A photometric screening method for dimeric naphthylisoquinoline alkaloids and complete on-line structural elucidation of a dimer in crude plant extracts, by the LC-MS/LC-NMR/LC-CD triad.

An efficient evaluation procedure for the chemical screening and on-line structural elucidation of dimeric naphthylisoquinoline alkaloids has been developed. The method is based on the lead tetraacetate oxidation of the central binaphthalene core of the alkaloids. UV spectra of the extracts after addition of the oxidant show, in the presence of naphthylisoquinoline dimers, the appearance of a characteristic long-wavelength absorption indicative of dinaphthoquinones. The efficiency and relevance of the method has been demonstrated in the discovery of a constitutionally and configurationally new dimeric naphthylisoquinoline alkaloid, named ancistrogriffithine A (4), from the previously uninvestigated Asian liana Ancistrocladus griffithii. After verification of this screening result by LC-ESI-MS/MS, the constitution and the relative configuration of the compound were elucidated on line, by LC-NMR and LC-CD on the extract. Using an LC-NMR-WET-ROESY experiment, itwas possible for the first time to determine the relative axial configuration of a natural biaryl compound on line, by observing long-range ROE interactions. Finally, an oxidative degradation right on the extract delivered the absolute configuration of 4, without isolation of the alkaloid. Ancistrogriffithine A is the as yet only dimeric naphthylisoquinoline from an Asian Ancistrocladaceae plant.

Lifting of the degeneracy in semibullvalenes by remote and direct substituents: a quantitative study using variable-temperature carbon-13 NMR spectroscopy.

A series of six 1,5-(ethylmethyl)semibullvalenes (1a <==> 1a', 2 <==> 2', 3 <==>3') and two 4(2)-substituted semibullvalenes (4 <==> 4'), each undergoing Cope equilibria between nondegenerate valence tautomers, was investigated by carbon-13 NMR spectroscopy at a range of temperatures in several different solvents. Gompper's treatment of substituent perturbation was extended, specifically accounting for the effects of the substituents on chemical shifts, to allow the determination of the thermodynamic parameters for these skewed equilibria. These new treatments were used to determine the population difference (f - f ') between the valence tautomers and the perturbation thermodynamic quantities DeltaH(P), DeltaS(P), and DeltaG(P). The slow-exchange limit was reached for the parent 1,5-(ethylmethyl)semibullvalenes 3a <==> 3a' from which it was established that the preferred valence tautomer is 3a with the ethyl group on the cyclopropane ring. Despite considerable effort, the slow-exchange limit could not be reached in any of our other remotely substituted semibullvalenes. Provided that the ethyl group always prefers the cyclopropyl position as in 3a, the 1-ethyl-5-methylsemibullvalenes 1a, 2, and 3 are more stable by DeltaH(P) = 0.7-1.7 kJ mol(-1) than their valence tautomers 1a', 2', and 3'. In the directly substituted semibullvalenes (4 left harpoon ovet right harpoon 4'), the preferred valence tautomers 4a and 4b have the bromine atom or the nitrile group on the vinyl position (C(4)) rather than on the cyclopropane ring (C(2)) and are more stable than 4a' and 4b' by DeltaH(P) = 4.8 and 7.0 kJ mol(-1), respectively.

Observation of exchangeable protons by high-performance liquid chromatography-nuclear magnetic resonance spectroscopy and high-performance liquid chromatography-electrospray ionization mass spectrometry: a useful tool for the hyphenated analysis of natural products.

The first high-performance liquid chromatography-nuclear magnetic resonance (HPLC-NMR) investigation of exchangeable protons of low-molecular-mass natural products is reported. Model alkaloids or crude plant extracts were dissolved in 2H2O-1H2O-MeCN (deuterium oxide-water-acetonitrile) or 2H2O-MeCN and, after direct injection or chromatographic separation, examined in a 60-microl NMR flow probe. Exchangeable amino protons initially detected by HPLC-electrospray ionization mass spectrometry were subsequently identified and investigated by stop-flow 1H-NMR, two-dimensional (2D) total correlation spectroscopy (TOCSY), and 2D nuclear Overhauser effect spectroscopy (NOESY). These experiments extend the applicability of HPLC-NMR for the investigation and structure elucidation of natural products.

A high-precision carbon-13 shift thermometer for the temperature range 100-300 K

The first carbon-13 shift thermometer for the temperature range of 100-300 K is based on the very rapid equilibration of a pair of semibullvalene valence tautomers. The temperature dependence of the equilibrium constant is reflected in strongly temperature-dependent shift differences Deltadelta between averaged signals, e.g., d(Deltadelta)/dT = 0.051 ppm K-1 at 300, 0.087 ppm K-1 at 200, and 0. 175 ppm K-1 at 110 K for the quaternary carbon atoms C2 and C6. At 37 temperatures T, which were measured with calibrated platinum resistance thermometers, shift differences Deltadelta were taken from nondecoupled carbon-13 spectra recorded from solutions of 1 in mixtures of chlorodifluoromethane and deuterated dimethyl ether without spinning. The least-squares fit of these Deltadelta vs T data to a polynomial equation of the fourth degree (Eq. [5], r2 = 0. 9999) allows the calculation of temperatures from measured shift differences with a standard deviation of 0.46 K and an estimated error of about 1 K. The heating effects of WALTZ-16 decoupling and the influence of solvents on Deltadelta are investigated. A comparison with existing NMR thermometers demonstrates the superior performance of the new carbon-13 shift thermometer with respect to precision and the accessible temperature range. Copyright 1998 Academic Press.