Quality analysis of hawthorn leaves (the leaves of Crataegus pinnatifida Bge. var major N.E.Br) in different harvest time.

INTRODUCTION: Harvest time plays an important role on the quality of medicinal plants. The leaves of Crataegus pinnatifida Bge. var major N.E.Br (hawthorn leaves) could be harvested in summer and autumn according to the Pharmacopoeia of the People's Republic of China (Pharmacopoeia). However, little is known about the difference of the chemical constituents in hawthorn leaves with the harvest seasonal variations. OBJECTIVE: The chemical constituents of hawthorn leaves in different months were comprehensively analysed to determine the best harvest time. METHODS: Initially, the chemical information of the hawthorn leaves were obtained by ultra-high-performance liquid chromatography and quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS). Subsequently, principal component analysis (PCA) was applied to compare the chemical compositions of hawthorn leaves harvested in different months. Then, an absolute quantitation method was established using high-performance liquid chromatography-charged aerosol detector (HPLC-CAD) to determine the contents of five compounds and clarify the changes of these components with the harvest seasonal variations. Meanwhile, a semi-quantitative method by integrating HPLC-CAD with inverse gradient compensation was also established and verified. RESULTS: Fifty-eight compounds were identified through UHPLC-Q-TOF-MS. PCA revealed that the harvest season of hawthorn leaves had a significant effect on the chemical compositions. The contents of five components were relatively high in autumn. Other four main components without reference standards were further analysed through the semi-quantitative method, which also showed a high content in autumn. CONCLUSIONS: This work emphasised the effect of harvest time on the chemical constituents of hawthorn leaves and autumn is recommended to ensure the quality.

[Rapid prediction of flavonoid content in Epimedium sagittatum by infrared spectroscopy].

Epimedii Folium is a well-known Chinese herbal medicine with the effect of nourishing kidney and strengthening Yang. Its main active ingredients are flavonoids. In this study, 60 samples of Epimedium sagittatum were collected for the determination of total flavonoids(TF) including the total amount of epimedin A, epimedin B, epimedin C, and icariin(abbreviated as ABCI) specified in the Chinese Pharmacopoeia as well as rhamnosylicariside Ⅱ and icariside Ⅱ. The calibration parameters of "first derivativemultiva-riate scattering correction in 1 900-650 cm~(-1) band(4-point smoothing)" and "first derivativestandard normal variable correction in 4 000-650 cm~(-1) full band(4-point smoothing)" were confirmed respectively. The quantitative model was established via Fourier infrared spectroscopy plus attenuated total reflection(FTIR-ATR) accessory combined with partial least squares(PLS) method and then used to predict the flavonoid content of 11 validation sets. The average prediction accuracy for ABCI in calibration set and validation set was 98.985% and 96.087%, respectively. The average prediction accuracy for TF in calibration set and validation set was 98.998% and 94.771%, respectively. These results indicated that FTIR-ATR combined with PLS model could be used for rapid prediction of flavonoid content in E. sagittatum, with the prediction accuracy above 94.7%. The establishment of this method provides a new solution for the detection of a large number of E. sagittatum samples.

[Content correlation of eight phenolic acids and flavonoids in different medicinal parts of PA-type Perilla germplasms].

In this study, 10 PA-type Perilla germplasms were selected to detect the content of two phenolic acids, i.e., rosmarinic acid(RA) and caffeic acid(CA), and six flavonoids, including scutellarin-7-O-diglucuronoside(SDG), luteolin-7-O-diglucuronoside(LDG), apigenin-7-O-diglucuronoside(ADG), scutellarin-7-O-glucuroside(SG), luteolin-7-O-glucuroside(LG), and apigenin-7-O-glucuroside(AG) in leaves, stems, and fruits. The total content of phenolic acids and flavonoids in leaves was 3.991-12.028 mg·g~(-1) and 12.309-25.071 mg·g~(-1), respectively, which was much higher than that in stems(0.586-2.015 mg·g~(-1) and 0.879-1.413 mg·g~(-1), respectively) and fruits(0.004-2.222 mg·g~(-1) and 0.651-1.936 mg·g~(-1), respectively). RA was detected in five fruit samples, and RA content between leaves and fruits showed a significant negative correlation in the other five samples. For flavonoids, only LG and LDG could be detected in stems, and SG and SDG were not detected in fruits, while other flavonoids were not detected in some samples. The content of total flavonoids and LG in leaves and fruits was significantly positively correlated, and the content of LG in stems and fruits was significantly positively correlated. In 10 stem samples, seven met the standard that the content of RA in the stem should be not less than 0.1% specified in the Chinese Pharmacopoeia(2020 edition). Only one fruit sample reached the standard of RA content in the fruit not less than 0.25% specified in the Chinese Pharmacopoeia.

New steroidal glycosides from the roots of Asparagus cochinchinensis.

Steroidal saponins were the main active constituents of the traditional medicinal herb Asparagus cochinchinensis. A phytochemical investigation of A. cochinchinensis roots led to the isolation of nine new steroidal glycosides (1-9) and seven known analogues (10-16). Their structures were established by spectroscopic analyses as well as necessary chemical evidence.

[Contents determination of eight phenolic compounds in Perilla frutescens leaves of different cultivation years and harvesting periods].

A method was established for content determination of two kinds of phenolic acids, including rosmarinic acid)(RA) and caffeic acid(CA), and six kinds of flavonoids including scutellarein-7-O-diglucuronide(SDG), luteolin-7-O-diglucuronide(LDG), apigenin-7-O-diglucuronide(ADG), scutellarin-7-O-glucuronide(SG), luteolin-7-O-glucuronide(LG), and apigenin-7-O-glucuronide(AG) in Perilla frutescens leaves. The content of eight chemical components was measured based on ten P. frutescens germplasms of different chemotypes of volatile oil, different cultivated years, and different harvesting periods. The results showed that there was a great difference between the two kinds of constituents of different germplasms. The total content of the two phenolic acids was 2.24-34.44 mg·g~(-1), and the total content of the six flavonoids was 11.55-34.71 mg·g~(-1). Then according to content from most to least, the order of each component was RA(2.13-33.97 mg·g~(-1)), LDG(1.31-14.80 mg·g~(-1)), SG(1.97-8.45 mg·g~(-1)), ADG(2.68-7.60 mg·g~(-1)), SDG(1.16-5.87 mg·g~(-1)), LG(0.78-1.91 mg·g~(-1)), AG(0.56-1.00 mg·g~(-1)), and CA(0.11-0.68 mg·g~(-1)). The chemical contents of the 5 PA-type germplasms in 2017 were mostly higher than those in 2018 showing a large variation with the cultivation years. These contents of two kinds of phenolic acids of 9 germplasms fluctuated with the harvesting time. The content decreased before early flower spike(the 3~(rd) to 18~(th) in August) at first and began to increase in flowering and fruiting period(the 18~(th) in August to 2~(nd) in September). However, these contents had slowly decreasing trend after 2~(nd) in September till 17~(th) in the same month. Interestingly, the content raised again in the maturity of fruits. The variation tendency of contents in six kinds of flavonoids components was inconsistent in different germplasms with the variation of harvesting time. The content of flavonoids in part of germplasms was negatively correlated with the fluctuation of phenolic acids. There was no correlation between phenolic acids and chemical type of the volatile oil. This paper may provide a reference for the high-quality germplasm of P. frutescens cultivation.

[Qualitative analysis of Gynostemma longipes for medicinal usage].

The Qinling-Daba Mountains area is the main producing areas of Gynostemma longipes for medicinal usage, and samples of wild whole plants in Pingli, Shaanxi Province and Qingchuan, Sichuan Province were collected. The ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UHPLC-Q-TOF-MS~E) was used to profile the chemical compositions and analyze the similarities and differences of G. longipes samples in these areas. Based on the accurate molecular weight and fragment information obtained from Q-TOF-MS~E, the structures of the main components were identified by combining with the mass spectra, chromatographic behaviors of reference standards and related literatures. The results showed that the components of wild G. longipes from different places among Qinling-Daba Mountains area were similar. Forty-five chemical components were identified in the whole plant of G. longipes from Pingli, Shaanxi Province, including 43 triterpenoid saponins and 2 flavonoids which contain all main peaks in its fingerprint. The main components are dammarane-type triterpenoid saponins, such asgypenoside ⅩLⅨ, gypenoside A and its malonylated product of glycosyl.

[Planting density of Perilla frutescens PA-type].

Two medicinal PA type Perilla germplasms were planted at five planting densities(D1,2 500 plants/Mu;D2,5 500 plants/Mu;D3,8 500 plants/Mu;D4,11 500 plants/Mu;D5,14 500 plants/Mu;1 Mu≈667 m~2). A total of 17 traits, including leaf shape, plant type, yield, volatile oil content and composition, were recorded and studied. With the planting density increased, the leaves appeared narrow, plants small, the deciduous leaves increased, and the leaf yield per plant was low, but the leaf yield per Mu increases significantly with the planting density, and was basically stable after reaching D4. The extraction rate of volatile oil from leaves at planting density D2-D5 was about 0.1% higher than that of D1, and there was no significant difference in the relative content of perillaldehyde, among 5 density. In order to achieve high leaf yield, it is recommended to plant at a density of D4, 11 500 plants/Mu(plant spacing is 15 cm, and row spacing is 40 cm); while comprehensive leaf yield and leaf morphology are recommended to be planted at a density of D2, 5 500 plants/Mu(plant spacing is 30 cm, and row spacing is 40 cm). At the same time, dense planting resistance of two germplasms were different. Number of secondary branches, first section with leaves and plant types were most important feature for the evaluation of the density tolerance of PA-type Perilla. This study provided a reference for the suitable density of PA-type Perilla, and laid a foundation for further study of the tolerance characteristics of different Perilla.

[Effects of light quality on growth and icariin flavonoid content of Epimedium pseudowushanense under different light intensity].

In this study, the growth index including plant height, compound leaf area, specific leaf area, leaf water content, number of branches, and leaf biomass per plant and the icariin flavonoids such as epimedin A, epimedin B, epimedin C and icariin of Epimedium pseudowushanense were determined on 30 d and 60 d under light intensity(18.2±2.5) µmol·m~(-2)·s~(-1)(L1) and(90.9 ±2.5) µmol·m~(-2)·s~(-1)(L2), and white light as control, red light, blue light and yellow light were used as three light quality treatments, to study the effect of light quality on the growth and flavonoids accumulation of E. pseudowushanense. The E. pseudowushanense was sui-table for growth under L1 light intensity, the blue light treatment significantly reduced the leaf area, but had little effect on the stem height, the red light treatment and the yellow light treatment had no obvious effect on the stem height and leaf area, but the yellow light treatment significantly increased the germination of new branches, and had a sustained promoting effect, and the biomass was significantly higher than the white light treatment at 60 d. The content of icariin flavonoids in red light, blue light and yellow light treatment was higher than that in white light treatment at 30 d and 60 d under L1 light intensity, while yellow light treatment promoted the synthesis of icariin flavonoids to the largest extent, which was 1.8 and 1.9 times of white light treatment(30 d and 60 d).Under L2 light intensity, the effect of strong light on promoting stem germination became the main factor, while the yellow light treatment showed no significant effect on promoting stem germination, and the red light treatment exhibited a significant effect on reducing leaf area. Icariin flavonoids under red light, blue light and yellow light treatment were all lower than that under white light treatment, that is, the effect of white light treatment on the synthesis of icariin flavonoids is better than red light, blue light and yellow light treatment. When the time of strong light treatment was longer, the degradation range of icariin flavonoids in other light treatment appeared, while red light treatment promotes the synthesis of icariin flavonoids. Therefore, the influence of light quality on E. pseudowushanense is quite different under different light intensity, no matter from growth index or flavonoid content index. The results support that the biomass and icariin flavonoid content can be increased by providing appropriate red and yellow light.

[Change of icariin content in five Epimedium species by heating process].

The change of icariin( ICA) content in thirty-three samples of five Epimedium species listed in the Chinese Pharmacopoeia( 2015 edition),including E. brevicornu,E. sagittatum,E. pubescens,E. koreanum,and E. wushanense has been investigated in this study. The results indicated that the optimized process procedure was baking at 150 ℃ for 30 min,and 3'''-carbonyl-2″-ß-L-quinovosyl icariin( CQICA) could not be translated into ICA and ICA could be converted under this heating process condition. ICA increased remarkably after the heating process by 1-3 times in E. brevicornu,E. wushanense and E. koreanum,and increased lightly in E. brevicornum and E. pubescens,while ICA slightly increased or decreased in E. sagittatum and E. wushanense.

[Study on morphological classification and chemical-type of Perilla frutescens cultivated germplasm].

Fifty cultivated Perilla seeds were collected all over the country and planted in Beijing experiment field for morphology and chemical-type researches. Twenty morphological characteristics were selected and observed, and the essential oil from leaves was extracted by steam distillation and analyzed by GC-MS to confirm chemical-types. There were significant diversities in plant height, leaf color and morphology, and fruit color and weight. Clustering analysis was carried out based on these morphological characteristics. Six types were divided with their chemical-type designated. Type Ⅰ: Six germplasms, attributed to P. frutescens var. crispa, with dwarf plants, thin creased purple leaf, named Crispa, their chemical types were diversified, including EK, PAPK, PA and PK. Type Ⅱ: Six germplasms, attributed to P. frutescens var. crispa, plants were taller than type I and with thin and creased green leaf, named Big Crispa, all PK type. Type Ⅲ: Seventeen germplasms, attributed to P. frutescens var. frutescens with leaf color upside green and underside purple, tall plant and wide distribution all over the China, named Ordinary Frutescens, all PK. Type Ⅳ: Four germplasms, attributed to P. frutescens var. acuta with tall plant and small seed, named Acuta, all PK. Type Ⅴ: Seven germplasms, attributed to P. frutescens var. frutescens with green leaves, tall plants and long clusters, named Long-spike Frutescens, all PK. Type Ⅵ: Ten germplasms, attributed to P. frutescens var. frutescens with big, thick and creased leaf, named Thick-leaf Frutescens, including PK, PP, PL and PA. The morphological classification of this paper would lay the foundation for the taxonomic naming and following evaluation of the Perilla germplasm resources.This study also showed that there was no correspondence but a certain correlation between volatile oil chemical-types and subspecies classification and morphological characteristics of Perilla.

[Investigation of aflatoxins,mycobiota,and toxigenic fungi during post-harvest handling of Ziziphi Spinosae Semen].

Ziziphi Spinosae Semen is one of the Chinese herbal medicine being susceptible to aflatoxins contamination. To investigate the sources of aflatoxins contamination and toxigenic fungi species on Ziziphi Spinosae Semen,32 samples were collected from multiple steps during the post-harvest processing in this study. Aflatoxins in these samples were determined by immunoaffinity column and HPLC coupled with post-column photochemical derivatization. The dilution-plate method was applied to the fungi isolation. The isolated fungi strains were identified by morphological characterization and molecular approaches. The results showed that aflatoxins were detected in 28 samples from every step during the processing of Ziziphi Spinosae Semen. Three samples were detected with aflatoxin B_1 and 2 samples with both aflatoxin B_1 and total aflatoxin exceeding the limit of Chinese Pharmacopoeia. Especially the samples from the washing step,with the highest detected amounts of AFB_1 and AFs were reached 94. 79,121. 43 µg·kg~(-1),respectively. All 32 samples were contaminated by fungi. The fungal counts on the newly harvested samples were 2. 20 × 10~2 CFU·g~(-1). Moreover,it increased as tphreocessing progresses,and achieved 1. 16×10~6 CFU·g~(-1) after washing. A total of 321 isolates were identified to 17 genera. Aspergillus flavus was the main source of aflatoxins during the processing and storage of Ziziphi Spinosae Semen. One isolate of A. flavus was confirmed producing AFB_1 and AFB_2. The fungal count was significantly increased by composting,and Aspergillus was the predominant genus after shell breaking. The contamination level of aflatoxins was increased by composting and washing.

[Comparative analysis on chemical constituents in Bupleurum chinense,B. marginatum,B. marginatum var. stenophyllum and B. smithii var. parvifolium].

UPLC-Q-TOF-MS was used to analyze the chemical differences in Bupleurum. chinense,B. marginatum,B. marginatum var. stenophyllum and B. smithii var. parvifolium. Chromatographic separation was carried out on an Acquity HSS T3 C_(18) column( 2. 1 mm ×100 mm,1. 8 µm,Waters) with the mobile phase composed of 0. 1% formic acid in water-acetonitrile in the gradient elution. A hybrid quadrupole time-of-flight tandem mass spectrometry( Q-TOF-MS~E) was used for mass spectrometric analysis. Finally,25 peaks were identified based on their exact mass data and fragmentation characteristics. B.chinense,B.marginatum,B. marginatum var. stenophyllum and B. smithii var. parvifolium were obviously clustered into 3 types through processing by principal component analysis( PCA). There was almost no difference between B. chinense and B. marginatum. However,the compounds existed in B. chinense were different from those in B. marginatum var. stenophyllum,and B. smithii var. parvifolium.

Nonspecific Symbiosis Between Sophora flavescens and Different Rhizobia.

We explored the genetic basis of the promiscuous symbiosis of Sophora flavescens with diverse rhizobia. To determine the impact of Nod factors (NFs) on the symbiosis of S. flavescens, nodulation-related gene mutants of representative rhizobial strains were generated. Strains with mutations in common nodulation genes (nodC, nodM, and nodE) failed to nodulate S. flavescens, indicating that the promiscuous nodulation of this plant is strictly dependent on the basic NF structure. Mutations of the NF decoration genes nodH, nodS, nodZ, and noeI did not affect the nodulation of S. flavescens, but these mutations affected the nitrogen-fixation efficiency of nodules. Wild-type Bradyrhizobium diazoefficiens USDA110 cannot nodulate S. flavescens, but we obtained 14 Tn5 mutants of B. diazoefficiens that nodulated S. flavescens. This suggested that the mutations had disrupted a negative regulator that prevents nodulation of S. flavescens, leading to nonspecific nodulation. For Ensifer fredii CCBAU 45436 mutants, the minimal NF structure was sufficient for nodulation of soybean and S. flavescens. In summary, the mechanism of promiscuous symbiosis of S. flavescens with rhizobia might be related to its nonspecific recognition of NF structures, and the host specificity of rhizobia may also be controlled by currently unknown nodulation-related genes.

[Effects of plant growth regulators and nitrogen on germination of Epimedium pseudowushanense after its stems and leaves reaped].

In order to explore appropriate measures to promote germination after the harvest of Epimedium pseudowushanense, 6-BA, urea, ammonium bicarbonate and GA3 were chosen to spray on the root and rhizomes, and then the biological indicators such as branches, leaf length, leaf width, plant height and so on, were measured in different periods, and the contents of epimedin A, epimedin B, epimedin C and icariin in the dry leaves were detected by HPLC. Results showed that 6-BA 90 mg·L⁻¹(B1), 6-BA 60 mg·L⁻¹(B2),6-BA 30 mg·L⁻¹+urea 300 mg·L⁻¹ (C1), 6-BA 60 mg·L⁻¹+urea 300 mg·L⁻¹(C2),6-BA 60 mg·L⁻¹+ ammonium bicarbonate 300 mg·L⁻¹(C4) significantly increased bud germination in the early period, and the plants quickly set up new system of photosynthesis, the branches in a month of which were higher than the control group respectively by 165.9%, 115.76%, 103.86%, 104.50%, 81.67%.However the branches developed the next year and the dry weight of leaves per plant in group B1 and B2 were much lower than that in control group. The groups that use 6-BA and nitrogen at the same time reaped a good yield of leaves even though the treatment had no significant influence on the branches developed the next year. The dry weight of leaves of C1, C2, C4 treatments were 36.80%, 32.84%, 45.97% more than the control group respectively. Therefore, C1, C2 and C4 treatments are the more appropriate to promote recovery after harvest. Furthermore, different groups, except 10 mg·L⁻¹ 6-BA treatment significantly reduced the content of epimedin C, other groups didn't have any significant effect on the contents of such flavonoids.

Anti-influenza triterpenoid saponins (saikosaponins) from the roots of Bupleurum marginatum var. stenophyllum.

This study to investigate antiviral components from the roots of Bupleurum marginatum var. stenophyllum led to the isolation of five novel saikosaponins, namely 6″-O-crotonyl-saikosaponin a (1), tibesaikosaponin I (2), tibesaikosaponin II (3), tibesaikosaponin III (4), tibesaikosaponin IV (5), along with 9 known analogues (6-14). Their structures were established by spectral data analyses (IR, MS, 1D and 2D NMR) and by comparison of spectral data with those of the related known compounds. Antiviral testing of all compounds against influenza A virus A/WSN/33 (H1N1) in 293TGluc cells showed that nepasaikosaponin k (12), saikosaponin n (13) and saikosaponin h (14) behaved more potent inhibitory activity and selectivity than the positive control, Ribavirin. The preliminary structure-activity relationship studies suggest that the 13, 28-epoxy group, the type of sugar chain and the type of olefinic bonds are significant for antiviral activity and selectivity.

Polyoxypregnane Glycosides from the Roots of Marsdenia tenacissima and Their Anti-HIV Activities.

A continuous phytochemical study on the roots of Marsdenia tenacissima led to the isolation and identification of 13 new polyoxypregnane glycosides named marstenacissides B10-B17 (1, 2, 4, 7, 8, 11, 12, and 14) and marstenacissides A8-A12 (3, 9, 10, 13, and 15) in addition to two known polyoxypregnane glycosides marsdenosides M and L (5 and 6). Their structures were established by spectroscopic techniques and by comparison with the reported data in the literature. Moreover, the anti-HIV activities of these isolates and the previous isolated marstenacissides A1-A7 and B1-B9 were assessed, some of which exhibited slight or negligible effects against HIV-1.

Comparison of chemical profiles between the root and aerial parts from three Bupleurum species based on a UHPLC-QTOF-MS metabolomics approach.

BACKGROUND: Bupleuri Radix (Chaihu) represents one of the most successful and widely used herbal medicines in Asia for the treatment of many diseases such as inflammatory disorders and infectious diseases over the past 2000 years. In the Chinese Pharmacopoeia, Chaihu is recorded as the dried roots of Bupleurum chinense DC. and B. scorzonerifolium Willd. (Umbelliferae). However, the widespread demand for the herb has tended to far outstrip the supply. Whether the aerial parts, which account for 70 ~ 85% of the dry weights of Bupleurum species, could be used as an alternative for the root has become an important scientific issue for the sustainable utilization of Bupleurum species. On the other hand, in some areas including the southeast of China as well as in Spain, the aerial parts of Bupleurum species have already been used in the folk medications. Therefore, to clarify whether the root and aerial parts of Bupleurum species are "equivalent" in the types and quantities of chemical constituents which subsequently influence their biological activities and therapeutic effects is of great importance for both the rational and sustainable use of this herb. METHODS: In the present study, the chemical profiles between the root and aerial parts of Bupleurum species from different species and collected from various locations were analyzed and compared by the ultra-high performance liquid chromatography quadrupole/time of flight-mass spectrometry (UHPLC-QTOF-MS). RESULTS: A total of 56 peaks were identified in the root and/or aerial parts from different batches of Bupleurum species, by comparison of references standards or with those reported in the literature. Principal Component Analysis (PCA) was conducted for displaying the differentiating clustering between these two parts. CONCLUSION: The results disclosed the distinct variations between them, which indicated that the aerial parts could not be used as an alternative of root from a chemodiversity perspective. The differentiating markers resulted from the PCA analysis could also be utilized for the differentiation between them. Further validation of their biological differences is anticipated in the future study.

Three New Sesquiterpene Glycosides from the Rhizomes of Trillium tschonoskii.

Three new sesquiterpene glycosides, possessing a rare aglycone with a sulfonyl between C-1 and C-15 positions, named 3-(3'E-7'R,8'-dihydroxy-4',8'-dimethyl-3'-nonenyl)-2,5-dihydro-1,1-dioxo-thiophen 7'-O-ß-d-glucopyranosyl-(1→4)-O-ß-d-glucopyranosyl-(1→4)-O-ß-d-glucopyranoside (1), 3-(3'E-7'R,8'-dihydroxy-4',8'-dimethyl-3'-nonenyl)-2,5-dihydro-1,1-dioxo-thiophen 7'-O-ß-d-glucopyranosyl-(1→4)-O-ß-d-glucopyranoside (2), and 3-(3'E-7'R,8'-dihydroxy-4',8'-dimethyl-3'-nonenyl)-2,5-dihydro-1,1-dioxo-thiophen 7'-O-ß-d-glucopyranosyl-6'-O-acetyl-(1→4)-O-ß-d-glucopyranosyl-(1→4)-O-ß-d-glucopyranoside (3), respectively, were isolated from the rhizomes of Trillium tschonoskii. Their structures were established on the basis of spectroscopic data, including HR-ESI-MS, IR, 1D and 2D NMR. The cytotoxic properties of the three compounds were investigated using human hepatic L02 cells.

[HPLC-MS/MS determination of residual amount of 4 plant growth retardants in 6 dried root and Rhizome Herbs].

HPLC-MS/MS was applied to the determination of residual amount of plant growth retardant such as paclobutrazol, daminozide, chlormequat and mepiquat chloride in dried root and rhizome herbs. The sample was extracted twice with acetonitrile containing 0.1% formic acid. The separation was performed on a Waters Atlantis HILIC column with an elution system consisting of acetonitrile-5 mmol•L⁻¹ ammonium acetate solution with 0.1% formic acid, methanol and acetonitrile. The MS spectrum was acquired in positive mode with multiple reactions monitoring (MRM). The linear range was 6-1 500 µg•kg⁻¹, and the optimized method offered a good linear correlation (r>0.997 8), excellent precision (RSD<11%) and acceptable recovery (from 79.3% to 103.3%). Four kinds of plant growth retardant have detected in some ofhenise herbs like Ophiopogonis Radix, Angelicae Sinensis Radix, Achyranthis Bidentatae Radix, Alismatis Rhizoma, Chuanxiong Rhizama and Notoginseng Radix et Rhizama, is among the more severe cases, dwarf lilyturf, multi-effect azole detection quantity is 63.4~1 351.66 µg•kg⁻¹, and Daminozide was detected in Ophiopogonis Radix, Angelicae Sinensis Radix, Chuanxiong Rhizama, Alismatis Rhizoma.

[Morphological characteristics of Chinese Perilla fruits].

The morphological traits of 55 Chinese Perilla fruit samples (size, 100 grains weight, color, hardness, surface ridge height) are described and the statistically analyzed. It can be divided into 6 categories by cluster analysis, namely: Ⅰ, big grain (diameter 1.5 mm above and 100 grains weight above 0.16 g), low ridge, hard; Ⅱ, big grain, low ridge, soft; Ⅲ, big grain, high ridge, soft, fruit; Ⅳ, big grain. high ridge, gray brown or dark brown; Ⅴ, small grain (diameter 1.5 mm below and 100 grain weight 0.16 g below), low ridge, hard, dark brown; Ⅵ, small grain, low ridge, hard, yellow brown. The 38 fruit samples were planted, among which 31 ones were P. frutescens var. frutescens, 4 ones P. frutescens var. crispa and 3 ones P. frutescens var. acuta. By chemotype classification, they were 29 PK type, 3 PA type, 2 PL type, 2 PP type, 1 EK type and 1 PAPK type. According the description of herb Perillae Fructus in China Pharmacopoeia, the plant originates from P. frutescens var. frutescens. In contrast, not all fruits of P. frutescens var. frutescens have accord features. The fruits with white pericarp are mainly from P. frutescens var. frutescens with purple leaves. The materials with small grain, low ridge, hard, yellow brown or dark brown, are likely to be PA type and mainly P. frutescens var. crispa.