Identification of sedative-hypnotic compounds shared by five medicinal Polyporales mushrooms using UPLC-Q-TOF-MS/MS-based untargeted metabolomics.

BACKGROUND: Five Polyporales mushrooms, namely Amauroderma rugosum, Ganoderma lucidum, G. resinaceum, G. sinense and Trametes versicolor, are commonly used in China for managing insomnia. However, their active components for this application are not fully understood, restricting their universal recognition. PURPOSE: In this study, we aimed to identify sedative-hypnotic compounds shared by these five Polyporales mushrooms. STUDY DESIGN AND METHODS: A UPLC-Q-TOF-MS/MS-based untargeted metabolomics, including OPLS-DA (orthogonal projection of potential structure discriminant analysis) and OPLS (orthogonal projections to latent structures) analysis together with mouse assays, were used to identify the main sedative-hypnotic compounds shared by the five Polyporales mushrooms. A pentobarbital sodium-induced sleeping model was used to investigate the sedative-hypnotic effects of the five mushrooms and their sedative-hypnotic compounds. RESULTS: Ninety-two shared compounds in the five mushrooms were identified. Mouse assays showed that these mushrooms exerted sedative-hypnotic effects, with different potencies. Six triterpenes [four ganoderic acids (B, C1, F and H) and two ganoderenic acids (A and D)] were found to be the main sedative-hypnotic compounds shared by the five mushrooms. CONCLUSION: We for the first time found that these six triterpenes contribute to the sedative-hypnotic ability of the five mushrooms. Our novel findings provide pharmacological and chemical justifications for the use of the five medicinal mushrooms in managing insomnia.

A tri-herb formulation protects against ethanol-induced mouse liver injury and downregulates mitogen-activated protein kinase phosphatase 1.

BACKGROUND: A tri-herb formulation comprising Ganoderma (the dried fruiting body of Ganoderma lucidum), Puerariae Thomsonii Radix (the dried root of Pueraria thomsonii) and Hoveniae Semen (the dried mature seed of Hovenia acerba) -GPH for short- has been using for treating liver injury; however, the pharmacological basis of this application of GPH is unknown. This study aimed to investigate the liver protective effects and mechanisms of action of an ethanolic extract of GPH (GPHE) in mice. METHODS: To control the quality of GPHE, the contents of ganodermanontriol, puerarin and kaempferol in the extract were quantified by ultra-performance liquid chromatography. An ethanol (6 ml/kg, i.g.)-induced liver injury ICR mouse model was employed to investigate the hepatoprotective effects of GPHE. RNA-sequencing analysis and bioassays were performed to reveal the mechanisms of action of GPHE. RESULTS: The contents of ganodermanontriol, puerarin and kaempferol in GPHE were 0.0632%, 3.627% and 0.0149%, respectively. Daily i.g. administration of 0.25, 0.5 or 1 g/kg of GPHE for 15 consecutive days suppressed ethanol (6 ml/kg, i.g., at day 15)-induced upregulation of serum AST and ALT levels and improved histological conditions in mouse livers, indicating that GPHE protects mice from ethanol-induced liver injury. Mechanistically, GPHE downregulated the mRNA level of Dusp1 (encoding MKP1 protein, an inhibitor of the mitogen-activated protein kinases JNK, p38 and ERK), and upregulated expression and phosphorylation of JNK, p38 and ERK, which are involved in cell survival in mouse liver tissues. Also, GPHE increased PCNA (a cell proliferation marker) expression and reduced TUNEL-positive (apoptotic) cells in mouse livers. CONCLUSION: GPHE protects against ethanol-induced liver injury, and this effect of GPHE is associated with regulation of the MKP1/MAPK pathway. This study provides pharmacological justifications for the use of GPH in treating liver injury, and suggests that GPHE has potential to be developed into a modern medication for managing liver injury.

Integration of sub-organ quantitative imaging LA-ICP-MS and fractionation reveals differences in translocation and transformation of CeO2 and Ce3+ in mice.

Information on the risk of exposure to cerium oxide (CeO2) nanoparticles (NPs) is limited. To assess risk, we must know where and how such NPs are distributed to the body after exposure, both short- and long-term. In this work, an integrated approach of quantitative LA-ICP-MS bioimaging and fractionation was employed to study the translocation and transformation of CeO2 and Ce3+ in mouse spleen and liver. The complementary information retrieved by the two techniques above on the accumulation of Ce and dissolution/aggregation were found consistent. In brief, a detailed fine scanning of a region of interest in the organ was performed after fast-screening at low spatial resolution. In the spleen, after short-term high-dose exposure, CeO2 NPs was found mainly in the marginal zone and caused an up-regulation of Zn in the white pulp. After long-term low-dose exposure, CeO2 was found in the marginal zone and white pulp. In the liver, CeO2 NPs were mainly distributed in the Kupffer cells and lobule periphery. The high spatial resolution LA maps of H&E-stained liver sections allowed imaging close to cell level; this enabled an estimation of Ce content in Kupffer cells. Furthermore, fractionation by ultrafiltration was also employed to differentiate the ionic and NP species in the organs. This fractionation showed aggregation of Ce ions in spleen, supporting the LA-ICP-MS results. Transmission electron microscopy revealed that long-term CeO2 exposure triggered an immune response to infection in the spleen and confirmed the differential deposition of Ce in the marginal zone. The integrated analyses based on ICP-MS together with histology and TEM investigation suggests that long-term low doses of CeO2 NPs may cause toxicity in the liver and impair functions of the immune system.

The evaluation on embryotoxicity of Dipsaci Radix with mice and embryonic stem cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Dipsaci Radix (DR), the dried root of Dipsacus asper Wall. ex Henry, has been used to treat pregnant disorders for thousands of years, and currently has been ranked as the first selective herb for prevention of miscarriage clinically; however, there is no sufficient evidence so far to assess its safety. The purpose of this study was to examine the impacts of DR aqueous extracts on embryonic development with mice and embryonic stem cells (ESCs). MATERIAL AND METHOD: In a segment II study, pregnant ICR mice were randomly assigned into 5 groups, i.e. mice were orally treated with DR aqueous extracts at dosages of 0 (distilled water, as negative controls (G1 group)), 2, 8, 32 g/kg/d (G2, G3, G4 group), and vitamin A (as positive controls (G5 group)) respectively. Maternal and embryo-fetal parameters were evaluated after cesarean section. The fetal skeletal development was further assessed with the alizarin red S and H&E staining and fluorescent imaging. Meanwhile, IC50 values for both ESCs and 3T3 cells were detected with MTT assays. RESULT: Compared to G1 group, the maternal body-weight in G3 and G4 groups was significantly lower (P<0.05-0.001), and the fetal malformation rate increased in G2, G3 and G4 groups as a dose-dependent manner, although a statistical significance was only reached in G4 group (P<0.001). The morphologic and histochemistry abnormalities of fetal skeletal development such as delayed osteogenesis and mineralization in the cartilaginous tissue were found after DR treatments (32 g/kg/d). There was no significant difference between IC50 ESC (6.826 ± 0.311 mg/ml) and IC50 3T3 (5.132 ± 0.142 mg/ml, P>0.05). CONCLUSION: DR aqueous extracts at the dosage of 8 or 32 g/kg/d (4.3 or 17.2 folds of recommended daily-dosage for adult human respectively) might cause adverse impacts in maternal healthy and embryo-fetal development. It suggests that high-dose and long-term administration of DR preparations should be unsafe to pregnant women.

Highly sensitive and selective organophosphate screening in twelve commodities of fruits, vegetables and herbal medicines by dispersive liquid-liquid microextraction.

The article describes a simple sample pretreatment procedure for the analysis of ten organophosphorus pesticides using dispersive liquid-liquid microextraction (DLLME) followed by gas chromatography-mass spectrometry (GC-MS) in three distinctively different types of matrices: fresh fruits, fresh vegetables and dried herbs. The method was carefully developed, focusing on the chemistry of various dispersive solvents, to achieve simultaneous, comprehensive extraction and preconcentration in a great span of selected matrices. According to matrix-matched validation study, the set of optimized DLLME conditions has been proven robust to determine target OPPs within a wide linear range from 0.1 to 1000 µg L(-1). With limited usage of organic extractants, remarkable enrichment factors up to 100-fold were obtained, enabling ultra-trace pesticide quantification down to sub-ppt levels at 0.12-4.92 ng kg(-1). Practical application of the method was illustrated by quantitative recovery (70-119%) and good precision (2.6-10% R.S.D.) in a representative range of three fruits and four vegetable commodities featured by the CODEX Alimentarius classification as well as their unique matrix compositions. A careful selection of dried herbs was further classified based on their morphological structures to validate analytical ruggedness of the method. Compared with existing methods for food analysis vis-à-vis OPPs, the present method is superior in terms of high sample throughput, minimal solvent consumption, and small sample size requirement. An additional, significant aspect of this universal DLLME method is that it models sample pretreatment methods with wide coverage of analytical matrices that are more effective, more comprehensive, and more flexible than those currently being used.

Development, optimization, and use of an APCI source with temperature-controlled vaporization of solid and liquid samples.

A trend is observed in mass spectrometry, in which solid samples without prior dissolution and chromatographic separation are brought directly into the ion source and are ionized, e.g., by corona discharge (Atmospheric Solids Analysis Probe) or plasma (Direct Analysis in Real Time). The Direct Inlet Probe-atmospheric-pressure chemical ionization (APCI) ion source presented here, which was coupled to a high-resolution quadrupole time-of-flight-mass spectrometer, differs from most of the other ion sources in having temperature-programmed heating of the sample. The resulting possibility to reduce ion suppression and ion-molecule reactions in the ion source was shown by the separation of two fatty acid methyl esters as a result of their boiling point difference. Using caffeine as sample, certain source parameters such as the auxiliary gas flow, the drying gas flow, and the position of the probe tip in the ion source were optimized. The ability to perform quantitative analyses was shown by the linear concentration response (R(2) = 0.9984) observed when analyzing different caffeine concentrations. An extract of a Chinese medicinal herb was used to examine the reproducibility (relative standard deviations of the most abundant m/z signals were ≤8.1 %). It was also possible to distinguish milled samples of Radix Angelicae sinensis and Radix Angelicae gigas from each other and to identify the coumarins they contain without sample preparation. Supplying synthetic air instead of nitrogen to the ion source makes APCI in the negative mode possible as well; this was proven by the analysis of n-nonyl-ß-D-maltoside.

Review of biological and pharmacological activities of the endemic Taiwanese bitter medicinal mushroom, Antrodia camphorata (M. Zang et C. H. Su) Sh. H. Wu et al. (higher Basidiomycetes).

Antrodia camphorata is an extremely rare fungus native to the forested regions of Taiwan. It is also a traditional Chinese medicine, and Taiwanese aborigines applied it for treating liver diseases and protecting from food and drug intoxication. Scientific studies have demonstrated that A. camphorata crude extracts and pure compounds possess a variety of beneficial functions, such as anti-hypertensive, anti-hyperlipidemic, anti-inflammatory, anti-oxidant, anti-tumor, and immuno-modulatory activities. Recent studies have shown that many of these biological and pharmacological activities can be attributed to various active constituents, including polysaccharides, terpenoids, steroids, lignans, benzoquinone derivatives, benzenoids, and maleic and succinic acid derivatives. A. camphorata has been considered as a novel phytotherapeutic agent. However, detailed mechanistic studies or even clinical trials on A. camphorata are still rare. With the help of modern analytical techniques, it is not surprising that many novel constituents are being identified or fractionated from A. camphorata mycelium and fruiting bodies. This review summarizes the latest published results from A. camphorata research, focusing on the biological and pharmacological activities of the crude extract and known constituents of A. camphorata.

Quantification of acylglycines in human urine by HPLC electrospray ionization-tandem mass spectrometry and the establishment of pediatric reference interval in local Chinese.

Urinary organic acids, plasma amino acids and acylcarnitine profile analyses are the main tools used to diagnose inborn errors of metabolisms (IEMs). However, without metabolic decompensation, these parameters are often not helpful. On the other hand, in cases of IEM, acylglycines are consistently raised even when patients appear to be in remission. This study aims to set-up a simple liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method for the determination of urine acylglycines, complementary to organic acid and acylcarnitine profiles, for the diagnosis of IEM. In addition, local reference intervals for various acylglycines are established by using this method. Acylglycines were isolated by solid-phase extraction, derivatized with n-butanol, separated by HPLC, and detected by ESI-MS/MS. Acylglycines were quantified with deuterated internal standards. Mean recoveries of acylglycines ranged from 90.2 to 109.3%. Within- and between-run imprecisions for all acylglycines have CVs less than 10%. Linear regression coefficients were greater than 0.99. Reference intervals were established according to CLSI guidelines by analyzing 204 samples from apparently healthy individuals less than 18 years of age. The distributions of AG in the "normal" urine were skewed towards the right. After log transformation, all the results were normally distributed. Partitioning into age group reference intervals was not indicated, according to the Harris and Boyd approach. In this context, a single reference interval for each acylglycine could be used. This method of urine acylglycines analysis is a powerful diagnostic tool, complementary to urine organic acids and plasma acylcarnitine profiling, for detecting certain inborn errors of metabolism.

The effects of Boehmeria nivea (L.) Gaud. on embryonic development: in vivo and in vitro studies.

AIM OF THE STUDY: Boehmeria nivea (L.) Gaud. was commonly used to treat miscarriages clinically. The aim of this study was to examine its safety for embryonic development. MATERIALS AND METHODS: Pregnant mice were randomly assigned into 5 groups, i.e. mice were oral-treated with distilled water (G1), with Boehmeria nivea extract of 2, 8 or 32 g/kg/day (G2, G3 or G4), and with 3 doses of vitamin A of 200,000 IU/kg as positive controls (G5). Meanwhile, IC(50) values for both embryonic stem cells (ESCs) and 3T3 cells were detected by cytotoxicity assays. RESULTS: (1) The resorptions and malformed fetuses in G5 were significantly higher than G1 (P<0.001), whereas the maternal body-weight and uterus-weight were lower than G1 (P<0.05); (2) there was no difference in the fetal body-weight, maternal relative body-weight gain, liver-, kidney- or heart-weight, relative organ-weight, and histological examination among five groups; (3) there was no difference in IC(50) values between ESCs and 3T3 cells, but high concentration of Boehmeria nivea extract might significantly lower ESCs' viability (P<0.05). CONCLUSION: Boehmeria nivea extract at 32 g/kg/day did not cause significant embryotoxicity or maternal toxicity in mice, although it might cause cytotoxicity in cultured ESCs at a high dose.

Development of a LC-MS/MS method for the determination of antrodin B and antrodin C from Antrodia camphorata extract in rat plasma for pharmacokinetic study.

A selective and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of antrodin B and antrodin C in rat plasma. Both target compounds, together with the internal standard (diazepam), were extracted from rat plasma samples by liquid-liquid extraction with ethyl acetate. Chromatographic separation was carried out on an Agilent XDB-C(8) column with an isocratic mobile phase consisting of acetonitrile and water (70:30, V/V) at a flow rate of 0.5 mL/min. The mass spectrometric detection was performed by selected reaction monitoring (SRM) mode via atmospheric pressure chemical ionization (APCI) source operating in positive ionization mode. The assay exhibited a linear dynamic range of 47.6-4760 ng/mL for antrodin B and 56.6-5660 ng/mL for antrodin C. The intra- and inter-day precision was less than 5.3% and the accuracy was less than 2.7% for both analytes. The validated method has been applied to the pharmacokinetic study of antrodin B and antrodin C in rats following oral administration of Antrodia camphorata extract.

Toxicity assessment of nine types of decoction pieces from the daughter root of Aconitum carmichaeli (Fuzi) based on the chemical analysis of their diester diterpenoid alkaloids.

Various processed types of FUZI (the daughter roots of the highly toxic plant Aconitum carmichaeli Debx, FZ) decoction pieces (the herbal materials processed according to the specifications of Chinese medicine manuals; " YINPIAN" in Chinese transliteration) are widely used in traditional medicine to treat various diseases, but their toxicities are not known. Nine types of FZ decoction pieces, including one raw slice and eight processed forms, were therefore prepared, each in 7 to 10 batches, to assess for their toxicity. Altogether 84 FZ samples were quantified on the amount of highly toxic diester diterpenoid alkaloids, i.e., aconitine, mesaconitine and hypaconitine by a newly developed HPLC method with HPLC-DAD and LC-MS techniques. The comparison of the processed FZ to raw slices of the root showed that the amount of each analyte in the processed FZ was drastically decreased. The sum of the three toxic compounds in the 8 types of processed FZ was only 3.91-34.80 % of this value in the FZ raw slice. This implies that the toxicity of processed FZ was decreased significantly. The amounts of toxic components in the 8 types of processed FZ varied significantly, often by a power of ten, indicating that the dosage of these herbs, when prescribed for clinical uses, should be cautiously set in order to avoid poisoning incidents.

Improved quality assessment of proprietary Chinese medicines based on multi-chemical class fingerprinting.

Our present study constitutes the first successful attempt to employ eight distinctive chemical groups of compounds for the quality evaluation of a complex traditional Chinese herbal medicine (TCHM) material: bazhen yimu (BZYM). Due to the complexity of its matrix, which is composed of nine different herbal ingredients, five representative chemical groups encompassing representative bioactive markers were initially chosen as targets for the quality assessment of this preparation. Furthermore, with the aid of LC-ESI-MS, three additional chemical groups were characterized. In summary, a total of nineteen markers belonging to eight different chemical groups were selectively displayed in the chromatographic fingerprint of BZYM preparation. With this fingerprint, the overall quality of any BZYM preparation can be comprehensively authenticated. The chromatographic separation was performed on an HP C(18)AQ column with a gradient elution of ACN and aqueous solution containing 0.1% phosphoric acid at the optimal detection wavelength of 230 nm. The established method was rigorously validated with respect to the ICH guidelines and represents the most extensive and facile HPLC quality control technique for this formulation. Compared with the conventional method of using a single or only a few markers of the same chemical group, this technique provides a new dimension for TCHM quality control.

Simultaneous quantification of eight bioactive components of Houttuynia cordata and related Saururaceae medicinal plants by on-line high performance liquid chromatography-diode array detector-electrospray mass spectrometry.

An on-line high performance liquid chromatography (HPLC)-diode array detector (DAD)-electrospray ionization mass spectrometry (ESI-MS) method has been developed to quantify simultaneously eight bioactive chemical components in Houttuynia cordata Thunb and related Saururaceae medicinal plants. Simultaneous separation of these eight compounds was achieved on a C(18) analytical column with gradient elution of acetonitrile and 0.2% acetic acid (v/v) at a flow rate of 0.6 mL/min and being detected at 280 nm. These eight compounds were completely separated within 90 min. Good linear regression relationship (r(2)>0.9978) within test ranges was shown in all calibration curves. Good repeatabilty for the quantification of these eight compounds in H.cordata was also demonstrated in this method, with intra- and inter-day variations less than 3.0%. The method established was successfully applied to quantify eight bioactive compounds in closely related species of H.cordata, which provides a new basis for quality assessment of H.cordata.

Improved chromatographic assessment of Chinese medicinal products by multi-chemical classes analysis.

An improved strategy for the quality evaluation of proprietary Chinese medicines (PCM) in complex matrices is presented. It involves the simultaneous separating and displaying of multi-chemical classes and analytes within a single chromatographic elution. The developed fingerprints have the properties of better coverage of multi-chemical classes among the composing herbs. This comprehensive and more facile approach was illustrated for Lemai Keli, a commonly used PCM listed in the Chinese Pharmacopoeia with highly complex analytical matrices. Ten marker compounds belonging to monoterpene glycosides, chalcone glycosides, phenolic acids and phenolic carboxylic aldehyde groups of the analytes together with three common chromatographic peaks were successively separated and displayed within the chromatographic window as a unique HPLC-PAD fingerprint. Rigorous method validation data demonstrated the suitability of the current method for quality assessment purposes in a highly complex PCM matrix. Additional chemometric treatment further demonstrated the uniqueness of the developed fingerprint, with a distinctive chemical profile compared to a highly similar PCM formulation (Kangen Karyu) from which it was considered to be impossible to differentiate when using the conventional single marker approach.

Improved chromatographic fingerprints for facile differentiation of two Ganoderma spp.

This paper addresses a comprehensive and comparative study of six phytochemical extraction methods for triterpenes from the fruiting body of Ganoderma spp. Quantitative analysis of extracts was performed by HPLC with photodiode array detection. In general, pressurized liquid extraction and microwave-assisted extraction under optimized conditions produce better yields, and the former also significantly reduces the total time of extraction and manipulation of a sample, as well as the amount of solvent used in comparison with conventional soxhlet, reflux, ultrasonic, and methanol-CO(2) supercritical fluid extractions. Based on the improved extraction protocol, the fingerprinting profiles for two species of Lingzhi were established using the consistent chromatographic features of 12 authentic samples. Eleven common peaks of ganoderic/ganoderenic acids were identified using LC-ESI-MS-MS. These specific triterpene groups were adopted as chemical markers for Lingzhi. Using chemometric analysis, the developed fingerprinting was successfully applied to differentiate between the two species under the Ganoderma genus and is applicable as a method for quality evaluation of this valuable medicinal fungus and its related proprietary products.

Comparative study on the aristolochic acid I content of Herba Asarifor safe use.

Herba Asari (Xixin, Manchurian Wildginger, Asarum spp.) is a traditional Chinese medicinal herb commonly used as a crude drug and an ingredient in patent medicines. The herb contains aristolochic acid I (AA-I), which has recently caused several incidents of poisoning in Hong Kong. Therefore, the safe use of Asarum is questionable. The present study was undertaken to assess the levels of AA-I using liquid chromatography-mass spectrometry (LC/MS) indifferent medicinal parts of Herba Asari and some proprietary Chinese medicines (PCM) containing it as an ingredient. The AA-I content in the aerial and root portions were compared, in the form of water and methanolic extracts. The results showed that all the aerial portions of Herba Asari generally contain higher levels of AA-I than the roots (in water extract: 0.0870.06 microg/g of root and 0.3270.021 microg/g of aerial), and the methanolic extracts typically contained more AA-I than the water extracts. Moreover, all the three PCM studies showed negligible amounts of AA-I(containing 0.0370.006 microg/g). Therefore, the root portion of Herba Asari was recommended for prescription as a decoction instead of grinding it into powder for oral administration.

Optimization and validation of a chromatographic method for the simultaneous quantification of six bioactive compounds in Rhizoma et Radix Polygoni Cuspidati.

A reverse-phase HPLC method was developed for simultaneous quantification of six bioactive compounds in Rhizoma et Radix Polygoni Cuspidati. These compounds--polydatin (1), resveratrol (2), rhein (3), emodin (4), chrysophanol (5) and physcion (6)--were analysed from 24 authentic samples of the herb using UV HPLC. Based on the UV absorption characteristics of the six compounds, absorption wavelengths of 306 nm were chosen to quantify compounds 1 and 2, and 290 nm for compounds 3-6. A reliable and reproducible quantitative HPLC method for analysing authentic samples of Rhizoma et Radix Polygoni Cuspidati from different cultivation regions was developed. The results showed that the concentration of compound 1 in samples from Sichuan was almost 2 fold higher than that of samples acquired in Guangxi. Furthermore, compounds 3 and 5 were not found in all the samples tested. Thus, instead of using polydatin (1) and emodin (4) as markers for quality assessment, as in conventional practice, these findings show that compounds 2 and 6 are more suited to act as marker compounds for a more specific assessment of the quality of this herb.

Quality assessment of Rhizoma et Radix Notopterygii by HPTLC and HPLC fingerprinting and HPLC quantitative analysis.

This paper describes an improved quality assessment method for Rhizoma et Radix Notopterygii (the rhizome and root of Notopterygium incisum Ting ex H.T. Chang or Notopterygium forbesii Boiss). The method was established by using fingerprinting and quantitation of marker compounds (isoimperatorin, notopterol and bergapten) in this herbal medicine. The authentication of Rhizoma et Radix Notopterygii using high performance thin-layer chromatography (HPTLC) fingerprinting was achieved by comparing the colors and Rf values of the bands in TLC fingerprints with those of the marker compounds. The HPLC fingerprints of 16 batches of herbal samples from different regions of China showed similar chromatographic patterns. Five peaks were selected as characteristic peaks, and three of these were identified by using LC-MS-MS techniques. The relative retention times of these characteristic peaks in the HPLC fingerprint were established as an important parameter for identification of Rhizoma et Radix Notopterygii. Finally, the pharmacologically active marker compounds isoimperatorin, notopterol and bergapten in this herb were quantitatively determined using a validated reverse-phase HPLC method.

[Orthogonal experiment using SFE-CO2 in extraction of essential oil from fresh Houttuynia cordata and analysis of essential oil by GC-MS].

OBJECTIVE: To optimize the extraction procedure of essential oil from H. cordata using the SFE-CO2 and analyze the chemical composition of the essential oil. METHOD: The extraction procedure of essential oil from fresh H. cordata was optimized with the orthogonal experiment. Essential oil of fresh H. cordata was analysed by GC-MS. RESULT: The optimize preparative procedure was as follow: essential oil of H. cordata was extracted at a temperature of 35 degrees C, pressure of 15,000 kPa for 20 min. 38 chemical components were identified and the relative contents were quantified. CONCLUSION: The optimum preparative procedure is reliable and can guarantee the quality of essential oil.

Application of atmospheric pressure chemical ionisation mass spectrometry in the identification and differentiation of Panax species.

An HPLC-MS method using an atmospheric pressure chemical ionisation (APCI) source has been developed to assist in the differentiation of three ginseng species: Panax quinquefolium (American ginseng), P. ginseng (Chinese ginseng) and P. notoginseng (sanqi) species. The differentiation method relies on the identification of ginsenosides Rf and F11 and notoginsenoside R1. R1 is observed in both P. notoginseng and Chinese ginseng, whilst F1, is found exclusively in the American species. The presence of these compounds permits the definitive identification of the species to be made. The APCI ionisation source has been employed to tackle the matrix interference in analysing Chinese medicinal materials and to minimise the associated matrix effects that are commonly encountered with other ionisation modes. Moreover, the method allows direct interface to conventional HPLC systems. More importantly, chemical reference standards of ginsenosides are not required in this method. This technique provides an alternative approach to analysing high molecular weight polar compounds that typically encountered in complex matrices of Chinese medicinal materials.