bioactive marker compound were comparatively evaluated for its effect on CYP P450 enzymes. The extract of fruit pulp was used for HPTLC, where the R(f) value of the marker was found to be 0.43. The calibration plot was linear in the range of 2-14 µg of GA and correlation co-efficient of 0.99965. The mean quantity of GA was found to be 2.5% w/w. The CYP P450 concentration of the rat liver microsome sample used in the study was found to be 0.417 nmol/mg protein. The in vitro effect of various concentrations of extracts and fractions showed a linear concentration-dependent inhibition of cytochrome P450 up to 60 µL. The study showed more inhibition of fraction when compared to the extract and GA. Still, the inhibition showed by fraction is less when compared with standard Ketoconazole. Thus, this study indicated the in vitro cytochrome P450 inhibition potential of T. Chebula.

HPLC method development and validation of cytotoxic agent phenyl‐heptatriyne in Bidens pilosa with ultrasonic‐assisted cloud point extraction and preconcentration

Extraction and pre-concentration of a bioactive marker compound, phenyl-1,3,5-heptatriyne from Bidens pilosa, prior to HPLC has been demonstrated using both organic and ecofriendly solvents. Non-ionic surfactants, viz. Triton X-100, Triton X-114 and Genapol X-80, were used for extraction. No back-extraction or liquid chromatographic steps were required to remove the target phytochemical from the surfactant-rich extractant phase. The optimized cloud point extraction procedure has been shown to be a potentially useful methodology for the preconcentration of the target analyte, with a preconcentration factor of 4-99. Moreover, the method is simple, sensitive, rapid and consumes lesser solvent than traditional methods. An isocratic chromatographic separation and quantitation was accomplished on a C(18) column with acetonitrile-acidified aqueous as mobile phase at a flow rate of 1.0 mL/min, UV detection at 254 nm and specificity with photo diode-array detector (PDA) and MS. Under the optimum experimental conditions recovery was satisfactory (99.18-100.33%) without interference from the surfactant. The method seems to be reliable with intraday precision and interday precision below 2.0%. Good linearity was obtained in the working range from 7.5 to 30 µg/mL with correlation coefficient >0.99. The limits of detection and quantitation were 1.84 and 6.13 µg/mL, respectively. The method was validated following international guidelines and successfully applied for quantitative assays of cytotoxic compound phenyl-1,3,5-heptatriyne in Bidens pilosa.

Extract and fraction from Ulmus wallichiana Planchon promote peak bone achievement and have a nonestrogenic osteoprotective effect

This study aimed to determine the skeletal effects of total ethanolic extract (TEE) and its butanolic fraction (BF) from the stem-bark of Ulmus wallichiana, which is rich in C-glycosylated flavonoids, in growing rats (for peak bone [PB] achievement) and in ovariectomized (OVx) rats (for menopausal bone loss). TEE (750 mg kg(-1) d(-1)) and BF (50 mg kg(-1) d(-1)) were given orally for 10 weeks to weaning female Sprague-Dawley rats and for 12 weeks to adult OVx rats of the same strain, respectively. In studies with OVx rats, sham operated + vehicle, OVx + 17beta-estradiol, and OVx + vehicle groups served as various controls. Bone mineral density (BMD), biomechanical strength, bone histology, formations of osteoprogenitor cells, osteoid formation, and bone turnover/resorption markers were studied. Bioactive marker compounds in TEE and BF were analyzed by high-performance liquid chromatography. One-way analysis of variance was used to test significance of effects. In growing rats, both TEE and BF increased BMD, bone strength, and bone formation rate, suggesting higher PB achievement. OVx rats treated with either TEE or BF exhibited increased BMD at various anatomical positions and improved bone strength and trabecular architecture compared with the OVx + vehicle group. Serum osteocalcin and urinary type 1 collagen degradation product levels in OVx rats treated with either TEE or BF were significantly lower than those of the OVx + vehicle group. Neither TEE nor BF exhibited uterine estrogenicity. Analysis of marker compounds revealed significant enrichment of two bioactive markers in BF over TEE. Derived from U wallichiana, BF at much a lower dose than TEE was effective in PB achievement and prevention of OVx-induced bone loss.

Authentication of the anti-tumor herb Baihuasheshecao with bioactive marker compounds and molecular sequences

Baihuasheshecao ( Hedyotis diffusa), a Chinese herb for cancer treatment, is frequently adulterated by a related species Hedyotis corymbosa. DNA sequencing of the complete internal transcribed spacer region was applied to differentiate H. diffusa from H. corymbosa and other closely related species. The molecular data showed that four out of seven herb samples of Baihuasheshecao were adulterants. Chemical analyses by TLC and HPLC were used to authenticate H. diffusa and H. corymbosa. Two marker compounds were identified exclusively in H. diffusa: 6- O-( E)- p-coumaroyl scandoside methyl ester (compound 1) and 10( S)-hydroxypheophytin a (compound 2). Both compounds showed moderate anti-proliferation effect on PC3 human androgen-independent prostate cancer cells, while compound 2 also showed strong anti-proliferation effect on LNCaP human androgen-sensitive prostate cancer cells. Accordingly, these bioactive marker compounds could be applied to verify the authenticity and assess the quality of Baihuasheshecao.

Purification of a Water Extract of Chinese Sweet Tea Plant (Rubus suavissimus S. Lee) by Alcohol Precipitation

The aqueous extraction process of the leaves of Rubus suavissimus often brings in a large amount of nonactive polysaccharides as part of the constituents. To purify this water extract for potential elevated bioactivity, an alcohol precipitation (AP) consisting of gradient regimens was applied, and its resultants were examined through colorimetric and HPLC analyses. AP was effective in partitioning the aqueous crude extract into a soluble supernatant and an insoluble precipitant, and its effect varied significantly with alcohol regimens. Generally, the higher the alcohol concentration, the purer was the resultant extract. At its maximum, approximately 36% (w/w) of the crude extract, of which 23% was polysaccharides, was precipitated and removed, resulting in a purified extract consisting of over 20% bioactive marker compounds (gallic acid, ellagic acid, rutin, rubusoside, and steviol monoside). The removal of 11% polysaccharides from the crude water extract by using alcohol precipitation was complete at 70% alcohol regimen. Higher alcohol levels resulted in even purer extracts, possibly by removing some compounds of uncertain bioactivity. Alcohol precipitation is an effective way of removing polysaccharides from the water extract of the sweet tea plant and could be used as an initial simple purification tool for many water plant extracts that contain large amounts of polysaccharides.

Production of Th1- and Th2-dependent cytokines induced by the Chinese medicine herb, Rhodiola algida, on human peripheral blood monocytes

Ethnopharmacological relevance Rhodiola algida, an herb ingredient used in Chinese medicine, has been clinically proven to be effective in enhancing human immune responses. Aim of study This study attempted to identify the potential immunomodulatory effect of Rhodiola algida extract in human immune system in vitro, and to examine its underlying molecular effects. Materials and methods Firstly, the bioactive marker compound salidroside was used for standardization of Rhodiola algida extract by reversed-phase HPLC. Secondly, the regulation of human immune responses was investigated in human peripheral blood monocytes. A series of cytokines known to play important roles in the human immune responses were examined. Results The current study provided quantitative assay for the marker compound, salidroside, in the Rhodiola algida extract for ensuring the quality consistency of Rhodiola algida used in the following experiments. Biological assay indicated that Rhodiola algida stimulates human peripheral blood lymphocytes and its underlying immunomodulatory effects probably through its regulation of IL-2 in Th1 cells and IL-4, IL-6, IL-10 in Th2 cells. Conclusion The findings may enable us to further explain the pharmacological properties in Chinese medicine and make Rhodiola algida a very promising immunomodulating agent.

Cultivar Variations of Australian‐Grown Danshen (Salvia miltiorrhiza): Bioactive Markers and Root Yields

Danshen (Salvia miltiorrhiza Bunge, Lamiaceae) is a commonly used and highly valued Chinese medicinal herb grown widely in China. In the present work, we studied cultivar variations of Australian-grown Danshen in order to select optimal cultivars for local herbal production. Root yields of seven cultivars, V1-V7, were monitored in a one-year field trial, and bioactive markers, including cryptotanshinone, tanshinone I, tanshinone IIA, and salvianolic acid B, were quantitatively determined using a validated RP-HPLC method. Significant variations were found in root yields, root production efficiencies, and contents of the bioactive marker compounds. Linear correlations were observed among the contents of three tanshinones but not among those of tanshinones and salvianolic acid B. Among the cultivars, V6 was the best cultivar for production of tanshinones, and V4 and V5 were best for production of salvianolic acid B. The findings indicate that it is possible to achieve optimal root yields, and high contents of tanshinones and salvianolic acid B by selecting specific Danshen cultivars.

A preliminary pharmacophylogenetic investigation in Schisandraceae

The family Schisandraceae (Magnoliiadae) contains approximately sixty species which are disjunctly distributed in the southeast of Asia and North America. It was divided into two genera, Schisandra and Kadsura, represented by 29 species in China, 19 in Schisandra and 10 in Kadsura. This paper reviews current knowledge about the chemistry, ethnopharmacology and pharmacology of the family in an attempt to present a preliminary study into the pharmacophylogenetics of the family as a whole. Dibenzocyclooctadiene lignans (I) are considered to be the main chemical components of the family. Despite their traditionally recognized hepatoprotective function, they also exhibit anti-oxidant, anti-cancer and anti-HIV potential. Those dibenzocyclooctadiene lignans (I) possessing hydroxyl or angeloyloxy groups at C-6 or C-9 in the ethylidene-cyclooctane ring tend to exhibit a higher anti-cancer activity. Spirobenzofuranoid dibenzocyclooctadienes (II), mostly present in Kadsura, contain a special tetrahydrofuran ring spanning the biphenyl linkage and these demonstrate particular anti-PAF activities. This supports the traditional use of Kadsura to improve blood circulation and "remove dampness". Spirobenzofuranoid dibenzocyclooctadienes (II) could be considered as the bioactive marker compounds in Kadsura and hence markers for assessing quality. The distribution of all known lignans in the family showed that Kadsura is relatively advanced in evolution. Cycloartanone triterpenes occur in both Schisandra and Kadsura. Those with the A-ring open (II) tend to exhibit greater anti-cancer and anti-HIV activity. 7/7/5/6 triterpene lactones (IV), showing strong cytotoxicity, have only recently been discovered in Kadsura longipedunculata and as such have potential as anticancer agents. Recently, novel nortriterpenoids possessing a unique skeleton were found in S. lancifolia and S. micrantha; some exhibited clear anti-cancer or anti-HIV activity and are the subject of separate studies.

A new, convenient method for determination of mangiferin, an anti-diabetic compound, inMangifera indicaL.

A simple, precise, and rapid HPTLC method has been established for quantitative determination of the bioactive marker compound mangiferin in Mangifera indica . Chromatography was performed on pre-activated silica gel 60F 254 plates with ethyl acetate-methanol-water-formic acid 10:1.0:1.0:0.5 ( v/v ) as mobile phase. Detection and quantification were performed densitometrically at 270 nm with mangiferin as external standard. The method was validated for selectivity, linearity, precision, accuracy, and robustness. Optimization of a suitable solvent for extraction was also achieved. Intra-day and inter-day RSD were 0.67% and 0.95%, respectively. Instrumental precision and repeatability were 0.43 and 0.81, respectively. Accuracy was checked by studying recovery at three levels; average recovery was 92.5%. The method was used for estimation of mangiferin in the stem bark and leaves of Mangifera indica . This method for quantitative monitoring of mangiferin in Mangifera indica is rapid, simple, and accurate and...

Acute and Subacute Toxicity Studies on the Polyherbal Antidiabetic Formulation Diakyur in Experimental Animal Models

The present study investigated the acute and subacute toxicity of Diakyur, a polyherbal antidiabetic formulation, in experimental animal models. Diakyur contains aqueous extract dry powders of Cassia auriculata, Gymnema sylvestre, Mucuna pruriens, Syzygium jambolanum, Terminalia arjuna, Salacia reticulata, and a crude powder of Cassia javanica. The raw materials were standardized by gravimetric, HPLC and High performance thin layer chromatography (HPTLC) methods for their respective bioactive marker compounds. In an acute toxicity study, Diakyur was administered orally at doses ranging from 100-12800 mg/kg p.o. and the animals were observed for any toxic symptoms up to 72 hr. The results indicated there were no toxic symptoms up to the dose level of 12800 mg/kg p.o. In a subacute toxicity study, Diakyur was tested at the dose of 1600 mg/kg p.o. once daily for 28 days. The animals were sacrificed on the 29th day and various blood biochemical parameters were measured. The liver, kidney, heart, adrenals, pancreas and uterus were processed for histopathological study. The results of the 28 day subacute toxicity study did not show evidence of any changes in body weight, food and water intake, hematological parameters, liver and kidney function tests when compared with the control animals. The vital organs of animals treated with Diakyur for 28 days did not show any histopathological evidence of pathological lesions. From the results it is concluded that Diakyur at the dose of 1600 mg/kg p.o. is safe for long-term treatment in diabetic conditions.