MPP+-Induced Changes in Cellular Impedance as a Measure for Organic Cation Transporter (SLC22A1-3) Activity and Inhibition

The organic cation transporters OCT1-3 (SLC22A1-3) facilitate the transport of cationic endo- and xenobiotics and are important mediators of drug distribution and elimination. Their polyspecific nature makes OCTs highly susceptible to drug–drug interactions (DDIs). Currently, screening of OCT inhibitors depends on uptake assays that require labeled substrates to detect transport activity. However, these uptake assays have several limitations. Hence, there is a need to develop novel assays to study OCT activity in a physiological relevant environment without the need to label the substrate. Here, a label-free impedance-based transport assay is established that detects OCT-mediated transport activity and inhibition utilizing the neurotoxin MPP⁺. Uptake of MPP⁺ by OCTs induced concentration-dependent changes in cellular impedance that were inhibited by decynium-22, corticosterone, and Tyrosine Kinase inhibitors. OCT-mediated MPP⁺ transport activity and inhibition were quantified on both OCT1-3 overexpressing cells and HeLa cells endogenously expressing OCT3. Moreover, the method presented here is a valuable tool to identify novel inhibitors and potential DDI partners for MPP⁺ transporting solute carrier proteins (SLCs) in general.     

Relationships between Inhibition,Transport and Enhanced Transport via the Organic Cation Transporter 1

The organic cation transporter 1 (OCT1, SLC22A1) transports a large number of structurally diverse endogenous and exogenous substrates. There are numerous known competitive and non-competitive inhibitors of OCT1, but there are no studies systematically analyzing the relationship between transport, stimulation, and inhibition. Here, we tested in vitro OCT1 inhibition by OCT1 substrates and transport of OCT1 inhibitors under uniform analytical conditions. Beyond inhibition testing with two model substrates, we tested nine additional OCT1 substrates for their mutual inhibition. Inhibition of ASP⁺ uptake by most OCT1 substrates was weak. The model substrate sumatriptan, with its moderately stronger inhibitability, was used to confirm this. Interestingly, OCT1 substrates exhibiting stronger OCT1 inhibition were mainly biaromatic β-agonistic drugs, such as dobutamine, fenoterol, ractopamine and ritodrine. Biaromatic organic cations were both, strong inhibitors and good substrates, but many OCT1 substrates showed little pairwise inhibition. Surprisingly, sumatriptan did significantly enhance dobutamine uptake. This effect was concentration dependent and additional experiments indicated that efflux inhibition may be one of the underlying mechanisms. Our data suggests, that OCT1 substrates are mainly weak OCT1 inhibitors and among those inhibiting well, noncompetitive inhibition could be responsible. Weak competitive inhibition confirms that OCT1 inhibition screenings poorly predict OCT1 substrates. Additionally, we showed that the OCT1 substrate sumatriptan can enhance uptake of some other OCT1 substrates. OCT1 transport stimulation was already observed earlier but is still poorly understood. Low OCT1 uptake inhibition and strong OCT1 efflux inhibition could be mechanisms exploitable for enhancing transport.     

Interaction Profiles of Central Nervous System Active Drugs at Human Organic Cation Transporters 1–3 and Human Plasma Membrane Monoamine Transporter

Many psychoactive compounds have been shown to primarily interact with high-affinity and low-capacity solute carrier 6 (SLC6) monoamine transporters for norepinephrine (NET; norepinephrine transporter), dopamine (DAT; dopamine transporter) and serotonin (SERT; serotonin transporter). Previous studies indicate an overlap between the inhibitory capacities of substances at SLC6 and SLC22 human organic cation transporters (SLC22A1–3; hOCT1–3) and the human plasma membrane monoamine transporter (SLC29A4; hPMAT), which can be classified as high-capacity, low-affinity monoamine transporters. However, interactions between central nervous system active substances, the OCTs, and the functionally-related PMAT have largely been understudied. Herein, we report data from 17 psychoactive substances interacting with the SLC6 monoamine transporters, concerning their potential to interact with the human OCT isoforms and hPMAT by utilizing radiotracer-based in vitro uptake inhibition assays at stably expressing human embryonic kidney 293 cells (HEK293) cells. Many compounds inhibit substrate uptake by hOCT1 and hOCT2 in the low micromolar range, whereas only a few substances interact with hOCT3 and hPMAT. Interestingly, methylphenidate and ketamine selectively interact with hOCT1 or hOCT2, respectively. Additionally, 3,4-methylenedioxymethamphetamine (MDMA) is a potent inhibitor of hOCT1 and 2 and hPMAT. Enantiospecific differences of R- and S-α-pyrrolidinovalerophenone (R- and S-α-PVP) and R- and S-citalopram and the effects of aromatic substituents are explored. Our results highlight the significance of investigating drug interactions with hOCTs and hPMAT, due to their role in regulating monoamine concentrations and xenobiotic clearance.     

Transport Turnover Rates for Human OCT2 and MATE1 Expressed in Chinese Hamster Ovary Cells

MATE1 (multidrug and toxin extruder 1) and OCT2 (organic cation transporter 2) play critical roles in organic cation excretion by the human kidney. The transporter turnover rate (TOR) is relevant to understanding both their transport mechanisms and interpreting the in vitro–in vivo extrapolation (IVIVE) required for physiologically-based pharmacokinetic (PBPK) modeling. Here, we use a quantitative western blot method to determine TORs for MATE1 and OCT2 proteins expressed in CHO cells. MATE1 and OCT2, each with a C-terminal V-5 epitope tag, were cell surface biotinylated and the amount of cell surface MATE1 and OCT2 protein was quantified by western analysis, using standard curves for the V5 epitope. Cell surface MATE1 and OCT2 protein represented 25% and 24%, respectively, of the total expression of these proteins in CHO cells. The number of cell surface transporters was ~55 fmol cm⁻² for MATE1 and ~510 fmol cm⁻² for OCT2. Dividing these values into the different J_max values for transport of MPP, metformin, and atenolol mediated by MATE1 and OCT2 resulted in calculated TOR values (±SE, n = 4) of 84.0 ± 22.0 s⁻¹ and 2.9 ± 0.6 s⁻¹; metformin, 461.0 ± 121.0 s⁻¹ and 12.6 ± 2.4 s⁻¹; atenolol, 118.0 ± 31.0 s⁻¹, respectively. These values are consistent with the TOR values determined for a variety of exchangers (NHEs), cotransporters (SGLTs, Lac permease), and uniporters (GLUTs, ENTs).     

Synchrotron radiation ablation of polymers having double bonds in their main chains

Polyacetylene (PA), poly(cis- and trans-1,4-butadiene)s (cis- and trans-PBs), and poly(p-phenylene vinylene) (PPV) were ablated by synchrotron radiation (SR), aiming to deposit thin, uniform films of each on a substrate. When PA was irradiated by SR, gaseous phenyl compounds were produced, and a thin amorphous film was deposited on the substrate, exhibiting no characteristics of PA. In the cases of PPV and trans-PB, the source materials were reproduced in the form of thin film on the substrate by SR ablation. When alkali halides, e.g. NaCl and KBr, were used as deposition substrates, PPV was deposited, in an ordered way, on their cleavage surfaces. However, the deposited film of trans-PB by SR ablation was non-crystalline, because it was produced as a copolymer by 1,4- and 1,2-addition polymerizations of ablated butadiene-based fragments. In comparison, thin films of these polymers were also prepared by thermally evaporating them in a vacuum. When trans-PB and PPV were thermally evaporated, thin films with chemically and structurally identical features to the source polymers were produced, respectively. In contrast, a deposited film from cis-PB by SR ablation consisted of carbon compounds, showing no sign of hydrocarbon compounds in it, while trans-PB was produced from cis-PB by thermal vapor deposition.     

SLC22 Transporters in the Fly Renal System Regulate Response to Oxidative Stress In Vivo

Several SLC22 transporters in the human kidney and other tissues are thought to regulate endogenous small antioxidant molecules such as uric acid, ergothioneine, carnitine, and carnitine derivatives. These transporters include those from the organic anion transporter (OAT), OCTN/OCTN-related, and organic cation transporter (OCT) subgroups. In mammals, it has been difficult to show a clear in vivo role for these transporters during oxidative stress. Ubiquitous knockdowns of related Drosophila SLC22s—including transporters homologous to those previously identified by us in mammals such as the “Fly-Like Putative Transporters” FLIPT1 (SLC22A15) and FLIPT2 (SLC22A16)—have shown modest protection against oxidative stress. However, these fly transporters tend to be broadly expressed, and it is unclear if there is an organ in which their expression is critical. Using two tissue-selective knockdown strategies, we were able to demonstrate much greater and longer protection from oxidative stress compared to previous whole fly knockdowns as well as both parent and WT strains (CG6126: p < 0.001, CG4630: p < 0.01, CG16727: p < 0.0001 and CG6006: p < 0.01). Expression in the Malpighian tubule and likely other tissues as well (e.g., gut, fat body, nervous system) appear critical for managing oxidative stress. These four Drosophila SLC22 genes are similar to human SLC22 transporters (CG6126: SLC22A16, CG16727: SLC22A7, CG4630: SLC22A3, and CG6006: SLC22A1, SLC22A2, SLC22A3, SLC22A6, SLC22A7, SLC22A8, SLC22A11, SLC22A12 (URAT1), SLC22A13, SLC22A14)—many of which are highly expressed in the kidney. Consistent with the Remote Sensing and Signaling Theory, this indicates an important in vivo role in the oxidative stress response for multiple SLC22 transporters within the fly renal system, perhaps through interaction with SLC22 counterparts in non-renal tissues. We also note that many of the human relatives are well-known drug transporters. Our work not only indicates the importance of SLC22 transporters in the fly renal system but also sets the stage for in vivo studies by examining their role in mammalian oxidative stress and organ crosstalk.     

New Cases of Hypochromic Microcytic Anemia Due to Mutations in the SLC11A2 Gene and Functional Characterization of the G75R Mutation

Divalent metal-iron transporter 1 (DMT1) is a mammalian iron transporter encoded by the SLC11A2 gene. DMT1 has a vital role in iron homeostasis by mediating iron uptake in the intestine and kidneys and by recovering iron from recycling endosomes after transferrin endocytosis. Mutations in SLC11A2 cause an ultra-rare hypochromic microcytic anemia with iron overload (AHMIO1), which has been described in eight patients so far. Here, we report two novel cases of this disease. The first proband is homozygous for a new SLC11A2 splicing variant (c.762 + 35A > G), becoming the first ever patient reported with a SLC11A2 splicing mutation in homozygosity. Splicing studies performed in this work confirm its pathogenicity. The second proband harbors the previously reported DMT1 G75R mutation in homozygosis. Functional studies with the G75R mutation in HuTu 80 cells demonstrate that this mutation results in improper DMT1 accumulation in lysosomes, which correlates with a significant decrease in DMT1 levels in patient-derived lymphoblast cell lines (LCLs). We also suggest that recombinant erythropoietin would be an adequate therapeutic approach for AHMIO1 patients as it improves their anemic state and may possibly contribute to mobilizing excessive hepatic iron.     

Linear and nonlinear optical properties of trans- and cis-poly(1-ethynylpyrene) based sonogel hybrid materials

The synthesis of sol-gel materials induced by ultrasonic action (sonolysis) is implemented as an alternative method for the fabrication of highly pure organic-inorganic composites with good monolithic and optical properties. The resulting SiO₂ glass exhibits high porosity and allows the inclusion of organic compounds in the colloidal sol-state. In this work, optical properties of trans-poly(1-ethynylpyrene) (trans-PEP) and cis-poly(1-ethynylpyrene) (cis-PEP) (M_w=24,000 g/mol) incorporated in this kind of gels were studied by absorption and fluorescence spectroscopies. Absorption spectra of the polymers showed that trans-PEP possesses a higher degree of conjugation than its homologue cis-PEP in sol-gel. Intramolecular interactions occur between adjacent pendant pyrene units (associated pyrenes) present in each polymer, giving rise to static excimer emissions, strongest in cis-PEP because of the shorter distances between aromatic rings. The results were compared to those previously reported for these polymers in solution. Besides, trans- and cis-PEP exhibited nonlinear optical properties like third harmonic generation (THG), which were measured in sol-gel phase for spin-coated film samples.     

Catalytic hydrogenation of linoleic acid over platinum-group metals supported on alumina

Catalytic activity and selectivity for hydrogenation of linoleic acid (cis-9,cis-12 18:2) were studied on Pt, Pd, Ru, and Ir supported on Al₂O₃. Stearic acid (18:0) and 10 different octadecenoic isomers (18:1) in the products could be separated completely by using a new capillary column coated by isocyanopropyl trisilphenylene siloxane for gas-liquid chromatography. The monoenoic acid isomers and dienoic acid isomers in the products on the various catalysts showed different distributions. The catalysts exhibited nearly equal selectivity for stearic acid formation. The 12-position double bond in linoleic acid has higher reactivity than the 9-position double bond in catalytic hydrogenation on platinum-group metal catalysts. In addition to hydrogenation products of linoleic acid, geometrical and positional dienoic acid isomers (trans-9,trans-12; trans-8,cis-12; cis-9,trans-13; trans-9,cis-13; cis-9,trans-12 18:2), due to isomerization of linoleic acid during hydrogenation, were contained in the reaction products. Ru/Al₂O₃ exhibited the highest activity for isomerization of linoleic acid with the noble metal catalysts. Conjugated octadecadienoic acid isomers have been observed in products of the reaction on Pt/Al₂O₃, Ru/Al₂O₃, and Ir/Al₂O₃. Catalytic activities of noble metals for positional and geometric isomerization of linoleic acid during hydrogenation decreased in the sequence of Ru ≥ Pt > Ir » Pd.     

Thermal, optical, electrochemical properties and conductivity of trans- and cis-poly(1-ethynylpyrene): a comparative investigation

The thermal, optical and electrochemical properties of trans-poly(1-ethynylpyrene) (trans-PEP) and cis-poly(1-ethynylpyrene) (cis-PEP) have been studied as a function of polymer backbone configuration and internal stacking. Absorption spectra of the polymers showed that trans-PEP possesses a higher degree of conjugation than its homologue, cis-PEP. Intramolecular interactions occur between adjacent pendant pyrene units (associated pyrenes) present in each polymer, giving rise to static excimer emissions, strongest in cis-PEP because of the shorter distances between aromatic rings. Data resulting from excitation spectra and fluorescence decay profiles proved that such interactions take place in the ground state. Cyclic voltammetry of trans- and cis-PEP exhibited irreversible behaviors with different oxidation potentials as a result of their dissimilar geometry.     

On the existence of cis/trans peptide mixtures in poly(N-methylglycine)

Unperturbed dimensions have been computed for poly(trans-N-methylglycine), poly(cis-N-methylglycine) and poly(cis/trans-N-methylglycine) by a Monte Carlo simulation technique using potential energy calculations. Computed characteristic ratios for the above polypeptides vary within a narrow range supporting the view that both cis and trans units could be present in the polymer in different proportions under different solvent conditions.     

Deregulated SLC2A1 Promotes Tumor Cell Proliferation and Metastasis in Gastric Cancer

Gastric cancer (GC) is one of the common reasons of cancer-related death with few biomarkers for diagnosis and prognosis. Solute carrier family 2 (facilitated glucose transporter) member 1 protein SLC2A1, also known as glucose transporter type 1 (GLUT1), has been associated with tumor progression, metastasis, and poor prognosis in many human solid tumors. However, little is reported about its clinical significance and biological functions in GC. Here we observed a strong up-regulation of SLC2A1 in patients with GC and found that SLC2A1 was significantly correlated with depth of invasion and clinical stage. Additionally, over-expression of SLC2A1 in GC cells promotes cellular proliferation and metastasis in vitro and enhances tumor growth in vivo as well as enhancement of glucose utilization. Meanwhile, elevated SLC2A1 also contributes to tumor metastasis in vitro. Our results indicate SLC2A1 exhibits a pivotal role in tumor growth, metastasis and glucose metabolism, and also suggest SLC2A1 as a promising target for gastric cancer therapy.     

New multifunctional sulfonamide-containing azobenzene chromophores: Synthesis and photochromic properties

The work presents synthesis and characterization of novel sulfadimethoxine and sulfabenzamide azo derivatives as well as the kinetic study of their trans-cis-trans isomerization. Spectral properties and kinetic constants were determined for the chromophores dispersed in poly(methyl methacrylate-co-butyl methacrylate) deposited on glass substrates in form of transparent, thin films. The maximum absorption bands were observed within wavelength in range of 446-457 nm for N-disubstituted aminoazobenzene derivatives and 361-362 nm for phenolic chromophores. For the phenolic azo derivatives of sulfadimethoxine and sulfabenzamide the photoisomerization induced with UV light followed the first order kinetics, while for other compounds the process combined of two parallel reactions with different rate constants was observed. The rate constants of thermal relaxation in the dark were usually few times smaller than those determined for photoinduced trans-cis isomerization. The change of real part of refractive index upon illumination determined by ellipsometry was within the range of 0.0043-0.0138.     

Use of HPLC in the identification of cis and trans-diaquabis(2,2′-bipyridine)ruthenium(II) complexes: crystal structure of cis-[Ru(H2O)2(bpy)2](PF6)2

[Ru(H₂O)₂(bpy)₂](PF₆)₂ complex was obtained by reacting HPF₆ in a [Ru(CO₃)(bpy)₂] aqueous solution. The complex can exist as cis and trans isomers and usually has been used in the preparation of several ruthenium–bipyridine species. Despite the possibility to have contamination of a specie in another, there is no analytical control involving the characterization of both complexes. Based on this we have proposed the use of high-performance liquid chromatography (HPLC) as an analytical technique to control the purity of cis and trans isomers. The separation was performed using a CLC-ODS column. The cis isomer eluted at 9.4 min while trans isomer eluted at 4.3 min. In aqueous solution the trans and cis isomer configurations were confirmed by NMR spectra (¹H). The attribution of cis isomer was also made based on the X-ray crystal structure (monoclinic, P2_1/c, a=12.320(2), b=13.852(2), c=34.220(3) Å, β=91.89(1)°, Z=8) which is reported. The six-coordinated ruthenium atom is chelated by two bipyridines and two molecules of H₂O.     

Prostaglandins of rat testis

The purpose of the study was to determine whether prostaglandins were present in mammalian testis, a tissue that has a large concentration of polyenoic fatty acids that are potential precursors of prostaglandins. Acid-soluble lipids of rat testis were extracted, purified, and fractionated by thin layer and column chromatographies.³H-Prostaglandins were added as internal reference standards to monitor recoveries and facilitate identification. Initial identification of prostaglandin species was done by chromatography. Further identification was done by elution of the prostaglandin zones followed by rechromatographies (both thin layer and column), measurements of UV absorption spectra, and by gas liquid chromatography. The results of these analyses indicate that prostaglandin E₁, 11α,15(S)-dihydroxy-9-oxo-β-trans-prostenoic acid; prostaglandin E₂, 11α-15(S)-dihydroxy-9-oxo-5-cis-13-trans-prostadienoic acid; and prostaglandin F₁, 9α,11α,15(S)-trihydroxy-13-trans-prostenoic acid occur in rat testicular tissue and that prostaglandin F_2α, 9α, 11α, 15(S)-trihydroxy-5-cis-13-trans-prostadienoic acid and prostaglandin E₂, 11α,15(S)-dihydroxy-9-oxo-5-cis-13-trans-prostadienoic acid may be the primary species of this tissue. Prostaglandin B₁, 15(S)-hydroxy-9-oxo-8(12),13-trans-prostadienoic acid and prostaglandin B₂, 15(S)-hydroxy-9-oxo-5-cis,8(12),13-trans-prostatrienoic acid also were detected, and some evidence was obtained for the presence of prostaglandin metabolites.     

Photo-activated metal carbonyl complexes as stereoselective catalysts for the homogeneous hydrogenation of dienes

Significantly increased activity of Cr(CO)₆ was achieved for the stereoselective homogeneous hydrogenation of methyl sorbate andtrans,trans-conjugated fatty esters at ambient temperature and pressure by exposing the catalyst to UV irradiation (3500 Å) in a solvent mixture of cyclohexane-acetonitrile (20:1). In this solvent mixture, methyl sorbate was converted quantitatively at ambient conditions into methylcis-3-hexenoate, and methyltrans-9,trans-11-octadecadienoate into methylcis-10-octadecenoate (99.9%). These products are expected by 1,4-addition of hydrogen. Under these conditions no hydrogenation of methyl linoleate occurred. Under the same conditions, cycloheptatriene-Cr(CO)₃ showed lower activity than Cr(CO)₆, and Mo(CO)₆ and mesitylene-Mo(CO)₃ showed no significant activity toward conjugated substrates. When Cr(CO)₆ and Mo(CO)₆ were irradiated at 2537 Å they caused the geometric isomerization of methyl sorbate without hydrogenation, but had no effect on methyl linoleate. A hydrogenation mechanism is proposed for Cr(CO)₆ that involves CH₃CN- and H₂-Cr(CO)₃ complexes as intermediates for the stereoselective 1,4-addition of hydrogen totrans,trans-conjugated dienes.     

Effect of dentinal surface preparation on the bonding of self-adhesive luting cements

The purpose of the present study was to evaluate the bond strength and the interaction morphology of self-adhesive resin luting cements (SLCs) to dentin prepared with different methods. Four SLCs were used: RelyX U100^®, RelyX U200^®, Clearfil SA Luting^®, and SmartCem2^®. A flat dentin surface of 40 human molars was exposed and each tooth was sectioned in four tooth-quarters, which were distributed into four groups according to the SLC used to cement indirect resin composite restorations. The tooth-quarters of each group were then distributed in four subgroups according to the method used for dentin preparation: flat-ended cylindrical fine-grit diamond, flat-ended cylindrical median-grit diamond, flat-ended cylindrical plain-cut tungsten carbide, or abraded with #600-grit SiC paper (control). The restored tooth-quarters were sectioned to obtain beams (0.8?mm²) and submitted to the microtensile bond strength test (n?=?10). The results were analyzed using two-way ANOVA/Tukey (α?=?0.05). Forty-four additional teeth were used for micromorphological investigation of the SLC/dentin interface and of the topographic aspect of the dentin surfaces after application of the SLCs. Only the bond strength of RelyX U200 was significantly influenced by the surface preparation. No interference was identified on the micromorphological aspect of the bonding interfaces. The topographic investigation of the dentinal surfaces showed that the SLCs were not able to effectively remove the smear layer and etch the underlying dentin, irrespective of the preparation method. So, the interference of the dentin preparation on the bond strength of SLCs is material dependent, but don’t influence the micromorphologic aspect of the interaction zone.     

Protein Abundance of Drug Transporters in Human Hepatitis C Livers

Transmembrane drug transport in hepatocytes is one of the major determinants of drug pharmacokinetics. In the present study, ABC transporters (P-gp, MRP1, MRP2, MRP3, MRP4, BCRP, and BSEP) and SLC transporters (MCT1, NTCP, OAT2, OATP1B1, OATP1B3, OATP2B1, OCT1, and OCT3) were quantified for protein abundance (LC-MS/MS) and mRNA levels (qRT-PCR) in hepatitis C virus (HCV)-infected liver samples from the Child–Pugh class A (n = 30), B (n = 21), and C (n = 7) patients. Protein levels of BSEP, MRP3, MCT1, OAT2, OATP1B3, and OCT3 were not significantly affected by HCV infection. P-gp, MRP1, BCRP, and OATP1B3 protein abundances were upregulated, whereas those of MRP2, MRP4, NTCP, OATP2B1, and OCT1 were downregulated in all HCV samples. The observed changes started to be seen in the Child–Pugh class A livers, i.e., upregulation of P-gp and MRP1 and downregulation of MRP2, MRP4, BCRP, and OATP1B3. In the case of NTCP, OATP2B1, and OCT1, a decrease in the protein levels was observed in the class B livers. In the class C livers, no other changes were noted than those in the class A and B patients. The results of the study demonstrate that drug transporter protein abundances are affected by the functional state of the liver in hepatitis C patients.     

The trans‐10,cis‐15 18:2: a Missing Intermediate of trans‐10 Shifted Rumen Biohydrogenation Pathway?

The “trans-10 shifted” biohydrogenation pathway is frequently established in the rumen when high starch diets are fed to ruminants, resulting in the accumulation of trans-10 18:1 in ruminant products. It has been proposed that the “trans-10 shifted” biohydrogenation pathway of α-linolenic acid generates two intermediates, the trans-10,cis-15 18:2 and trans-10,cis-12,cis-15 18:3, although none of these have been found in the rumen. We analyzed digestive contents and meat samples from two trials, where animals were fed: a compound feed diet supplemented with 8 % oil blend containing linseed oil (samples A); and a forage based diet supplemented with 6 % linseed oil (samples B). The use of the new SLB-IL111 chromatographic column allowed the detection of two different 18:2 isomers in each sample trial, which could not be resolved when the CP-Sil 88 column is used. The two 18:2 isomers were characterized by mass spectrometry using 4,4-dimethyloxazoline derivatives. However, because they were subject to higher temperatures and present different chromatographic properties compared with the fatty acid methyl esters, we also used the “covalent adduct chemical ionization” technique to confirm the identity of both 18:2 isomers. We detected and identified the 10,15-18:2 in samples A and the 11,15-18:2 in samples B. The geometry of both isomers was tentatively assigned as trans,cis taking in account their elution order and biologic plausibility. As far as we know, this is the first time that the trans-10,cis-15 18:2 has been found in ruminant digestive contents and meat samples associated with the “trans-10 shifted” biohydrogenation pathway of α-linolenic acid.     

Polymorphism in single-acid triglycerides of positional and geometric isomers of octadecenoic acid

The polymorphism of 25 glycerol trioctadecenoates with double bonds ranging from Δ4-Δ17 was investigated by differential scanning calorimetry. Triglycerides withcis bonds in odd positions Δ7-Δ13 exhibited three intermediate melting (β′-) forms, but those withcis bonds in even positions, exceptcis Δ4, lacked β′-forms. Among thetrans compounds, only Δ11, 13, and 14 showed β′-forms. Thecis andtrans Δ5 triglycerides were unusual, because they readily assumed low melting (α-) forms that were not easily converted to high melting (β-) forms. β-Form mp of compounds in each series (cis ortrans) alternated depending upon double bond position; an even position correlated with high mp. Heats of fusion (ΔH_f) for β-forms, likewise, fluctuated with double bond position but nonuniformly;trans Δ6 had the highest ΔH_f (43 cal/g),cis Δ12 the lowest (21 cal/g).