Chemical and enzymatic treatment of endothelin

Endothelin-1 (ET), the most potent vasoconstrictor yet discovered, is a peptide containirig 21 amino acids with two intrachain disulfide bridges. With the aim of obtaining two-chain derivatives, Et was submitted to chemical and enzymatic treatments. Reaction of ET with CNBr in 70% HCOOH gave, in addition to the expected [Hse⁷ lactone]-7,8-seco-ET and unreacted material, a by-product whose molecular weight was 25 m.u. greater than that of ET. When the reaction mixture, after lyophilisation, was immediately quenched with NH3-saturated dry MeOH, two products could be recovered in a 5:1 ratio, both obtained by nucleophilic attack of the homoserine lactone: the expected [Hse⁷-NH₂]-7,8-seco-ET and [Hse⁷]ET, resulting from competitive intramolecular reaction of the deprotonated α-amino group of the Asp⁸ residue. The Lys⁹-Glu¹⁰ bond turned out to be very resistant to enzymatic attack both by Lys-C-endopeptidase and trypsin. The 9,10-seco-ET derivative could be obtained by treatment with Lys-C-endopeptidase only by using a high enzyme/ET ratio and after a prolonged incubation time. Cleavage of the Lys⁹-Glu¹⁰ bond could not be achieved by treatment with trypsin, even with a high enzyme/substrate ratio. The main product was 13, 14-seco-ET, deriving from the action of chymotripsin (present as an impurity in the trypsin preparation) on Tyr¹³. The structure of these peptides was confirmed by amino-acid sequence analysis and fast atom bombardment mass spectrometry (FAB-MS). Nicking of the ET structure at different positions had different impact on the biological properties of the resulting derivatives. © Munksgaard 1995.     

四逆汤煎煮过程中乌头类生物碱的溶出和水解平衡

目的 研究四逆汤中乌头类生物碱的溶出平衡和水解平衡。方法 利用电喷雾质谱分析，比较了制附子、四逆汤、四逆汤药渣和含有3种双酯型生物碱的混合对照品体系中的乌头碱类二萜生物碱。结果 在煎煮过程中双酯型生物碱溶解并发生水解反应，而脂类生物碱则难溶于水。乌头碱、中乌头碱、次乌头碱水溶性相近，但是次乌头碱在水中的热稳定性更高，C19二萜骨架上C3取代基(—OH或—H)的变化影响生物碱的稳定性。结论四逆汤中的乌头碱类生物碱的种类及含量由其溶解性和化学稳定性共同决定。     

一阶导数光谱法测定丙吡胺尿药浓度及其缓释片的相对生物利用度

本文报道了用导数光谱法测定磷酸丙吡胺的尿药浓度,本方法能消除尿中杂质干扰,测定结果准确。6名受试者交叉口服磷酸丙吡胺缓释片和普通片,测得尿药浓度计算消除速度常数K,排泄速率(Δx/Δt)_(max) 所需时间,证明缓稀片药物排泄慢,药效维持时间长,且缓释片的相对生物利用度与普通片基本相同。     

Orthogonal solid-phase synthesis of a monobiotinylated analog of Neuropeptide Y

Analogs of Neuropeptide Y (NPY) were synthesized with conventional Boc/benzyl protective group strategy. Instead of Asn⁷ in the native scquence, Boc-Lys(Alloc)-OH was incorporated. At the end of the synthesis the Alloc group was selectively removed by palladium-catalyzed hydrostannolysis and biotin coupled to the e-amino group of Lys⁷. After cleavage and characterization with plasma desorption mass spectrometry the N^e-7-biotinyl-[Lys⁷]-NPY and the nonbiotinylated analog [Lys⁷]-NPY were investigated as ligands to the NPY receptor from rat cerebral cortex. Both analogs were found to be high affinity ligands to the NPY receptor and bound with essentially the same affinity as unmodified NPY.     

城市污灌废水的致突变性检测及色谱/质谱分析

应用Ames致突变试验,对比研究了武汉市易家墩、黄孝河污灌区污灌废水与土壤的致突变性。同时应用色谱/质谱(GC/MS)分析技术。测定了主要污水中有机污染物的化学组成。结果表明:罗家渠废水的致突变活性最强;受各条污水灌溉的土壤的致突变活性顺序与相应废水的强弱顺序一致;罗家渠废水中致突变致癌物主要是萘、联苯等。     

Determination of metabolites of a novel platinum anticancer drug JM216 in human plasma ultrafiltrates

The present study describes the application of on-line liquid chromatography-electrospray ionisation in conjunction with a high resolution magnetic sector mass spectrometer to identify metabolites of a platinum(IV) anticancer drug JM216 bis(acetato)amminedichloro(cyclohexylamine)platinum(IV)] in human plasma. Four metabolites were identified following incubation of JM216 in human plasma: JM118 [amminedichlorocyclohexylamineplatinum(II)], a platinum(II) complex; JM383 [bis(acetato)amminedihydroxo(cyclohexylamine)platinum(IV)]; JM518 [bis(acetato)amminechloro(cyclohexylamine)hydroxoplatinum(IV)] and its isomer JM559. The platinum complexes mass spectra were dominated by the natriated [M + Na]⁺ ion. Elemental compositions of these natriated ions were confirmed by accurate mass measurement on a magnetic sector mass spectrometer in the course of LC/MS analysis. This study demonstrates the capability of direct LC-ESI/MS with accurate mass measurement for analysis of platinum complexes in biological samples. Our results suggest that LC-ESI/MS is a powerful technique for structure elucidation of novel metabolites, and could make valuable contributions to drug metabolism research.     

Uptake and biotransformation of sevof lurane in humans: A comparative study of sevof lurane with halothane, enflurane, and isoflurane

Study Objective: To compare the volatile anesthetic sevoflurane with halothane, enfurane, and isof urane on the uptake and biotransformation in humans.

Design: Prospective pharmacokinetic study of sevofurane administration in human subjects.

Setting: Inpatient surgery clinic at a university medical center.

Patients: Thirty-two Japanese patients, free of systemic diseases, undergoing minor elective surgery with endotracheal general anesthesia.

Interventions: The patients were assigned randomly to one of four groups: halothane, enflurane, isofurane, or sevofurane. One of the four volatile anesthetics being investigated [equivalent to 1.1 minimum alveolar concentration (MAC): halothane, 0.85%; enfurane, 1.85%; isofurane, 1.27%; and sevofurane, 1.88%; in inspired concentrations throughout the first hour of anesthesia] was administered for 60 minutes.

Measurements and Main Results: In all patients, serum and urinary fluoride concentrations were measured. The concentrations of all gases were measured separately with a mass spectrometer. The cumulative uptake of each anesthetic agent during a certain period was calculated as an integration of the uptake rate per minute. The results for one-hour inhalation of sevofurane (1.1 MAC) showed an uptake (corrected for body surface area and MAC) of 490 ml/m²/MAC and estimated degradation rate of 3.3%. For purposes of comparison, similar studies of halothane (uptake, 653 ml/m²/MAC; degradation rate 15.7%), enfurane (1150 ml/m²/MAC; 1.3%), and isofurane (439 ml/m²/MAC; 0.6%) were also conducted. Sevofurane had a peak serum inorganic fluoride concentration of 19.3 μmol/L, and no abnormality in hepatic or renal functions was observed in any of the subjects during the two weeks postoperatively.

Conclusions: Accurate determinations of uptake and degradation rate for sevoflurane and three other volatile anesthetics in Japanese patients were obtained. These findings have established that, despite its relatively large MAC *1.71%), sevoflurane has a small uptake due to its low solubility. However, the degradation rade was shown to be as high as 3.3%, resulting in a higher serum fluoride concentration than seen after administration of isoflurane, halothane, and (possibly) enflurane.     

氢化物原子荧光光度法测定食品中砷的不确定度评定

目的探讨氢化物原子荧光光度法测定食品中砷的不确定度评定方式。方法依据JJF1059-1999建立数学模型,合成不确定度。结果当食品中砷含量为0.22mg/kg时,合成不确定度为0.008mg/kg,扩展不确定度为0.02mg/kg。结论本评定过程清晰合理,主要影响因素不遗漏、不重复,适用于原子荧光光度法的不确定度评定。     

Determination of the ratios of the aromatic amino acid residues by first- or second-derivative UV spectrometry for a simple characterization of peptides

A method for the simultaneous determination of the ratios of the three aromatic amino-acid residues in peptides was set up in acidic conditions. Binary and ternary mixtures of these amino acids were prepared, and first- and second-derivative spectra then calculated from their 0.1 nm resolution spectra between 240 and 320 nm. Certain spectral bands were chosen to differentiate tyrosine from tryptophan on the first-derivative spectra, and phenylalanine from tyrosine and tryptophan on the second-derivative spectra. Variation of the amplitude of the chosen bands was shown to be a linear function of the ratio of the aromatic amino acids in the mixture. This technique was validated with peptides whose sequence was known. The difference between theoretical and experimentally determined ratios was lower than 10%. Since the results are obtained as ratios, neither the concentration nor the nature of the peptide has to be known. The feasibility of application using a photodiode array detector with high resolution in reversed-phase high-performance liquid chromatography is discussed. © Munksgaard 1995.     

荧光分析法测定血清中微量硒

为测定血清中微量硒,较低温度消化后,用2,3-二氨基萘为试剂,以荧光法测定,取得了满意的结果。方法简便快速,结果稳定准确。血清中加入标准硒,8次测定的回收率为96.6％～102.4％,平均值±标准差为99.6±1.8％。