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Circadian rhythm disorder (CRD) in space flight can lead to memory impairment, performance decrements and adverse health outcomes, the main manifestations of which are circadian desynchronization, sleep loss and insomnia. Sleep deprivation (SD) provide the means to evaluate these effects and the risks associated with CRD on ground. Gastrodia elata Blume (GEB) has beneficial effects on the treatment of sleep disturbances and memory loss. Fresh GEB (FG), an unprocessed raw tuber of GEB, has been used as functional health food in Asian countries for a long time. However, the research report of FG to ameliorate memory impairment caused by insomnia or lack of sleep is meager. In this study, ICR male mice were sleep-deprived continuously and water extract of FG (WFG) was orally administrated (3 and 9 g/kg/d, i.g) during the SD process lasted for 25 days, except control and model groups gavage administration with water, positive control group with modafinil (MOD, 0.1 g/kg/d, i.g). We studied the effect of WFG on CRD-induced learning and memory impairment using a set of behavioral analyses including the object location recognition test (OLRT), novel object recognition test (NORT), and the passive avoidance test (PAT). In addition, oxidative stress parameters were assessed by measuring the malondialdehyde (MDA) and superoxide dismutase (SOD) reactivity in serum and hippocampus. Our results revealed that SD decreased discrimination index (DI) in OLRT and NORT, with shorter latency into the dark chamber in PAT. Both WFG and MOD treatment can reverse these changes (P0.05). We concluded that WFG treatment improve CRD-induced learning and memory impairment and oxidative stress damage which makes FG a promising candidate as herbal health product of memory decline in CRD.

Free and conjugated forms of ferulic acid (FA) are generally available in higher plant taxa such as Chinese Angelica (CA, the roots of Angelica sinensis (Oliv.) Diels). These various forms of FA were found readily inter-convertible and the extractable level of each depended on solvent properties and acidity. Extraction efficiency using various pH solutions namely, water, 70% methanol, methanol-formic acid (95:5) and methanol-2% NaHCO3 in water (95:5) was compared. Extractable FA were found varying in samples under neutral solvents extraction, whilst relatively consistent for slightly acidic and alkali solvents which were therefore chosen as the optimal media to extract and determine the reproducible levels of free and total available FA. An accurate and rapid high performance liquid chromatographic (HPLC) analysis was conducted using an Alltima C18 column (5μm, 4.6mm i.d.×250mm) with a guard column (C18, 5μm, 4.6mm i.d.×7.5mm) at 30℃, eluted with a mixture of 1.0% acetic acid and acetonitrile in a gradient program at a flow rate of 1.0mL/min and detected at 320nm. Altogether 20 different types of CA samples including whole root, root head, rootlets, whole root slice, Angelica processed by Chinese yellow wine, and charred Angelica were quantified for free and total available FA. Total available FA was found more abundant than free counterpart with an average ratio of 3.15 (n=20) in the range of 1.29 to 8.23 for these CA samples. The extraction protocol was proven reliable to quantitatively convert all conjugated FA into its free forms and thereby accurately determined by HPLC method for quality assessment.

The combined photocatalysis and ozonation (UV/Ag-TiO2+O3) process has been investigated for its potential use for ballast water treatment by usingDunaliella salina(D. salina) as an indicator microorganism. Inactivation curves were obtained, and the mechanism was studied. Compared to individual unit processes, the combined UV/Ag-TiO2+O3tests yield enhanced inactivation efficiency. Scanning electron microscopy (SEM) measurements showed that presence of O3during UV/Ag-TiO2exposure expedited the destruction of cell structure, which was further confirmed by the formation of malondialdehyde (MDA) and decrease in chlorophyll-a (chl-a) concentration. The results suggest that UV/Ag-TiO2+O3was efficient for inactivation ofD. salinain simulated ballast water, and the accelerated destruction of cell structure was an important reason for the enhanced inactivation efficiency.

In natural product research, it is a common experience that fractionation of biologically-active crude extracts can lead to the loss of their original activity. This is attributed to synergistic effects, where two or more components are required to be present together for full activity of the sample. Our previous study showed that a volatile oil of Angelica sinensis radix (VOAS) inhibited endothelial cell proliferation in culture. Here we have used a bioactivity-guided fractionation method to preserve any synergistic effects of VOAS combining countercurrent chromatography (CCC), the MTS cell viability assay and gas chromatography (GC). Using a two-phase CCC solvent system (heptane–ethyl acetate–methanol–water at a volume ratio of 27:23:27:23%), forty-five fractions were isolated, nine of which exhibited anti-endothelial properties. GC analysis showed two bioactive alkylphthalides, Z-ligustilide and n-butylidenephthalide (BP) were the major compounds detected in the bioactive fractions, and were absent in non-bioactive fractions. Our results indicate that Z-ligustilide and BP are the main constituents responsible for the anti-endothelial properties of VOAS. This rapid and reliable approach in preserving sample activity while isolating and identifying its active compounds suggests that this protocol can be a powerful tool for drug discovery from natural products.

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.

Objective To isolate, incubate, and identify 4-chlorophenol-degrading complex bacteria, determine the tolerance of these bacteria to phenolic derivatives and study their synergetic metabolism as well as the aboriginal microbes and co-metabolic degradation of mixed chlorophenols in river water. Methods Microbial community of complex bacteria was identified by plate culture observation techniques and Gram stain method. Bacterial growth inhibition test was used to determine the tolerance of complex bacteria to toxicants. Biodegradability of phenolic derivatives was determined by adding 4-chlorophenol-degrading bacteria in river water. Results The complex bacteria were identified as Mycopiana, Alcaligenes, Pseudomonas, and Flavobacterium. The domesticated complex bacteria were more tolerant to phenolic derivatives than the aboriginal bacteria from Qinhuai River. The biodegradability of chlorophenols, dihydroxybenzenes and nitrophenols under various aquatic conditions was determined and compared. The complex bacteria exhibited a higher metabolic efficiency on chemicals than the aboriginal microbes, and the final removal rate of phenolic derivatives was increased at least by 55% when the complex bacteria were added into river water. The metabolic relationship between dominant mixed bacteria and river bacteria was studied. Conclusion The complex bacteria domesticated by 4-chlorophenol can grow and be metabolized to take other chlorophenols, dihydroxybenzenes and nitrophenols as the sole carbon and energy source. There is a synergetic metabolism of most compounds between the aboriginal microbes in river water and the domesticated complex bacteria. 4chlorophenol-degrading bacteria can co-metabolize various chlorophenols in river water.

The herbal materials of Asian ginseng (the root of Panax ginseng ), American ginseng (the root of Panax quinquefolius ) and Notoginseng (the root of Panax notoginseng ) were differentiated by conventional Fourier transform infrared spectroscopy (1D-FTIR) and two-dimensional (2D) correlation FTIR applying a thermal perturbation. Altogether 30 samples were collected and analyzed. Their entire 1D-FTIR spectra in the range of 4000–400 cm −1 and 2D-FTIR spectra in the region of 850–1530 cm −1 were generally similar based on the peaks position and intensities. This indicated the chemical constituents in these species of herbs were not distinctively different. However, variation in peak intensity were observed at about 1640 cm −1 , 1416 cm −1 , 1372 cm −1 and 1048 cm −1 in the 1D-FTIR spectra among these species for their ease differentiation. Clustering analysis of 1D-FTIR showed that these species located in different clusters. Much difference in their second derivative FTIR pattern among the three species also provided information for easy differentiation. These species of herbs were further identified based on the positions and intensities of relatively strong auto-peaks, positive or negative cross-peaks in their 2D-FTIR spectra. The findings provide a rapid and new operational procedure for the differentiation of these notable herbs. The visual and colorful 2D-FTIR spectra can provide dynamic structural information of chemical components in analyte and demonstrated as a powerful and useful approach for herbs identification.

A highly precise and accurate method, based on high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS), was developed for the qualitative and quantitative comparison of the main constituents in the rhizome of Ligusticum chuanxiong (LC) and three related umbelliferous medicinal plants. A comprehensive validation of the developed method was conducted, and the method was highly sensitive, reproducible and accurate. The unique properties of the present method were validated by analyzing 20 related herbal samples including 5 LC samples, 5 Cnidium officinale samples (CO), 5 Angelica sinensis samples (AS) and 5 Angelica acutiloba samples (AA). Twelve compounds including phenolic constituents, alkylphthalides and phthalide dimers were identified by online ESI-MS and by comparison with literature data and standard compounds, and six of them were quantified by HPLC-DAD simultaneously. The results demonstrated that identical compound types were identified as the main constituents of LC, CO, AS and AA herbs. The results also support the alternative application of these medicinal plants in Chinese and Japanese folk medicines. In the present study, it was found that the variation in the abundance of senkyunolide A was significant in these related herbs; it is therefore feasible to choose senkyunolide A as a characteristic compound for quality evaluation and chemical authentication of these herbs.

A high-performance liquid chromatographic (HPLC) fingerprint of Chinese Angelica (CA) was developed basing on the consistent chromatograms of 40 CA samples (Angelica sinensis (Oliv.) Diels). The unique properties of this HPLC fingerprints were validated by analyzing 13 related herbs including 4 Japanese Angelicae Root samples (JA, A. acutiloba Kitagawa and A. acutiloba Kitagawa var. sugiyame Hikino), 6 Szechwan Lovage Rhizome samples (SL, Ligusticum chuanxiong Hort.) and 3 Cnidium Rhizome samples (CR, Cnidium officinale Makino). Both correlation coefficients of similarity in chromatograms and relative peak areas of characteristic compounds were calculated for quantitative expression of the HPLC fingerprints. The amount of senkyunolide A in CA was less than 30-fold of that in SL and CR samples, which was used as a chemical marker to distinguish them. JA was easily distinguished from CA, SL and CR based on either chromatographic patterns or the amount of coniferyl ferulate. No obvious difference between SL and CR chromatograms except the relative amount of some compounds, suggesting that SL and CR might have very close relationship in terms of chemotaxonomy. Ferulic acid and Z-ligustilide were unequivocally determined whilst senkyunolide I, senkyunolide H, coniferyl ferulate, senkyunolide A, butylphthalide, E-ligustilide, E-butylidenephthalide, Z-butylidenephthalide and levistolide A were tentatively identified in chromatograms based on their atmospheric pressure chemical ionization (APCI) MS data and the comparison of their UV spectra with those published in literatures.

Activity of Chinese Danggui (DG), the processed root of Angelica sinensis (Oliv.) Diels, is linked to the ferulic acid content but the stability of ferulic acid during extraction for medicinal use is not known. The stabilities of ferulic acid and coniferyl ferulate were evaluated in the extracts of DG using a variety of extraction solvents. These included various combinations and proportions of methanol, water, formic acid, 1 M aqueous hydrochloric acid and 2% sodium hydrogen carbonate (NaHCO3) in water. Coniferyl ferulate was found liable to hydrolyze into ferulic acid in neutral, strongly acidic and basic solvents, where heat and water could facilitate this hydrolysis. However, the hydrolysis was relatively resisted in weakly organic acid. Based on the stability evaluation, two new terms, namely: free ferulic acid and total ferulic acid, were suggested and defined. Free ferulic acid refers to the natural content of ferulic acid in herbs. Total ferulic acid means the sum of free ferulic acid plus the amount of related hydrolyzed components. Meanwhile, the high-performance liquid chromatographic (HPLC) method was developed to assay free ferulic acid and total ferulic acid in DG using methanol-formic acid (95:5) and methanol-2% NaHCO3 in water (95:5) as extraction solvents, respectively. Ten DG samples were investigated on their contents of free and total ferulic acid. The results indicated that the amount variety of free ferulic acid was larger than that of their counterparts, and the ratio of total ferulic acid to free ferulic acid was 4.07 +/- 2.73 (mean +/- SD, n = 10). The chemical assay of DG using total ferulic acid content would be a better choice to assess the herbal quality and was recommended.

Semistatic 96h LC50 values of 19 nitroaromatics were determined for the Carp (Cyprinus carpio), and the toxic symptoms were described. QSAR were developed using the LC50 values and six descriptors, 1XV, 1Kα, Σσ−, ELUMO; log P and I. Through muItiplicate regression analysis, one best equation was yielded: −logLC50=3.522+0.926Σσ−-+0. 1731,n= 19, r=0. 907,s=0. 271.The equation was used to estimate LC50 for 16 analogues. Inferred from the remarkable correlation between ELUMO and 1, the indicator variable (1) relates to electron effect or electron affinity of molecular essentially in the series of compounds. According to from the equations, for the compounds studied here,the dinitro compounds (their I, Σσ− and −ELUMO are all high) may be reactive, and the mononitro compounds may be less reactive or nonreactive.

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.

An HPLC-PAD-MS method has been developed for the qualitative and quantitative analysis of the major chemical constituents in Polygonum multiflorum Thunb. Chromatographic separation was conducted on an Alltima C18 column using water:acetonitrile:acetic acid as the mobile phase. Altogether nine compounds of various classes, including stilbene glucosides, anthraquinone glucosides and anthraquinone derivates, were identified by online ESI/MS. Their identities were ascertained by comparison with data derived from the literature and/or standard compounds. Five components were quantified by HPLC-PAD and the method was fully validated. All the linear regressions were acquired with R2 > 0.99 and the quantification limits (with a signal-to-noise ratio of 3) ranged from 0.63 and 1.57 ng. Repeatability was evaluated by intra- and inter-day assays and RSD value was within 2.47%. Recovery studies for the quantified compounds were found to be within the range 96.32-102.53% with RSD less than 2.35%. The overall procedure is rapid and reproducible and is considered suitable for qualitative and quantitative analysis of a large number of samples.

A high-performance liquid chromatography-diode array detector-mass spectrometry (HPLC-DAD-MS) analytical method was developed for detection of the nucleosides and nucleobases in two species of Lingzhi, the dried sporophore of Ganoderma lucidum and G. sinense. The method, combining advantages of both DAD and MS, was successfully used to qualitatively identify for six nucleosides namely, adenosine, cytidine, guanosine, inosine, thymidine, uridine and five nucleobases namely, adenine, guanine, hypoxanthine, thymine and uracil in Lingzhi samples. Quantitative analyses showed that uridine was the most abundant nucleoside in these Lingzhi samples and the contents of nine target analytes were found to be different in pileus and stipes of the fruiting bodies and among the different species of G. spp. The established method might apply as an alternative approach for the quality assessment of Lingzhi.

An HPLC-DAD fingerprinting profile of Rhizoma Et Radix Polygoni Cuspidati was established basing on the consistent chromatographic features of 24 authentic herb samples. The major types of chemical constituents, stilbenes and anthraquinones, were analyzed and included in the fingerprint. Eight common peaks of Polygonum Cuspidatum were identified by using HPLC-MS. The developed fingerprint was applied to differentiate Rhizoma Et Radix Polygoni Cuspidati from Radix Polygoni Multiflori and Radix Et Rhizoma Rhei. Although the three herbs belong to the family of Polygonaceae, the results indicated that these could be differentiated by using the established method.

Using the HPLC/DAD/ESI/MS method, the qualitative and quantitative analysis of senkyunolide A (SA) in the rhizomes of Ligusticum chuanxiong (Rhizoma chuanxiong; CX) and roots of Angelica sinensis (DG) was established. As a result, it was found that SA is a characteristic standard compound for the quality evaluation and chemical differentiation between CX and DG. Methanol was chosen in the preparation of standard solutions and extraction of samples based on the stability data. The identity of SA in CX and DG was unambiguously determined based on the quasimolecular ions in ESI-MS. A comprehensive validation of the method, including sensitivity, linearity, reproducibility and recovery, was conducted using the optimized chromatographic conditions. The linear calibration curve was acquired with R2>0.999 and limit of detection (S/N=3) was estimated to be 12.5 mug/g. The reproducibility was evaluated by repeated sample injection and replicated analysis of samples with the relative standard deviation (RSD) value found within 0.68%. The recovery rates of SA varied within the range of 96.91-101.50% with RSD less than 2.38%. In the present work, the contents of SA were quantified within 3.94-9.14 mg/g and 0.108-0.588 mg/g for 12 batches each of CX and DG. The results demonstrated that SA is a useful standard compound for the quality evaluation and chemical differentiation between CX and DG. The analytical procedure is precise and reproducible and thus suitable for the analysis of a large number of samples.

50% effective inhibition concentration 48h-EC50 of 40 substituted benzenes to the algae (Scenedesmus obliquus) was determined. The energy of the lowest unoccupied molecular orbital (ELUMO) was calculated by the quantum chemical method MOPAC6.0-AM1. By using ELUMO and the hydrophobicity parameter log KOW the quantitative structure–activity relationship model (QSAR) was developed: log 1/ EC 50 =0.272 log KOW − 0.659ELUMO + 2.54, R2=0.793, S.E.=0.316, F=71.07, n=40. A series of equations were obtained about the measured EC50 values of different subclasses of compounds. For those compounds containing double –NO2, their toxicity may be related chiefly to the intracellular reduction of –NO2 obtaining electron, while for anilines and phenols, KOW contributes most to the QSAR and ELUMO very little.

Since nitrobenzenes constitute a class of industrial chemicals that are present in Songhua River and probably in many other industrialized countries as well, it is useful to gain insight into their potential hazard to aquatic organisms. For this reason, it was decided to determine data on the toxicity for bacteria in the Songhua River. Furthermore, the toxicity to Ph. phosphoreum was determined in the Microtox assay, in order to further evaluate the usefulness of this assay for hazard assessment. Quantitative structure-activity relationships (QSARs) have been developed for aromatic nitro compound toxicity to aquatic species, but no data on the toxicity of nitrobenzenes to environmental bacteria were used. In this study, the toxicity of various substituted nitrobenzenes to bacteria in Songhua River and to Ph. phosphoreum has been investigated, establishing quantitative structure-activity relationships with n-octanol-water partition coefficient (log P), the energy of the lowest unoccupied molecular orbital (E{sub LUMO}) and the sum of substituent constant ({Sigma}{sigma}-). 12 refs., 2 tabs.