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Interleukin-8 (IL-8) is regarded as an important mediator of inflammation because of its potent and specific chemotactic activity on neutrophils. In the present investigation, human umbilical vein endothelial cells (HUVEC) stimulated with thrombin were found to produce IL-8, in a dose- and time-dependent manner. After stimulation with 10 U/ml thrombin for 24 hr, the level of IL-8 in the conditioned medium was 14 ng/ml, or enough to elicit PMN chemotaxis in vitro. Northern blot analysis revealed that thrombin as well as IL-1 beta elevates the level of IL-8 mRNA preceding the formation of IL-8 protein. A synthetic peptide SFLLRN [human thrombin receptor-activating peptide (TRAP)] was found to mimic the action of thrombin. Preincubation with anti-thrombin compounds such as hirudin and antithrombin-III-heparin almost completely suppressed the action of thrombin without affecting the actions of other stimuli including IL-1 beta, phorbol 12-myristate 13-acetate (PMA) and TRAP. Diisopropylfluorophosphate-treated thrombin did not stimulate IL-8 production. Calphostin-C, a protein kinase C (PKC) inhibitor, attenuated the production of IL-8 by thrombin, TRAP and PMA, but left the action of IL-1 beta unchanged. These results strongly suggest that catalytic activation of thrombin receptor by thrombin results in PKC-dependent IL-8 production accompanied by an increase in IL-8 mRNA level.

To produce a thrombi-targeting plasminogen activator, we expressed a fused gene that contains a modified pre-sequence of Mucor pussilus rennin (MPR) followed by a chimeric gene of single-chain urokinase-type plasminogen activator (scu-PA): :annexin V (AV). The fused gene was ligated into an integrative vector, under the control of the alcohol oxidase 1 (AOX1) promoter (p), and transformed into Pichia pastoris. Transformants were monitored for the secretion of fibrinolytic activity. The highest expressing clone, HB225, secreted as much as 600 international units (IU) of fibrinolytic activity per ml of culture medium under optimal conditions. It contained three tandem copies of the full-size vector disruptively integrated into the AOXI sequence. Western blot analysis revealed that the secreted chimera was highly susceptible to proteolysis. Addition of excess amino acids (aa) to the culture medium minimized the degree of proteolysis. Two major species of chimera, 85 and 65 kDa, were then isolated from the culture medium. The former was the intact form consisting of a single-chain and showing full enzyme activity after activation by plasmin. The latter was an enzymatically processed form consisting of two chains held by a disulfide bond, having full enzyme activity without activation. Both chimeras exhibited calcium-dependent phospholipid (PL)-binding affinities similar to the parent AV.

Liquid-phase photofluorination (LPPF) of 2-phenyl-2-methoxyhexafluoropropane 1 gave the desired F -2-cyclohexyl-2-methoxypropane, 2 , in 21% yield. An unusual rearranged product, F -2-cyclohexylmethyl isopropyl ether, 4 , and a degradation product, F -2-cyclohexylpropane, 3 , were also produced in 10 and 20% yields, respectively. A plausible reaction mechanism for the formation of the rearranged product 4 was presented. The Cl-containing ethers with cyclic structures, 2-chlorohexafluorocyclopentenyl cyclopentyl ether, and 2-chlorohexafluorocyclopentenyl 2,2,3,3,4,4,5,5-octafluoro- n -pentyl ether were also perfluorinated by the LPPF method to give the corresponding Cl-containing F -ethers in fair yields. The total retention of the chlorine atom through perfluorination and the ability to have a benzene substituent on the substrate are benificial features of this novel perfluorinating method.

The tertiary amines derived from hexafluoropropene dimers were subjected to liquid-phase photofluorination (LPPF). The corresponding perfluorotertiary amines, which were difficult to synthesize by the conventional fluorination methods such as electrochemical fluorination and indirect fluorination using high-valency metal fluorides, were obtained in good yields. Prepared by this method were F -( N,N -dimethyl-2- methylpentyl) amine 3 , F -( N,N -diethyl-2-methylpentyl) amine 4 , F -3-(1-pyrrolidino)-2-methylpentane 11 , F - ( N,N -dimethyl-1,1-dimethyl-butyl) amine 13 . Discussion on the regioselectivity in the reaction of F -2-methyl-2-pentene (D-II) with secondary amines is also included in connection with the necessity of preparing a regioselective adduct,3-(1-pyrrolidino)- F -2-methyl-2- pentene, for the synthesis of 11 .

Thrombin, besides being a potent coagulation factor, exerts influence on endothelial and leukocyte functions and may thus be involved in the regulation of inflammatory reactions. The present study investigated whether thrombin stimulates the production of growth-related cytokine/melanoma growth-stimulatory activity (GRO alpha/MGSA) in endothelial cells. Human umbilical vein endothelial cells (HUVEC) stimulated with thrombin were found to product GRO alpha/MGSA in a dose- and time-dependent manner. This action of thrombin was completely suppressed by preincubation with either hirudin or antithrombin-III (AT-III)-heparin. Interestingly, the thrombin receptor-activating peptide SFLLRN mimicked the action of thrombin. In addition, staurosporine, a protein kinase C (PKC) inhibitor, attenuated the production of GRO alpha/MGSA by thrombin, SFLLRN and phorbol 12-myristate 13-acetate (PMA), but left the action of interleukin-1 beta (IL-1 beta) unchanged. These results suggest that catalytic activation of thrombin receptor by thrombin results in GRO alpha/MGSA production, at least in part, via a pathway involving PKC in HUVEC.

An intravenous infusion of Fluosol enhanced significantly the t-PA thrombolysis of the arterio-venous shunt made by insertion of 125I-fibrin clot in rabbits. The plasma radioactivity released through thrombolysis increased in both time and dose-dependent manner after the administration of t-PA. Fluosol in combination with t-PA increased the plasma radioactivity, compared with the t-PA treatment alone at the corresponding dosage. The coronary blood flow was markedly reduced to almost zero after the thrombin injection into narrowed LCX with a clamp in open-chest dogs. An intravenous infusion of Fluosol or Pluronic F-68 solution at a dose of 15 ml/kg significantly shortened the thrombolysis time by intracoronary infusion of urokinase alone. While, little change in the QTc interval of ECG and the plasma CPK-MB activity was observed in the Fluosol group in combination with urokinase, suggesting a myocardial protective action of Fluosol possibly due to its oxygen carrying effect.

This paper describes a novel, safe, effective technique, with a new reactor design, for the synthesis of perfluorochemicals. This method, the liquid-phase photofluorination with undiluted fluorine, is applicable to the preparative-scale synthesis of isomerically-pure branched F-alkanes, F-ethers and F-tert-amines which are difficult to prepare by the classical fluorination methods ( i.e. electrochemical or CoF 3 ). The technique is based on the controlled inverse addition of the materials to be fluorinated to a well-stirred F 2 -saturated inert solvent, with simultaneous UV irradiation. The application of this method to the synthesis of F-ethers is described: the following were made by this method, F-2-propoxy-2-methyl-pentane, F-2-isobutoxy-2-methylpentane, F-n-heptylpropylether, F-2-pentoxy-2-methoxypropane, F-3-propoxy-2-methylpentane, F-2-ethyl-2-isopropyltetra-hydrofuran, F-3-isobutoxy-2-methylpentane, F-2-methoxy-4-ethyl-4-methyl-hexane, and F-2-methyl-(1-ethyl-1-methylpropyl)oxetane.

We have developed an adriamycin-oxidized dextran conjugate (ADM-OXD) which was found to possess not only higher antitumor activity but also less animal toxicity than ADM. It appeared that improvement of the therapeutic index of ADM-OXD was due to its bio-distribution. Therefore, we examined the tissue distribution of [14C]-OXD and [14C] ADM in tumor bearing rats and mice. ADM-OXD gave much higher blood levels than ADM. It was also found that ADM-OXD was more highly concentrated in the tumor tissues than ADM in both rats and mice. However, heart tissues showed lower concentration of ADM-OXD. On the other hand, ADM-OXD tended to remain in the liver, kidney and spleen. Approximately 50% of the administered ADM-OXD was excreted into the urine within 6 hours after its administration.

In order to study the elimination rate of perfluorochemical (PFC) emulsion from the blood and the PFC contents in the main organs of the tumor-bearing animals, Wistar strain rats were intramuscularly implanted with Walker 256 carcinosarcoma and 9L glioma cells were innoculated into the femoral muscle of Fischer rats. After 7 d the Wistar rats received an injection of 4 g PEC/kg of a PFC emulsion. The Fischer rats received a similar injection after 11 d. The rats injected with the PFC emulsion were periodically sacrificed for a week. There was no significant difference in PFC contents in the blood and main organs between normal rats and the tumor-bearing rats with either Walker 256 carcinosarcoma or 9L glioma. The PFC contents in Walker tumor were about 10 times as much as those of 9L tumor. The innoculated tumors did not substantially affect the PFC retention in the blood and the accumulation of PFC in main organs after intravenously injected with the PFC emulsion.

The thermal stability of a cysteine-free alkaline protease (Alp) secreted by the eukaryote Aspergillus oryzae was improved both by the introduction of engineered twin disulfide bridges (Cys-69/Cys-101 and Cys-169/Cys-200), newly constructed as part of this study, and by the addition of calcium ions. We performed an extensive kinetic analysis of the increased thermal stability of the mutants as well as the role of calcium dependence. The thermodynamic activation parameters for irreversible thermal inactivation, the activation free energy (deltaG), the activation enthalpy (deltaH), and the activation entropy (deltaS) were determined from absolute reaction rate theory. The values of deltaH and deltaS were significantly and concomitantly increased as a result of introducing the twin disulfide bridges, for which the increase in the value of deltaH outweighed that of deltaS, resulting in significant increases in the value of deltaG. The enhancement of the thermal stability obtained by introducing the twin disulfide bridges is an example of the so-called low-temperature stabilization of enzymes. The stabilizing effect of calcium ions on wild-type Alp is similar to the results we obtained by introducing the engineered twin disulfide bridges.

Tissue distribution of the radioactivities after intravenous administration of [14C] adriamycin ([14C] ADM) or [14C] ADM linked to oxidized dextran ([14C] ADM-OXD) in mouse bearing Lewis lung carcinoma (LLC) and rat bearing Walker 256 carcinosarcoma was studied. ADM conjugated with OXD increased plasma half-life and gave high area under the plasma concentration-time curve (AUC). The AUC values were 13.0 and 5.8 times higher than those of the [14C] ADM group in mice and rats, respectively. In the tumor tissues, AUC values of the [14C] ADM-OXD group were also respectively 1.6 and 1.9 times higher than those of the [14C] ADM group. However, the AUC values in the heart of the [14C] ADM-OXD group were about half those of [14C] ADM group in both animals. Thus the distribution of ADM was changed by the conjugation with OXD. The excretion profile of ADM was also changed by the conjugation. During 6h after administration, [14C] ADM-OXD was mainly excreted into rat urine at 45.2% of the original dose, but in the [14C] ADM group recovery in urinary excretion was 4.2%. Using [14C] ADM-OXD and ADM-[14C] OXD, the respective tissue distribution of ADM and OXD portions in the ADM-OXD was studied in rats bearing Walker 256. The radioactivities of both [14C] ADM-OXD and ADM-[14C] OXD groups increased in tumor and liver within 1h after administration. In the liver, both radioactivities were retained for 24h, which suggested that ADM and OXD were retained as conjugated form, however, different behavior was observed between the two groups in tumor tissues. Peak [14C] ADM-OXD radioactivity was reached 6h after administration and then decreased, while, ADM-[14C] OXD was quickly cleared from the tumor after 1h administration. The results suggested that ADM was released from ADM-OXD and the remaining ADM-OXD, which was low in ADM content, was then cleared from the tumor tissues.

An anticancer agent, N-[[[4-(5-bromo-2-pyrimidinyloxy)-3-chlorophenyl]amino]carbonyl]-2 - nitrobenzamide (HO-221, 1), shows poor oral absorption and is only slightly soluble in water (0.055 microgram/mL at 37 degrees C). The coprecipitates with polyvinylpyrrolidone or a vinylpyrrolidone and vinylacetate copolymer (copolyvidone) showed a marked increase of the dissolution rate and attainment of temporary supersaturation of 1. The oral bioavailability of these preparations in dogs at a dose of 1 of 5 mg/kg was approximately 60%, which was 3.5 times greater than that of a micronized preparation. Further, the enteric coprecipitate with hydroxypropyl methylcellulose phthalate 200731, which showed a dissolution profile similar to that of the copolyvidone preparation at pH 6.5 but no dissolution at pH 1.2, revealed the almost complete oral absorption. Because intraduodenal administration of the copolyvidone coprecipitate showed a higher absorption than that of per oral administration, it was suggested that the partial precipitation of crystallites in the nonenteric coprecipitates occurred before reaching the absorption site, the small intestine.

The protease-resistant isoform of prion protein (PrP) has been implicated in the pathogenesis and transmission of Creutzfeldt-Jakob disease (CJD), scrapie and other related diseases, but the relationship between the infectious agent and PrP awaits elucidation. In the present study, we have examined levels of infectivity together with accumulation of the protease-resistant form of PrP (PrPCJD) in various tissues of CJD agent-inoculated mice. Accumulation of PrPCJD occurred only in tissues, including brain, salivary gland and spleen, in which infectivity was readily detectable throughout the course of the experiment. The brain showed the highest levels of both infectivity and PrPCJD accumulation, with well correlated kinetics. On the other hand, the high titres of infectivity detected in salivary gland and spleen early after inoculation of the agent were obviously distinguishable from PrPCJD. Furthermore, in the salivary gland, the kinetics of infectivity and the accumulation of PrPCJD reversed; infectivity declined as PrPCJD accumulated in the tissue. Our findings indicate that PrPCJD accumulation is associated with replication of the agent; however, PrPCJD is unlikely to be the agent itself.

N-[[[4-(5-Bromo-2-pyrimidinyloxy)-3-chlorophenyl]amino]carbonyl] - 2-nitrobenzamide (HO-221) is presently under development as an oral anticancer agent with a novel mode of action. However, HO-221 exhibits extremely poor bioavailability after oral administration because it is only slightly soluble in water (0.055 micrograms/ml at 37 degrees C). Our previous study revealed that the micronization of HO-221 to the submicron region improved this oral bioavailability. In this study, the oral pharmacokinetics of this micronized HO-221 was investigated in rats, dogs and monkeys. After oral administration, the agent was moderately absorbed with the Tmax of 6.5-8.0, 17.3-20.0 and 12.0 h, and eliminated with the terminal half-lives of 11.9-15.0, 66.8-78.3 and 42.3 h in rats, dogs and monkeys, respectively. The bioavailability was incomplete (3.7-21.4%). In rats, the plasma concentration did not increase proportionally with increasing oral doses. In dogs, food enhanced the bioavailability 2.2-fold with a standard meal and 3.6-fold with a high fatty meal as compared with fasting conditions.

N-[[[4-(5-Bromo-2-pyrimidinyloxy)-3-chlorophenyl]amino]carbonyl]-2 -nitrobenzamide (HO-221) is being developed as an anticancer agent with a novel mode of action. HO-221 shows poor oral absorption and is only slightly soluble in water (0.055 micrograms/ml at 37 degrees C). In this study, it was shown that the reduction in particle size of HO-221 to the submicron region (0.453 microm, mean by volume) could be achieved by a wet milling in a decaglycerin monolaurate aqueous solution with small glass beads. The wet milling suspension obtained showed improved dissolution rate and oral absorption in rats. A solid dosage form could also be made from that suspension with addition of sucrose palmitate which prevented aggregation caused by the hydrophobic interaction. The solid dosage form thus obtained showed twice as much oral absorption in dogs as the preparation made by dry milling.

Quantitative structure-activity relationships of a series of perfluorochemicals (PFCs) for use as oxygen transporters were investigated in terms of their in vivo half-lives (t1/2). The logt1/2 values can be correlated by a linear combination of lipophilic (e.g., critical solution temperature (CST)), physicochemical (e.g., molecular weight and vapor pressure (Pv)) and topological (e.g., number of fluorines (NOF)) parameters on regression analyses. The most important excretion-rate-determining factor was the CST. The Pv alone of PFCs did not correlate well with logt1/2. However, a combination of CST and Pv produced the most meaningful and important correlation, indicating that Pv plays a pivotal role in the excretion and can be considered a kind of correction factor to the CST. The t1/2 can thus be predicted from the combination of CST and Pv. It can also be predicted from structural information : the smaller the NOF, the shorter the t1/2. The incorporation of hetero atom (s) such as nitrogen and oxygen in the PFC molecule, which has been considered to produce better emulsion stability but results in persistence in the body, had relatively little effect in terms of t1/2. These results suggested that, taking the emulsifiability of hetero atom-containing PFCs into consideration, a cyclic F-ether and F-tertiary amine with fused ring (s), containing fewer fluorines than acyclic analogues, might be a good candidate compound as an oxygen transporter.

Electrochemical fluorination of quinolizidine gave F-quinolizidine in 16-23% isolated yields. 4-Methylquinolizidine was also fluorinated electrochemically to give the corresponding amine stereoisomers along with their fragmented and rearranged products in isolated yields of 28-34% and 2-3%, respectively. Introduction of an F-methyl group into the quinolizidine in place of the methyl group prior to electrochemical fluorination did not stabilize the 4-methyl quinolizidine structure and rather allowed the formation of F-quinolizidine to a greater extent. Oxidation of 4-(F-methyl)-F-quinolizidine with fuming sulfuric acid in the presence of a catalytic amount of HgSO 4 gave 6-oxo-4-(F-methyl)-F-quinolizidine in 60% yield, the F-nmr spectra of which gave further support to the structure of the starting material. The physical and spectral properties of these F-chemicals are described.

Electrochemical fluorination of N-methyldecahydroquinoline afforded mainly a mixture of cis and trans N-(F-methyl)-F-decahydroquinoline, their rearranged isomers and F-propyl-F-cyclohexane arising from the cleavage at carbon-nitrogen bonds, in a ratio of approximately 2:4:6:3. N-Methyldecahydroisoquinoline was also fluorinated electrochemically to give a mixture of cis and trans N-(F-methyl)-F-decahydroisoquinoline, their rearranged isomers and 1-(F-ethyl)-2-(F-methyl)-F-cyclohexane in a ratio of approximately 4:4:6:1. No correlation was found between the cis and trans ratio of starting materials and that of the corresponding perfluorinated amines. Fluorination of N-methyl-1,2,3,4-tetrahydroquinoline gave much lower yields.

The excretion mechanism of perfluorochemicals (PFC) via the lung has been studied by physicochemical and histochemical methods. Monocytes that had phagocytized PFC particles were found in the lung capillaries, and alveolar macrophages that had phagocytized PFC particles were found in the alveoli. Vacuolated macrophages were observed microscopically in the alveolar space of rats injected with PFC emulsions. The PFC content in alveolar macrophages reached a maximum level of 1.44 mg/l x 10(8) cells 72 h after injection. It was confirmed by means of a wavelength dispersive x-ray analyzer that fluorine atoms were present in the vacuolated alveolar macrophages. The peroxidase activity of the PFC-phagocytizing macrophages suggest that monocytes phagocytize PFC particles in the circulation and migrate to the alveolar space. This study shows that the monocyte/macrophage system is related to one mechanism for excretion of PFC after intravenous injection of PFC emulsions.

ペルフルオロオクチルブロミド(PFOB)は臭素原子を1個もつペルフルオロ化合物であり,X線遮蔽能をもつことから,造影剤としての応用が期待されている。Longらは,PFOB乳剤を担がん動物に投与し,腫瘍部位の造影に成功したと報告した。今回,著者らはAH130細胞とVX2 carcinomaの2種類の実験がんを用い,腫瘍造影剤としてのPFOB乳剤の可能性について検討した。AH130細胞を大腿部筋肉内に移植したラットにPFOB乳剤を投与し,投与後24時間目にX線撮影したところ,腫瘍の明瞭な造影籐が観察された。また,VX2 carcinomaを肝に移植した家兎に25w/v%PFOB乳剤を10ml/kg投与し,CT scanを行なった。肝正常組織にくらべ,腫瘍周辺部が高いdensityとして造影された。PFOB含量を測定した結果,周辺部に高濃度のPFOBが検出された。電子顕微鏡による組織学的検討からも,腫瘍周辺部に多数のマクロファージが存在し,そのマクロファージの中にPFOB粒子と思われる穎粒が多数散見され,特徴的な腫瘍のCT像がPFOB粒子の集積によることが明らかにされた。腫瘍造影剤としてのPFOBの有用性が確められた。

Electrochemical fluorination of 2-methoxy-1,1,1-trifluoro-2-(F-methyl) octane gave the corresponding perfluorinated ether in 27% yield, along with cyclic by-products in 9%. A mixture of partly fluorinated tertiary amines, consisting of 1-dipropylamino-F-1-propene and 1-dipropylamino-2-hydryl-F-propane, did not afford a superior yield of tri(F-propyl)amine compared to the unfluorinated tripropylamine. 1-Diethylamino-2-(F-methyl)-F-1-pentene was fluorinated to give the corresponding F-tertiary amine in fairly good yields, together with 1-di(F-ethyl)amino-2-hydryl-2-(F-methyl)-F-pentane and their fragmented products. The study indicates that blocking of the α-carbon atom of an ether with F-methyl groups seems to reduce fragmentation, resulting in good yields of an unrearranged product. However, partial fluorination of a tertiary amine prior to electrochemical fluorination rather allows high yields of undesired by-products, as far as our experiments were concerned.

Electrochemical fluorination of N-cyclopentylpyrrolidine gave the corresponding F-amine together with a ring-opened compound N-(F-pentyI)-F-pyrrolidine in the ratio of 1 to 1, in 55% yield. N-cyclohexylpyrrolidine, N-cyclopentylpiperidine, and N-cyclohexylpiperidine were also eletrochemically fluorinated in the same manner to give the corresponding F-amines, their isomers with rearranged structures, and ring-opened ones, in the ratio of ca - 4:2:1, and 2:1:1, respectively in 51 to 53% yields. Supporting spectral data are presented.

It has been assumed that a mononuclear phagocyte system is related to the excretion of PFC emulsions: PFC particles are phagocytized by blood monocytes to be expelled through the lung alveoli. This monocyte-related mechanism may well explain excretion at an early stage when PFC particles are abundant in the blood stream. It does not, however, fully explain the manner by which PFC cells are released from the RES cells into the blood stream and into the adipose tissue. To explain this, the following mechanism has been proposed and discussed based on some experimental results. PFC emulsion particles taken up by the RES organs, are stripped at their surfactant layers in the cells and move across the cell membranes to the blood vessels and into other tissues such as adipose, at a rate that depends on the lipophilicity of the PFC's. In the blood stream, PFCs are delivered by lipoproteins to the lung and excreted into the expired air. Pharmacokinetical analysis with a compartmental model for the excretion also supported this proposed mechanism.

To choose a perfluorochemical as a candidate for the artificial erythrocyte or organ perfusate, screening of perfluorochemicals was performed from the standpoints of persistence of them in the circulation and deposition in the body tissues of rabbits, mice and rats. In view of elimination from the body, perfluorodecalin was found to be the most suitable compound among five different perfluorochemicals examined. As for emulsifiers, egg yolk phospholipid was found to be the first choice on account of its ability to sustain perfluorochemicals for an adequate period in the circulation as well as the feasibility for preparing emulsion among the three emulsifiers examined. It was found also that the smaller the particle size of emulsion, the better suited for an artificial erythrocyte of perfusate.

Adriamycin linked to oxidized dextran (ADM-OXD) via Schiff's base formation was compared with free adriamycin with regard to antitumor activity, acute toxicity and plasma pharmacokinetics in rats following i.v. administration. ADM-OXD showed higher activity against Walker carcinosarcoma 256 than free adriamycin. On the other hand, the acute toxicity of ADM-OXD was about three times less than that of free adriamycin. In contrast to free adriamycin, a very high plasma level of adriamycin was found after i.v. administration of ADM-OXD. The area under the plasma concentration curve with ADM-OXD was about 160-fold higher than with free adriamycin. Thus, the improvement of the therapeutic index in the case of ADM-OXD might be due to the difference in the disposition of ADM-OXD and free adriamycin in rats.

As a part of studies on perfluorochemicals for candidate as an artificial blood, the elimination of six kinds of the substance was studied to find the way and type of excretion in rats given their emulsions intravenously. (1) These compounds are solely excreted through expiration, none through urinary and biliary with an exception of FC-43 (perfluorotrifbutylamine. (2) The elimination curves of these compounds from the body appear to be exponential). (3) The elimination rate of each substance was fairly related to its vapor pressure, the higher, the more rapidly eliminated. (4) Within six compounds examined sofar, FDC (perfluorodecalin) is selected on account of its rapid elimination.

Preparative liquid-phase direct perfluorination of organic compounds with elemental fluorine has not previously been developed, presumably owing to the occasional violence of the reaction and its tendency to give oligomers. We have devised a safe, effective technique based on controlled inverse addition of the organic substrate to a well-stirred saturated (about 0.02 M) solution of elemental fluorine in an unreactive solvent, accompanied by UV-irradiation. All work to date has been conducted in reactors constructed of Teflon, brass, copper and nickel, using F-hexane as the solvent, at a temperature of typically −5 to −20°C. 100% fluorine at atmospheric pressure is continuously supplied to maintain saturation; illumination is by a medium-pressure Hg arc in a sapphire immersion well. The substrates to be fluorinated are, in most cases, partly fluorinated amines and ethers derived from commercially-available F-olefins. Perfluorinated products are obtained without the accompanying isomerization usual in the electrochemical (ECF) process, in good yields and at fluorination rates also comparable to those reported in ECF reactors of similar volume. Typical results will be presented, along with supporting spectral data and physical properties.

The negative chemical ionization mass spectra of representative perfluorinated alkanes, cycloalkanes, ethers and tertiary amines have been examined, using Ar at about 0.5 torr as the reagent gas. The compounds chosen are typical of those under study as components of fluorochemical emulsion blood substitutes. Many such PFC's, particularly those with cyclic or branched structures, give intense molecular ions; most give simple spectra with a few major fragment ions at high mass, in marked contrast to the EI spectra which are dominated by m/e 69 (CF 3 +) and 131 (C 3 F 5 +) of no value for structure elucidation. NCI-GC/MS is more sensitive than conventional EI-GC/MS and promises to be more generally useful for structure determination. Specific examples from the various classes will be presented, and their NCI and EI mass spectra compared.