The Quantitation of Heroin and Selected Basic Impurities Via Reversed Phase HPLC. I. The Analysis of Unadulterated Heroin Samples

Abstract

Methodology was developed employing reversed phase liquid chromatography for the simultaneous determination of heroin, O³-monoacetylmorphine, o⁶-monoacetylmorphine, acetylcodeine, noscapine and papaverine in unadulterated illicit powders. An HS-5 C18 column was used with a gradient system using methanol and a hexylamine phosphate buffer at pH 2.2. This method, suitable for automated analysis, used a multi-mode detection scheme via the use of a photodiode array detector. In order to arrive at the optimum chromatographic conditions in terms of selectivity and stability, a study was performed on the effect of various mobile phase parameters on log k′ for heroin, various impurities, and common adulterants. The mobile phase parameters included amine concentration, organic modifier type, and eluent pH.     

Quantitative Analysis of Adulterants in Illicit Heroin Samples via Reversed Phase HPLC

Abstract

Methodology is presented using reversed phase liquid chromatography for the simultaneous quantitation of phenobarbital, methaqualone, phenolphthalein, nicotinamide, and N-phenyl-2-naphthylamine in heroin samples. A Partisil 5 ODS-3 column was used with a gradient system using methanol and a sodium dodecylsulfate-phosphate buffer.     

Quantitation of Creatinine in Urine and Plasma Samples by Reversed Phase HPLC

Abstract

Creatinine determination in urine and plasma affords an index of the renal function. Reversed-phase high pressure liquid chromatography was used for the separation and quantitation of creatinine in normal and arsenic exposed human urine samples. Acetonitrile/water (1:1) was the mobile phase. The method was compared with the Jaffé alkaline picrate reaction. Results show that the HPLC procedure has high reproducibility and samples are stable at the storage conditions. Plasma samples required depro-teinization and extraction with CH₃CN prior to HPLC analysis, while urine samples required only centrifugation.     

反相高效液相色谱法分析无机阴离子研究

本文研究了提高ＲＰ－ＨＰＬＣ测定无机阴离子的选择性的方法，设计了利用对离子试剂和背景试剂兼作ｐＨ调节剂，建立了几种分离分析系统和实际样品分析方法。     

Analysis of Plasma Angiotensins by Reversed Phase HPLC and Radioimmunoassay

Abstract

The analysis of biologically active angiotensin peptides in blood plasma by high performance liquid chromatography in a weakly non-polar reversed phase (C₂) chromatographic system combined with quantification of chromatographically isolated peptides by radioimmunoassay has been developed. This system is able to resolve each of seven closely-related peptides of the angiotensin group. The chromatographic system was applied to plasma samples which have been prepared for chromatographic analysis by C₁₈ cartridge extraction. Samples were reconstituted in HPLC solvent prior to injection into the HPLC system. Separated angiotensin were collected by fraction collector and the volatile components of the solvent system were blown off under an air stream. The content of several of the various angiotensin peptides in the fractions was then determined by radioimmunoassay using an appropriate antiserum. Antiserum to angiotensin II (octapeptide) was used to quantify the biologically active components angiotensin II, angiotensin III (hepta-peptide) and C-terminal hexapeptide. Recovery of angiotensin II in the C₁₈ cartridge extraction has been assessed at 85.0 ± 0.9% (mean ± SEM) using I¹²⁵-labelled angiotensin II, and 82.2 ± 4.45% using synthetic unlabelled angiotensin II. Recovery of standard preparations of angiotensin II in the HPLC system have been estimated at 67.5 ± 6.08%. The application of this technique to evaluating some components of the angiotensin in normal plasma is presented.     

Reversed Phase Chromatographic Analysis of 13 Amino Acids in Honey Samples

A simple method using reversed phase high-performance liquid chromatography (RP-HPLC) was developed for the simultaneous analysis of 13 amino acids. Amino acids were pre-column derivatized with 9-fluorenylmethyl chloroformate (FMOC-Cl) before analysis by RP-HPLC. Experimental parameters affecting the derivatization and chromatographic separation were investigated. Amino acids were derivatized with FMOC-Cl under alkaline condition in 0.1 mol/L borate buffer pH 10.0 at room temperature. The FMOC-amino acid derivatives were separated on an Atlantis C18 column under the gradient elution of 0.05 % trifluoroacetic acid and acetonitrile and UV detection at 265 nm. Linear ranges were 0.2–100.0 μg/mL with the correlation coefficients greater than 0.992. Limits of detection and limits of quantitation were in the range of 0.05–2.0 and 0.2–5.0 µg/L, respectively. The intra-day precision (n = 3) of retention time was less than 1 %, while for the peak area was less than 4 %. The inter-day precision (n = 3 × 3) of retention time was less than 2 % and the peak area was less than 8 %. This method was applied in honey samples and the results showed that proline is the major amino acids in honey samples.

     

Analysis of Allopurinol and Oxipurinol in Plasma by Reversed Phase HPLC

Abstract

A reversed phase HPLC assay is described for the quantitation of allopurinol and oxipurinol in human plasma. The strategies for the development of this method are discussed. The analysis is performed on a C-18 column using an acidic aqueous mobile phase. Sample preparation consists of protein precipitation with a mixture of trichloracetic acid and perchloric acid. The detection limit of the assay for both substances is in the region of 30ng/ml. This method has been applied to an investigation of the relative bioavailability of two commercial preparations of 300mg allopurinol tablets in eight healthy volunteers.     

高压液相色谱内表面反相填料的发展及其应用

内表面反相填料（ＩｎｔｅｒｎａｌｓｕｒｆａｃｅｒｅｖｅｒｓｅｄｐｈａｓｅＩＳＲＰ）是１９８５年首次研制出来的一种高压液相色谱填料，其微孔内表面接疏水基团，外表面接亲水基团，当含蛋白质或其它大分子的样品进入柱子后，蛋白等大分子不被系统保留，而小分子物质则可以进入微孔得到分离。这一填料的研制和发展使血清样品直接进样成为可能，在药物分析方面有独特的优点。本文综述了ＩＳＲＰ的特点、合成方法、工作原理及其应用。     

Analysis of Temocillin and Impurities by Reversed Phase Liquid Chromatography: Development and Validation of the Method

A robust, specific, precise and sensitive high-performance liquid chromatographic method has been described for purity control of temocillin. Chromatographic separation was achieved using a Symmetry C₁₈ (150 × 4.6 mm, 5 µm) column kept at 30 °C. The mobile phase consisted of a gradient mixture of mobile phases A (5 g/L solution of Na₂HPO₄·2H₂O, pH 7) and B (ACN-MeOH-H₂O, 50:10:40 v/v/v) pumped at a flow rate of 1.0 mL/min. UV detection was performed at 235 nm. The developed method was validated according to the ICH guidelines for its robustness, selectivity, sensitivity, precision and linearity. An experimental design was applied for the robustness study. Linearity was assessed both at impurity level in the range from LOQ to 10 % and assay level from 25 % to 150 % (0.6 mg/mL = 100 %). It is the first liquid chromatographic method described for the separation of temocillin and its potential impurities. It was possible to identify four degradation products from the forced degradation studies. The degradants do not interfere with the main peak and other known impurities showing that the method is specific and stability-indicating.     

An Improved Ion-Pair Reversed Phase LC Method for Analysis of Major Impurities of Phosphorothioate Oligonucleotide Cantide

An improved ion-pair reversed phase LC (IP-RP-LC) method was developed using triethylammonium/hexafluoroisopropanol (TEA/HFIP) buffer as the mobile phase as an alternative to capillary gel electrophoresis (CGE) and ion-exchange chromatography (IEC) for routine quality control of the synthetic 20-mer phosphorothioate oligonucleotide (PS-ODN) Cantide. The most commonly observed impurities included an oligomer with a deletion at the 5′-end (5′n-1) and a monophosphodiester (P=S/O). The method described here was sufficiently selective to simultaneously separate and identify these impurities. To the best of our knowledge, this is the first report on the separation of P=S/O analogs from PS-ODN using IP-RP-LC. Relatively high TEA concentrations and higher pH values were recommended. The separation performance was maintained even in the presence of multiple P=S/O fragments containing different PO residues. The method was successfully applied to the analysis of failure sequences of authentic crude Cantide as well as the purified product. In addition to 5′n-1 and P=S/O analogs, some minor impurities such as 3′n-1 oligomers were also partially resolved.     

Simultaneous Determination of Heroin, Amphetamine and their Basic Impurities and Adulterants Using Microemulsion Electrokinetic Chromatography

Heroin and amphetamine are two most abused drugs that can be highly complex, containing various impurities, byproducts, adulterants and diluents due to differences in the agricultural and manufacturing procedures. Therefore, the comprehensive analysis of above illicit drugs seized or purchased undercover by law enforcement authorities is important for legal and intelligence gathering purposes, and clinical and pharmaceutical purposes as well1. In last decade, micellar electrokinetic chromatogr…     

Indirect Ultraviolet Visualization and Quantitation of Methenamine with Reversed Phase Ion Pairing Liquid Chromatography

Abstract

This study reports for the first time, the successful indirect ultraviolet visualization of a polycyclic polyamine, methenamine, a urinary antibacterial agent. The ability to detect a non-UV active compound quantitatively using a UV monitor is demonstrated and the optimized experimental conditions are discussed. The calibration curve obtained was linear for over two orders of magnitude extending down to a detection limit of 250 picomoles.     

Determination of Polyphenylarenes and Polynuclear Aromatic Hydrocarbons by Reversed Phase HPLC

Abstract

Standard mixtures of polycyclic compounds are analyzed by reversed phase microcapillary liquid chromatography. Acetonitrile/water and acetonitrile/THF/water are employed as stationary phases, and capacity ratios are reported. Polyphenylarenes can be distinguished from polynuclear aromatic hydrocarbons by a large shift in capacity ratios when changing the mobile phase. The shift in capacity ratios is most significant for 1,3,5-triphenylbenzene, which is demonstrated by a gradient elution. The influence of THF and water on retention behaviour of solutes is described.     

Simultaneous Determination of Artemether and Azithromycin in Suppositories by Reversed Phase HPLC

Chromatographic parameter assessments for RP-HPLC-UV method development for the simultaneous analysis of artemether and azithromycin for the pharmaceutical analysis of a rectal coformulation currently under development for the treatment of malaria infected children. Using methanol based mobile phase for the analysis of both artemether and azithromycin provided a more robust method in terms of resolution and peak symmetry. The method validated for suppository used 80% methanol and 20% phosphate buffer 15 mM at pH 9. The UV detection was at 210 nm. The accuracy profiles indicated a method validation between 80–120% for both active pharmaceutical ingredients. The preparation process of the suppository was validated based on theoretical values of artemether and azithromycin present in the formulation; active pharmaceutical ingredients were homogenously distributed within the suppository.     

高纯橙皮素的制备及HPLC纯度分析

建立了一种基于反相硅胶纯化的高纯橙皮素制备方法,充分优化了上样量、流速及洗脱液极性等因素,采用红外光谱法和核磁共振对纯化后产物进行定性分析,高效液相色谱法检测纯化前后橙皮素的纯度,最终产物的纯度达99.87%。该方法快速简便、制备量大、分离效率高,每天制备效率可达到克级,为制备橙皮素标准物质提供了技术支撑。     

正反相模式下键合纤维素手性固定相对手性化合物的拆分

在自行合成的键合型纤维素-(3,5-二甲基苯基氨基甲酸酯)(CDMPC-CSP)手性固定相上,在正相模式下对苯偶姻和甲霜灵外消旋体进行了拆分,考察了流动相中醇以及温度在手性识别中对手性分离的影响。同时考察了反相模式下外消旋体的拆分。     

Leaching of Triazine Pesticides in Loamy Soil and their Determination by Reversed Phase HPLC

Abstract

The separation and identification of triazine pesticides (ametryn, atrazine, cyanazine and simazine) was carried out on Nova Pak C₁₈ column (150 × 3.9mm). The mobile phase used was acetonitrile-water (65:35, v/v) adjusted to pH 4.5 with acetic acid. The flow rate of the mobile phase used was 1.0mL/min. The detection of the pesticides was carried out at 250 nm. The values of the separation factor (α) were in the range of 1.49–5.32 and the values of the resolution factors (R _s) were ranged from 1.18 to 2.99 for the separated pesticides. The developed HPLC method was used to determine the concentrations of the reported pesticides in the loamy soil samples. The recovery of the pesticides from soil samples was found to be about 50%. The relative standard deviation and limit of detection were in the range of 0.01–0.02 and 0.5–1.0 μg/mL respectively.     

向日葵仁中绿原酸和咖啡酸的反相高效液相色谱法测定

建立了同时测定向日葵仁中绿原酸和咖啡酸的反相高效液相色谱法 ;采用美国Alltech公司的ALLTIMAC18 柱(250mm×4.6mmID,5μm) ,流动相为乙腈 -2 %(φ)乙酸溶液 (体积比15∶85) ,流速为1.0mL/min ,紫外检测波长327nm ;向日葵仁中绿原酸(chlorogenicacid)和咖啡酸(caffeicacid)在18min内得到很好的分离 ,该法简便、快速、结果准确、重复性好。     

高效液相色谱法同时快速测定凉茶中11种非法添加化学药物

采用核-壳亚3μm色谱柱建立了高效液相色谱同时测定凉茶中11种非法添加药物(对乙酰氨基酚、阿司匹林及其降解物水杨酸、磺胺甲唑、磺胺嘧啶、磺胺二甲嘧啶、氧氟沙星、环丙沙星、洛美沙星、土霉素、强力霉素)的方法。样品经甲醇-乙腈(1∶1)超声提取,安捷伦Poroshell 120 EC C18(100 mm×4.6mm,2.7μm)色谱柱分离,以0.1%三氟乙酸三乙胺溶液(p H 3.0)-甲醇-乙腈为流动相,梯度洗脱,流速1.2 m L/min,柱温35℃,采用二极管阵列检测器检测,外标法定量。11种成分的线性范围为0.5~25 mg/L,相关系数均不小于0.998 2,检出限为1~5 mg/L,平均回收率为86.9%~108.0%,相对标准偏差(RSD)为0.5%~1.4%。按上述方法对286批凉茶样品进行检测,发现9批阳性样品,检出成分为对乙酰氨基酚、阿司匹林及其水解物水杨酸、磺胺甲唑。阳性样品进一步采用液质联用法定性确证。该方法操作简便、灵敏、准确,适用于凉茶中11种化学成分的测定。