Development and validation of a stability-indicating HPLC method for the determination of degradation products in dipyridamole injection

Summary The development and subsequent validation of an isocratic high-performance liquid chromatographic (HPLC) procedure employing ultraviolet (UV) detection for the determination of degradation products in Dipyridamole Injection is reported. The development of this assay involved the evaluation of several factors including buffer type, ionic strength, pH, organic composition, and column type. The described method is simple, reproducible, accurate, and selective. The precision, relative standard deviation (RSD), amongst five sample preparations for total degradation products was not more than (NMT) 10.2 %, while the individual degradation products were NMT 12.1%. Intermediate precision, as determined from fifteen sample preparations, generated by two Analysts on different HPLC systems over three days, exhibited an RSD for total and individual degradation products of 8.2 % and NMT 27.5 %, respectively. The mean absolute recovery of dipyridamole using the described method is 102.1±1. 9%, (mean±SD, n=12) over the concentration range of 0.03 % to 5.0 % of its label claim of 5 mg mL⁻¹. The limit of detection and limit of quantitation were 0.1 and 0.3 μg mL⁻¹, respectively. The linearity of the peak response was verified with respect to dipyridamole concentration over a range of 0.3 and 50 μg mL⁻¹ (0.03 % to 5.0 % label claim). The Standard and Assay Preparations are stable for up to 48 hours at room temperature. The selectivity was evaluated by subjecting the finished product (Dipyridamole Injection) to thermal, acidic, basic, oxidative and fluorescent radiation stress conditions. No interference in the analysis of degradation products was observed, showing the method is stability-indicating.     

超高效合相色谱法快速检测塑料食品接触材料中13种紫外吸收剂

建立了一种同时检测塑料食品接触材料中13种紫外吸收剂的超高效合相色谱法。以甲醇为溶剂对塑料食品接触材料样品进行超声提取，经C18固相萃取柱净化，过0.22 μm有机滤膜，采用超高效合相色谱仪分析。选择ACQUTY UPC² HSS C18 SB色谱柱（150 mm×3.0 mm，1.8 μm），以超临界二氧化碳为流动相，异丙醇为改性剂进行梯度洗脱，在最优色谱条件下，13种紫外吸收剂能够在4 min内实现有效分离。结果表明，在各自线性范围内，13种紫外吸收剂的线性关系良好，标准曲线相关系数不低于0.9985，检出限（S/N=3）为0.05~0.15 mg/kg，加标回收率为86.8%~115.7%，相对标准偏差为0.73%~5.61%。该方法快速简便，准确可靠，同时大大减少了有机溶剂的消耗，可用于塑料食品接触材料中13种紫外吸收剂的快速检测。     

Simultaneous sample preparation for high-performance liquid chromatographic determination of Vitamin A and β-carotene in emulsified nutritional supplements after solid-phase extraction

Determination of small amounts of the fat-soluble species Vitamin A (VA) (2.5 μg/g) and β-carotene (9 μg/g) from emulsified nutritional supplements containing 50 kinds of co-existing compounds and a fat content between 2000 and 8000 times higher was performed by solid-phase extraction (SPE) and high-performance liquid chromatography (HPLC) with fluorescence detection set at ex. 350 nm and em. 480 nm, and visible detection at 450 nm using an Inertsil ODS 80A (5 μm) analytical column. Mobile phases of methanol-ethanol (50:50) and acetonitrile-ethanol (70:30) were used for the both vitamins. A Bond Elut C₁₈ cartridge was chosen for SPE after comparison with eight other types of SPE cartridge. Retention time of VA and β-carotene was 7 and 8 min, respectively, giving a limit of detection of ca. 0.1 ng per injection at a signal-to-noise ratio 3:1. Recoveries of VA and β-carotene were over 90% by the standard addition method. Relative standard deviation of VA and β-carotene were ca. 2.9 (n=5) and 2.3% (n=5), respectively.     

Synthesis of (+)-goniothalesdiol and (+)-7-epi-goniothalesdiol

A total synthesis of (+)-goniothalesdiol, a 3,4-dihydroxy-2,5-disubstituted tetrahydrofuran isolated from Goniothalamus borneensis (Annonaceae), and its 7-epimer is reported using oxycarbonylation methodology for construction of polyhydroxylated substituted heterocycles. Diastereoselectivity of addition of organometallic reagents to 2,3-O-isopropylidene-d-threose derivatives using theoretical calculations based on the semiempirical PM5 was studied.     

QuEChERS/超高效液相色谱-串联质谱法同时测定水产品中四溴双酚A与六溴环十二烷

建立了同时测定水产品中四溴双酚A(TBBPA)和六溴环十二烷(HBCD)的超高效液相色谱-串联质谱方法。样品前处理采用QuEChERS方法,均质生物样品用水分散后加乙腈提取,经C_(18)分散固相萃取净化。采用CORTECS~?C_(18)色谱柱(4.6 mm×100 mm×2.7μm)分离,以甲醇-水为流动相进行梯度洗脱,流速为0.7 mL/min。采用电喷雾离子源,在负离子模式下以多反应监测(MRM)方式检测。结果表明,TBBPA和3种HBCD单体在0.5～500μg/L范围内线性良好,相关系数(r)大于0.998,检出限(LOD,S/N=3)和定量下限(LOQ,S/N=10)分别为0.04～0.16μg/kg和0.12～0.55μg/kg。在5、20、50μg/kg加标水平下的平均回收率为74.0%～121%,相对标准偏差(RSD)为0.20%～23%。该方法操作简单、快速、重现性好,适用于水产品中TBBPA和HBCD的快速检测。     

Assigning the C-15 configuration of 15-hydroxy-, 15-methoxy-, 15-ethoxy-hexahydrofurofuran neoclerodane diterpenoids

Examination of ¹H and ¹³C NMR spectra allows the establishment of rules for assigning the correct configuration at C-15 of 15-hydroxy-, 15-methoxy-, 15-ethoxy-hexahydrofurofuran neoclerodane diterpenoids. The structure of several diterpenoids has been assigned or ammended.     

Current mass spectrometry strategies for selenium speciation in dietary sources of high-selenium

This document reviews the most relevant mass spectrometry approaches to selenium (Se) speciation in high-Se food supplements in terms of qualitative and quantitative Se speciation and Se-containing species identification, with special reference to high-Se yeast, garlic, onions and Brazil nuts. Important topics such as complexity of Se speciation in these materials and the importance of combining Se-specific detection and molecule-specific determination of the particular species of this element in parallel with chromatography, to understand their nutritional role and cancer preventive properties are critically discussed throughout. The versatility and potential of mass spectrometric detection in this field are clearly demonstrated. Although great advances have been achieved, further developments are required, especially if speciatedcertified reference materials (CRMs) are to be produced for validation of measurements of target Se-containing species in Se-food supplements.     

Glass and polymeric membrane electrodes for the measurement of pH in milk and cheese

While there is a considerable interest in the food industry in determining various analytes using ion-selective electrodes (ISEs), only few reports describe their use for direct measurements in food. In this study, the suitability of glass electrodes and ionophore-based solvent polymeric ISEs for the determination of pH in Process cheese, Cheddar cheese and milk was investigated. The liquid junction potential between a 3 M KCl bridge electrolyte and diluted as well as undiluted Process cheese was found to be negligible. Reference electrodes with ceramic plug and sleeve-type junctions performed well, although precautions needed to be taken to prevent plugging at the junctions. While the protein rennet casein posed no problems in pH measurements, the extraction of neutral lipophilic compounds or hydrophobic peptides into solvent polymeric membranes was evident, resulting in some loss of selectivity for monovalent cations upon exposure to cheese. However, it was found that ISEs based on tridodecylamine (R₃N) as ionophore and o-nitrophenyl octyl ether (oNPOE) as plasticizer can be used to accurately measure the pH of milk and, after desensitization of the electrodes in a cheese emulsion, of diluted Process cheese. Since pH measurements with a glass electrode showed that emulsions of cheese moderately diluted to a cheese content of 70% have the same pH as undiluted cheeses, it is possible to determine the pH in cheese with ionophore-based ISEs. R₃N membranes also performed well in undiluted milk.     

Separation of phytosterol oxidation products by combination of different polarity gas chromatography capillary columns

The number of characterized phytosterol oxidation products (POPs) from both ring- and side-chain structures has increased during recent decades, resulting in difficulties in the separation of POPs on different gas chromatography (GC) capillary columns. The main objective of this study was to separate a mixture of 29 purified and characterized oxidation products from sito-, campe- and stigmasterol using GC capillary columns with different polarity. For the first time in the area of POPs analysis, the separation efficiency of the combination of two capillary GC columns with different polarities was investigated. A non-polar 5% phenyl coated (DB5-MS) and a mid-polar 35% phenyl coated (DB35-MS) column was combined with a pressfit connector. The main improvement was enhanced base line separation for many of the analyzed POPs, compared with the separations achieved using the individual columns. However, three pairs of POPs co-eluted: 24-hydroxysitosterol/campesterol-5beta,6beta-epoxide, stigmasterol-5beta,6beta-epoxide/campesterol-5alpha,6alpha-epoxide and stigmasterol-5alpha,6alpha-epoxide/campestanetriol.     

Actin-binding marine macrolides: total synthesis and biological importance

Marine organisms produce a fascinating range of structurally diverse secondary metabolites, which often possess unusual and sometimes unexpected biological activities. This structural diversity makes these marine natural products excellent molecular probes for the investigation of biochemical pathways. Recently, a number of novel and stereochemically complex macrolides, having a large macrolactone (22- to 44-membered) ring, that interact with the actin cycloskeleton have been isolated from different marine sources. Actin, like tubulin, is a major component of the cytoskeleton and has important cellular functions. Although the details of these interactions are still under investigation, these marine macrolides are becoming increasingly important as novel molecular probes to help elucidate the cellular functions of actin. Owing to their potent antitumor activities, these compounds, for example the aplyronines, also have potential for preclinical development in cancer chemotherapy. Their appealing molecular structures, with an abundance of stereochemistry, and biological significance, coupled with the extremely limited availability from the marine sources, have stimulated enormous interest in the synthesis of these compounds. This review summarizes the biological properties of these unusual marine natural products and features the recently completed total syntheses of swinholide A, scytophycin C, aplyronine A, mycalolide A--all of these being potent cytotoxic agents that target actin--and a diastereoisomer of ulapualide A. Rather than detailing each individual step of these multistep total syntheses, the different synthetic strategies, key reactions, and methods adopted for controlling the stereochemistry are compared.