Metabolite profiling of Andrographis paniculata (Burm. f.) Nees. young and mature leaves at different harvest ages using 1H NMR-based metabolomics approach.

Andrographis paniculata (Burm. F.) Nees. is considered as the herb of the future due to its precious chemical compounds, andrographolide (ANDRO), neoandrographolide (NAG) and 14-deoxyandrographolide (DAG). This study aims to profile the metabolites in young and mature leaf at six different harvest ages using 1HNMR-based metabolomics combined with multivariate data analysis. Principal component analysis (PCA) indicated noticeable and clear discrimination between young and mature leaves. A comparison of the leaves stage indicated that young leaves were separated from mature leaves due to its larger quantity of ANDRO, NAG, DAG, glucose and sucrose. These similar metabolites are also responsible for the PCA separation into five clusters representing the harvest age at 14, 16, 18, 20, 22 weeks of leaves extract. Loading plots revealed that most of the ANDRO and NAG signals were present when the plant reached at the pre-flowering stage or 18 weeks after sowing (WAS). As a conclusion, A. paniculata young leaves at pre-flowering harvest age were found to be richer in ANDRO, NAG and DAG compared to mature leaves while glucose and choline increased with harvest age. Therefore, young leaves of A. paniculata should be harvested at 18 WAS in order to produce superior quality plant extracts for further applications by the herbal, nutraceutical and pharmaceutical industries.

Quality Evaluation of Andrographis paniculata Capsules Based on Rapid and Accurate LC-ESI-MS/MS Assay of Three Diterpenoids.

Andrographis paniculata is an important traditional Chinese herbal material widely used for treatment of fever and diarrhea. The three diterpenoids [andrographolide (AP), dehydroandrographolide (DP) and neoandrographolide (NP)] in A. paniculata were characterized as the key active components for its extractive and related medicinal preparations. A rapid and accurate quantitative analysis of these three diterpenoids in A. paniculata capsules was accomplished by means of a valid and reliable high-performance liquid chromatography with tandem mass spectrometric (HPLC-MS/MS) analytical method. An optimized gradient elution method, which under the mobile phase consisting of methanol and water, was established successfully. An XDB C18 column (3.5 µm, 2.1 mm × 50 mm) was successfully applied to separate these three diterpenoids at the flow rate of 0.55 mL/min. A triple quadrupole mass spectrometer with multiple reaction monitoring (MRM) mode using an electrospray ionization source was served for analytical detection. The linearity, specificity, accuracy, recovery, precision, stability and repeatability were demonstrated to fully confirm the validity of this study. Good linear regression relationships for the three analytes were obtained over the range of 0.50-1000 ng/mL. Intra-day and inter-day precisions were evaluated with relative standard deviation < 7.62%, and the extraction recovery varied from 93.8% to 102.0%. The rapid and accurate HPLC-MS/MS analytical method was developed for the quality evaluation of A. paniculata capsules successfully. The content of AP, DP and NP obtained were 3.90-4.08, 4.77-5.04, 4.32-4.48 mg/g in A. paniculata capsules, respectively. Moreover, it is expected that this analytical technique can be applied to detect three effective components rapidly and accurately in other different medicinal preparations of A. paniculata as a method for their quality control.

Determination of Bitterness of Andrographis Herba Based on Electronic Tongue Technology and Discovery of the Key Compounds of Bitter Substances.

Andrographis Herba (AH), the dry aerial segments of Andrographis paniculata (Burm.f.) Nees, is a common herbal remedy with bitter properties in traditional Chinese medicine (TCM) theory. Although bitterness is one of the features representing Chinese medicine, it has not been implemented as an index to assess the quality and efficacy of TCM because of peoples' subjectivity to taste. In this study, 30 batches of AH with different commercial classifications (leaves, stems, or mixtures of both) were collected. Bitterness of AH was quantified by electronic tongue technology. Meanwhile, chemical compositions were characterized through establishing high-performance liquid chromatography fingerprints. The result indicated that the radar curves of the bitterness from different AH commercial classifications displayed different taste fingerprint information. Based on six taste factors, a Principal Component Analysis (PCA) score three-dimensional (3D) plot exhibited a clear grouping trend (R²X, 0.912; Q², 0.763) among the three different commercial classifications. Six compounds (Peaks 2, 3, 4, 6, 7, 8) with positive correlation to bitterness were discovered by a Spearman correlation analysis. Peaks 2, 6, 7, 8 were identified as andrographolide, neoandrographolide, 14-deoxyandrographolide, and dehydroandrographolide, respectively. The electronic tongue can be used to distinguish AH samples with different commercial classifications and for quality evaluation.

Monitoring of sixteen fragrance allergens and two polycyclic musks in wastewater treatment plants by solid phase microextraction coupled to gas chromatography.

A methodology based on headspace solid phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS) determination was developed for the monitoring and evaluation of the removal efficiency of 16 common fragrance allergens and two polycyclic musks in wastewater treatment plants (WWTPs). An experimental design with a full factorial model was applied to evaluate the effects of the experimental parameters on the extraction (e.g., salt content, time and extraction temperature). After determining the optimum conditions (2.4 g NaCl, 45 min at 90 °C), an external calibration was performed and quality parameters of the proposed method were evaluated. Method detection limits in the range of 0.01-1.7 µg L(-1) were obtained. Satisfactory inter-day precision values between 4% and 23% (n=5) were obtained for most compounds. The method was applied to the monitoring of the target analytes in samples from two WWTPs. Seven target compounds were detected at the primary effluent of both plants at µg L(-1) levels. Limonene, linalool and eugenol were quantitatively eliminated during the secondary treatments of both WWTPs, while lilial, benzyl salicylate, galaxolide, and tonalide were still detected at the effluent waters.

[Research on UPLC-PDA fingerprint of andrographis paniculata and quantitative determination of 4 major constituents].

Andrographis paniculata from different parts and origins were analyzed by UPLC-PDA fingerprint to provide refererice for related preparation technology. Using the peak of andrographolide as reference, 27 common peaks were identified, and digitized UPLC-PDA fingerprints for 23 batches of andrographis paniculata were established in this research. Principal component analysis (PCA) was carried out after feature extraction. The contents of andrographolide, neoandrographolide, deoxyandrographolide, dehydroandrographolide were determined by external standard method. The Plackett-Burman design combined with pareto chart was used to analyze the factors influencing the robustness of the method. It was found that the medicinal part has a more remarkable influence on the quality of andrographis paniculata than the origin. The contents of the 4 lactones the differ greatly in the different parts of andrographis paniculata, and the pH of the mobile phase is an important factor that influenced the robustness of the method.

Extraction of three bioactive diterpenoids from Andrographis paniculata: effect of the extraction techniques on extract composition and quantification of three andrographolides using high-performance liquid chromatography.

Andrographis paniculata (Burm.f.) wall.ex Nees (Acanthaceae) or Kalmegh is an important medicinal plant finding uses in many Ayurvedic formulations. Diterpenoid compounds andrographolides (APs) are the main bioactive phytochemicals present in leaves and herbage of A. paniculata. The efficiency of supercritical fluid extraction (SFE) using carbon dioxide was compared with the solid-liquid extraction techniques such as solvent extraction, ultrasound-assisted solvent extraction and microwave-assisted solvent extraction with methanol, water and methanol-water as solvents. Also a rapid and validated reverse-phase high-performance liquid chromatography-diode array detection method was developed for the simultaneous determination of the three biologically active compounds, AP, neoandrographolide and andrograpanin, in the extracts of A. paniculata. Under the best SFE conditions tested for diterpenoids, which involved extraction at 60°C and 100 bar, the extractive efficiencies were 132 and 22 µg/g for AP and neoandrographolide, respectively. The modifier percentage significantly affected the extraction efficiency.

[Study on chemical constituents of Drosera peltata var. multisepala].

Chemical investigatation of Drosera peltata var. multisepala led to the isolation of eleven compounds using various chromatographic techniques. The structures of these compounds were elucidated as isoshinanolone-4-O-beta-D-glucoside (1), isoshinanolone (2), epi-isoshinanolone (3), plumbagin (4), droserone (5), droserone-5-O-glucoside (6), quercetin (7), kaempferol (8) , gossypetin-8-O-glucoside (9), 3,3'-dimethoxy ellagic acid (10), and ellagic acid (11) by their physicochemical properties and spectral data analysis. Compound 1 was a new compound. Compounds 3, 8, 10, and 11 were isolated from this plant for the first time.

Quantitative analysis of four major diterpenoids in Andrographis paniculata by 1H NMR and its application for quality control of commercial preparations.

A quantitative proton nuclear magnetic resonance technique (qHNMR) has been successfully introduced to quantify andrographolide, dehydroandrographolide, deoxyandrographolide and neoandrographolide in Andrographis paniculata, a commonly used important traditional Chinese medicine. Creative use of trifluoroacetic acid-d, which satisfactorily resolved the overlapping signals of these compounds in crowded regions of δ 4.5-5.6 ppm in (1)H NMR spectrum, made their quantification possible. Optimization of other experimental conditions, including internal standard, NMR pulse sequence, and NMR relaxation delay time, finally established the (1)H NMR based quantification approach, which was validated with satisfactory accuracy, precision, repeatability, and recovery. Except for deoxyandrographolide and neoandrographolide in two compound recipes, this method was successfully applied to quantify the four major components in fourteen raw herb materials and five commercial preparations, providing quantification results in good agreement with those determined by HPLC. The inherent advantages of qHNMR, such as its rapidity and simplicity, make itself a feasible alternative to HPLC for the quality control of A. paniculata raw material and herbal preparations.

In vitro biological activities of the essential oil from the 'resurrection plant' Myrothamnus moschatus (Baillon) Niedenzu endemic to Madagascar.

The essential oil of the 'resurrection plant' Myrothamnus moschatus (Baillon) Niedenzu endemic to Madagascar, where it is used in traditional medicine, was investigated for the first time for some biological activities. GC-FID and GC-MS analyses revealed trans-pinocarveol (35.6%), pinocarvone (20.0%), ß-selinene (8.5%) and perillyl acetate (6.0%) as the major essential oil constituents. The oil strongly inhibited MDA-MB 231 (human breast adenocarcinoma) cells, with an IC(50) of 15 µg mL(-1), and the growth of Candida albicans, with an inhibition diameter of 18 mm, while its antioxidant activity, measured by DPPH and ABTS assay, proved to be negligible.

Chemical composition and antioxidant activity of the essential oil of aerial parts of Petasites albus from Iran: a good natural source of euparin.

The chemical composition of the essential oil of Petasites albus (known as 'Baba Adam' in Iran) was investigated by capillary gas chromatography and gas chromatography-mass spectrometry for the first time. Twenty components were identified, accounting for 99.7% of the oil composition. The major compounds were euparin (73%), α-eudesmol (13.2%) and ß-selinene (4.5%). Euparin, the main component of the essential oil, was isolated and characterised by spectroscopic techniques. The antioxidant activities of the essential oil and euparin were evaluated by using the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay and are shown to exhibit a moderate antioxidant activity.

[Analysis of the chemical constituents of essential oil from Anaphalis aureopunctata by GC-MS].

OBJECTIVE: The chemical components of essential oil from Anaphalis aureopunctata were analyzed by GC-MS. METHODS: Essential oil was extracted by stem distillation (SD). The chemical components of essential oil were analyzed by GC-MS. RESULTS: The main components in the oil were Eudesma-4(14), 11-diene, alpha-Bisabolol,6,10,14-trimethyl- 2-Pentadecanone,Caryophyllene oxide,alpha-Selinene and so on. CONCLUSION: This is the first time to adopt GC-MS to analyze the chemical components of volatile oil of Anaphalis au-reopunctata, and this study can provide science basis for further research development of Anaphalis aureopunctata.

Antimicrobial activity of confertifolin from Polygonum hydropiper.

Confertifolin (6,6,9a-trimethyl-4,5,5a,6,7,8,9,9a-octahydronaphtho[1,2-c] furan-3 (1H)-one) was isolated from the essential oil of Polygonum hydropiper L. (Polygonaceae) leaves using column chromatography. Confertifolin showed activity both in bacteria and fungi. The lowest MIC for bacteria was observed against Enterococcus faecalis (31.25 microg/mL). Significant MIC for fungi was observed against Scopulariopsis sp (7.81 microg/mL), Curvularia lunata (7.81 microg/mL), Epidermophyton floccosum (7.81 microg/mL), Trichophyton mentagrophytes (16.62 microg/mL), Trichophyton rubrum (MTCC 296) (16.62 microg/mL), Aspergillus niger (31.25 microg/mL), Botrytis cinerea (31.25 microg/mL) Magnaporthe grisea (62.5 microg/mL), Trichophyton simii (125 microg/mL) and Trichophyton rubrum (clinical isolate) (125 microg/mL).

[Study on lignans of Polygonum capitatum].

OBJECTIVE: To study the lignan of Polygonum capitatum. METHODS: The compounds were isolated and purified by using of SiO2, Sephadex LH-20 column chromatography and pre-HPLC. The structures of the compounds were determined on the basis of their physical properties and the spectral data. RESULTS: Four compounds were isolated and identified as schizandriside (1), (-)-isolariciresinol-2a-O-beta-D-xylopyranoside (2), (-) -5 '-methoxyisolariciresinol-2a-O-beta-D-xylopyranoside (3) and nudiposide (4). CONCLUSION: Compounds 1 - 4 are isolated from the genus for the first time.

Composition of the essential oil of Helichrysum chasmolycicum growing wild in Turkey.

The chemical compositions of the essential oil obtained from the aerial parts of Helichrysum chasmolycicum were analyzed by gas chromatography and gas chromatography-mass spectrometry. From the 57 identified constituents, representing 66.55% of the oil, the main constituents of the oil were beta-caryophyllene (27.6%), beta-selinene (8.9%), alpha-selinene (8.4%), caryophyllene oxide (7.3%), and carvacrol (2.4%). The essential oil was almost totally characterized by sesquiterpene hydrocarbons such as beta-caryophyllene and alpha- and beta-selinene.

Total assignments of 1H and 13C NMR spectra of isocatalpanol and a derivative of tecomaquinone.

Isocatalpanol and tecomaquinone I were obtained from roots of Lippia sidoides, a medicinal plant from northeast Brazil. Reduction of tecomaquinone I with NaBH4 yielded a new derivative. Structural elucidation was done on the basis of spectral data, mainly by high-field NMR and electron ionization mass spectrometry.

Determination and variation of three active diterpenoids in Andrographis paniculata (Burm.f.) Nees.

Simple and rapid methods have been developed for the extraction and simultaneous determination of the three active diterpenoids, andrographolide (AP1), 14-deoxy-11,12-didehydroandrographolide (AP3) and neoandrographolide (AP4), from various samples of Andrographis paniculata (Burm.f.) Nees. Methanol extracts from the dried leaves, stems and crude products were analysed by isocratic HPLC using a methanol and water mobile phase with monitoring at 220 nm. There was a large variation of the three active diterpenoids in different A. paniculata products obtained from Thai markets. The results indicated that the amounts of these active compounds consumed, based on the recommended daily doses, from materials obtained from the different suppliers will be different. In addition, the stability of these three active compounds was also examined in dry herbs stored at room temperature. The results showed that andrographolide was more stable than the others. In contrast, the content of 14-deoxy-11,12-didehydroandrographolide increased and the neoandrographolide content fluctuated during storage time. The combination of different levels of these compounds in the source materials and the changes during storage could have a significant effect on the efficacy of this traditional herbal medicine in clinical treatment.

Identification and characterization of Podophyllum emodi by API-LC/MS/MS.

An API-LC/MS/MS method was developed for the identification of the medicinal herb Podophyllum emodi based on the profile of its aryltetrahydronapthalene and related lignan marker compounds. This was done by matching the structural information from the tandem mass spectrometric data with those lignan marker compounds already reported for the herb. The method could be employed in the absence of reference standards for the markers and was particularly useful in view of the scarcity of supply of these chemical standards. It has been used successfully to differentiate Podophyllum emodi from two commonly used medicinal herbs of a different genus but having similar appearance, Radix clematidis and Radix gentiana, as well as a closely related herb, Podophyllum peltatum.

Determination of the aryltetralin lignan content of podophyllum resins and roots/rhizomes.

A simple, quantitatively reproducible method for determining the aryltetralin lignan content of the resins and roots/rhizomes of Podophyllum hexandrum and P. peltatum is described. The method confirms that P. hexandrum resins and roots/rhizomes contain approximately four times the quantity of lignans as do those of P. peltatum and also that there is a significant variation in the lignan content of P. hexandrum resins.