Multiple-analyte fluoroimmunoassay using an integrated optical waveguide sensor.

A silicon oxynitride integrated optical waveguide was used to evanescently excite fluorescence from a multianalyte sensor surface in a rapid, sandwich immunoassay format. Multiple analyte immunoassay (MAIA) results for two sets of three different analytes, one employing polyclonal and the other monoclonal capture antibodies, were compared with results for identical analytes performed in a single-analyte immunoassay (SAIA) format. The MAIA protocol was applied in both phosphate-buffered saline and simulated serum solutions. Point-to-point correlation values between the MAIA and SAIA results varied widely for the polyclonal antibodies (R2 = 0.42-0.98) and were acceptable for the monoclonal antibodies (R2 = 0.93-0.99). Differences in calculated receptor affinities were also evident with polyclonal antibodies, but not so with monoclonal antibodies. Polyclonal antibody capture layers tended to demonstrate departure from ideal receptor-ligand binding while monoclonal antibodies generally displayed monovalent binding. A third set of three antibodies, specific for three cardiac proteins routinely used to categorize myocardial infarction, were also evaluated with the two assay protocols. MAIA responses, over clinically significant ranges for creatin kinase MB, cardiac troponin I, and myoglobin agreed well with responses generated with SAIA protocols (R2 = 0.97-0.99).

Planar integrated optical methods for examining thin films and their surface adlayers.

Thin film integrated optical waveguides (IOWs) have gained acceptance as a method for characterizing ultrathin dielectrical films and adlayers bound to the film surface. Here, we present the expressions that govern IOW methods as well as describe the common experimental configurations used in attenuated total reflection, fluorescence and Raman applications. The applications of these techniques to the study of adsorbed or surface-bound proteins to polymer and glass waveguides are reviewed.

Femtomolar sensitivity using a channel-etched thin film waveguide fluoroimmunosensor.

A dual channel, evanescent fluoroimmunoassay format is used to detect femtomolar analyte concentrations (i.e. less than 1 part per trillion [w/w]) on an etched channel siliconoxynitride thin film integrated optical waveguide. Two assays are used to demonstrate the dose-response behaviour of the sensor: (1) a direct assay of a fluorescently-labeled protein ligand binding to an immobilized protein receptor, and (2) an indirect sandwich assay of a non-fluorescent protein ligand binding to an immobilized protein receptor, as detected by the binding of a fluorescently-labeled secondary receptor protein. A red-emitting cyanine dye (Cy-5), which minimized background fluorescence and scatter losses of the waveguide, was used in both assays. To our knowledge, this is the first report of femtomolar sensitivity in an immunosensing instrument.

The effects of field preservation on alkaloid content of fresh coca leaves (Erythroxylum spp.).

In order to test the effects of commonly used preservation agents on the alkaloid content of herbarium specimens, fresh leaves of Erythroxylum coca, E. novogranantense, and E. novogranatense var. truxillense were air-or heat-dried or treated with six different liquid preservatives. The leaves were then extracted and analyzed quantitatively for cocaine content. Leaves which were soaked in preservatives showed appreciable pre-extraction of cocaine and probably of other alkaloids. The results compare well with a similar experiment conducted on flavonoid content of the leaves of a palm Jessenia bataua. If portions of herbarium specimens are to be useful for phytochemical screening using microtechniques, at least part of the collection must be air- or heat-dried to retain the chemical constituents.

Amazonian coca.

A general overview of various aspects of Amazonian coca (Erythroxylum coca var. ipadu) is presented. This plant is considered a distinct variety of coca which has been developed as a cultivated plant in the upper Amazon basin. It differs from typical Andean coca in morphological, physiological and chemical features as well as in the method of preparation and use by Amazonian tribes. The main topics here discussed are the history, distribution, botany, chemistry, origin, methods of preparation and use, and the effects of Amazonian coca.

Coca pests and pesticides.

The major pests of coca are listed and discussed along with methods used to control them in the past and present. Results of analyses for pesticide residues in samples of commercial Peruvian coca leaves are presented. Levels of pesticides found in these samples are too low to be considered a medical risk to coca chewers.

Cocaine in blood of coca chewers.

Coca leaves (Erythroxylum coca Lamarck) and powder (5 - 10 g) were taken orally by human subjects in the same way as South American natives do. The cocaine, as measured by mass fragmentography, was immediately detected in the blood, reached peak concentrations from 10 - 150 ng/ml plasma at 0.38 - 1.95 hours, and persisted in the plasma for more than 7 hours. Half-lives of the elimination of cocaine were calculated and ranged from 1.0 to 1.9 hours. The absorption half-lives ranged from 0.2 to 0.6 hours. The shape of the curves fits with the subjective effects reported. There is no reason to believe that the stimulating effect achieved by the use of either coca leaves or powder is not due to cocaine.