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Currently, black pepper commands the leading position among all the spices as a spice of great commercial importance in all the world trade and finds its way into the dietary habits of millions of people worldwide. Black pepper is biologically known as Piper nigrum and contains piperine as the main active chemical constituent. This paper highlights various general methods for extracting piperine from the crude drug such as maceration extraction, hydrotropic extraction, accelerated solvent extraction, thin-layer chromatography, and extraction with ethanol & dichloromethane Ionic fluid-based ultrasonic-assisted extraction, etc. In this review, piperine and its analogs exhibit numerous pharmacological activities and synthetic schemes of insecticidal activity, anti-cancer activity, anti-inflammatory activity, anti-diabetic activity, anti-hyperlipidemic activity, antifungal activity, narcotic activity, etc. and its structure-activity relationship. The biochemistry of piperine has also been summarized in the presented article. This very exhaustive review details the complete information about piperine, its derivatives, and further processing. Furthermore, the current study summarises recent research that has linked piperine to its use as a treatment for a variety of ailments.

Abdominal aortic aneurysm (AAA) rupture is one of the most common causes of mortality in cardiovascular diseases, but currently there is no approved drug for AAA treatment or prevention in the clinic. Naringenin (NGN) has been reported to have anti-AAA effects. However, water solubility and in vivo absorption of NGN are not satisfactory, which leads to its low bioavailability, thus affecting its pharmacological effects. In this project, the improving effects of isonicotinamide (INT) co-crystal and hydroxy propyl methyl cellulose (HPMC) or polyvinyl pyrrolidone (PVP) on the solubility, in vivo absorption, and anti-AAA effects of NGN were evaluated.In the current study, co-crystals of naringenin-isonicotinamide (NGN-INT) were prepared, and effects of PVP or HPMC on precipitation rate, supersaturation, and bioavailability of NGN were explored. In addition, with or without HPMC supply, the effects of NGN-INT co-crystal on anti-AAA efficacy of NGN were investigated on an elastase-induced AAA mouse model, and the results were compared with the efficacy of the NGN crude drug.Our results demonstrate that NGN-INT formulation, compared to the NGN crude drug, enhanced the dissolution rate of NGN and significantly increased Csubmax/suband AUCsub(0-∞)/subof NGN by 18 times and 1.97 times, respectively. Addition of PVP or HPMC in NGN-INT co-crystal further increased bioavailability of NGN in NGN-INT. The in vivo pharmacodynamic study showed that NGN-INT with HPMC significantly improved the inhibitory effects of NGN against AAA.NGN-INT significantly improved the absorption and aortic protective effects of NGN. The supersaturation-prolonging effect of HPMC further enhanced bioavailability and anti-AAA effects of NGN-INT.

Syo-seiryu-to (SST) is a traditional herbal medicine that has been used clinically to treat allergic rhinitis (AR) in Japan. SST improves acute symptoms, such as sneezing and rhinorrhea, as well as chronic symptoms, such as nasal obstruction, in patients with AR. However, its therapeutic mechanisms remain unknown. We examined the effects of SST and eight constituent crude drugs on phorbol 12-myristate-13-acetate (PMA)-induced gene up-regulation of IL-33 and histamine H1 receptor (H1R), which are responsible for the pathogenesis of AR. We found that SST and its crude drugs, except for Pinellia tuber, significantly and dose-dependently suppressed PMA-induced both IL-33 and H1R mRNA up-regulation in vitro. The half-maximal inhibitory concentration values of the seven crude drugs to inhibit PMA-induced IL-33 mRNA up-regulation were correlated with those related to H1R mRNA up-regulation, suggesting that they act on a common signal molecule. These results suggest that SST improves nasal congestion that is induced by IL-33-related eosinophil infiltration and inhibits sneezing and rhinorrhea that are induced by H1R-mediated histamine signaling in the nasal mucosa of AR patients through its inhibition of a common molecule in the gene expression pathways of IL-33 and H1R. The results could explain the advantages of traditional herbal medicine, in which mixing various crude drugs not only acts on a common target to enhance its pharmacological action, similar to the effect of a high concentration of a single crude extract but also has the benefit of reducing the side effects of each crude drug.

The extract from the rhizome of Costus spectabilis (Costaceae) has been used to treat many illnesses including eye infections and cataract. The present study aimed to establish the pharmacognostic features of the rhizome by evaluating its macro-morphological characteristics, micro-morphological features using its anatomical section, physicochemical constants and elemental profile of its powder according to standard methods. Macro-morphology revealed features typical of a rhizome. Microscopical examination shows vascular bundles scattered throughout the ground tissue of starch-filled cellulosic parenchyma, parenchymatous cortex, endodermis and Oleoresin cells. Chemomicroscopy of the powder showed cellulosic cell walls, lignified cell walls, starch grains, fats and oil. The powder was found to have a moisture content of 12.3%, Total ash of 4.3%, acid insoluble ash of 2.0%, water soluble ash of 1.2%, water soluble extractive of 11.0% and alcohol soluble extractive of 6.5%. Elemental analysis showed the presence of Zn, Cu, Mg, Fe, Pb, Ni, Mn and Cd at concentrations of 24.62, 2.63, 1449.21, 113.23, 36.50, 31.90, 86.93 and 1.23 mg/kg in the powder respectively. The study has established some pharmacognostic features and elemental composition of the rhizome of C. spectabilis. The information could serve useful in providing quality control parameters and standardization of the crude drug. Keywords: Macro-morphology; Microscopical; Physico-chemical; Standardization; Elemental analysis

Objectives: The purpose of the present study was to explore the Pharmacognostic parameters for the standardization of Rosa centifolia Petals. Material and Methods: The flowers of Rosa centifolia were authenticated and shade dried. Rosa centifolia petals were collected then macromorphological, physiochemical assessment, and the micrometric study was carried out. The dried powder form of the Rosa centifolia Petals was then extracted with different solvent systems alcoholic, ethyl acetate, and petroleum ether, and their extractive values were calculated. Most of the phytochemicals were found in ethanol and ethyl acetate fractions. Thin Layer chromatography (TLC) of the ethanol, ethyl acetate, and petroleum ether extract was performed for important phytochemicals flavonoids and polyphenol. Flavonoids and phenolics showed their presence in all extracts with one spot in each extract for ethanol, ethyl acetate, and petroleum ether. Result: The Physicochemical exploration showed values for moisture content, moisture sorption capacity, ash values, and extractive values which are within the limits of World Health Organization standards for the crude drug from medicinal plants. Micromeritic analysis of petal powder reveals good flowability. Ethanolic extractive values were found to be higher when compared to extractive values of, ethyl acetate and petroleum ether. Preliminary Phytochemical examination for the sample indicated the presence of carbohydrates, glycosides, alkaloids, flavonoids, and amino acids. Rf value for flavonoid and phenolic on TLC were found to be for ethanol 0.78 and 0.77, for ethyl acetate 0.81 and 0.78, for petroleum ether 0.81 and 0.78 respectively. Conclusions: The current research would be useful to supplement the information regarding pharmacognostical characteristics, physicochemical evaluation, micrometric analysis, and phytochemical exploration in the Ayurvedic system of medicine for its identification and medicinal use. Keywords: Rosa centifolia, Macromorphological description, Physicochemical evaluation, Phytochemical screening, TLC screening.

The spike (S) protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) contains a cleavage motif R-X-X-R for furin-like enzymes at the boundary of the S1/S2 subunits. The cleavage of the site by cellular proteases is essential for S protein activation and virus entry. We screened the inhibitory effects of crude drugs on in vitro furin-like enzymatic activities using a fluorogenic substrate with whole-cell lysates. Of the 124 crude drugs listed in the Japanese Pharmacopeia, aqueous ethanolic extract of Cnidii Monnieris Fructus, which is the dried fruit of Cnidium monnieri Cussion, significantly inhibited the furin-like enzymatic activities. We further fractionated the plant extract and isolated the two active compounds with the inhibitory activity, namely, imperatorin and osthole, whose IC50 values were 1.45 mM and 9.45 µM, respectively. Our results indicated that Cnidii Monnieris Fructus might exert inhibitory effects on furin-like enzymatic activities, and that imperatorin and osthole of the crude drug could be potential inhibitors of the motif cleavage. Supplementary Information The online version contains supplementary material available at 10.1007/s11418-021-01519-9.

A high-performance thin-layer chromatography method for the simultaneous quantitative determination of salicylic acid, kaempferol, gallic acid, and protocatechuic acid in hydro-alcoholic fractions of Cinnamomum verum J. Presl bark was developed for the first time in this species. For achieving good separation, a mobile phase of toluene‒ethyl acetate‒formic acid (5:4:1, V/V) was used. Densitometric determination was carried out at specified wavelengths for different standard compounds in reflection/absorption mode. The calibration curves were linear in the range of 100‒600 ng per spot for salicylic acid, kaempferol, gallic acid, and protocatechuic acid. During the analysis, defatted hydro-alcoholic fraction from C. verum J. Presl bark showed the presence of salicylic acid (0.098% w/w), kaempferol (0.24% w/w), gallic acid (0.01% w/w), and protocatechuic acid (0.27% w/w). The proposed method is simple, precise, specific, and accurate. The statistical analysis of the data obtained proves that the method is reproducible and selective and can be used for the routine analysis of the reported phenolic compounds in crude drug and extracts. The simultaneous quantification of these biomarkers has not yet been reported in C. verum J. Presl which may be utilized for the proper standardization of the drug. Research also provides new herbal options for different medicinally valuable secondary metabolites in polar categories with different therapeutic potentials. Exploration of bioactive metabolites in C. verum J. Presl bark can also increase its industrial value, which may promote its cultivation and may create a good commercial option for our farmers.

Derris scandens, Albizia procera, and Diospyros rhodocalyx have traditionally been used as herbal remedies for pain relief in Thailand. The ethanolic extracts of these plants obtained by Soxhlet extraction were analyzed by the developed high-performance liquid chromatography-diode-array detection method. Lupeol, the anti-inflammatory triterpene, was selected as a chemical marker for this investigation. All extracts together with that compound were further evaluated for their potential on anti-inflammatory activity using 5-lipoxygenase inhibition assay. Lupeol in each extract was quantified and expressed in the range of 21.44 ± 0.89-40.72 ± 0.40 mg per 100 g of crude drug and the enzyme inhibitory activity of all tested extracts presented as half-maximal inhibitory concentration values ranged between 63.71 ± 2.09 and 91.09 ± 1.40 μg/mL. This study shows that the developed analytical method is effective for analyzing triterpene lupeol in these plants and also reveals the relationship between a lupeol content and the anti-inflammatory effect.

Extraction is a significant step in the processing of the crude drug to get its chemical constituents out and keep them of high and exportable quality. The plants have various chemical constituents that are responsible for various activities in which antioxidant activity is the important one. There is another step that is crucial in the extraction process that is the selection of the suitable for extraction. Various solvents are used for extraction. They too range from the highly non-polar solvents like benzene and chloroform to the highly polar solvents like Ethanol and distilled water. So, in this work, the focus has been put on to find the effect of the extraction method and the extraction solvent on the antioxidant profile of the guava leaves. The extracts of the solvent Water in the method of ultrasound showed the highest inhibition of the free radicals and the least was with the extracts of the pet ether and using soxhlation. This is indicative that the method of extraction is critical, and the solvent of extraction played a vital role in the content of chemical constituents and the pharmacological activity too. The results showed that the ultrasound method was beneficial in extracting the soft drugs like leaves, and the distilled water was effective in extracting the chemicals from the guava leaves.

The rhizome of Cnidium officinale (Umbelliferae) (known as Senkyu in Japan; COR) has been used as a crude drug in Japanese Kampo formulas, such as Jumihaidokuto (to treat eczema and urticaria) and Kakkontokasenkyushin'i (to treat rhinitis). COR contains phthalides, which are thought to be potent principal constituents. Few studies have been reported about the comparison of anti-inflammatory activity of COR constituents. We aimed to identify the constituents in COR and compare their anti-inflammatory activity. COR was extracted with methanol and fractionated into ethyl acetate (EtOAc)-soluble, n-butanol-soluble, and water-soluble fractions. Primary cultured rat hepatocytes were used to assess anti-inflammatory activity by monitoring the interleukin (IL)-1β-induced production of nitric oxide (NO), an inflammatory mediator. The EtOAc-soluble fraction significantly suppressed NO production without showing cytotoxicity in IL-1β-treated hepatocytes, whereas the n-butanol-soluble fraction showed less potency, and the water-soluble fraction did not significantly affect the NO levels. Four constituents were isolated from the EtOAc-soluble fraction and identified as senkyunolide A, (3S)-butylphthalide, neocnidilide, and cnidilide. Among these phthalides and (Z)-ligustilide, senkyunolide A and (Z)-ligustilide efficiently suppressed NO production in hepatocytes, whereas the others showed less potency in the suppression of NO production. Furthermore, senkyunolide A decreased the levels of the inducible nitric oxide synthase (iNOS) protein and mRNA, as well as the levels of mRNAs encoding proinflammatory cytokines (e.g., tumor necrosis factor α) and chemokine C-C motif ligand 20. These results suggest that senkyunolide A may cause the anti-inflammatory and hepatoprotective effects of COR by suppressing the genes involved in inflammation.

The ethanol/water-soluble extracts of the roots of Smilax moranensis M. Martens & Galeotti, Smilacaceae, which have been appreciated since pre-Hispanic times and traditionally used to treat type 2 diabetes in Mexico, were fractionated by the application of the affinity-directed method to identify glucose-6-phosphatase and α-glucosidase inhibitors. Mass spectrometry was used to identify the inhibitor as free ligands after released from the enzymatic complex by denaturing acidic conditions. The affinity study led to the identification of chlorogenic acid as a glucose-6-phosphatase inhibitor, which is the most abundant metabolite present in the analyzed crude drug. In addition, the affinity studies led to the identification of a flavonolignan glycoside as an α-glucosidase inhibitor. In silico analysis with α-glucosidase MAL12 from the yeast Saccharomyces cerevisiae allowed to determine the best conformational rearrangement for the α-glucosidase inhibitors.

Endotoxin is an unintentional contaminant that has numerous activities and can affect various biological experiments using cells. In this study, we measured the endotoxin activity of samples from a plant extract library (PEL) and determined their degrees of contamination. Endotoxin was detected in approx. 48% (n = 139) and approx. 4% (n = 5) of field-collected and crude drug samples, respectively, and in concentrations >5.0 EU/mL in some samples. The concentrations of endotoxin that affect cells in vitro vary depending on the target cell type. Although the degree of contamination varied in the present study, it was considered to have little effect on the cell experiments. More than 150 PEL samples had problems with reaction courses or recovery rates of Limulus amoebocyte lysate (LAL) tests. In the LAL tests, using three plant extracts [Sanguisorba officinalis L. (Rosaceae), Oenothera biennis L. (Onagraceae), and Lythrum salicaria L. (Lythraceae)], the polyphenolic compounds in the plant extracts affected LAL test and their effects differed depending on the plant species. When the 16 single polyphenol compounds were added to the LAL tests, the compounds with caffeoyl and pyrogallol moieties were found to affect the LAL reaction course and recovery rate. Furthermore, none of the compounds had any effects at concentrations of 1 µM. Because the plant extracts contained analogs of various polyphenolic compounds, they were presumed to actually act synergistically. Our findings demonstrated that attention must be paid to the recovery rate and reaction process of LAL tests with samples containing polyphenolic compounds.

A high-performance thin-layer chromatography method for the simultaneous quantitative determination and validation of scopoletin and gallic acid in the methanolic fraction of Jatropha glandulifera L. was developed for the first time. For achieving good separation, the mobile phase of toluene‒ethyl acetate‒glacial acetic acid (7.5:2.5:0.1, V/V) was used. Densitometric determination was carried out at 254 nm for scopoletin and gallic acid, in reflection/absorption mode; the calibration curves were linear in the range of 100‒600 ng per spot. During the analysis, the methanolic fraction of J. glandulifera L. showed the presence of scopoletin (0.08%) and gallic acid (0.04%). The proposed method is simple, precise, specific, and accurate. The obtained data can be used for the routine analysis of reported biomarkers in crude drug and extracts. The simultaneous quantification and method validation of these biomarkers have not yet been reported in J. glandulifera L. which may be utilized for the proper standardization of the plant.

This study aimed to characterize the morphological and anatomical traits of six Ocimum species namely Ocimum tenuiflorum L. Syn. O. Sanctum L. (green var. known as Radha Tulasi), O. tenuiflorum L. (purple var. known as Krishna Tulasi), O. basilicum L.( green var. known as Sugandhi Tulasi), O. basilicum var. pupurascens L.( purple var. known as Durlabha Tulasi), O. canum Sims. (Kanaka Tulasi) and O. gratissimum L. (Rama Tulasi). Chlorophyll estimation of young and matured leaves was also estimated. Cholorophyll a, Chl. b and total chlorophyll was maximum in both young and matured leaves of O. gratissimum L. compared to other species. Carotenoid pigments was maximum of 203.94 mg/ml in young leaves of O. gratissimum L. and 244.92 mg/ml in matured leaves of O. sanctum L. Comparative morphological characters and anatomical variation were studied for identification of the desired species in drug manufacturing industries before making formulations. This study will certainly help to ensure the quality of the crude drug as these are collected by untrained workers and supplied in dry condition to the drug manufacturing industries. Therefore, there is a chance of adulteration because of vested interest or ignorance as many plants are known by common names or in vernacular names. It was found that the six species can be identified and authenticated using these experimental findings before preparation of compound Ayurveda, Siddha and Unani (ASU) drugs in Indian System of Medicines.

Graphic abstract Unprocessed dried tuberous root of Pinellia ternata (Pinellia Tuber) has been used as a component of traditional Japanese Kampo medicine, while this crude drug is usually used after processing with Ginger in traditional Chinese medicine (TCM). It is known that the raphides contained in unprocessed Pinellia Tuber can induce severe acrid irritation of the oral and laryngopharynx mucosa when it is boiled insufficiently. However, the mechanism of the inducing irritation by the raphides nor that of the detoxification by heat or processing with Ginger have been left unclear, mainly because of the difficulties in the extraction and the purification of the raphides and in the assays of acridity. In this study, we developed an easy protocol that can be used to collect raphides from unprocessed Pinellia Tuber and an assay protocol that can be used to evaluate the acridity of the raphides in vitro. The raphides of Pinellia Tuber were discovered to have a lipophilic character and to be collected easily by the extraction using petroleum ether. It was also found that the denaturation of the raphides could be assayed by the dispersity of them in petroleum ether layer of the water/petroleum ether partition, and the acridity of the raphides was found to be in correlation with the assayed denaturation. The raphides were denatured by heat, methanol, or Ginger extract and the denaturing activity of Ginger on raphides was found to be attributable to its lipophilic and thermostable components, which may explain the meaning of the processing of Pinellia Tuber with Ginger in TCM, and may lead to the development of an easier and safer protocol to administer Pinellia Tuber. In addition, it was found that rinsing the mouth with salad oil can effectively relieve irritation of the oral mucosa caused by unprocessed Pinellia Tuber, probably due to the lipophilicity of the raphides. Electronic supplementary material The online version of this article (10.1007/s11418-020-01425-6) contains supplementary material, which is available to authorized users.

The present study was designed to develop and validate a high-performance thin-layer chromatography (HPTLC) system for the simultaneous quantitative determination of berberine and rutin in Tinospora cordifolia extract and their pharmaceutical preparations. Chromatographic development was done using a blend of n-hexane, ethyl acetate, glacial acetic acid and methanol (10:1.1:1.1:2.5, v/v) as the mobile phase. Detection was completed densitometrically at 254 nm. The RF estimation of berberine and rutin was observed to be 0.67 ± 0.02 and 0.47 ± 0.02, respectively. The developed HPTLC method was validated according to ICH guidelines; the method was specific, linear and accurate and can be used to determine berberine and rutin in marketed herbal preparations. The Tinospora cordifolia plant extract was further evaluated for antioxidant activity using HPTLC, and berberine was found to be more active than rutin during DPPH (2,2-diphenyl-1-picrylhydrazyl) scavenging activity. The method was found simple, rapid, accurate, specific and robust for the analysis of berberine and rutin in crude drug using the same method.

St. John’s wort has been used for centuries in traditional medicine of many cultures, and nowadays it is well-known as a clinically important antidepressant drug. Considering the rising market demand for Hyperici herba, quality control of crude drug is of paramount importance. In this paper we performed HPLC-DAD chemical profiling of St. John’s wort tea samples obtained at local markets, pharmacies and health food stores in the Balkan Peninsula countries, Austria and Turkey. Furthermore, water alcoholic extracts of the collected samples were evaluated in terms of their antioxidant potential, as well as the ability to inhibit biologically important enzymes such as acetylcholinesterase, monoamine oxidases A and B (MAO-A and MAO-B), α-amylase and α-glucosidase. Significant variability within the samples in the quantities of hypericin, hyperforin, rutin, quercetin, gallic, chlorogenic, caffeic and p-hydroxybenzoic acid was noticed. Chemotaxonomic modelling enabled the identification of three clusters of samples based on the quantities of rutin, hypericin and hyperforin. Generally, the extracts exhibited a significant potential to inhibit MAO-A (median IC50 = 10.01 μg/mL) and α-glucosidase (median IC50 = 12.40 μg/mL). The results of antioxidant potential evaluation suggest strong neutralization of hydroxyl and nitroso radicals, but moderate inhibition of lipid peroxidation process. Overall, the conducted study emphasizes the possibility of St. John’s wort teas quality control, taking into account unavoidable variability of chemical profile as well. The stated is of great importance when grinding degree of herbal material excludes taxonomic identification of biological origin as an option within quality control.

In this study, a quantitative 1H NMR method (1H-qNMR) for determining the contents of imperatorin, byakangelicin, and oxypeucedanin in A. dahurica in traditional Chinese medicine (TCM) has been established. Dried plant material was extracted exhaustively with methanol by an ultrasonication-assisted extraction method. The 1H-qNMR measurements were performed on a 600 -MHz spectrometer with hydroquinone as the internal standard reference in deuterated dimethyl sulfoxide (DMSO-d6) solvent. Quantification was carried out using the 1H resonance signals at 6.55 ppm for hydroquinone and 7.68, 7.38-7.39, and 6.38-6.39 ppm for imperatorin, byakangelicin, and oxypeucedanin, respectively. The linearity, limit of detection (LOD), limit of quantitation (LOQ), precision, reproducibility, stability, and recovery of the methodology were evaluated, and results were good. The newly developed method has been applied to determine the three coumarins in A. dahurica.

Background Herbal plants are potent in curing various ailments of ancient times as they have comparatively lesser side effects. The demands for natural drugs, mostly from plant sources, are increasing over the past few decades. Because of their potent antioxidant activity, Acacia species are used to treat a variety of diseases. One of the species Acacia ferruginea, an endangered medicinal plant, is widely used in the traditional medicine system, and it is considered that standardization would be beneficial. The present study investigates the physicochemical parameters, preliminary phytochemical screening, trace metals by SEM–EDS, and fluorescence properties of various extracts (non-polar to polar) of leaf and bark parts. Standard spectrophotometric methods (UV–Vis, FT-IR, fluorescence spectroscopy) are employed to analyze the functional groups, and the DPPH and total antioxidant methods are used to assess antioxidant potential. Results The ethyl acetate extract of leaves and ethanol extract of the bark are found to be the highest in yield, 16.32% and 2.54%. Results reveal that the total ash percentage and moisture content are of bark and the water-soluble ash of leaves is higher (10.3 ± 0.85, 7.6 ± 0.34, 3.22 ± 0.24%). The bark polar extract contained more macro-elements such as Na, K, Mg, Ca, S, and Cl. Phytochemical analysis reveals the polar extracts of leaves and bark show saponins, flavonoids, steroids, phenolic compounds, and non-polar extracts show mild positive. The total alkaloids, phenolics, and terpenoids (1.58 ± 0.08%; 0.56 ± 0.11; 0.75 ± 0.15) are found to be higher in A. ferruginea leaves. The FT-IR result shows the presence of alkanes, alkenes, aromatic compounds, aldehydes, phenolics and does not contain any toxic substances since there is no peak observed in the region between 2220 and 2260 cm−1. The in vitro antioxidant activity of the species demonstrated that both the leaf and bark parts have prominent antioxidant properties. Conclusions The results obtained from the preliminary standardization of A. ferruginea are very helpful in the determination of the quality and purity of the crude drug. The refurbished findings of A. ferruginea are promising, and further research is important to identify the bioactive compounds, thereby developing nutritional supplements and medications through therapeutic compound isolation. Graphical Abstract

To evaluate the medicinal properties of a glycyrrhizin (GL)-deficient strain of Glycyrrhiza uralensis, we investigated the anti-allergic effect of the hot water extract obtained from its roots on contact hypersensitivity in mice, and compared it with that of the hot water extract of a commercial crude drug, Glycyrrhiza Radix. The hot water root extract of the GL-deficient strain contained glucoglycyrrhizin (GGL) and rhaoglucoglycyrrhizin (RGL) instead of GL, and it showed anti-allergic activity against contact hypersensitivity in a fashion similar to that of the crude drug extract. We further confirmed the presence of glycyrrhetinic acid (GA), a major metabolite of GL, in mice serum after oral administration of the hot water root extract of a GL-deficient strain. We demonstrated that GGL underwent hydrolysis by intestinal microflora of mice to form GA. These results suggest that a GL-deficient strain of G. uralensis is a useful medicinal resource since the glycosides of GA work in a fashion similar to that of GL when orally administered.

Background As the dry rhizome of Anemone raddeana Regel, Rhizoma Anemones Raddeanae (RAR), which belongs to Ranunculaceae, is usually used to treat wind and cold symptoms, hand-foot disease and spasms, joint pain and ulcer pain in China. It is well known that the efficacy of RAR can be distinctly enhanced by processing with vinegar due to the reduced toxicity and side effects. However, the entry of vinegar into liver channels can cause a series of problems. In this paper, the differences in the acute toxicity, anti-inflammatory and analgesic effects between RAR and vinegar-processed RAR were compared in detail. The changes in the chemical compositions between RAR and vinegar-processed RAR were investigated, and the mechanism of vinegar processing was also explored. Methods Acute toxicity experiments were used to examine the toxicity of vinegar-processed RAR. A series of studies, such as the writhing reaction, ear swelling experiment, complete Freund’s adjuvant-induced rat foot swelling experiment and cotton granuloma, in experimental mice was conducted to observe the anti-inflammatory effect of vinegar-processed RAR. The inflammatory cytokines of model rats were determined by enzyme-linked immunosorbent assay (ELISA). Liquid Chromatography-Quadrupole-Time of Flight mass spectrometer Detector (LC-Q-TOF) was used to analyse the chemical compositions of the RARs before and after vinegar processing. Results Neither obvious changes in mice nor death phenomena were observed as the amount of vinegar-processed RAR in crude drug was set at 2.1 g/kg. Vinegar-processed RAR could significantly prolong the latency, reduce the writhing reaction time to reduce the severity of ear swelling and foot swelling, and remarkably inhibit the secretion of Interleukin-1β(IL-1β), Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) proinflammatory cytokines. The content of twelve saponins (e.g., Eleutheroside K) in RAR was decreased after vinegar processing, but six other types (e.g., RDA) were increased. Conclusions These results revealed that vinegar processing could not only improve the analgesic and anti-inflammatory effects of RAR but also reduce its own toxicity. Trial registration Not applicable.

BACKGROUND: Corncob is one of crude drug which containing phenolic compounds that can be used as an active ingredient for sunscreen preparations. In this study, extracts of dried corn cob made into a gel formulation using SRC (Semi Refined Carrageenan) and glucomannan as a gelling agent. AIM: This study aims to determine the optimal concentration of SRC (Semi Refined Carrageenan) and glucomannan to the physical stability of the gel. METHODS: Gel made into four formulas with a ratio of 1: 4 and the concentration of each formula was 1%, 1.5%, 2%, 2.5%. Each formula was evaluated for 6 weeks of storage that includes organoleptic test, homogeneity, pH, viscosity, dispersive power, freeze-thaw test and centrifugation. RESULTS: The results showed the fourth formula did not change the organoleptic test and homogeneity test. In the test separation of the freeze-thaw method, the fourth formula was stable, while in the centrifugation test formula 1 and 2 was been separation. pH and viscosity results obtained from statistics with a two-way ANOVA showed significant differences in each formula. CONCLUSION: From the results of this study concluded that the formula 3 with a concentration of 2% was the optimal concentration as a gelling agent.

Phellodendron bark ("Obaku") is an important crude drug used in Kampo-medicine. Recently, powder formulation of phellodendron bark was approved as an "efficacious treatment for bruise, sprain, and periodontal diseases", and it has been marketed as an OTC agent. To obtain this approval, the examination of quality control-related characteristics is necessary. Therefore, we established a quantitative method for jatrorrhizine, palmatine, and berberine determination. In this study, we compared the contents of the three constituents obtained from the extracts of Japanese and Chinese phellodendron bark and found remarkable difference.

Gymnema sylvestre (Madhunashini) is one of the most important medicinal plants used as a crude drug for its preventive and therapeutic properties. Among other constituents of Gymnema, gymnemic acid is a major component responsible for biological and pharmacological actions. The present study deals with the influence of different media strength and initial medium pH on the growth of hairy roots and gymnemic acid production from Gymnema sylvestre. Higher strength of the media (1.5X) favoured the biomass production (114.64 g/L FW and 12.63 g/L DW) and gymnemic acid content (11.7 mg/g DW) in the tested range of 0.25, 0.50, 0.75, 1.0, 1.5 and 2.0 X strength. Among the different hydrogen ion concentration (pH) of 4.0, 4.5, 5.0, 5.5, 5.8, 6.0 and 6.5, initial medium pH of 6.0 favoured the biomass production (102.41 g/L FW and 11.52 g/L DW) and medium pH of 5.8 favoured the gymnemic acid production (11.30 mg/g DW).

We evaluated whether the origins of crude drugs derived from arthropods and annelids could be identified using molecular biological techniques. DNA was extracted from 20 crude drugs prepared from different animals using a commercial kit with added phenol treatment. The target regions used to identify origin were the mitochondrial 16S ribosomal RNA (rRNA), 12S rRNA, and cytochrome oxidase subunit I (COI) gene regions. Extracted DNA was amplified by polymerase chain reaction, and then sequenced by the Sanger method. The aligned sequences were compared with all available sequences using BLAST to estimate the origins of the crude drugs. The origin of crude drugs used in this study could be estimated using this method. The COI region was the best for identifying origin among three regions examined, based on the success rate of PCR amplification and analysis. Moreover, the 12S rRNA region was also useful for origin identification, with the exception of the earthworm. However, the origin of some crude drugs could not be strictly identified due to matches to various species in all three regions. One likely cause was that the species of origin of a crude drug has not been registered in DNA databases. We found that even the same crude drug from the same pharmaceutical company had different origins by production lot or import source country. Therefore, this method is useful not only for DNA-based origin identification but also quality control of production lots.

1H NMR-based metabolomics has been applied in research on food, herbal medicine, and natural products. Although excellent results were reported, samples were directly extracted with a deuterated solvent (e.g., methanol-d4 or D2O) in most reports. As primary metabolites account for most of the results, data for secondary metabolites are partially reflected. Consequently, secondary metabolites tend to be excluded from factor loading analysis, serving as a significant unfavorable feature of 1H NMR-based metabolomics when investigating biologically active or functional components in natural products and health foods. Reversed-phase solid-phase extraction column (RP-SPEC) was applied for sample preparation in 1H NMR-based metabolomics to overcome this feature. The methanol extract from Saposhnikoviae radix (SR), an important crude drug, was fractionated with RP-SPEC into 5% methanol-eluting fractions, and the remaining fraction was collected. Each fraction was subjected to 1H NMR-based metabolomics and compared to results from conventional 1H NMR-based metabolomics. Based on principal component analysis (PCA) and partial least squares projections to latent structures discriminant analysis (PLS-DA), the 5% methanol fraction and conventional method reflected the amount of saccharides such as sucrose on the PC1/PLS1 axes, and wild and cultivated samples were discriminated along those axes. The remaining fraction clearly distinguished SR from Peucedanum ledebourielloides root. The compounds responsible for this discrimination were deemed falcarindiol derivatives and other unidentified secondary metabolites from the s-plot on PLS-DA. The secondary metabolites from original plants were, therefore, presumed to be concentrated in the remaining fraction by RP-SPEC treatment and strongly reflected the species differences. The developed series is considered effective to perform quality evaluation of crude drugs and natural products.

Chemotherapy-induced peripheral neuropathy (CIPN) is one of the dose-limiting side effects of cancer chemotherapy. Although the control of CIPN is important, it is difficult to manage with currently available therapeutic drugs. Therefore, there is a need for novel therapeutic agents for treating CIPN. Goshajinkigan (GJG) is a Kampo formula composed of ten crude drugs. While GJG has been used for the treatment of CIPN, the active constituents of GJG and their underlying mechanisms of pharmacological effects are still unknown. Our previous study revealed that repetitive oral administration of the water extract of Plantaginis Semen, a crude drug ingredient of GJG, inhibited the mechanical allodynia induced by an intraperitoneal injection of paclitaxel in mice. To elucidate the active compounds of Plantaginis Semen, activity-guided separation of the water extract of Plantaginis Semen was performed. From the active fraction, four iridoids (1–4) were identified. Repetitive oral administration of aucubin (1) at 100 or 30 mg/kg and 100 mg/kg of the fraction crude 3 [primarily comprised of pedicularis-lactone (3)], showed anti-allodynic activity, suggesting 1 and 3 could be some of the active compounds responsible for the anti-allodynic property of Plantaginis Semen and GJG. Our study establishes that oral administration of 1 has potent anti-allodynic effect in addition to the activity of intraperitoneally administered 1 reported previously. Identification of active anti-allodynic compounds found in Kampo formulations will support the development of novel therapies for the management of CIPN in cancer patients. Electronic supplementary material The online version of this article (10.1007/s11418-019-01327-2) contains supplementary material, which is available to authorized users.

Glycyrrhizae Radix is an important crude drug in Japan and is the most frequently prescribed drug in Kampo medicines for the treatment of a wide range of diseases. Glycyrrhizin (GL), the major active ingredient of Glycyrrhizae Radix, has various pharmacological actions but causes adverse effects such as pseudoaldosteronism. In a previous study, the GL content of shoseiryuto was found to be unexpectedly low, and Schisandrae Fructus in shoseiryuto reduced the pH value of the decoction and drastically decreased the extraction efficiency of GL from Glycyrrhizae Radix. In the present study, we investigated the extraction efficiency of GL from Glycyrrhizae Radix in decoctions comprising Glycyrrhizae Radix and five different fruit-derived crude drugs. Among the five fruit-derived crude drugs tested, Schisandrae Fructus markedly decreased both the pH value of the decoction and the extraction efficiency of GL. A comparison of the pH value of the decoction and the GL content of 12 Kampo prescriptions (containing at least Glycyrrhizae Radix and Schisandrae Fructus) showed that the GL content per daily dose was proportional to the compounding amount of Glycyrrhizae Radix, and that the extraction efficiency of GL from Glycyrrhizae Radix was strongly correlated with the pH value of the decoction. In addition, the pH value of the decoction was similar to the pH value documented in interview forms provided by pharmaceutical companies. These results suggested that the GL content in Glycyrrhizae Radix-containing Kampo products can be estimated from both the compounding amounts of Glycyrrhizae Radix and the pH value documented in their interview forms. Knowledge of GL content will help avoid adverse reactions due to Glycyrrhizae Radix.

Extraction of crude drugs by using different solvents provides polarity-based fractions containing specific type of secondary metabolites. Carissa carandas L. fruits were extracted and fractionated, petroleum ether extract was processed by a fatty acid methyl ester (FAME) technique for characterization by gas chromatography—mass spectrometry (GC—MS) analysis, and the remaining part was extracted with methanol for high-performance thin-layer chromatography (HPTLC) analysis, followed by simultaneous quantitative determination of protocatechuic acid and quercetin in methanolic fractions. A validated method for the simultaneous quantification of protocatechuic acid and quercetin was developed and is being reported for the first time in C. carandas L. fruits to the best of our knowledge. Petroleum ether and methanol fractions were found to be the best for the highest possible recovery of targeted analytes. Chromatographic elution of FAME compounds generated from petroleum ether extract was evaluated by GC—MS profiling. Nineteen fatty acid compounds were separated with the highest quantity of octacosanal (13.31%). On the other hand, a polar fraction was processed by HPTLC profiling. For achieving good separation, a mobile phase of toluene-ethyl acetate-formic acid (6:3:1; V/V) was used. Densitometric determination was carried out at 310 nm in the reflection/absorption mode. The calibration curves were linear in the range of 100–600 ng per spot for protocatechuic acid and quercetin. During the analysis, the dried raw material from C. carandas L. fruits showed the presence of protocatechuic acid (0.04%) and quercetin (0.05%). The proposed method is simple, precise, specific, and accurate. The statistical analysis of the data obtained proves that the method is reproducible and selective, which can be used for the routine analysis of the reported phenolic compounds in crude drug and extracts. The simultaneous quantification of these phenolic compounds has not yet been reported in C. carandas L. fruits which may be utilized for the proper standardization of the drug.

To investigate the mechanism of Radix Kansui (RK) stir-fried with vinegar (VRK) decreased hepatotoxicity in mice. According to a random number table, 40 mice were randomly divided into negative control group (0.5% carboxymethylcellulose sodium, 20 mL/kg), positive control group (0.1% mixture of carbon tetrachloride in soybean oil, 20 mL/kg), RK group (the ethyl acetate extracts of RK, 250 g crude drug/kg) and VRK group (the ethyl acetate extracts of VRK, 250 g crude drug/kg) with 10 mice per group. All mice were administered orally by gavage daily for 7 continuous days. The morphology of liver tissues was examined to assess the liver injury by a transmission electron microscope. Hepatocyte apoptosis in vivo was determined by terminal deoxynucleotidyltransferase-mediated dUTP-biotin nickend labeling (TUNEL) assay. Immunohistochemical technique was adopted to detect the expression of particular antiapoptotic and proapoptotic proteins in the mitochondrial pathways, including B-cell lymphoma (Bcl-2) and caspase-3, as well as the expression of inflammatory mediators, including nuclear factor kappa B (NF- κ B) and intercellular adhesion molecule-1 (ICAM-1). Liver injury and hepatocyte apoptosis were observed in RK mice, and the liver injury were significantly reduced in VRK-treated mice. In immunohistochemistry study, compared with the negative control group, RK inhibited dramatically the Bcl-2 protein expression and significantly increased the expression of caspase-3, NF- κ B and ICAM-1 (all P

Dried achene or anthocarpous accessory fruits of Rosa multiflora Thunb., Rosae fructus ("Eijitsu" in Japanese), have been used in clinical practice to improve constipation within traditional Japanese medicine. Recently, it has been claimed that the efficacy of this crude drug is decreasing, and multiflorin A, the purgative component, was not detected within the tested samples. In order to clarify the causes of this issue, we investigated Rosa section Synstylae (Rosaceae), including R. multiflora, growing in Japan and South Korea with a focus on the secondary metabolite, multiflorin A. We recognize that there are two chemotypes based on the presence (Type I) or absence (Type II) of multiflorin A. Type I contains quercitrin, multinoside A, multiflorin B, and multinoside A acetate as major index compounds. Type II contains hyperin, isoquercitrin, quercetin 3-O-glucuronide, and 3'-methoxy-isoquercitrin as the major index compounds. The chemotype of Rosa section Synstylae (Rosaceae) plants collected in Japan (excluding Tsushima Island) were all classified as Type I with exception of two species, R. luciae and R. sambucina. On the other hand, both Type I and Type II were detected within Rosae fructus obtained from R. multiflora collected in South Korea and Tsushima Island, Japan. The results indicate that Rosae fructus from R. multiflora (Type I) from Japan, excluding Tsushima Island, should be employed clinically, which we describe as purgative.

A high-performance thin-layer chromatography (HPTLC) method for the simultaneous quantitative determination and validation of ursolic acid, β-sitosterol, lupeol, and quercetin in the methanolic fraction of Ichnocarpus frutescens L. was developed for the first time. For achieving good separation, a mobile phase of tolueneethyl acetate-formic acid (8:2:0.1, v/v) was used. Densitometric determination was carried out at 500 nm for ursolic acid, 550 nm for β-sitosterol, 650 nm for lupeol, and 310 nm for quercetin in reflection—absorption mode, and the calibration curves were linear in the range of 100–600 ng per spot. During the analysis, the methanolic fraction of I. frutescens L. showed the presence of ursolic acid (0.34%), β-sitosterol (0.27%), lupeol (0.27%), and quercetin (0.26%). The proposed method is simple, precise, specific, and accurate. The obtained data can be used for routine analysis of reported biomarkers in crude drug and extracts. The simultaneous quantification and method validation of these biomarkers have not yet been reported in I. frutescens L., which may be utilized for the proper standardization of the plant.

The stem of Nauclea officinalis have been utilized as a crude drug in China, so other parts of Nauclea officinalis are abandoned, resulting in a waste of traditional Chinese medicine resources. To determine the distribution and content of the alkaloids, phenolic acids and iridoid in different organs (stem, branch, leaf and bark) of this plant, a reliable method has been established using LC-MS/MS. Nine constituents, namely strictosamide, vincosamide, chlorogenic acid, sweroside, naucleamide B, protocatechuic acid, pumiloside, vanillic acid and cryptochlorogenic acid were simultaneously determined in six minutes. Meanwhile, the antipyretic, anti-inflammatory and analgesic activities were evaluated for comparative analysis of the pharmacological activity in different parts from Nauclea officinalis. The results showed that the content of active components in other organs of Nauclea officinalis was higher than that in stems, and pharmacological effects of branches and leaves were also better. The established approach could be helpful for the quality control of Nauclea officinalis, and also provide necessary information for the rational utilization of Nauclea officinalis resources.

Uncaria Hook (UH) is a dry stem with hook of Ucaria plant and is contained in Traditional Japanese and Chinese medicine such as yokukansan, yokukansankachimpihange, chotosan, Gouteng-Baitouweng, and Tianma-Gouteng Yin. UH contains active indole and oxindole alkaloids and has the therapeutic effects on ailments of the cardiovascular and central nervous systems. The recent advances of analytical technology led to reports of detailed pharmacokinetics of UH alkaloids. These observations of pharmacokinetics are extremely important for understanding the treatment’s pharmacological activity, efficacy, and safety. This review describes properties, pharmacology, and the recently accumulated pharmacokinetic findings of UH alkaloids, and discusses challenges and future prospects. UH contains major indole and oxindole alkaloids such as corynoxeine, isocorynoxeine, rhynchophylline, isorhynchophylline, hirsuteine, hirsutine, and geissoschizine methyl ether (GM). These alkaloids exert neuroprotective effects against Alzheimer’s disease, Parkinson’s disease, and depression, and the mechanisms of these effects include anti-oxidant, anti-inflammatory, and neuromodulatory activities. Among the UH alkaloids, GM exhibits comparatively potent pharmacological activity (e.g., agonist activity at 5-HT1A receptors). UH alkaloids are absorbed into the blood circulation and rapidly eliminated when orally administered. UH alkaloids are predominantly metabolized by Cytochrome P450 (CYP) and converted into various metabolites, including oxidized and demethylated forms. Regarding GM metabolism by CYPs, a gender-dependent difference is observed in rats but not in humans. Several alkaloids are detected in the brain after passing through the blood–brain barrier in rats upon orally administered. GM is uniformly distributed in the brain and binds to various channels and receptors such as the 5-HT receptor. By reviewing the pharmacokinetics of UH alkaloids, challenges were found, such as differences in pharmacokinetics between pure drug and crude drug products administration, food-influenced absorption, metabolite excretion profile, and intestinal tissue metabolism of UH alkaloids. This review will provide readers with a better understanding of the pharmacokinetics of UH alkaloids and their future challenges, and will be helpful for further research on UH alkaloids and crude drug products containing UH.

Licorice, the dried root or stolon of Glycyrrhiza glabra or G. ularensis, is commonly used worldwide as a food sweetener or crude drug. Its major ingredient is glycyrrhizin. Hypokalemia or pseudoaldosteronism (PsA) is one of the most frequent side effects of licorice intake. Glycyrrhizin metabolites inhibit type 2 11β-hydroxysteroid dehydrogenase (11βHSD2), which decomposes cortisol into inactive cortisone in the distal nephron, thereby inducing mineralocorticoid receptor activity. Among the several reported glycyrrhizin-metabolites, 18β-glycyrrhetyl-3-O-sulfate is the major compound found in humans after licorice consumption, followed by glycyrrhetinic acid. These metabolites are highly bound to albumin in blood circulation and are predominantly excreted into bile via multidrug resistance-associated protein 2 (Mrp2). High dosage and long-term use of licorice are constitutional risk factors for PsA. Orally administered glycyrrhizin is effectively hydrolyzed to glycyrrhetinic acid by the intestinal bacteria in constipated patients, which enhances the bioavailability of glycyrrhizin metabolites. Under hypoalbuminemic conditions, the unbound metabolite fractions can reach 11βHSD2 at the distal nephron. Hyper direct-bilirubin could be a surrogate marker of Mrp2 dysfunction, which results in metabolite accumulation. Older age is associated with reduced 11βHSD2 function, and several concomitant medications, such as diuretics, have been reported to affect the phenotype. This review summarizes several factors related to licorice-induced PsA, including daily dosage, long-term use, constipation, hypoalbuminemia, hyper direct-bilirubin, older age, and concomitant medications.

Citrus-type crude drugs (CCDs) are commonly used to formulate decoctions in Kampo formula (traditional Japanese medicine). Our previous study reported metabolomic analyses for differentiation of the methanol extracts of Citrus-type crude drugs (CCDs) using ultra-HPLC (UHPLC)/MS, and 13C- and 1H-NMR. The present study expanded the scope of its application by analyzing four CCD water extracts (Kijitsu, Tohi, Chimpi, and Kippi); these CCDs are usually used as decoction ingredients in the Kampo formula. A principal component analysis score plot of processed UPLC/MS and NMR analysis data indicated that the CCD water extracts could be classified into three groups. The loading plots showed that naringin and neohesperidin were the distinguishing components. Three primary metabolites, α-glucose, β-glucose, and sucrose were identified as distinguishing compounds by NMR spectroscopy. During the preparation of CCD dry extracts, some compounds volatilized or decomposed. Consequently, fewer compounds were detected than in our previous studies using methanol extract. However, these results suggested that the combined NMR- and LC/MS-based metabolomics can discriminate crude drugs in dried water extracts of CCDs.

The main objective of cancer treatment with chemotherapy is to kill the cancerous cells without affecting the healthy normal cells. In the present study, bioactivity-guided purification of the n-chloroform soluble fraction from the methanol extract of Roscoea purpurea resulted in the identification of two new labdane diterpenes: coronarin K (1) and coronarin L (2), along with eight known compounds, coronarin A (3), bisdemethoxycurcumin (4), kaempferol 3-O-methyl ether (5), kaempferol (6), fenozan acid (7), 3-(3-methoxy,4-hydroxyphenyl)-2-propenoic acid ferulic acid (8), caffeic acid (9), and gallic acid (10). The structural identification of new compounds (1 and 2) were determined by detailed analysis of 1D (1H and 13C) and 2D NMR (COSY, HSQC, and HMBC) spectroscopic data. The relative configurations of 1 and 2 were determined with the help of NOESY correlations and comparison of optical rotations with known labdane diterpenes, with established stereochemistry, while structure of known compounds was established by direct comparison of their NMR data with those reported in the literature. This is the first report of isolation of this labdane diterpenes and phenolic classes of secondary metabolites in R. purpurea. In the preliminary screening, the methanol extract and its fractions were tested for the cytotoxic activity against a panel of four cancer cell lines (A549, HCT-116, Bxpc-3, and MCF-7); extract and its chloroform fraction were found to be active against the lung cancer cell line, A-549, with IC50 value 1–5) were screened for their cytotoxicity against all the cell lines by MTT assay. Coronarin K, 1 showed significant cytotoxic potential against lung cancer cell lines (A-549), with IC50 value of 13.49 μM, while other compounds did not show activity below 22 μM.

Background Dryopteris ramosa has numerous potentials uses in the treatment of different maladies as old traditional medication. The fronds of D. ramose are edible and orally administered for producing antibiotic effect. They are also used as astringent and febrifuge, and as a pesticide. Methods Extraction of fronds of D. ramosa using solvents of increasing polarity, namely, ethyl acetate, methanol and water were tested for phytochemical (qualitative tests, GC-MS), antimicrobial (well method), antioxidant (DPPH), antifungal (tube dilution), cytotoxic activity (brine shrimps lethality assay) and LOX and COX inhibitory activities were performed using standard methods. Results The phytochemical analysis of the crude methanolic extract revealed that the fronds are rich in flavonoids, alkaloids, saponins, tannins, glycosides and triterpenoids. The total flavonoid content of the ethyl acetate fraction was 46.28 μg QE/mg extract. The GC-MS analysis revealed nine major compounds that constituted the crude drug and potentially had a role in reported activities. The crude extract was the most active amongst all the fractions against the bacterial and fungal strains used such that it inhibited the growth of P. aeruginosa with a zone of 13 mm and a MIC value of 16 μg/ml as compared to the standard cefixime, which inhibited the zone by only 10 mm and a MIC value of 32 μg/ml. The highest antioxidant potential in DPPH assay was shown by the crude extract with 91.948% free radical scavenging activity. The bring shrimps lethality potential of the crude extract was the highest, with a LD50 value of 47.635 μg/ml. The ethyl acetate fraction inhibits 91.36% of alpha glucosidase enzyme at a concentration of 0.5 mg/ml. In case of acetylcholine esterase inhibition assay, the methanol fraction inhibits 58.26% of the enzyme activity. Similarly, for butyrylcholine esterase inhibition, the maximum inhibitory effect was seen in the methanol fraction, with a percentage inhibition of 47.32%. Conclusion These test results support traditional medicinal uses of the plant. Dryopteris ramosa could be imperative for being used as a therapeutic agent and the medicinal importance of this plant should be further investigated.

Uncaria rhynchophylla (Miq.) Jacks (Rubiaceae) is a of component of the Chinese crude drug Gou-teng, which is widely used as a remedy for several ailments including cerebrovascular diseases, hypertension, eclampsia, and epilepsy. In chapter, we review the phytochemistry and pharmacological attributes of U. rhynchophylla, with a focus on the neuroprotective activity of the plant. The plant is reported to exert antioxidant, antiinflammatory, antihypertensive, antiarrhythmic, anticancer, anticonvulsive, and neuroprotective properties, which are related to the presence of various classes of bioactive compounds, predominantly indole alkaloids such as rhynchophylline and isorhynchophylline. In vitro and in vivo studies have provided strong evidence that the bioactive compounds present in U. rhynchophylla may potentially be useful in treating neuronal disorders through different modes of action. For instance, with respect to Alzheimer’s disease, constituents of the plant have been showed to play significant roles in inhibiting acetylcholinesterase, Aβ aggregation, counteracting oxidative stress and neuroinflammation, and increasing neuronal survival and synaptic plasticity. This chapter advocates for further investigations into U. rhynchophylla as a novel source of therapeutic agents to prevent and/or manage Alzheimer’s disease.

The essential oil (EO), the methanolic (MeOH), and the 70% ethanolic (70% EtOH) extracts obtained from the aerial parts of Ocimum campechianum Mill. (Ecuador) were chemically characterized through gas-chromatography coupled to mass spectrometry detector (GC&ndash, MS), high-performance liquid chromatography coupled to diode array-mass spectrometry detectors (HPLC-DAD-MS) and studied for their in vitro biological activity. The radical scavenger activity, performed by spectrophotometric 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2&prime, azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays, highlighted significant IC50 values for the EO, extracts and their main constituents (eugenol and rosmarinic acid). EO (and eugenol) showed noteworthy activity against Pseudomonas syringae pv. syringae and a moderate effect against clinical Candida strains, with possible synergism in association to fluconazole against the latter microorganisms. The extracts and pure molecules exhibited weak cytotoxic activity against the HaCat cell line and no mutagenicity against Salmonella typhimurium TA98 and TA100 strains, giving indication of safety. Instead, EO showed a weak activity against adenocarcinomic human alveolar basal epithelial cells (A549). The above-mentioned evidence leads us to suggest a potential use of the crude drug, extracts, and EO in cosmetic formulation and food supplements as antioxidant agents. In addition, EO may also have a possible application in plant protection and anti-Candida formulations.

Quantitative NMR (qNMR) is applied to determine the absolute quantitative value of analytical standards for HPLC-based quantification. We have previously reported the optimal and reproducible sample preparation method for qNMR of hygroscopic reagents, such as saikosaponin a, which is used as an analytical standard in the assay of crude drug section of Japanese Pharmacopoeia (JP). In this study, we examined the absolute purity determination of a hygroscopic substance, indocyanine green (ICG), listed in the Japanese Pharmaceutical Codex 2002, using qNMR for standardization by focusing on the adaptation of ICG to JP. The purity of ICG, as an official non-Pharmacopoeial reference standard (non-PRS), had high variation (86.12 ± 2.70%) when preparing qNMR samples under non-controlled humidity (a conventional method). Additionally, residual ethanol (0.26 ± 0.11%) was observed in the non-PRS ICG. Next, the purity of non-PRS ICG was determined via qNMR when preparing samples under controlled humidity using a saturated sodium bromide solution. The purity was 84.19 ± 0.47% with a lower variation than that under non-controlled humidity. Moreover, ethanol signal almost disappeared. We estimated that residual ethanol in non-PRS ICG was replaced with water under controlled humidity. Subsequently, qNMR analysis was performed when preparing samples under controlled humidity in a constant temperature and humidity box. It showed excellent results with the lowest variation (82.26 ± 0.19%). As the use of a constant temperature and humidity box resulted in the lowest variability, it is recommended to use the control box if the reference ICG standard is needed for JP assays.

Ethnopharmacological relevance Shuang-Huang-Lian preparation has captured wide attention since its clinical applications for the successful treatment of upper respiratory tract infection. However, its functional basis under actual therapeutic dose in vivo was still unrevealed. Aim of the study This study aimed to reveal the anti-flu substances and mechanism of Shuang-Huang-Lian water extract (SHL) on H1N1 infected mouse model by a strategy based on serum pharmaco-chemistry under actual therapeutic dose and network pharmacology. Materials and methods H1N1 infected mouse model was employed for evaluation of the anti-flu effects of SHL. A simultaneous quantification method was developed by UPLC-TQ-XS MS coupled switch-ions mode and applied to characterize the pharmacokinetics of the multiple components of SHL under actual therapeutic dose. The potential active ingredients were screened out based on their pharmacokinetic parameters. And then, a compound mixture of these active candidates was re-evaluated for the anti-flu activity on H1N1 infected mouse model. Furthermore, the anti-flu mechanism of SHL was also predicted by network pharmacology coupled with the experimental result. Results SHL significantly increased the survival rate and prolonged survival days on H1N1 infected mice at a dosage of 20 g crude drug/kg/day by reversing the increased lung index, down-regulating the inflammatory cytokines (TNF-α, IL-1β, IL-6) and inhibiting the release of IFN-β in bronchoalveolar lavage fluids (BALF). Concomitantly, the pharmacokinetic parameters of fourteen quantified and twenty-one semi-quantified constituents of SHL were characterized. And then, five compounds (baicalin, sweroside, chlorogenic acid, forsythoside A and phillyrin), which displayed satisfactory pharmacokinetic features, were considered as potential active ingredients. Thus, a mixture of these five ingredients was administered to H1N1-infected mice at a dose of 4.24 mg/kg/day. As a result, the therapeutical effects of the mixture were similar to SHL in terms of survival rate, lung index and the release of cytokines (TNF-α, IL-1β and IL-6) in BALF. Moreover, network pharmacology analysis indicated that the TNF-signal pathways might play a role in the anti-flu mechanism of SHL. Conclusions A mixture of five compounds (baicalin, sweroside, chlorogenic acid, forsythoside A and phillyrin) were the anti-flu substances of SHL. The strategy based on serum pharmaco-chemistry under actual therapeutic dose provided a new sight on exploring in vivo effective substances of TCM.

Shinkiku (Massa Medicata Fermentata) is a traditional crude drug used to treat anorexia and dyspepsia of elder patients in east Asia. Shinkiku is generally prepared by the microbial fermentation of wheat and herbs. Shinkiku is also used in Japanese Kampo medicine as a component of 半夏白朮天麻湯 (Hangebyakujutsutemmato). However, the quality of shinkiku varies by manufacture because there are no reference standards to control the quality of medicinal shinkiku. Thus, we aim to characterize the quality of various commercially available shinkiku by chemical and microbial analysis. We collected 13 shinkiku products manufactured in China and Korea and investigated the microbial structure and chemical constituents. Amplicon sequence analysis revealed that Aspergillus sp. was common microorganism in shinkiku products. Digestive enzymes (α-amylase, protease, and lipase), organic acids (ferulic acid, citric acid, lactic acid, and acetic acid), and 39 volatile compounds were commonly found in shinkiku products. Although there were some commonalities in shinkiku products, microbial and chemical characteristic considerably differed as per the manufacturer. Aspergillus sp. was predominant in Korean products, and Korean products showed higher enzyme activities than Chinese products. Meanwhile, Bacillus sp. was commonly detected in Chinese shinkiku, and ferulic acid was higher in Chinese products. Principal component analysis based on the GC-MS peak area of the volatiles also clearly distinguished shinkiku products manufactured in China from those in Korea. Chinese products contained higher amounts of benzaldehyde and anethole than Korean ones. Korean products were further separated into two groups: one with relatively higher linalool and terpinen-4-ol and another with higher hexanoic acid and 1-octen-3-ol. Thus, our study revealed the commonality and diversity of commercial shinkiku products, in which the commonalities can possibly be the reference standard for quality control of shinkiku, and the diversity suggested the importance of microbial management to stabilize the quality of shinkiku.

Cortex Moutan (Paeonia suffruticosa Andr.) is a common traditional Chinese medicine and has been widely used in clinic for 2000 years in China. As sources for this crude drug are always mixed with other species, many cultivars on herbal market may lead to quality instability. Multi-component quantitative analysis is an efficient method to reflect chemical profiles of herb medicine and is always taken as the main method for quality evaluation. So, the aim of this work is to develop analytical method to quantify paeonol, paeoniflorin, gallic acid, oxypaeoniflorin, benzoylpaeoniflorin and paeonolide in Cortex Moutan (CM) to evaluate the chemical qualities of CM from different species or cultivars. Besides, we also study the intestinal absorption characteristics of paeonol and paeoniflorin for further pharmacological evaluation. In the present study, all of the standard markers were performed on an Ecosil C18 (250 mm × 4.6 mm, 5 μM, Lubex Co., Guangzhou, China) with linear gradient elution of 0.2% formic acid water and acetonitrile. The proposed method was applied to analyze 50 batches of samples with acceptable linearity (R2, 0.9995 - 0.9999), precisions (RSD, 0.47% - 2.08%), repeatability (RSD, 039% - 2.63%), stability (RSD, 0.52% - 2.45%), and recovery (RSD, 0.72% - 3.03%) of the six compounds. Furthermore, the Hierarchical Cluster Analysis was applied to classify the 50 samples based on contents of the six compound markers. The results obtained from multi-component quantification of CM clearly indicated that CM originated from P. suffruticosa and P. ostii presented different chemical properties, and that samples from the two materials could be gathered into one branch, respectively, while CM sourced from cultivars of P. suffruticosa showed great variety on chemical quality. The results from Hierarchical Cluster Analysis implied that the established method could be used as a powerful tool for the quality evaluation of CM. The intestinal absorption study indicated that the intestinal absorption activities for paeoniflorin and paeonol showed an increasing absorption with time. Paeonol had lower absorption rate (6.69% - 15.93%) than that of paeoniflorin (19.0% - 30.70%). As a result, the established method is suitable for the quality evaluation of CM. The results of intestinal absorption characteristics of paeonol and paeoniflorin offer an insight for pharmacological evaluation and clinical efficacy research of CM.

Objective: The main objective of the current study was to perform pharmacognostical, physiochemical analysis and in vitro antioxidant activity of Tamarindus indica Linn. (Family: Fabaceae) fruit. Materials and Methods: The present study deals with pharmacognostical characters as identification parameters of the fruit, which were subjected to macro- and microscopical studies. Physiochemical analysis was performed as per World Health Organization–recommended parameters. Toxicological parameters such as heavy metals, aflatoxins, and pesticide residue and presence of microbial count were analyzed. This study also deals with the thin layer chromatography/high performance thin layer chromatography fingerprinting and antioxidant activity. Results: The microscopy study revealed the presence of epidermis, vascular bundles, parenchyma cells, mucilage fiber, starch grain, and nonlignified fibers. Physiochemical parameters such as loss on drying, moisture content, ash value, and extractive value were also determined. Heavy metal, aflatoxin, total microbial load, and pesticide residues were found to be variable but under the permissible limits. Conclusion: It is critical to analyze these parameters in each crude herbal drug before further processing to ensure their safety and efficacy for better approval at the international podium. This study revealed precise identification for the meticulous crude drug which will be valuable in detection and manage to adulterations of the raw material.

Objective: The objective of the present study was to investigate the hepatoprotective role of Radix Fici Hirtae on acute alcohol-induced liver injury in mice. Methods: The component of Radix Fici Hirtae was extracted using petroleum ether, chloroform, ethyl acetate and n-butanol and divided into three dose groups of high, medium and low according to the clinical man's normal dose of the 50 g crude drug/d (0.83 g/kg body weight). Saline in concentration of 10 mg/mL, 5 mg/mL and 2.5 mg/mL and a dose of mouse lavage (0.2 mL/10 g mouse body weight) were added to the solution. Histopathlogical analysis of liver was performed. Finally, liver protection was validated by examining the effect of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (AKP), and lactate dehydrogenase (LDH) on the hepatic function of mice in alcohol-induced liver injury model. Results: Except for group with saturated n-butyl alcohol, for the rest of the groups, pathological changes of hepatic lipid and inflammatory cells infiltration were alleviated and liver sinus was normal. As compared to model group, the concentrations of AST, ALT, AKP and LDH in chloroform groups and ethyl acetate groups were significantly decreased. Conclusions: Extracts of Radix Fici Hirtae are effective for the prevention of alcohol-induced hepatic damage in mice. The results revealed that extracts of Radix Fici Hirtae could be used as hepatoprotective agent. Keywords: Radix Fici Hirtae, Alcohol-induced liver injury, Hepatoprotective

Polygala Root (the root of Polygala tenuifolia WILLDENOW; Japanese name "Onji"), a well-known crude drug, traditionally used as an expectorant and sedative, has been attracting increased interest in recent years owing to its newly found pharmacological effect related to neuroprotection. However, there is no specific method for identifying and estimating the quality of this crude drug in the Japanese Pharmacopoeia, 17th edition. Therefore, in order to develop a TLC-based simple and convenient identification method using characteristic chemical marker(s) for the drug and its extract products, UV-sensitive constituents of Polygala Root were first investigated. A total of 23 aromatic compounds were isolated and characterized. Two new compounds, namely, polygalaonjisides A (1) and B (2), were characterized as syringic acid 4-O-(2'-O-β-D-apiosyl)-β-D-glucoside and 2-O-(β-D-glucosyl)-3'-O-benzoylsucrose, respectively. Based on these phytochemical results, a TLC method focusing on three marker spots with Rf value of approximately 0.4-0.5 due to tenuifolisides A and B and 3,6'-di-O-sinapoylsucrose was proposed as a simple and convenient test to identify Polygala Root or its single-extract products on the market. The data presented in this paper could be useful in stipulating a confirmation test to identify Polygala Root.

Ephedra Herb is a crude drug defined as the terrestrial stem of Ephedra sinica, E. intermedia, or E. equisetina. It is often used to treat headaches, bronchial asthma, nasal inflammation, and the common cold. In this study, we isolated characteristic non-alkaloidal constituents from the extracts and identified them in relation to the habitat of Ephedra Herb. Extracts were prepared from Ephedra Herb collected from Inner Mongolia and Gansu. High-performance liquid chromatography was performed to quantitatively analyse the amount of ephedrine alkaloids in each extract. We compared the chemical compositions of the extracts by thin layer chromatography (TLC) to find spot characteristics depending on the habitat. 1H-NMR, 13C-NMR, and 2D-NMR spectra of the samples were also examined. The ephedrine content of all extracts satisfied the quality standard stated in the Japanese Pharmacopoeia. Nonetheless, we found each notable constituent characteristic to the Ephedra Herbs from both habitats. In order to identify them, Ephedra Herb extracts were separated by column chromatography, resulting in the isolation of (±)-α-terpineol-β-D-O-glucopyranoside (1) and (E)-7-hydroxy-3,7-dimethyloct-2-en-1-yl-β-D-O-glucopyranoside (2) as the characteristic constituents in Ephedra Herb from Inner Mongolia. Epheganoside (3), a new eudesmane-type sesquiterpene glycoside, and scopoletin (4) were found to be the characteristic constituents in Ephedra Herb from Gansu. The results obtained from this study can be used to distinguish between the habitats of Ephedra Herb.

A high-performance thin-layer chromatography (HPTLC) method for the simultaneous quantitative determination of ursolic acid and β-sitosterol in the methanolic fraction of Paederia foetida L. leaves was developed for the first time. For achieving good separation, a mobile phase of toluene‒ethyl acetate‒formic acid (8:2:0.1, v/v) was used. The densitometric determination was carried out at 550 and 522 nm in reflection/absorption mode for ursolic acid and β-sitosterol. The calibration curves were linear in the range of 100-600 ng per spot for ursolic acid and β-sitosterol. During the analysis, the methanolic fraction of P. foetida L. leaves showed the presence of ursolic acid (0.12 ± 0.05%) and β-sitosterol (0.08 ± 0.12%). The proposed method is simple, precise, specific, accurate, less time-consuming, and cost-effective. The statistical analysis of the data obtained proves that the method is reproducible and selective and can be used for the routine analysis of the reported phenolic compounds in crude drug ...