阿胶、龟甲胶中脂溶性成分的高效液相色谱指纹图谱

采用高效液相色谱法研究并建立东阿阿胶、龟甲胶脂溶性成分的指纹图谱,为药用动物胶的质量控制提供了有效的方法。采用液-液-液三相静态萃取方法制备样品,以水-乙腈为流动相进行二元梯度洗脱,检测波长为205 nm,柱温25 ℃,分析时间为60 min。采集20批样品的色谱图并对其进行相似度和聚类分析。分别标定了阿胶、龟甲胶的共有峰,其相似度分析及聚类分析结果显示两种胶间存在着明显的差异。该方法稳定可靠,可以有效地区别不同种属的药用动物胶,为动物胶剂的鉴别及质量控制提供了依据。     

五味子提取物高效液相色谱分析方法的优化

针对五味子乙醇提取物的复杂体系,借助于复杂样品分析系统软件(CSASS),根据组分在4次简单线性梯度下42个峰的保留时间,快速准确地计算出各组分的液相色谱保留参数a,c值和峰形参数σ,W1/2。借助这些参数,对五味子色谱的分离情况进行高精度仿真预测。在此基础上,应用移动重叠分辨分离图和谱图仿真技术,发展了计算机辅助的五味子提取物的高效液相色谱全局优化方法。在优化条件下,五味子提取物的高效液相色谱分析可在40 min内完成,且常量成分和部分低含量成分都能够得到较好的分离。所建立的方法已成功地用于五味子中化合物保留时间及峰形的预测,并在此基础上对其色谱分离条件进行优化。     

Development and application of a high resolution liquid chromatographic method for the analysis of complex pigment distributions

Ternary and binary gradient systems have been developed for the high-performance liquid chromatographic analysis of complex pigment distributions typical of natural samples. Improved chromatographic resolution reveals significantly more pigment components in extracts from a sediment (Priest Pot, Cumbria, UK), a microbial mat (les Salines de la Trinital, South Catalonia, Spain) and a culture (C. phaeobacteroides) including novel bacteriochlorophyll derivatives. The methods developed are directly suited to LC-MS analysis and the automated acquisition of MS/MS data for pigments.     

High-efficiency high performance liquid chromatographic analysis of red wine anthocyanins

The analysis of anthocyanins in natural products is of significant relevance in recent times due to the recognised health benefits associated with their consumption. In red grapes and wines in particular, anthocyanins are known to contribute important properties to the sensory (colour and taste), anti-oxidant- and ageing characteristics. However, the detailed investigation of the alteration of these compounds during wine ageing is hampered by the challenges associated with the separation of grape-derived anthocyanins and their derived products. High performance liquid chromatography (HPLC) is primarily used for this purpose, often in combination with mass spectrometric (MS) detection, although conventional HPLC methods provide incomplete resolution. We have previously demonstrated how on-column inter-conversion reactions are responsible for poor chromatographic efficiency in the HPLC analysis of anthocyanins, and how an increase in temperature and decrease in particle size may improve the chromatographic performance. In the current contribution an experimental configuration for the high efficiency analysis of anthocyanins is derived using the kinetic plot method (KPM). Further, it is shown how analysis under optimal conditions, in combination with MS detection, delivers much improved separation and identification of red wine anthocyanins and their derived products. This improved analytical performance holds promise for the in-depth investigation of these influential compounds in wine during ageing.     

A high performance liquid chromatographic method for the quantitation of flecainide in plasma.

A routine high performance liquid chromatographic method for the rapid determination of fleca?nide (Flecaine), using a novel internal standard, N-methylfleca?nide, has been developed. After deproteinization of spiked samples, fleca?nide was totally recovered at neutral pH. Fleca?nide and the internal standard were separated on a reversed phase XL 3 microns ODS column using 10 mM phosphate buffer, pH 3.0: acetonitrile (70:30) as mobile phase, in less than 10 min. With spectrofluorometric detection, the limit of quantitation for fleca?nide was 10 ng/mL. Intra- and inter-assay precision variations were 0.24% and 1.4%.     

反相液相色谱法测定α-甲氧基萘的含量

用反相液相色谱法分析了α-甲氧基萘的含量和其中的有害杂质。色谱条件：色谱柱Nucleosil－C18;流动相V（甲醇）：V（水）=90：10;紫外检测器,面积归一化法定量。方法的变异系数为0．29％。     

Validation and evaluation of a high performance liquid chromatographic method for the determination of aluminium in wine

A new method is reported for the determination of aluminium in wine by HPLC, involving derivatisation with 8-hydroxyquinoline (oxine) in the presence of micelles resulting in the formation of a fluorescent derivative. The complex is separated on a C18 column using a mobile phase of oxine - SDS - 35% acetonitrile, in a pH 7 buffer. The method was validated in the range 0.125-2 mg/l in a synthetic wine. The method was applied to the determination of aluminium in white, rosé and red wines and results compared with those obtained by atomic absorption (GFAA). Aluminium concentrations found by HPLC in white wines were greater than those found in red wines. Further investigation using a polyphenol-enriched white wine revealed a statistically significant inverse relationship between wine polyphenol content and the aluminium concentration determined by HPLC. The method may therefore be envisaged for the determination of unbound aluminium in wine.     

Development of a method using high‐performance liquid chromatographic fingerprint and multi‐ingredients quantitative analysis for the quality control of Yangxinshi Pian

A simple and reliable method of high‐performance liquid chromatography with diode array detection method was developed for fingerprint analysis and simultaneous determination of six compounds including puerarin, salvianolic acid B, berberine hydrochloride, palmatine chloride, dehydrocorydaline, and icariin in the Chinese medicine preparation Yangxinshi Pian. The separation was performed on an Agilent Eclipse XDB‐C₁₈ reserved‐phase column (250 mm × 4.6mm I.D., 5 μm) using gradient elution with 50 mmol/L monopotassium phosphate aqueous solution and methanol as mobile phase at a flow rate of 1.0 mL/min. The column operating temperature was set at 30°C, and the detection wavelength was 280 nm. The method was validated by linearity, precision, accuracy, stability, and recovery. For fingerprint analysis, 25 peaks were selected as the common peaks, and four kinds of similarities including cosine similarity (S), ratio of similarity (S′), projection content similarity (C), and content similarity (P) were applied to evaluate the quality consistency of different batches of Yangxinshi Pian. The results showed that the developed method was an efficient tool for quality evaluation of Yangxinshi Pian.     

高效液相色谱法测定水产品中的红霉素残留量

建立了以高效液相色谱法测定水产品中红霉素残留量的方法。提出了以乙腈为提取剂,经过液-液萃取、固相萃取、反萃取等步骤对样品进行分离净化的样品处理方法。测定下限为200μg/kg,回收率70%,批内精密度和批间精密度均小于10%。     

Improved high performance liquid chromatographic method for determination of carotenoids in the microalga Chlorella pyrenoidosa

Microalgae have become an important commercial source of carotenoids and microalgae-derived functional foods are consumed by people worldwide. Therefore, an HPLC method was developed to discern the variety and content of carotenoids in the microalga Chlorella pyrenoidosa. The microalga sample was powdered, extracted, saponified and subjected to HPLC analysis. A mobile phase of methanol-acetonitrile-water (84:14:2, v/v/v) (A) and methylene chloride (100%) (B) with the following gradient elution was developed: 100% A and 0% B in the beginning, maintained for 14 min, decreased to 95% A in 25 min, 75% A in 30 min, 74% A in 35 min, 45% A in 50 min and returned to 100% A in 55 min. A total of 32 carotenoids were resolved within 49 min by using a C30 column with flow rate at 1 mL/min and detection at 450 nm. An internal standard beta-apo-8'-carotenal was used to quantify all the carotenoids. All-trans-lutein was present in exceptionally large amount (125034.4 microg/g), followed by cis isomers of lutein (27975.3 microg/g), all-trans-alpha-carotene (2465.8 microg/g), zeaxanthin (2170.3 microg/g), cis isomers of beta-carotene (2159.3 microg/g), all-trans-beta-carotene (2155.0 microg/g), cis isomers of alpha-carotene (1766.7 microg/g), beta-cryptoxanthin (334.9 microg/g), neoxanthin and its cis isomers (199.7 microg/g), neochrome (65.2 microg/g), auroxanthin (38.5 microg/g) and violaxanthin and its cis isomers (38.1 microg/g).     

A high performance liquid chromatographic method for the screening of 17 diuretics in human urine

A simple and adequate HPLC method was developed for screening of human urine for the following 17 diuretic drugs: acetazolamide, bendrofluazide, bumetanide, canrenoic acid, chlorothiazide, chlorthalidone, clopamide, epitizide, etacrynic acid, furosemide, hydrochlorothiazide, indapamide, mefruside, piretanide, spironolactone, torasemide, and triamterene. The assay involves extraction from two 2 mL urine samples with ethyl acetate at pH = 5, washing with a phosphate buffer at pH = 6 and analysis by HPLC using a reversed phase C18 column and ultraviolet detection with a diode array detector for all drugs (except triamterene) using two eluents consisting of water, triethylamine, phosphoric acid and acetonitrile at different ratios and different pH values. Triamterene is determined by direct injection of diluted urine onto the column and is measured by fluorescence detection. The recoveries of the diuretic drugs were determined at two different concentrations and ranged from 43–110% (median: 87%) which is sufficient to detect abuse of these drugs. The repeatability of the assay ranged from 1–12% (median: 5.5%). Received: 13 July 1998 / Revised: 23 October 1998 / Accepted: 29 October 1998     

A high performance liquid chromatographic method for the screening of 17 diuretics in human urine

A simple and adequate HPLC method was developed for screening of human urine for the following 17 diuretic drugs: acetazolamide, bendrofluazide, bumetanide, canrenoic acid, chlorothiazide, chlorthalidone, clopamide, epitizide, etacrynic acid, furosemide, hydrochlorothiazide, indapamide, mefruside, piretanide, spironolactone, torasemide, and triamterene. The assay involves extraction from two 2 mL urine samples with ethyl acetate at pH = 5, washing with a phosphate buffer at pH = 6 and analysis by HPLC using a reversed phase C18 column and ultraviolet detection with a diode array detector for all drugs (except triamterene) using two eluents consisting of water, triethylamine, phosphoric acid and acetonitrile at different ratios and different pH values. Triamterene is determined by direct injection of diluted urine onto the column and is measured by fluorescence detection. The recoveries of the diuretic drugs were determined at two different concentrations and ranged from 43–110% (median: 87%) which is sufficient to detect abuse of these drugs. The repeatability of the assay ranged from 1–12% (median: 5.5%).     

A review of quantitative ion exchange,high performance liquid and gas chromatographic analysis of amino acids in physiological fluids

Methodologies for quantitative analysis of amino acids in physiological fluids based on classical ion exchange, high performance liquid chromatography, and gas chromatography are analyzed. As judged by the relative number of reports on these techniques, it appears that classical ion exchange continues to be the main method of choice for amino acid determinations and only limited advantage has been taken of the benefits (e.g. lower capital outlay and running costs and shorter analysis times) offered by other techniques. More importantly, however, there appears to be insufficient quantitative evaluation of the methodologies used. As a result, the validity of many reports based on data from amino acid analysis of physiological samples may be questioned.     

Development and validation of a high performance liquid chromatographic method for the simultaneous determination of potassium clavulanate and cefadroxil in synthetically prepared tablets

A simple, sensitive and rapid high performance liquid chromatographic method was developed and validated for the simultaneous determination of potassium clavulanate and cefadroxil in synthetically prepared tablets. Chromatographic separation and detection was carried out on a C-18 column using 0.05 M potassium dihydrogen phosphate buffer (pH 5.0) and acetonitrile in the ratio of 94: 06 (v/v) as mobile phase at wavelength of 225 nm. The method was linear in the concentration range of 3.75–22.5 μg/mL for potassium clavulanate and 15–90 μg/mL for cefadroxil. The flow rate was 1.0 mL/min and the total analysis time was less than 10 min. The mean recoveries was found to be greater than 99% with RSD less than 1.0%. The proposed method was validated by performing linearity, recovery, specificity, robustness, LOD/LOQ and within-day and between-day precision. The chromatographic results obtained from the synthetically prepared tablets show that the method is highly precise and accurate for the simultaneous quantitation of clavulanate potassium and cefadroxil.     

Development and validation of a simple stability-indicating high performance liquid chromatographic method for the determination of miconazole nitrate in bulk and cream formulations

A simple and stability-indicating high performance liquid chromatographic method was developed and validated for the determination of miconazole nitrate in bulk and cream preparations. The extraction step for cream samples consisted in a warming, cooling and centrifugation procedure that assures the elimination of the lipophilic matrix component, in order to avoid further precipitation in the chromatographic system. Separation was achieved on a ZORBAX Eclipse XDB - C18 (4.6 mm × 150 mm, 5 μm particle size) column, using a mobile phase consisting of water, methanol and acetonitrile, in a flow and solvent gradient elution for 15 min. The column was maintained at 25 °C and 10 μL of solutions were injected. UV detection was performed at 232 nm, although employment of a diode array detector allowed selectivity confirmation by peak purity evaluation. The method was validated reaching satisfactory results for selectivity, precision and accuracy. Degradation products in naturally aged samples could be simultaneously evaluated, without interferences in the quantitative analysis.     

Simplified high performance liquid chromatographic quantitation of antipyrine

Antipyrine is used as an index of hepatic drug metabolism. The authors devise a simple and rapid high performance liquid chromatography method to quantitate antipyrine in serum with small volume (50 microliters). Minimal sample preparation is needed and the assay is sensitive to concentrations of 0.2 micrograms/ml. Mobile phase consists of 35% methanol in 67 mM phosphate buffer pH 3.5 with antipyrine detected by monitoring the eluate at 225 and 254 nm. Within-run and run-to-run coefficients of variation are less than or equal to 2.8 and 4.6%, respectively. Recoveries from serum average 100.8%. It is concluded that this technique for antipyrine is simple, rapid, and well characterized.     

A high performance liquid chromatographic method for the determination of ethylenethiourea in urine and on filters

Urine samples are evaporated and pretreated with silica gel and alumina. Filters undergo ultrasonic treatment in water. Ethylenethiourea (ETU) is then determined by reversed-phase high-performance liquid chromatography with detection at 230 nm. The detection limit for ETU is 0.1 ng per injection and linear response is found for the range 0.3–110 ng; 0.2 μg 1^?1 ETU can be detected in urine. Recoveries from spiked filters (7 μg ETU/filter) varied from 79 to 94%. The methods are sensitive enough for application in occupational hygiene work.     

高效液相色谱法测定方便面中丙烯酰胺

建立了一种检测方便面中丙烯酰胺的液相色谱方法。萃取剂选择1g/L的蚁酸溶液,将萃取液以12000r/min,2℃离心18min,上清液再以14500r/min,0℃离心18min,可分离基质中淀粉、蛋白质、脂肪等干扰物;方法采用ZORBAX SB-C18(250mm×4.6mm,5μm)色谱柱,流动相为A(乙腈∶水=1∶24(V/V))和B(乙腈),流速0.5mL/min,在丙烯酰胺的最大吸收197nm处检测,提高了方法的检出限及回收率。该方法有良好的线性关系(r=1.00000),检出限为48.0μg/kg。回收率94.4%～109.8%,相对标准偏差为4.9%～5.6%。     

A high performance liquid chromatographic method for the determination of albuterol enantiomers in human serum using solid phase extraction and chemical derivatization.

A high performance liquid chromatographic method was developed for the simultaneous assay of R(-)- and S(+)-albuterol in human serum. The assay involves solid phase extraction as a sample clean-up step and derivatization of racemic albuterol to its diastereomeric thioureas with 2,3,4,6-tetra-O-acetyl-alpha-D-glucopyranosyl isothiocyanate. Chromatographic separation was accomplished under isocratic conditions using an octadecylsilane column and a mobile phase consisting of 29:71 acetonitrile:distilled water containing 0.1% triethylamine, pH 4.0 (adjusted with concentrated phosphoric acid) at a flow rate of 0.8 mL/min. The diastereomers were detected using a fluorescence detector set at 223 nm excitation and no emission filter. Racemic bamethane was used as internal standard. Drug to internal standard peak-height ratios were linear over a 2-20 ng/mL range for each enantiomer. The limit of detection of each analyte was 1.0 ng/mL (S/N = 3).