Primary Comments on Chemotaxonomy of Paris spp. Based on Saponins Analysis

Aim The several species of the genus Paris called ＂Chonglou＂ are famous traditional Chinese herbal medicines. We established the quantitative analysis method of the steroidal saponins in some species of the genus Paris and discussed their relations. Methods We detected the contents of 11 steroidal saponins in Paris samples with a Kromasel C18 （ 150 mm× 4.6 mm ID, 5μm） column which was subjected to gradient elution with acetonitrile-water （30：70- 60：40, V/V） at a flow rate of 1 mL· min^-1 by HPLC-ELSD and established chemical cluster tree using SPSS 11 software. Results All the samples could be separated and calibration curves of 11 saponins were prepared. We successfully detected the contents of 11 steroidal saponins in 14 Paris spp. in 30 min. The recovery for the assay of saponins was between 95 % and 97 %. The RSD of precision of 11 saponins and stability of samples were below 3 %. Chemical phylogenetic tree based on saponin contents indicated that 17 samples of Paris spp. clustered separately. Conclusion The established method is accurate and convenient, and suitable for the quantitative analysis of these 11 steroidal saponins in Paris spp.. The chemical phylogenetic tree is in accordance with Takhtajian classical taxonomy.

HPLC-ELSD法测定柴胡中柴胡皂苷a、d的含量

目的 :采用高效液相色谱法测定柴胡中柴胡皂苷 a、d的含量。方法 :采用 Alltech C1 8(4.6 mm× 2 5 0 mm ,5μm )色谱柱 ,甲醇 -乙腈 -水 -冰醋酸 (18∶ 40∶ 42∶ 0 .2 )为流动相 ,流速 1.0 0 m l/ min,Alltech EL SD- 5 0 0检测器 ,外标法定量。结果 :柴胡皂苷 a在 5 .0 2 5～ 15 .0 75μg范围呈线性 ,回收率为 96 .2 % ,RSD为 1.43% (n =9) ;柴胡皂苷 d在5 .6 70～ 17.0 10 μg范围呈线性 ,回收率为 95 .0 % ,RSD为 1.76 % (n =9)。结论 :本法简便可行 ,重现性好 ,可作为柴胡药材质量检测的方法。

Determination of Astragaloside IV in Yupingfeng Oral Solution by HPLC-ELSD Method

[Objective] This study aimed to establish a method for determining the content of Astragaloside IV in Yupingfeng oral solution.[Method] The HPLC-ELSD method was adopted.The chromatographic column was Venusil MP（4.6 mm × 150 mm,5 μm）.The mobile phase was acetonitrile-water（35∶65）.The ELSD evaporator tube temperature was 65 ℃.N2 was used as the carrier gas（pressure,30 psi）.[Result] When the content of Astragaloside IV ranged from 0.5 to 5.0 μg,the Astragaloside IV content showed a good linear relationship with peak area（r=0.999,n=6）.The average recovery was 96.36%,and the RSD was 2.46%.[Conclusion] This method is accurate and reliable,and can be applied in the quality control of Yupingfeng oral solution.

Preliminary Studies on High-performance Liquid Chro-matography Chemiluminescence Determination of theSaturated Fatty Acids(C_(16) C_(18))in Human Serum

The peroxyoxalate chemiluminescence(CL)detection of fatty acids in human se- rum combined with high-performance liquid chromatography (HPLC)is described.Some fatty acids in serum were extracted with a 1 :1(v/v)mixture of chloroform-n-heptane.2-(4-Hydrazinocarbonyl- phenyl)-4,5-diphenylimidazole (HCPI) was used as a fluorescent labelling reagent of the fatty acids. The labelling reaction was carried out at 30℃ for 1 h at pH 6.5 and the resulting reaction mixture was sudjected to HPLC. The labelled fatty acid C_(17)(P-C_(17))was used as the internal standard. The la- belled fatty acids C_(16) and C_(18) were separated within 18 min on an ODS-8OTM column (150 mm× 6 mm ID,5μm,Tosoh Japan).The calibrlation curves of fatty acids from the spiked control serum were Y_1=-0.003 7 + 0.0028X_1,r=0.994 for FA C_( 16) and Y_2=0.00 1 2 + 0.00098X_2,r=0.999 for FA C_( 18),respectively.The average recoveries of facids from the spiked contrl serum were 107.2%(n=8,RSD=4.3%)for FA C_(16) and 97.35%(n=8, RSD=4.0%)for FA C_(18),respectively.The lower detection limits of fatty acids after reaction were 12μmol per 20μl injection for FA C_(16) and 18 μmol per 20μl injection for FA C_(18),respectively(signal to noise ratio, S/N=2).The HPLC/CL method was applied to the determination of FA C_(16) and FA C_(18) in normal human serum and the results showed that the concentrations of fatty acids in normal human serum were 0.134 ± 0.009 μ mol/ml serum(n=5) for FA C_(16) and 0.052±0.028 μmol/ml serum(n=5)for FA C_(18),respectively.

Isolation and Identification of a Taxol-producing Endophytic Fungus Identified from Taxus media

[Objective] The aim was to isolate and identify a taxol-producing endophytic fungus from Taxus media. [Method] 32 strains of endophytic fungi were identified form the inner bark of T. media,and their fermentation products were detected by high performance liquid chromatography (HPLC). [Result] Through the screening,a strain of taxol-producing endophytic fungi M57 was obtained,which could produce 45-50 μg/L of taxol,and M57 was defined as Rhizopus sp. through morphological observation and 18S rDNA sequence analysis. [Conclusion] The finding of Rhizopus sp. M57 provided a promising strain for producing taxol with taxol-producing fungi fermentation process.

简述几种新型的饲料检测技术

饲料检测技术具有较强的实用性价值,在饲料检测中扮演着重要角色,因此本文对几种饲料检测技术的应用原理进行了介绍以便更好地选择适合的检测技术,提高检测的准确率和检测效率。1光学分析技术1.1原子吸收光谱法该法是光学分析技术中的一种,能够对目标物内元素的浓度进行检测,具有精准和快速等特点,具有良好的应用效果。此项技术,不仅可以对目标物内的铜元素进行总含量测定,还能借助火焰原子吸收光谱法来对镉元素的质量分数进行分析判断,从而为饲料产品的品质提供保障。该技术的不足之处是对真空紫外环境下的元素检测的准确性不是很好,使用不当还会干扰曲线分析。

Determination of Baicalin in Traditional Chinese Preparation by High Performance Liquid Chromatography with Chemiluminescence Detection

A new method for the determination of baicalin with HPLC-CL was developed. The method was based on the chemiluminescence reaction between KMnO4 and baicalin sensitized from HCHO. The linear range was 3.7?0-6~9.8?0-5 mol/L with detection limit of 1.7?0-6 mol/L and the relative standard deviation was 2.5 % (Cs=6.6?0-5 mol/L, n=5). The method has been applied to the determination of baicalin in oral administration, injection, Scutellariae radix and granules with good results.

Analysis of Sucrose Esters with Long Acyl Chain by Coupling of HPLC-ELSD with ESI-MS System

The analysis of sucrose esters with long acyl chain by improved high performance liquid chromatographic method with evaporative light scattering detection （HPLC-ELSD） and electrospray ionization mass spectrum （ESI-MS） is investigated. The improved HPLC-ELSD method for the separation and quantitation of commercial and synthesized sucrose esters is described. Samples are analyzed by means of a reversed-phase （RP） HPLC using a Hypersil C8 column （250 mm× 4.6 mm, 5 μm particle size） with methanol-tetrahydrofuran （vo）ume ratio of 90 ： 10） and water under gradientcondition as the mobile phase, in which the flow rate is 1.0 ml·min^-1 and the column temperature is set at 40℃. This procedure provides a complete separation and determination ot monoester, diester, triester and higher esters with different acyl chain lengths in each fraction by a single run, in combination with the ESI-MS technology. With this method, it is possible to determine the approximate compositions of monoto polyesters in one analysis and quantitate pure positional isomers precisely using an external standard method. It is found that the method of ESI-MS coupling with HPLC system for the analysis of sucrose esters is straight forward, rapid and inexpensive, and can be readily applied in synthesis, purification and structure studies.

Optimization of dilute acid hydrolysis of Enteromorpha

Acid hydrolysis is a simple and direct way to hydrolyze polysaccharides in biomass into fermentable sugars. To produce fermentable sugars effectively and economically for fuel ethanol, we have investigated the hydrolysis of Enteromorpha using acids that are typically used to hydrolyze biomass： H2SO4, HC1, H3PO4 and C4H404 （maleic acid）. 5%（w/w） Enteromorpha biomass was treated for different times （30, 60, and 90 min） and with different acid concentrations （0.6, 1.0, 1.4, 1.8, and 2.2%, w/w） at 121~C. H2SO4 was the most effective acid in this experiment. We then analyzed the hydrolysis process in H2SO4 in detail using high performance liquid chromatography. At a sulfuric acid concentration of 1.8% and treatment time of 60 min, the yield of ethanol fermentable sugars （glucose and xylose） was high, （230.5 mg/g dry biomass, comprising 175.2 mg/g glucose and 55.3 mg/g xylose）, with 48.6% of total reducing sugars being ethanol fermentable. Therefore, Enteromorpha could be a good candidate for production of fuel ethanol. In future work, the effects of temperature and biomass concentration on hydrolysis, and also the fermentation of the hydrolysates to ethanol fuel should be focused on.

Studies on the Correlation between Germination and cAMP Content of Different Color Chlamydospores in Ustiloginoidea virens

Ustilaginoidea virens is the causal agent of a serious disease of rice. To reveal the relationship between germination and the 3'-5'-cyclic adenosine monophosphate(c AMP) content from the dormant(black or green-black) and non-dormant(yellow)chlamydospore in Ustiloginoidea virens,this study adopts ultrasonic-bath method and high-performance liquid chromatography(HPLC) method, for extraction c AMP content of the different color chlamydospore. The results demonstrated that, as for the freshly chlamydospores collected from the false smut balls during their germination, c AMP content of yellow chlamydospore appeared a slight growth during 0-12 h, and showed a rapid declining as the germination time extended(12-48 h). Above all, in yellow chlamydospores, the germination rate and the content of c AMP presented a very notably negative correlation(|r|=0.861 9>r0.01=0.834), but the correlation between germination rate and content of c AMP was not obvious in black chlamydospores. The germination rate and the content of c AMP presented a notably positive correlation under different storage period(0, 2, 4, 6, 8,10 months) of the two color chlamydospores(the yellow of chlamydospore: |r | = 0.785 1 > r0.05= 0.707; the black of chlamydospore: |r| = 0.957 9 > r0.05= 0.707).

茯苓止痒丸的质量标准研究

目的：探讨茯苓止痒丸的质量控制方法。方法：采用薄层色谱法（TLC）定性鉴别茯苓止痒丸中的苍术、黄柏及甘草，用高效液相色谱法（HPLC）测定茯苓止痒丸中的盐酸小檗碱含量。结果：苍术、黄柏、甘草具有鉴别特征，阴性无干扰；盐酸小檗碱在0.4152~2.076mg范围内呈良好的线性关系，r=0.9999，平均加样回收率为99.37%，RSD=0.36%。结论：薄层色谱法及高效液相色谱法简便准确，重现性好，可作为茯苓止痒丸的质量控制标准。