Separation,determination of six impurities in methotrexate drug substance using ultra-performance liquid chromatography

Methotrexate(MTX) is an antineoplastic therapeutic medicine as antimetabolite of folic acid. In this paper, a sensitive and rapid ultra-performance liquid chromatographic(UPLC) method was developed and validated for the separation and determination of impurities in MTX drug substances. The UPLC method was accomplished on an Agilent Zorbax Extend C-18(50 mm 4.6 mm, 1.8 mm) with a gradient elution system composed of sodium dihydrogen phosphate in water(20 mmol/L, pH 3.0) and acetonitrile. The flow rate was 2.2 mL/min. The method was validated. The calibration curves displayed good linearity(r 0.999) within the tested concentration ranges. The limit of detection(LOD) and limit of quantification(LOQ) of the six analytes were all less than 0.774 mg/mL and 1.03 mg/mL. The relative standard deviation(RSD) for intra- and inter-day precision of the six analytes was less than 9.8%, including at the LOQ. The average recovery ranged from 95.2% to 103% except at the LOQ, where recovery ranged from 82.7% to 117%. The validated method was successfully used to determine the relative abundance of six impurities in the MTX drug substances.     

Capillary Electrophoretic Determination of Jinggangmycin A in Formulations Using Direct UV Detection

A simple, fast and reliable method was developed for the analysis of jinggangmycin A （validamycin A） in commercial formulations. The running buffer used was acetate buffer （100 mmol/L, pH 4.7） with 15 kV as the applied voltage. The detection was achieved by using direct UV mode at 200 nm and the detection limit was 0.2 μg/mL. Linearity in the concentration range of 5-500 μg/mL was excellent （RE 〉 0.999）. The run-to-run repeatability （n = 3）, as expressed by the relative standard deviation （RSD） for migration times and peak areas were less than 0.5% and 3.0% respectively. The mean recovery ranged from 97.2% to 101.4%.     

Analysis of insecticide thiacloprid by ion chromatography combined with online photochemical derivatisation and fluorescence detection in water samples

This study describes a novel application of ion chromatography coupled with post-column photochemically induced fluorimetry derivatisation in alkaline medium and fluorescence detection(IC-hv-FD) for the determination of neonicotinoid pesticide, thiacloprid. In an aqueous medium, this compound showed fluorescence with an excitation maximum at 236 nm and an emission maximum at 353 nm. The 10 mmol/L NaOH with 10%(v/v) acetonitrile solution pumped at flow rate of 1.0 mL/min was used for the chromatographic elution to isocratically separate thiacloprid on an Ion Pac1 AS 11(250 mm 4 mm i.d; 13 mm particle size, Dionex) anion-exchange column. The linear concentration range of application was 0.04–10.0 mg/L, with a relative standard deviation(RSD, n = 7) of 1.7%(for a level of 2.0 mg/L) and detection limit(LOD, S/N = 3) of 9.9 mg/L. The procedure was applied with satisfactory results to the analysis of thiacloprid in ground and lake water samples. Pesticide average spiked recoveries ranged between 95.5% and 114.0%.     

Determination of Sophocarpine,Matrine, and Sophoridine in KUHUANG Injection by GC-MS

A GC-MS procedure was developed to determine sophocarpine (SC), matrine (MT), and sophoridine (SRI) in KUHUANG injection. The chromatographic separation was performed on an HP-5MS column (30 m × 0.35 mm inner diameter, 0.25 μm film thickness) with helium as the carrier gas. The oven temperature was programmed from 170 to 205°C at a rate of 0.8 K/min, and n-tetracosane was used as the internal standard (IS). There was a good linear relationship between the peak area ratio of analyte to IS and the concentration of analyte in the ranges 0.0360–0.100 mg/mL for SC, 0.0461–0.346 mg/mL for MT, and 0.0473–0.355 mg/mL for SRI. The recoveries were 86.1–100.9% for SC, 87.9–104.0% for MT, and 92.5–107.2% for SRI. The interday and intraday relative standard deviations (RSDs) of determinations were less than 4.6, 2.7, and 5.1% for the assay of SC, MT, and SRI, respectively. The limits of detection (LOD) were 0.018 mg/mL for SC, 0.0231 mg/mL for MT, and 0.0287 mg/mL for SRI. The limits of quantitation (LOQ) were 0.036 mg/mL for SC, 0.0461 mg/mL for MT, and 0.0473 mg/mL for SRI. The results indicate that the developed method can be used to improve the quality control of KUHUANG injection. __________ From Zhurnal Analiticheskoi Khimii, Vol. 60, No. 10, 2005, pp. 1087–1093. Original English Text Copyright ? 2005 by Wu, Chen, Cheng. The text was submitted by the authors in English.     

高效液相色谱法测定家兔血清中盐酸山莨菪碱含量

rapid method for the separation and determination of anisodamine hydrochloride in serum of rabbit has been developed with Nucleosil column of 4. 6× 250mm and CH3OH-H2O-(C2H5)3N (30:70 : 0.125)as mobile phase. Anisodamine hydrochloride was determined by reversed phase HPLC at 214nm.Atropine sulfate was used as an internal standard. The calibration curve was Y= 0.01945X+0. 05623,r= 0. 9998,n= 6.It was rectilinear within the range between 0. 75μg/mL-50.0μg/mL,and the recovery was from 97. 0% to 98. 2%.     

Determination of Nickel and Palladium in Environmental Samples by Low Temperature ETV-ICP-OES Coupled with Liquid-liquid Extraction with Dimethylglyoxime as Both Extractant and Chemical Modifier

A novel method for the determination of nickel and palladium in environmental samples by low temperature ETV-ICP-OES with dimethylglyoxime(DMG)as both the extractant and chemical modifier has been developed.In this study,it was found that nickel and palladium can form complexes with dimethylglyoxime(0.05%,mass fraction)at pH 6.0 and can be extracted into chloroform quantitatively.The complexes can be evaporated into plasma at a suita-ble temperature(1400 ℃)for ICP-OES detection.Under the optimized conditions,the detection limits of nickel and palladium are 0.48 and 0.40 ng/mL,respectively,while the RSD values are separately 5.0% and 3.1%(ρ=50 ng/mL,n=7).The proposed method was applied to the determination of the target analytes in environmental samples with satisfactory results.     

A novel spectrophotometric determination of copper(Ⅱ) with bromosulphonazo Ⅲ

A novel ehromogenic reaction involving copper(Ⅱ) and bromosulphonazo Ⅲ (Br-SAZⅢ) in hexamethylenetetramine-hydrochloric buffer solution was investigated. The results showed that a blue complex of copper(Ⅱ) and bromosulphonazo Ⅲwas formed with a molar ratio of 1:1. The apparent molar absorptivity was 3.3 × 105Lmol-1cm-1 and themaximum absorption peak was at 616.8 nm. The proposed procedure was used for quantitative estimation of Cu(Ⅱ) inthe concentration range of 0-1.024 μg/mL with the detection limit (3σ) of 7.03 × 10-4 μg/mL (n = 20). The relativestandard deviations (RSDs) were 0.56-4,68%. Under the optimized conditions, total copper in the vegetables and tea wassuccessfully determined.     

A novel spectrophotometric determination of copper（Ⅱ） with bromosulphonazo Ⅲ

A novel chromogenic reaction involving copper（Ⅱ） and bromosulphonazo Ⅲ （Br-SAZⅢ） in hexamethylenetetramine- hydrochloric buffer solution was investigated. The results showed that a blue complex of copper（Ⅱ） and bromosulphonazo Ⅲ was formed with a molar ratio of 1：1. The apparent molar absorptivity was 3.3×10^5Lmol-1cm-1 and the maximum absorption peak was at 616.8 nm. The proposed procedure was used for quantitative estimation of Cu（II） in the concentration range of 0-1.024μg/mL with the detection limit （3σ） of 7.03 × 10^-4 μg/mL （n = 20）. The relative standard deviations （RSDs） were 0.56-4.68%. Under the optimized conditions, total copper in the vegetables and tea was successfully determined. 2007 Li Yuan. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.     

Two-stage pH Control Mode in Batch Fermentation of a Novel Bioflocculant from Corynebacterium Glutamicum

The effect of pH of the fermentation medium on cell growth and the production of a novel bioflocculant(named REA-11) by Corynebacterium glutamicum CCTCC M201005 were investigated. The maximum biomass(2.23 g/L) and flocculating activity(142.2 U/mL) were simultaneously obtained at the 14th hour when the pH value of the culture medium was maintained at 7.0 during the whole fermentation process. The production of REA-11 kept on a trend of increase till the later phase of fermentation process, which resulted in the ultimate flocculating activity of the culture broth to enhance to nearly 100 U/mL at pH 6.0. A two-stage pH control mode was adopted in REA-11 production in which the pH value of the culture medium was controlled at 7.0 during the first 14 h, then decreased to 6. 0 that was maintained until the end of the fermentation process. With the two-stage pH control mode, the maximum flocculating activity reached 178. 8 U/mL which was 30% higher than that obtained under the condition of pH 7.0 and the biomass enhanced about 15%. Compared with the fermentation process without pH control, REA-11 production and cell growth via the two-stage pH control mode increased 80% and 25 %, respectively.     

A novel liquid chromatographic method with fluorescence detection for quantitation of tadalafil and dapoxetine hydrochloride in pharmaceutical dosage form and human plasma北大核心CSCD

Tadalafil( TAD)and dapoxetine HCl( DAP)are recently co-formulated and both show native fluo-rescence. Therefore,a novel,accurate,specific and sensitive reversed-phase high performance liquid chroma-tographic method with fluorescence detection was developed and validated for their separation and quantitation in dosage form and human plasma using avanafil as an internal standard( IS). Separation was achieved using isocratic elution within 7. 0 min on C18 column and acetonitrile-0. 15% triethylamine( 40∶60,v/v;pH 4)as a mobile phase. The flow rate was 1. 0 mL/min and the detection was time-programmed at 330,410 and 370 nm for TAD,DAP and IS,respectively,after excitation at 236 nm. The linear ranges from 0. 01 to 30. 00 μg/mL for each drug with the limits of detection of 4. 20 and 7. 20 ng/mL for TAD and DAP,respectively. The method was validated in accordance to the International Conference on Harmonization( ICH)guidelines and was successful-ly applied to spiked human plasma with mean recoveries of 98. 17% and 98. 83% for TAD and DAP respectively.     

α-Amino acid derived enaminones and their application in the synthesis of N-protected methyl 5-substituted-4-hydroxypyrrole-3-carboxylates and other heterocycles

A new and simple synthesis of novel N-protected methyl 5-substituted-4-hydroxypyrrole-3-carboxylates, which exist in equilibrium with their 4-oxo tautomers, has been developed in two steps starting from N-protected α-amino acids. The key intermediates are enaminones, which can also be isolated, characterized, and used for the construction of other functionalized heterocycles, before they spontaneously decompose to pyrrole products. 4-Hydroxypyrroles are prone to partial aerial oxidation but can be efficiently alkylated or reduced to stable polysubstituted pyrrolidine derivatives.     

Specific intramolecular aromatic CH insertion of diazosulfonamides

The chemoselectivity in the intramolecular CH insertion of various diazosulfonamides has been experimentally studied. The results reveal that the aliphatic 1,4-, 1,5-, or 1,6-C(sp³)?H insertions of diazosulfonamides are not accessible, while the aromatic 1,5-C(sp²)?H insertion can be realized specifically by adjusting the diazo-adjacent group. In addition, the general chemoselectivities in the intramolecular CH insertions of diazosulfonyl compounds are summarized. Generally, diazosulfones undergo both aromatic 1,5-C(sp²)?H and aliphatic 1,5- and 1,6-C(sp³)?H insertions, while diazosulfonates undergo aliphatic 1,5- and 1,6-C(sp³)?H insertions. However, diazosulfonamides only undergo aromatic 1,5-C(sp²)?H insertion.     

N-Heterocyclic carbene-palladacyclic complexes: synthesis,characterization and their applications in the C-N coupling and α-arylation of ketones using aryl chlorides

N-Heterocyclic carbene-palladacyclic complexes 3 were successfully achieved in a one-pot procedure under mild conditions. The structure of 3a was unambiguously confirmed by X-ray single crystal diffraction and it was an active catalyst in the Buchwald-Hartwig amination and α-arylation of ketones even at very low catalyst loadings (0.01?mol%).     

Iodine-mediated oxidative Pictet-Spengler reaction using terminal alkyne as the 2-oxoaldehyde surrogate for the synthesis of 1-aroyl-β-carbolines and fused-nitrogen heterocycles

An efficient iodine-mediated oxidative Pictet-Spengler reaction in dimethyl sulphoxide (DMSO) using terminal alkynes as the 2-oxoaldehyde surrogate for the synthesis of aryl (9H-pyrido[3,4-b]indol-1-yl)methanones is described. The scope of the protocol includes the total synthesis of Fascaplysin, Eudistomins Y₁ and Y₂. The methodology is extended for preparing pyrrolo[1,2-a]-quinoxaline and indolo[1,5-a]quinoxaline derivatives. The utility of 1-aroyl-β-carbolines was demonstrated by performing palladium-catalyzed β-carboline directed ortho-C(sp2)-H functionalization of the phenyl ring with thiomethyl (SMe) group using DMSO as source and for accessing 4-aryl-canthin-6-ones.     

Facile synthesis of bicyclic 1-arylpyrazol-5-ones

In this Letter, we described a facile method for constructing fused bicyclic 1-arylpyrazol-5-one ring system. We employed various methylene-containing carboxylic acids as the substrates and proved that the pyrazolone ring closure requires activated methylene group in intermediate II. Accordingly, a series of structurally diversified, fused bicyclic 1-arylpyrazol-5-ones was prepared in moderate to high yields using the requisite substrates.     

A carbanion induced synthesis of highly congested pyrazole and imidazole containing heterocycles

An efficient approach to the synthesis of highly congested di, penta and hexacyclic pyrazoles as well as imidazole fragment containing novel heterocyclic molecule has been developed through a carbanion induced transformation of suitably functionalized 2H-pyran-2-ones, benzo[h]chromene and thiochromeno[4,3-b]pyrans. Due to the presence of fluorescence, we report their prime application metal sensor as off/on switching in ferric ions.     

Synthesis of pyrrolo[1,2-a]pyrazines and pyrazino[1,2-a]indoles by Curtius reaction in Morita-Baylis-Hillman derivatives

Synthesis of substituted pyrrolo[1,2-a]pyrazines and pyrazino[1,2-a]indoles from the Morita-Baylis-Hillman derivatives of acrylates via saponification followed by Curtius reaction is described.     

正丁胺对激光染料荧光谱影响的研究

用正丁胺作为碳源,采用射频辉光放电制备碳膜,选用激光染料R6G和聚乙二醇混合液作为蒸气源,采用单源热蒸发,在蒸发室与染料同时沉积得到混合膜,用拉曼光谱和红外光谱分析了碳膜的结构和键合方式,分析表明:碳膜中存在胺基团和氢原子.混合膜的荧光谱测量结果表明,认为正丁胺对染料荧光谱的影响是因为胺基和氢原子的存在.     

Selective synthesis of 4,5-dihydroimidazo- and imidazo[1,5-a]quinoxalines via modified Pictet–Spengler reaction

An efficient tandem approach for the selective synthesis of 4,5-dihydroimidazo[1,5-a]quinoxalines 6a–g and imidazo[1,5-a]quinoxalines 7a–h by the reaction of 2-imidazolyl anilines 4a–c with aryl aldehydes 5a–k under mild reaction conditions is described. Introduction of electron releasing alkyl groups in substrates 4a–b was found to be instrumental for the success of the reaction.     

Understanding the Diels-Alder reactivity of 1,2-azaborine analogues

The Diels-Alder reactivity of 1,2-heteroborines (H₄C₄B(H)X, X?=?NH, PH, AsH; O, S, Se) has been computationally explored by means of Density Functional Theory (DFT) calculations. The influence of the HB?=?X fragment on the reactivity of the system has been quantitatively analyzed in detail by means of the so-called Activation Strain Model (ASM) of reactivity. It is found that the interaction between these species and the dienophile is significantly stronger than that computed for their all-carbon isoelectronic counterpart, benzene. In addition, the strain energy plays a key role in the observed reactivity trends. The role of the aromaticity strength of these heteroarenes on the reactivity is also assessed.