METHOD FOR BIOLOGICAL TESTING AND/OR DEVELOPING PHARMACEUTICALS FOR TREATMENT OF DISORDERS

Abstract: A method for establishing a treatment protocol for correcting an abnormal condition or disease in a living organism. The method comprises the steps of analyzing body fluids from normal and abnormal individuals to generate analysis patterns of the normal and abnormal fluids, comparing the patterns to determine differences in the patterns, and establishing a treatment protocol for the abnormal individual through chemical or metabolic therapy which treating will normalize the abnormal pattern.

Description: METHOD FOR BIOLOGICAL TESTING AND/OR DEVELOPING PHARMACEUTICALS FOR TREATMENT OF DISORDERS T is application is in part a continuation of my copending Application Serial No. 670,483, filed November 13, 1984 which in turn is in part a continuation of my copending Applica ion Serial No. 579,401, f led February 17, 1984 ( now U.S. Patent No. 4,511,659), which is n part a continuat on-in-part of my Application Serial Number 472,387 fi led March 4, 1983 (now abandoned), and 0 in part a continuation of my copending Application Serial Number 425,183 filed September 28, 1982 which is, in turn, a continuation of my Application Serial Number 111,917 filed January 4, 1984 ( now patent 4,404,065) . This invention relates to analytical systems, i.e ** - * apparatus and methods for qualitati ely and quantitatively testing materials n solut on. The invention has particular utility for the detection and determination of biologically active materials such as neurotransm tters and other neurochemical substances n 0 brain tissue, cerebrospinal fluid, plasma, serum, saliva, urine and the like, such as catecholamines, their precursors, cofactors and their metabolites. The invention is uniquely capable of generating analytical values and patterns of a large number of compounds of 5 biological, diagnostic and pharmaceutical significance and of describing complex metabol c pathways of such substances from single small samples of biological materials and the use of such values and patterns for rational development of pharmaceuticals and w ll be 0 described in connection with such utility although other uses are contemplated. There is an extensive body of literature relating abnormalities in neurotrans itters, precursors, and metabolites to degenerati e, neu ropsych at ri c and 5 behavioral disorders, hypertension and certain carcinomas. See, for example, Sch ldkraut et al in The Brain, Biochemistry and Behavior, Proceedings of the Sixth, Arnold 0. Beck an Conference in Clinical Chemistry, pages 47-68. Although the potential role of these compounds in a number of significant disorders has been established, their routine analysis has not yet achieved widespread clinical use. Two problems n the clinical uti lity of neurotrans itter measurements are related to the economic and technical limitations of current technology. First, there is felt to be a high degree of i nter laboratory and intersample uncertainty in quantitative values. Second, it has been difficult to measure enough of the known metabol cally related compounds of a particular neurotransmitter to fully describe its biochemical significance in an individual sample, or to detect, identify and measure unusual neurotransmitters -- an i portant aspect of basic research in various disease states that is presently very expensive and specia lized. Whi le a number ' of i nt ra labo rato ry technique intercompaπ ' sons for a variety of neurotransm tters have been carried out, there has been no comprehensi e study w thin and among different techniques and laboratories for neurotrans i ters in typical samples of interest. In the absence of such studies, given the complexity of the analytical problem and the historically wide variation whenever an analyte has been subjected to rigorous interlaboratory test ng, the current values for normal and abnormal neurotransmitter levels must be taken with unspecified and probably wide limits of confidence. A lthough the analysis of single neurotransmitters or metabol tes from a complex biochemical pathway has been ■ shown to correlate with a number of disorders uti lizing statistical analysis over a large number of samples, the analytical level of a s ngle neurotransmitter in an individual sample, with a few exceptions, has had relatively low cl nical diagnostic uti lity. Essentially the state of the field of b ochemical correlates of disorders s that whi le between large populations of normal and abnormal indi duals a correlation generally can be determined for a particular b ochemical, the scatter that results from both analytical and biochemical phenomena typically does not permit the level of a particular biochemical to be uti lized diagonostically for a particular s ngle individual. Nor may a single biochemical value be ut lized for the rational prescription o r development of a pharmaceutical for that indi idual. Th s is not parti cularly surpris ng in that both the levels and effects of a particular neurotransmitter are modified by a number of other neurotransmitters n the same, or parallel metabolic pathways. If, for instance, 5-HT (serotonin) is to be used as a diagnostic tool for depression, suicidal tendencies, or schizophrenia, it would be necessary and perhaps pro ide a route to definitive diagnosis and pharmaceutical specification or development, to simultaneously determine the approximately 40 other compounds that derive from tryptophan and significantly effect the indolaminergic neuronal system's activity. In recent years, LCEC (Liquid Chro atσgraphy with Electrochemical Detection) has become a common tool for the deter ination of catecholamines biogenic amines and thei r metabolites in biolog cal fluids. Because of sensit ity l mitations (typically 20-50 pg) and the complexity of biological samples, both separation and concentration steps typically have been necessary. Heretofore, plasma catecholamine analysis typically required three steps. First, the sample s collected and the catecholamines separated and concentrated, for example, using the alumina extraction procedure of Anton and Sayre ( See A.H. Anton and D.F. Sayre, J. Pharmacol, Exp. Ther., 138 ( 1962 ) , p. 360-375 ) . The analytes, norepinephrine, epinephrine and dopamine, along ith the internal standard DHBH (dihydroxybenzylam ne ) , then are separated chromatograph cally, and finally detected e Iect rochemi ca I l . Typical sample size requirements are 1.0 ml plasma o r serum. In routine clinical use, there have been numerous problems with conventional techniques (alumina adsorption, ion exchange and extraction ) due to a large number of poorly understood variables in the overall analysis system of sample acquisition, storage, preparation and sensor response. These problems have quite likely confused the relationships that may exist between levels and distribution of the catecholamines and various physiological and behavioral phenomena and disease states. In the analysis of complex biological materials such as blood, serum and cerebrospinal fluids which may contain numerous different constituents, the important (e.g. abnormal) metabolites such as neurotransmitters to be identified may be present in only parts per tri llion. Whi le a ch romat og rap i c column can achieve macro separation of the various constituents it may not provide adequate spatial ( in time) separation of the extremely small portion of metabolites of interest from the much larger percentage of the many other compounds coeluted from the column at the same time as the metabolites of interest. Many of these nterfering coeluted materials are electrochem cally active but electrochemically irreversible while many metabol tes such as neurotransmitters are both electroche ically active and electrochemically reversible. It has been found that the analytical problems of reliable measurements of neurochemi ca Is and related compounds are complicated by the fact that interferences with conventional or prior technologies are disorder related. Th s was discussed in my prior publication, ( Matson et al, Clinical Chemistry, Vol. 30, No. 9, 1984) for dopamine, dopac and seratonin measurements in directly analyzed brain extract and cerebrospinal fluid for normal, schizophrenics and Alzhei ers. Recent work has indicated that even for the widely used and accepted technique of alumina extraction for plasma catecholamines that inferences may be disorder spec fic. Higher values for Norepinephrine (NE) and Epiπephrine (EP) were observed following alumina extraction and analysis of a single energy convent onal electrochemical detector than for a three cell redox detector on neonatal stress blood samples. Analysis of the neonate extracts on the sixteen channel chemical imaging system revealed several unexpected compunds that are potential interferences including dihydroxyphenylacetic acid (DOPAC), 3 hydroxykynurenami ne (3-OHKYA), and 3-hydroxy-anth rani I c ac d (3-OHAN). These compounds have not been detected in normal adult plasma alumina extracts. In my aforesaid U.S. Patent No. 4,511,659, there is provided an electrochemical detection system comprising a plurality of coulomet ri ca I ly efficient electrochemical cells, in series, for sequentially oxidizing and reduc ng selected substances in a sample solution under controlled conditions prior to measurement on a downstream testing electrode or electrodes. More specifically, in accordance with the invention provided in my aforesaid U.S. Patent No. 4,511,659, a sample solution (e.g. a body fluid) is passed through a suitable chromatographic column and the eluant is streamed in contact with a series of electrochemically - isolated, in-line coulometric electrodes operated under conditions so as to establish a series of "gates" for the sequential oxidation and reduction of substances in the sample solution whereby to screen ( remove) selected interfering and electrochemically i rreversible substances contained in the sample solution, whi le passing selected electrochemically reversible products for detection and measurement on a downstream electrode. The gate electrode series is followed in-l ne by one or more, preferably an array of six or more cou lometric measuring electrodes, each formed of porous electrode base material such as fritted graphite, fritted carbon or other conducti e fritted material, for detecting and measuring the electrochemically reversible compounds of interest (e.g. neuro ransmitters) . As reported in my aforesaid U.S " . Patent No. 4,511,659, there are several beneficial effects of this approach to electrochemical analysis. Long-term drift in response is effecti ely eliminated by acqui ring essentially 100% of the signa l. The capabi lity of analyzing essentially 100% of a materia l allows the assay of compounds of unknown purity by relating them to the basic principles of electrochemical reaction embodied in Faraday's Law. Poisoning of the electrode, a dominant problem with electrochemical sensors, is effectively el minated by the use of a much larger relative surface area for reaction. And, finally, and most i portant to the eventual development of array and gate cells, a coulometric electrode by virtue of its essentially 100% efficiency allows sequential oxidation and/or reduction of compounds at success ive-in- line detectors. The improved sensitivity of the detection system as discussed in my aforesaid U.S. Patent No. 4,511,659, particularly where two or more active testing electrodes follow the screening electrodes has g en the abi lity to do direct injections of serum fi ltrates and has also allowed the generation of reproducible patterns of compounds ith catecholamine like electroche ical behavior of a large number of resolvable components. This provides the possibi lity of performing pattern recognition for the diagnosis or perhaps even predict e diagnosis, of various disorders or disease states. The present invention in broad aspect provides a system for resolving and detecting hundreds of compounds in a single sample at femtogram levels whereby to provide a small molecule inventory or metabolic pathway pattern of a individual. The small molecule inventory may be cons dered to ref lect the underlying act ity and distribution of the enzymatic pathways of an ndividual and hence reflect an operational measure of the genone determining those enzymes. The small molecule inventory of an individual may thus be used to determine the health state of the ind v dual and/or to diagnose disease states. Correlation of the patterns from a plurality of individuals provides an understanding of the mechanisms of disorders or disease states or conditions and in turn provides a rat onal route to pharmacological development leading to treatment, cure or suppression of such disorders, disease states or cond t ons . More particularly, in accordance with the present invention, sample solutions (e.g. body fluids) a r e electrochemically tested for both normal and abnormal indi iduals or conditions, patterns are generated for sa d normal and abnormal individuals or cond tions, the patterns compared and differences identified, and the ident fied differences uti lized to design a specific treatment protocol. In one aspect the present invention provides a method for developing pharmaceuticals for disorders in l ing organisms and the pharmaceut cals so produced. The method comprises the steps of analyzing body fluids from normal and afflicted (abnormal) indi iduals to generate analysis patterns representative of constituents of the metabolic pathways of said ind viduals; comparing said patterns to determine differences in said patterns representati e of (a) the presence or absence of specific compounds in said patterns, or (b) differences in the concentration of said compounds in said patterns; and introducing into a pharmaceutically acceptable carrier therefor one or a mixture of chemical compounds known to act at sites within said metabolic pathways to (a) block or potentiate those metabolic pathways identified as abnormal by the pattern, or (b) supply or retard those unique compounds identified as abnormal in said patterns. The invention in another aspect provides a method for developing pharmaceut cals for d sorders n living organisms and the pharmaceut cals so produced. The method comprises the steps of analyzing body fluids from normal and afflicted (abnormal) individuals to generate analysis patterns representati e of individuals' electrochemically ac ve small molecule inventory, comparing said patterns to determine differences in said patterns representat ve of (a) the presence or absence of specific electroche ically active compounds in said patterns, or (b) differences in the concentration of sa d compounds in said patterns; and introducing into a pharmaceutical carrier one or a mixture of chemical compounds known to act at sites within metabolic pathways of said organisms to (a) block or potentiate those metabolic pathways iden ified as abnormal by the pattern, or (b) supply or retard those unique compounds identified as abnormal in said patterns. The invention also provides a method for identifying pharmacological materials for treating living organisms, wh ch method compr ses the steps of analyzing body fluids from .normal and afflicted (abnormal) individuals to generate che ical analysis patterns representative of the electrochemically active small molecule inventory contained in said normal and abnormal body fluids; comparing said patterns and identifying differences in said patterns representative of (a) the presence or absence of specific compounds in said patterns, or (b) differences in the concentration of said compounds in the metabolic pathways of said individuals; and selecting from chemical compounds known to act at sites within sa d metabolic pathways one or a mi ture of compounds to (a) block or potentiate those metabolic pathways identified as abnormal by the pattern, or (b) supply or retard those unique compounds identified as abnormal in said patterns. In yet another aspect the invention provides a method of identifying an abormality in a body fluid of an indiv dual which comprises the steps of determining the presence and relative concentration of electrochemically active compounds contained in said body fluid by passing a sample containing sa d body fluid sequentially through a liquid ch ro atographi c column for achieving time-space separation of materials eluting from the column and an electrochemical detection apparatus for electrochemically testing a sample containing electrochemically active materials in solution, the detection apparatus compr sing a plurality of coulometric cells arranged in series, maintaining the testing electrodes of said coulometric cells at d fferent potentials, said cells operating at progressively varying potentials along the path of flow of the eluant through the cells, and recording the results of coulometric measurements from said cells so as to separate said measurements by measuring potential as well as by time of elution, and comparing the said relative concentrations thus obtained with know.n concentrations of normal or abnormal body fluids. Also provided in accordance w th the present invention is a method for identifying an abnormality in a body fluid of an individual which comprises the steps of determining the presence and relative concentration of electrochemically active compounds contained in said body fluid by passing a sample containing said body fluid sequentially through a liquid chromatograph ic column by step gradients for achieving time spaced separation of the materials elu ing from the column and an electrochemical detection apparatus for electrochemically testing the eluant from said column, the detection apparatus comprising a plurality of cou.lometric cells arranged in series, maintaining the coulometric electrodes of said cells at different potentials, said cells operating at progressively varying potentials along the path of flow of the eluant through the cells, recording the results of coulometric measurements from said cells so as to separate said measurements by measuring potential as well by time of elution, and comparing said measurements thus obtained with measurements of normal and abnormal body fluids to provide identificat on and relevant concentrations of constituents coeluting from the column. Yet another aspect of the invention provides a method of identifying abnormalit es in the small molecule inventory of an individual comprising the steps i n sequence of : determin ng the presence and relative concentration of electrochemically active constituents in the metabolic pathways containing such abnormalities by pass ng a sample containing said body fluid sequentially through a liquid chromatograph column for achieving time-space separation of the constituents eluting from the column and an electrochemical detection apparatus comprising a plurality of coulometric cells arranged n series; maintaining the coulometric electrodes of said cells at different potentials, said cells operating at progressively varying potentials along the path of flow of the eluant through the cells, said plurality of cells being sufficient n number to separate by measur ng poten ials those relevant constituents coeluted from the chromatograph ic column at any instant of time and recording the results of such coulometric measurements so as to separate said measurements by measur ng potential as well as by time of elution and comparing the said relevent concentrations thus obtained with known concentrations of normal or abnormal body fluids and thereby identify changes of rates through the various metabol c pathways of said relevant constituents as well as unknown metabolites and abnormal precursors for normal metabolites. The invention also provides a method of determining the relative concentration of pr ncipal constituents in the metabolic pathways of an animal by passing a sample containing body fluid from said animal sequentially through a liqu d chromatograph column for achieving time-space separation of the constituents eluting from the column and an electrochemical detection apparatus comprising a plurality of coulometric cells arranged in series; maintaining the coulometric electrodes of said cells * at different potentials, said cells operating at progressively varying potentials along the path of flow of the eluant through the cells, said plurality of cells being sufficient in number to separate by measuring potentials those relevant constituents coeluted from the chro atographi c column at any instant of time, and recording the results of such coulometric measurements so as to separate said measurements by measuring potential as well as by time of elution. Also pro ided in accordance ith the present invention is a method for treating a disorder in a living organism which method comprises the steps of analyzing body fluids from normal and afflicted (abnormal) individuals to generate analysis patterns representative of const tuents of the metabol c pathways of sa d individuals; comparing said patterns to determine differences in said patterns representative of (a) the presence or absence of specif c compounds in said patterns, or (b) differences in the concentrations of said compounds n said patterns; introducing into a pharmaceutically acceptable carrier therefor one or a mixture of chemical compounds known to act at sites within said metabol c pathways to (a) block or potentiate those metabolic pathways identified as abnormal by the pattern, or (b) supply or retard those unique compounds identified as abnormal in sa d patterns; and adm nistering to said living organism said identified compounds in said pharmaceutically acceptable carrier so as to (a ) block or potentiate those metabolic pathways identified as abnormal by the pattern, or (b) supply or retard those unique compounds identified as abnormal in said patterns. Finally, the invention provides a ethod of treating a disease state in an animal, comprising the steps in sequence of identifying abnormalities in a body fluid of said animal by determining the relative concentration of the principal constituents in the metabolic pathways containing such abnormalities by passing a sample containing said body fluid sequentially through a liqu d chromatograph column for achieving time-space separation of the constituents eluting from the column and an electrochemical detection apparatus comprising a plurality of coulometric cells arranged in series; maintaining the coulometric electrodes of said cells at different potentials, said cells operating at progressively varying potent als along the path of flow of the eluant through the cells, said plurality of cells being sufficient in number to separate by measuring potentials those relevant constituents coeluted from the chromatographic column at any instant of time and recording the results of such coulometric measurements so as to separate said measurements by measuring potential as well as by time of elution and comparing the said relevent concentrations thus obtained with known concentrations of norm*al or abnormal body fluids and thereby identifying changes of rates through the various metabolic pathways of said relevant constituents as well as unknown metabolites and abnormal precursors for normal metabolites, and identify those compounds responsible for said changes of rates, and administering to sa d animal said identified compounds so as to provoke a change of rate through said metabolic pathways of said individual substantially to approach that found in said normal fluids. Yet other objects of the invention will in part appear obvious and will in part appear hereinafter. The invention accordingly comprises the process comprising the several steps and relation of one or more of such steps with respect to each of the others, all of which- are exemplified in the following detailed description, and the scope of application as will be indicated in the clai s. For a fuller understanding of the nature and objects of the present invention, reference should be had to the follo ng detailed description taken in com ination w th the accompanying dra ings wherein: Fig. 1 is a side elevational view, in cross-sect on, of an electroche ical detection apparatus useful in accordance with the process of the present invention; F g. 2 is a block diagram of the electrical cont ols and functions of the electrochemical detec ion apparatus of Fig . 1 ; Fig. 3 is a time/potential plot of a chromatogram obtained in accordance with the present invention; Fig. 4 represents the major metabolic pathways of tyrosine (TYR) ; F g. 5. represents the major metabol c pathways of tryptophan (TRP); F g. 6 is a schematic diagrammat c dra ing of a detection system useful in accordance with the process of the present invention wherein a plurality of test (or measuring) cells are employed after the gate cells and the outputs of the test cells are processed by a central processing un t; Fig. 7 is a flow chart of an embodiment of disease detection and treatment design method accord ng to the present invention; Fig. 8 is a block diagram of an electrochemical apparatus adapted for the pract ce of the method shown in Fig. 7; and Fig. 9 is a voltage/time resolution pattern obtained in accordance with the present invention. Further understanding of the features and objects of the present invention wi ll be had from the following detai led description of one preferred embodiment of the invention which i llustrates an electrochemical neurotransmitter pattern generation and resolution and detection of the principal constituents or agents contained in cerebrospinal fluid and affecting motion sickness. It wi ll be understood, however, that the process of the present invention may be advantageously employed for detecting and resolving of various other electroactive metabolites n a biological sample solution, including but not limited ' to neurotransmitters, precursors and metabolites, hormones, transmitting compounds, peptides, certain vitamines, pterins, purines, certain drugs and metabolites and other materials that mediate metaboli sm. More particularly, the present invention in one aspect provides a method by which sample so lutions for both normal and abnormal indiv duals and conditions are tested, principal cons ituents or agents resolved and detected, patterns indicating the presence and concentration of such constituents or agents generated, and the differences between the patterns determ ned whereby compounds suspected of causing said abnormalities or conditions may be identified, and a treatment protocol established through chemical or metabolic therapy, to normalize the (abnormal) pattern, or block or potentiate those metabol c pathways identified as abnormal by the pattern, or supply or retard those unique compounds ident ified as abnormal in the pattern, or utilize changes in the patterns during therapy and the concentration of the therapeutic drugs and metabolites to modify the course of the therapy. The first step of the process is to analyze the sample solutions for normal and abnormal individuals or conditions to generate patterns for comparison. The electrochemical detection described in my aforesaid U.S. Patent o. 4,511,659 is particularly adapted for this purpose. As reported therein such apparatus is capable of responding to and differentiating between electrochemically reversible species in a sample solution and for discri inating against species in a sample solution that are electrochemically irreversible. Sample solutions from normal and abnormal individuals are flowed through a chromatograph ic column and the eluant is passed through a series of electrochemically isolated cells or "gates" operated at potentials resulting in sequent al oxidation and reduct on of various species contained n the sample solution. These gates a re followed by an array of measuring electrodes for measuring electrochemical activity of compounds of interest. The strategy of analysis d scriminates against both background current from the mobi le phase itself and removes (screens) compounds that are irreversible, while electrochemically reversible species of interest, e.g. the catecholamines are cycled back and forth from oxid zed to reduced state at various potentials whereby to enhance the sensitivity and specificity of downstream detection and measuring elect rodes . For convenience of illustration the function of the gate cells is described as "removing" or "screening" the electrochemically irreversible compounds from the eluant. This is not what actually happens; these compounds are merely electrochemically altered by the gate cells so that the potential existing on the following detecting electrode(s ) wi ll not change the oxidation state of the "removed" compound and therefore the "removed" compound will not be detected and coulometrically measured. It is as if the "removed" compound were not present in the eluant. It is no longer electroactive under the detection conditions. The gate/array cell pr ncipal was evaluated using a cell configured typically as shown in Figs. 1 and 2. In essence, a gate/array cell is designed to set up windows for determination of selected compounds in the catecholam nergic systems and in the indolam nergic system. These assays include Homogenistic Acid (HGA), 3-hydroxy-L-tyros ne (L-D0PA), Norepi neph ri ne (NE), ihydroxyphenylacetic acid (D0PAC), Epinephrine (EP), Dopamine (DA), 3-Methoxy, 4-Hydroxypheny Ig lyco I (MHPG), and 3-Methoxytyrami ne (3-MT) to describe the catecholaminegic system, and Tryptophan (TRP), 5-Hydroxyt ryptophan (5-HTP), 5-Hydroxyt ryptopha I (5-HT0L), Serotonin or 5-Hydroxytryptamine (5-HT), 5-Hydroxy Indole Acetic Acid (5-HIAA), and Homovanillic Acid (HVA) to describe the serotonergic system. The electrochemical gate/array cell should have a minimum of 8 gates or arrays to effect .he desired resolution and preferably will have a minimum of 16 gates or arrays. Preferab Iy the analysis is conducted in three steps. The first step comprises an open array analysis in which the potential on the array is set at relatively wide testing increments. Generally the open array evaluation is performed n two passes, a first pass in which the materials of interest are dissolved in a low organic modifier, e.g. a mobile phase having for example 0 to 1 percent eCH and CH,CN, and a second pass in which the mater als of interest are dissolved in a high organic modifier, typically 30 to 60 percent or more MeOH, and CH-CN in lower proportion. This provides a first cut. Thereafter, compounds of potential interest based on analysis of the data are evaluated as before, but using a tighter array setting the electrodes to relati ely low potential increments over a specific region. As before the analysis is done in two passes - i.e. in a low organic modifier and in a high organic modifier. Finally, selected components then are analyzed under REDOX conditions with oxidat on followed by reductive assay. Analyzing the sample in this manner provides extremely high resolution and spec ficity. Refer ing to the detai ls of Figs. 1 and 2, there is shown a preferred embodiment of the electrochemical detection apparatus useful in accordance with the process of the present invention, indicated generally at 10. Electrochemical detection apparatus 10 comprises a hollow, liquid-tight enclosure indicated generally at 12 and having an inlet 14 and an outlet 16 found in a pair of end plates 17 and 18, respectively. Enclosure 12 is formed of a liquid-impervious, rigid, electrically insulating chem cally inert material such as unplasticized polyvinylchloride, polytetra- fluoroethylene, fluorohydrocarbon resin or the like. Disposed within enclosure 12 are sixteen electrochemically isolated electrochemical cells 20, 20A, 22, 22A...34, 34A. Electroche ical cells 20...34A are hydraulically connected to one another via stub tub ng members 32 which together w th cells 20...34A define a fluid flow path between inlet 14 and outlet 16. Each electrochemical cell 20...34A comprises a three electrode system consisting of at least one working electrode 34a, 34b, 34c, 34d, 3^e, 34f, 34g, 34h ( T, . . .T., ) respectively; at least one counter i i o electrode 36a, 36b, 36c, 36d, 36e, 36f, 36g, 34h, respectively; and, at least one reference electrode 38a, 38b, 38c, 38d, 38e, 38f, 38g, 34h, respect vely. Electrochemical cells 20...34A are fixedly positioned within enclosure 12 by suitable means (not shown). Each working electrode 34a ... h (T 1 T 16 ) is in the form of a flat disc formed of a porous electrode base material such as fritted graphite or fritted carbon or other conductive fritted mater als. Most of these preferably have a relatively large area to volume ratio to give large half ti es (up to 90) at the flow rates contemplated and ith the electrochemically active materials of interest. Half time is the time requ red for half of a quantity of a compound to react at an electrode. Allowing a reaction to proceed for two half times causes 75% reaction, 5 half times 97% reaction. In order to maximize response of the porous working electrodes the electrode base material was pretreated in accordance ith the teachings of my copending U.S. application serial number f i led contemporaneously herewith (Attorneys' Docket No. ESA 140/142) to load the secondary porosity of the electrode base material with a selected non-electroact ve material (i.e. under anticipated sensor conditions) to reduce background noise and enhance selecti ity to aminergic compounds. The overall process was as follows: A piece of commercially available fritted graphite was impregnated by a mixture of 80% C-48 paraffin and 20% C.9163-ML density polyethylene containing 1.00 grams per liter sodium salt of naphthalene sulfonic acid at 110 C and cycling between vacuum and 1000 psi pressure. Electrodes were cut from the mpregnated fritted graphite material and these electrodes were treated to remove the impregnating material blocking the frits by heating at 100 C for five minutes under a vacuum of 750-760 'mm HgAbs. Thereafter the treated electrode material was rinsed with 50-50 tet rahydrofuran-hexane . The resultant electrode material which contained approximately 0.01% by weight of sorbed napthalene sulfonic acid and approximately-one percent by weight of high molecular weight organic material, were assembled i the flow cell detector of a type illustrated in F g. 1, of my aforesaid U.S. Patent 4,511,659. Also provided are electrical connections 40a, 40b, 40c, 40d, 40e, 40f, 40g 40h for connecting working electrodes 34a...h, -T... . . T,.,) respecti ely, to potential controls 42a, 42b, 42c, 42d, 42e, 42f, 42g, 42h, respectively, for applying selected working potentials to the various working electrodes 34a ... h; electrical connections 44a, 44b, 44c, 44d, 44e...44h, for connect ng counter electrodes 36a...h, respectively, to potential controls 46a, 46b, 46c, 46d, 46e...44h (T. . . . T 16 ) respectively for applying selected counter potentials to the various counter electrodes; and electrical connections 50a, 50b, 50c, 50d, 5Ce, 5Cf, 50g, 50h, respectively, for connecting refetence electrodes 38a... h, respectively to a potential control 52 for applying a reference potential to the various reference electrodes 3δa...h. Counter electrodes 36a...h and reference electrodes 38a. ,.h preferably comprise inert metal terminals such as platinum or palladium wire. Alternat ely, reference electrodes 38a... h may compr se silver/si lver chlor de reference electrodes or the like. The counter electrodes 36a...h and reference electrodes 38a...h preferably are arranged in pairs to bracket an assoc ated working electrode 34a ... h . In order to minimize or reduce diffusion effects on the counter and reference electrodes, the electroactive surfaces of the metal te inal counter and reference electrodes were coated with a permeable membrane in accordance th the teachings of my copending U.S. application ser al no. filed contemporaneously herewith ( Attorneys' Docket No. ESA 140/142). The overall process was as follows: Palladium wires were spray coated with Teflon S to bui ld up a coating of approximately 0.0002" t ickness. (Teflon S is a trademark of E.I. DuPont de emours Company for a synthetic tetrafluorohydrocarbon.) The coated wires thus prepared were dipped sequentially in commercially available Teflon etching solution, concentrated HN0,/H-S0, (50:50 by volume) and tet rahydrofuron in a sonifier bath for approximately 1 inute n each to develop a semipermeable membrane on the w re surface. The Teflon coated/etched palladium wires were cut and assembled as the counter and reference electrodes in accordance with the teachings of my aforesa d U.S. Patent o. 4,511,659. As ill become clear from the following description electrochemical cells 20...32A act as gate electrodes for discri inating and screening interfering species, whi le electrochemical array cells 34 and 34A contain the measuring electrodes. In order to achieve high noise discrimination against pressure spikes and voltage fluctuations the measuring electrode should have a relatively small electrode area and thus a relatively small number (e.g. 4 ) of reaction half-times as compared with the various upstream gate electrodes. In order to maximize bandspreading and reduce signal-to-noise ratio the electrochemical cell (20...34A) was pretreated to optimize the flow path therethrough in accordance ith the teachings of my copending U.S. application serial no. fi led contemporaneously herewith (Attorneys' Docket No. ESA 139) . As taught therein, a melted polymer mixture comprising 67 volume percent C / 0 hydrocarbon and 33 volume percent 0.916 density polyethylene, heated to 98 C was pumped into the cell at 800 psi. Thereafter, water heated to 98 C was passed through the cell at 0.1 ml/mi n at 20 psi. The cell was then cooled from 98 C to ambient (25 C) over a 15 minute time period by continuing the flow of cooler and cooler water. Thereafter, the cell was flushed at 1 ml/min for 30 minutes with 10/10/80 THF hexane acetoni tr i le . Further understanding of the principles and advantages of the present invention may be had by reference to the following examples which a re based upon electrochemical analysis employing an electrochemical apparatus made in accordance with Figs. 1 and 2 and comprising eight electroche ically isolated electrochemical cells 20...34A. The working electrodes 34a ... h each comprise fritted graphite discs, treated as above described, each having a wor ing area of 2 approximately 4cm ( 90 half times ) . Electrode 34h has a working area of .approxi ately 0.3cm (4 half times) . Counter electrodes 36a...h and reference electrodes 38a ... h comprise inert metal (palladium ) wire terminals treated as above described. C cerebrosp na I fluid samples from motion-sick, non-sick and motion-sickness res stant cats were ana lyzed. The initial hypotheses were: 1. Some compound or compounds are produced within individual animals during motion stimulus that causes the emetic response in susceptible animals; or 2. Some difference in the small molecule inven.tory or che ical patterns exists in susceptible vs. non-susceptible animals. Initially, conditions of maximum resolution and sensitivity of known compounds suspected of having a relationship to motion sickness were considered. Thereafter, conditions were defined for maximum resolution of unknown compounds or compounds as appear to be different among sick, non-sick and motion-sickness resistent animals. Cerebrospinal fluid samples were directly injected in a mobile phase comprising 5% MeOH, 2.5% CHjCN and C.1M aH 2 P0 adjusted to pH 3.00 with H,P0, and 650 mg/l octane sulfonic acid sodium salt uti lizing a Brownlee RP18 3 cm. Guard Column and nto a Waters Cl8μ Bondapak Analytical Column. Mobile phase modifications were carried out for the following samples, under the following operating parameters (all voltage values in millivolts) : RED0X analysis of the samples was performed (oxidation followed by step-wise reduction of the products for detection) under detector conditions chosen for maximum resolution of NE, EP, L-D0PA, D0PAC and DA on T . , HGA on T- and T. and indoles on T,, T-, and T fi and the results recorded in Table I . Detector: G -350, C c +400, T. '-30, T 2 -60, T 3 -140, T 4 -150, T 5 -200, T 6 -230, T-. -310, T 8 -350. TABLE I ng/mL in CSF Samp le No. 2-201* 2-203 2-239* 2-243 2-342 2-554 MHPG 8.20 7.10 8.00 8.80 7.60 5.30 NE (LT 0.15) 0.30 0.24 (LT 0.20) (LT 0.20) (LT 0.20) **HGA 1.02 4.80 5.30 4.40 4.80 4.40 L-DOPA 0.66 1.10 0.90 0.89 0.89 0.67 EP 1.83 2.10 1.87 2.00 1.75 1.60 DOPAC 7.97 7.56 10.10 8.23 9.60 7.40 DA (LT 0.38) (LT 0.30MLT 0.30 ) (LT 0.30KLT 0.30MLT 0.30 ) 5HTOL 1.07 1.25 1.30 1.21 1.65 1.14 5HIAA 162.00 162.00 190.00 213.00 213.00 168.00 5HTP — HVA 116.00 112.00 131.00 125.00 135.00 109.00 3MT (LT 0.90) (LT 0.90) (LT 0.90) (LT 0.90MLT 0.90) (LT 0.90) 5HT 2.67 0.53 2.00 ( LT 0.43 ) 0.60 ( LT 0.43 ) * Average two analysis **Coelutes with HGA k' = 1.71 T7+ T8 n* I** 5.20 44.00 74.00 41.00 45.50 41.50 k« = 0.93 nAT2 (GT 100) 24.00 7.00 8.50 4.50 3.50 k' = 0.93 nAT5 49.00 21.00 (LT 3.00) 3.00 (LT 3.00)(LT 3.00) k'= 0.93 nAT6 (GT 100) 47.00 21.00 12.00 9.00 LT = less than GT = g reater than Table II shows the result of standards analyzed under these conditions. TABLE II RESPONSE nA OF STANDARD 10 SD CV% T3+T5 MHPG 13.8 14.2 14.4 13.86 0.332 2.40 9 T3 NE 6.0 6.2 6.2 6.04 0.133 2.21 9 T6 HGA 9.5 9.0 9.5 9.30 0.422 4.53 10 T3 L-DOPA 10.0 10.0 9.5 9.80 0.258 2.63 10 T3 EP 4.0 3.6 3.6 3.62 0.175 4.84 10 T3+TT4 DOPAC 29.8 31.0 30.8 30.82 0.553 1.79 10 T3 DA 3.4 3.4 3.2 3.32 0.193 5.82 10 T7+T8 5HT0L 18.5 18.5 17.0 14.83 0.750 5.05 6 18.37 0.250 1.33 4 T7+8 5HIAA 91.0 92.0 92.0 78.75 1.290 1.64 6 91.38 0.650 0.71 4 T3+T4 HVA 7.2 7.2 7.2 7.30 0.254 3.48 10 In the initial high resolution/selec ivity Redox condition several anamolous peaks were noted some of which are listed by their k 1 and electrode number in Table I. Consequently, a more open 16 channel array from 0 to 1600 millivolts without a gate cell preceding t was utilized to define the more complete structure of profile of the preli inary samples. A series of form mobile phase compositions of increasing organic modifier of 0.5, 5.0, 1.5 and 30% ethanol was used. Approximately 300 components were isolated in the various mobi le phases. Several of these components appeared to vary as a function of e esis. Qualitative identification of these components was made by comparison of potent ia I/1 ime locations of compounds from Table III. ' TABLE III Partial List of Abbreviations for Chemical Compounds and Typical Ranges Found With Preliminary Gradient Work in Cerebrospinal Fluid (CSF) + indicates observed but not quantified NM indicates not analyzable by technique * indicates compound to be included in study indicates not present ng/mL Typical Values or Range in CSF ( Fe I i ne ventricular) Acet acetaminophen* ANA anthranilic acid NM ASC ascorbic acid* 100-300 BEND 8-endorphin* THB*DHB biopterin* 3-30 BUF bufotinene* CAF caffeic acid* C.1-1 CAT catecho I* ' 7-17 CCK8 cholecystokinin 8* CCK4 cholecystokin n 4 CYSH cysteine* DHBAC 3,4-di hydroxybenzoi c acid* 1-8 D0MA di hydroxymande I i c acid* 2-6 D0PEG d ydroxypheny I glycol* 0.2-1.0 DHPGA 3,4-di hydroxypheny l- glyoxyllic acid DOPAC di hydroxyphenylacet i c acid* 8-30 DHQ 4,8-di hydroxyqu no Ii ne* DMPA 3,4-dimethoxypheπylaceti c ac d DMPEA 3,4-dimethoxyphenyl- ethyla ine DA dopamine* 0.08-0.17 EP epinephrine* 0.05-0.32 EPIN epi ni ne* + FER ferulic acid* + FAN formylanthranilic acid FKY formyl kynurenine* GA gentistic acid* 0.8-2.0 GSH glutathione* 50-70 HIST hi stami ne* HIS hi sti dine* HGA homogentisic acid* 0.5-3.0 HQ hydroqui none HVA homovanillic acid* 50-250 HVOL homovanillyl alcohol* 0.5-1 HVRA homoveratπ ' c acid* HOR hordenine* + 3-OHAN 3-hydroxyanthranilic acid 0.1-0.2 5-HIAL 5-hydrox indo Ieacet a Idehyde* 5-HIAA 5-hydroxyindoleacetic ac d* 150-350 30HKYA 3-hydroxykynu enamine* 3-OHKY 3-hydroxykynureni ne* 0.3-1.0 6-OHMEL 6-hydroxymelatonin* 3-HPA 3-hydroxyphen lacet i c acid* 5-HT 5-hydroxytryptamine* 0.1-1.0 5-HTP 5-hydroxyt ryptophan* 0.2-0.6 5-HTP -ol 5-hydroxytryptophol* 1.5-3.0 I i ndo le* IAA ndoleacetic acid* IET i ndoIe ethano I IPA i ndo lepyruvi c acid IS i sati n* KYA kynurenic acid* KYN kynurenine* 1.0-2.0 LD L-dopa* 0.2-4.0 LENK leucine enkephalin* MEL melatonin* MN etanephri ne* LT 0.01 MENK methionine enkephalin* 5-MGA 5-methoxygent i s i c acid MHMA 3-methoxy-4-hydroxymandel c acid* MHPG 3-methoxy-4-hydroxyphenyl glycol* 10-40 MHPG- MHPG sulfate conjugate* 100-140 S0 4 5-MIAA 5-methoxyindoleacetic acid* 3-MT 3-methox tyram i ne* 0.020 3-MD 3-0-methy Idopa MGA methylgentistic acid* MHIST methylhistamine* HPA m-hydroxy Ipheny Iacet i c acid HMA m-hydroxymande I i c acid mHPPA m-hydroxypheny Ipy ruvi c acid mOCT m-octopami ne mTYRA m-tyramine* 1.0-5.0 mTYR m-tyrosine* 5.0-10-0 NA5HT N-acety I serotoni n* 0.5-2.5 NM5-HT N-methyl-5-hydroxytryptamine* DM5HI NN-dimethyl-5-hydroxyindole* DMTR NN-dimethyltryptamine DMTR-P NN-dimethyltryptamine phosphate E norepi nephri ne* 0.2-1.5 NMN normetaneph ri ne* LT 0.05 OHTA o-hyd roxypheny lacet i c acid OTYRA o-tyramine oTYR o-tyros i ne* 0CT octopamine* 0.05-1.5 pCOU p-coumaπ ' c acid* + pHMA p-hydrox mande I i c acid* pHPAA p-hydroxypheny laceti c acid* 4-10 pHPLA p-hydroxypheny I lact i c acid* 0-10 pHPPA p-hydroxyphenyIpy ruvi c acid* PAC phenylacetic acid NM PA phenylalanine NM PAE phenylethyla ine NM PLA phenyllactic acid NM PPA phenylpyruvic acid PAL py ri doxa I* PALP5 p ridoxa l-5-phosphate* PYRA p ridoxam ine* PYRA5P pyradoxamine-5-phosphate* PYR pyridoxine* + QA quinaldic acid SYN synephπ'ne* + TRPA tryptamine* 1.0-1.8 TRP tryptophan* 800 TYRA tyramine* 0.5-1.0 TYR tyrosine* 1500 U uric acid* 600 VA vani I li c acid* VMA vani I ly Imande I i c acid* 0.2-0.3 From this work the thirty-nine compounds listed in Table IV were decided upon as candidates for nvolvement in the e esis event. From the preliminary data, one mechanism was postulated for emesis and emesis blocking as a guide to the sample acquisition and analysis protocols to be used in a following study. The mechanism postulated was in the resting cat NE and HGA exist at very low free levels in prote bound pools of approximately 1 and 4 ng/mL, respecti ely. Stimulus causes the free NE to spike due to neuronal emission. Referring to Figs. 4 and 5, this emission is mediated by several other neurotransmitter pathways, including the serotonergic and adrenergic and possibly by B . compounds ( whi-ch are probable variable peaks in the second array condition ) . The free NE causes the emesis response and is then deteriorated by adsorption into the bound pool. Free HGA spi es s multaneously either by emission or release from the pool and persists acting as a blocking agent for further emesis from the same stimulus. Several other possible mechanisms could be postulated from the ini ial data and several other possible abnormalities in the cerebrospinal fluid data were observed in the preliminary study. Among these mechanisms were possible problems associated with the 150 P i sample size acquired. Therefore, in a follow-up study 50-80 Ul samples were used and conditions were set up to maximize the total information that could be obtained from a single sample. A step gradient procedure combined with a sixteen channel array cell was ut lized to describe the majority of the components of the tyrosine and tryptophan pathways on cerebrospinal fluid samples. Four separate mobi le phase solutions were prepared as fo I lows : Mobile Phase I comprising 0.5 volume percent MeOH, 1g/l disodium octane sulfonic acid, 0.1 M NaH-,P0, adjusted to pH 3.00 with H,P0,; Mobile Phase II comprising 0.5 MeOH, 150 mg/l disodium octane sulfonic acid, 0.1 M NaH-,P0 , adjusted to pH 3.00 with H,P0,; Mobi le Phase III comprising 1.0 volume percent isopropanol alcohol, 0.5 volume percent MeOH, 175 mg/l disodium octane sulfonic acid, 0.1 M NaH ? P0 , adjusted to pH 3.00 with H,P0,; and Mobile Phase IV comprising 5 volume percent isopropanol alcohol, 0.5 volume percent MeOH, 275 mg/l disodium octane sulfonic acid, 0.1 M N aH_,P0. d 4 adjusted to pH 3.00 with H,P0.. _. •+ The overall procedure is as follows:-- First the column was swept with Mobi le Phase IV. Mobi le Phase I was then flowed through the column for one minute. 200 microliters comprising a mixture of 1.0 mg/ml ascorbic acid and 0.5 mg/ml EDTA was injected into the column and Mobi le Phase I was flowed through the column for an additional fi e minutes. At the six minute mark, the mobi le phase was changed to obi le Phase II and flow continued for an additional eighteen minutes. Sample solution was i jected into the column at the 24 minute mark. At the 35 minute mark the mobi le phase was shifted to Mobi le Phase III and at the 60 minute mark the mobi le phase was shifted to Mobi le Phase IV. Mobi le phase flow was continued for twenty-five minutes, the mobi le phase was then shifted back to Mobi le Phase I and the sequence repeated with additional samples. A typical voltage/time resolution pattern for this method is shown in Fig. 9. A double blind analys s of 106 samples from emesis suscept ble, emesis moderately susceptible and emesis non-susceptible animals was performed. Thi rty-nine components under controlled test were analyzed in each sample and separated from 150 additional unknown components uti lizing the electroche ical imaging system of the present invention. Column temperature was maintained at 23.5 plus or minus 0.1 C. The results of the precision of the method as derived from coanalyzed standard is shown in Table IV. Of the thirty-nine components, appro i ately thi rty were present and unambiguously identified in all samples and submitted to extensive statistical analysis. TABLE IV C CV Compound DHPAA 4.1 MHPG-SO 7.4 THB 4.3 DOMA 6.16 DO PEG 4.81 pHMA 2.37 U 2.32 NE 4.60 VMA 5.37 MHMA 8.23 MHPG 3.71 OCT 6.2 HGA 8.3 EP 5.43 DHBAC 2.91 N 3.80 LD 6.31 DOPAC 5.21 SYN 10.4 GA 4.31 pHPLA 6.21 3-OHAN 9.85 DA 7.62 3-OHKY 7.95 TYR 6.34 PHPAC 6.-12 mTYR 6 .01 M 5. 7 2 5HT-01 6 .71 EPIN 5, 71 TYRA 6, 84 5-HIAA 3, 74 HVOL 4 80 HVA,VA 3.01 5-HTP 4 54 3-MT 6.12 5HT 4.03 CAF 5.12 TRP 5.01 The potential/t me loca ions of the analytes in the tyrosine ( TYR) and tryptophan (TRP) metabolic pathways (Figs. 4 and 5) were plotted on X-and-Y time and potential axes as shown in Fig. 9. Individual analytical values were obtained by comparison of the magnitude of a standard at the XY locations for the samples. From the data produced on 106 samples for 28 compounds, significant differences between emesis susceptible and emesis non-suscept ble individual animals were found in the Noradrenergic, Dopaminergic, Serotonergic and Purine metabolic systems. The compounds quantitated reflect the various pathways affecting the Dopamine metabolites and others. These values were considered in terms of an algorithm to reflect the possible concentrations of MHPLA (3-methoxy-4-hydroxy phenyl lactic acid) to the HVA concentration, 3-MT to increasing the DOPAC concentration, and DHPE (di hydroxypheπyI ethanol) and MHPE or HVOL (3-metho y-4-hydroxyphenyl ethanol) to decreasing the DOPAC concentration: From this information an appropriate c'ompound to block the motion induced emesis was deduced, and the compound was successfully tested on moderately emesis susceptible an mals. it can be seen the present invention permits determination of the precise identity of causitive agents, e.g. neurotransmitters responsi le for motion-sickness, and allows the manipulation of metabolic pathways which appear to have the effect of blocking motion s ckness. Determination of the precise identity of these compounds provides a valuable adjunct both to the diagnosis of the causative conditions or syndromes leading to a disorder, and also can provide the physician, veterinarian or pharmacologist ith an analysis tool for describing essentially the complete small molecule o r substance inventory of an individual or group of individuals, essentially the entire complex metabolic pathways of such substances, and the identity of naturally occurring substances or chemical adjuncts for treating a d sorder or related diseases. This permits, for example, those compounds identified as blocking agents, or precursors of said blocking agents, to be administered directly to an individual, or dietary modification which will cause the individual to produce internally the desired blocking agents may be administered. This also permits the exact definition of differences in metabolic pathways between classes of individuals and allows the selected administra ion of compounds wh ch enhance or retard the appropriate • segment of the metabolic system. This provides a key to the rational identification, development, manufacture and/or adm nistration of existing or new phar acolog cal drugs and treatment protocols. Thus, in order to treat a disorder resulting from a diagnosed che ical mbalance such as a neurological disorder the physician or veterinarian need merely admin ster to the patient sufficient blocking agents, substances, or precursors thereof to make up for a deficiency in the naturally occurring blocking agent or substance or administer the appropriate metabolic inhibiting substance to modify the metabolic pathways described using the chemical imaging system above described. oreover, the abi lity to determine the precise chemical nature of a neurological disorder, e.g. in comparison with a normal indiv dual, is an extremely important and desirable test, and may permit early ident fication and classification of individual disorders, and a treatment protocol with those naturally occurring compounds, substances or precursors which have a specific ty for blocking, mediating or otherw se eliminating the disorder. In other words, the present invention permits normalizing information from precursors and metabolites of the neurotransmitters or other chemicals identif ed as active in disorders such that algorithms for the compounds measured can be developed. These algorithms can be utilized to compensate for biological variability in individuals, products and metabolism and refine the assessment of the effect of an individual neurotransmitter or pathway. Thus in the emesis case en ire metabolic pathways when considered as a whole were shown to be different between susceptible and non-susceptible individuals, intervention guided by the chemical imaging pathway definition to make the susceptible pathways approach the non-susceptible was efficatious in suppressing emesis. It should also be recognized that the patterns obtained may show specific presence or absence of compounds in a particular disease state. For example, as discussed in my aforesaid U.S. Patent 4,511,659, a significant binary difference between cerebrospinal fluid samples from patients with dementia of the Alzheimer type and normals was found in the apparent oxidation states of the neurotrans itters serotinin or 5-hydroxy tryptamine (5HT), and its precursor 5-hydroxy tryptophan (5HTP) . Assuming these compounds to have causati e effect in Alzheimers, by blocking the generation of these compounds through metabolic pathways, the poten ial toxic effects of such compounds may be mediated or eliminated. Having identified those specif c compounds which appear to reflect an underlying condition, one skilled in the art rationally may determine the point in the metabolic pathway in which to intervene in order to eliminate or block the generation of potentially toxic materials. Blocking agents have an established and valued place in symptomatic treatment of various allergic reactions or disease. The present invention is based in part on the realizat on that there are certain naturally occurring blocking agents in li ing organisms. In certain instances disease or disorder is caused by an mbalance or insufficiency of blocking agents. In other instances such as stress, e.g. due to motion sickness, blocking agent production may be insufficient, or simply too slow to produce the desired blocking effect from the initial insult. The present invention in part provides a method for identifying those indi iduals susceptible to a particular disorder, and those particular blocking agents or precursors thereof, or other substances or compounds, which advantageously may be administered to that indiv dual whereby to enhance or retard a appropriate segment of that indi idual's metabol c pathway. Thus, for example, while classical blocking drugs are pharmacological antagonists that appear to act by occupying receptors on the effector cell, to the exclusion of agonist molecules without themselves init ating a response, blocking agents identified in accordance with the present invention necessarily need not react in a simi lar manner. The important criteria is that the so-called blocking agent or other substance employed in therapy produces in the individual a normalized or near normalized reaction. Unlike artificial drugs the blocking agents or other substances or compounds identified in accordance ith the present invention are naturally occurring in the body of the individuals. Accordingly the usual side effects, contraindications, etc. as normally may be experienced with administration of artificial drugs may not result in administration of blocking agents or other substances or compounds identified in accordance with the present invention. One skilled in the art will recognize the aforesaid invention is susceptible to modification. Thus, for example, while the electrochemical apparatus has been i llustrated as having sixteen electrochemically isolated gate/array cells, any number of electrochemically isolated electrochemical-gate/array cells may be employed in series to achieve the desired results. Moreover, reaction half-times may be modified by manipulating indi idual cell volumes which in turn may reduce certain kinds of noise. Also, two or more sensing electrode cells may be employed downstream of one another to detect and measure additional groups of compounds of interest. For example, after a four-electrode gate sequence to eliminate irreversible substances and define the upper and lower potential limits, an array of sensors (for example 10 to 16), if desired, including also additional gate electrodes, could be arranged in an increasing ox idative and then reductive modes. This will effectively display the current voltage curves for eluting compounds for both their oxidative and reducti e modes. The current from each electrode will produce one set of simultaneous equat ons as follows: I. i = A(C ) + B(C, ) = C(C ) a b c where A, B and C are constants defined by the potential and the nature of the compounds a, b and c and where c / - _ * - ...are the concentrations of a, b and a D c c. It would then be possible to solve and display, for as many coeluting components as there are sensors in the array, pro iding that there is not an absolute identity in the signature or current voltage curve. Employing an array of sensors in sequence would enhance sensitivity of the cells, and also enhance separation of compounds that co-e lute . Fig. 6 is a block diagram of a system for processing such dual signals. The elements illustrated in Fig. 6 are given the same numbers as the equivalent elements in Figs. 7 and 8. Through the use of peak signatures, there have been detected in the analysis of cerebrospinal fluid and brain tissue several peaks whose "signatures" were clearly not identical to that of the standard compound normally observed at that retention time. Prior art detection methods would not have allowed for the resolution of such co-eluting compounds and would have resulted in erroneous data. This new technology has clearly provided a much needed method for determining the existence of co-eluting compounds and has virtually eliminated the misidentification of neutrotransmitters. Additionally, two-d mensional pattern diagrams, i.e. electrochemical fingerprints of various selected materials, can be generated, similar to two-dimensional pattern diagrams achieved by standard chromatography techniques by suitably integrating signals on var ous electrodes. In such case, samples of known substances may be flowed through the electrochemical apparatus, and two-d mensional pattern diagrams in the form of vo Itammograms representing the selected substances generated using the apparatus and procedures above described. The resulting pattern diagrams may be stored in a suitable central processing un t for subsequent pattern matching and ident fication. The process thus described also may be advantageously employed for directly analyzing body fluids such as urine or blood for the purpose of diagnosing diseases or predisposition to diseases of a subject and establishing pharmaceut cal protocols (sequence, type and concentration of pharmaceuticals) for treating such diseases, thus providing unexpected and marked advantages over the many step c romatog raphi c analysis techniques reported by A.B. Robinson and L. Cauling, in the paper entitled "Techniques of Ortho-Molecular Diagnosis" in Clinical Che istry, Vol. 20, No. 8, 1974, pages 967-965, and by Miyagi et al, in U.S. Patent No. 4,338,811. Referring to Figs. 7 and 8, a sample fluid may be directly streamed into the electrochemical detection apparatus 10 as described in detail supra, where substances of interest may be separated and detected in the manner previously described. Individual peaks in the output signal from the detector 10 may then be integrated in an integrator 54, and the output signal from the integrator 54 applied to a central processing unit 56 which is referred to as a CPU hereinafter as described by Mijagi et al. In the CPU 56, a two-dimensional pattern d agram representing the relation between the peak areas and the retention times provided by the input signal is formed according to a preset program. When peak matching (shown in Fig. 7) is required, a reference ch romatogram is read out from a _. memory 58 such as a magnetic tape or a magnetic disk, and the two dimensional pattern diagram above described is formed after the CPU 56 judges whether or not the retention t me of each peak in the subject's chromatogra coincides with the retention time of the corresponding peak in the reference chromatogram and carries out peak matching when coincidence is not detected. Then, the data of the upper and lower limits of normal persons are read out from the memory 58 upper limit pattern diagram and lower limit pattern diagram of the normal persons in the CPU 56. The pattern diagram of the data of the subject is then compared geographically with these upper and lower limit pattern diagrams in the CPU 56 so as to judge whether the subject is normal or abnormal. The result of judgment is displayed on a display unit 60 and recorded by a hard copy unit 62. As described hereinbefore, the process of the comparison and judgment may be d splayed when so des i red . As shown in Fig. 8, the process of disease diagnosis is completed when the subject is judged to be normal. When, on the other hand, the subject is judged to be abnormal, the CPU 56 retr eves disease data in a manner as described hereinbefore from the file of classified diseases stored in the memory 58. On the basis of the thus retrieved disease data, the CPU 56 forms the upper limit pattern diagram and lower limit pattern diagram of the disease and compares geograph cally these pattern diagrams with the subject's pattern diagram so as to select the corresponding disease or analogous disease. Further the CPU 56 calculates the correlation coeffic ent between the pattern diagram of the subject and the average-valve curve of the selected disease and identifies the chemical deficiency of the subject, and thus a treatment protocol. The result of selection of the corresponding disease or analogous disease, the result of calculation of the correlation coefficient and/or the process of disease diagnosis by comparison of the pattern diagrams, and the treatment protocol, are displayed on the display unit 60 and recorded by the hard copy unit 62. When it is necessary to modify some of the data on the basis of doctor's other observations n the course of judgment of the normality or abnormality or in the course of disease selection, necessary data a r e applied from a manipulator panel 64 to the CPU 56 for the purpose of modification of the data . The nvention has been described for use in analyzing Tyrosine and Tryptophan derived neurotrans itters, precursors and metabolites vis-a-vis thei r role in motion-induced emesis. It wi ll be understood, however, that the invention advantageously may be used to diagonose and characterized disorders, and to characterize and establish pharmaceutical protocols for a variety of environmentally induced, nherited and/or degenerative disorders such as Alzheimer's Disease, Parkinson's Disease, Huntington's Disease, ALS, MS and seni le dementias; behavioral disorders such as depression, suicide, hyperacti ity, weight control, schizophrenia, learning disabi l t es, memory retention, alcohol and substance abuse and dependency and sleeplessness; emesis in pregnancy or following chemot erapy; post operative anesthesia response; and hypertension and c a rd i o-vascu la r involvement which are gi en as exemplary. One ski lled in the art ll recognize that the apparatus and method above-described advantageously may be employed for resol ing mixtures of a large number of electroactive organic substances among which are mentioned unsaturated hydrocarbons, azides, triazines and phenoth i az i nes, amino acids, amines and am des, phenols, aromatic OH, quinolines, quinones, i ines, olefins, ketones, aldehydes, esters, olefinic esters, ethers, o rganomet a I I i c s, diazo compounds, nitro compounds, and halogens. Electroactive organometallic compounds n association with macro molecules also can be resolved using the electrochemical apparatus and method above desired. Sti ll other changes and advantages wi ll be obvious to one s i lled in the art.

Claims: CLAIMS 1. A method for develop ng pharmaceuticals for disorders in li ing organisms which method comprises the steps of analyzing body fluids from normal and afflicted (abnormal) individuals to generate analysis patterns representative of constituents of the metabolic pathways of said individuals; comparing said patterns to determine differences in said patterns representative of (a) the presence or absence of specific compounds in said patterns, or (b) differences in the concentration of said compounds in said patterns; and introducing into a pharmaceutically acceptable carrier therefor one or a mixture of chemical compounds known to act at sites ithin said metabolic pathways to (a) block or potentiate those metabolic pathways identified as abnormal by the pattern, or (b) supply or retard those unique compounds identified as abnormal in said patterns. 2. A method according to clai 1, wherein said body fluids comprise electrochemically active compounds, and including the steps of passing said body fluids sequentially through a liquid chromatograph ic column for achieving time-space separation of the materials eluting from the column and an electrochemical detection apparatus whereby to generate electrochemical patterns of said electrochemically active compounds. 3. A method according to claim 1, wherein said normal fluid contains a blocking agent for said abnormality, and including the step of examining said chemical analysis patterns to identify said blocking agent . 4. A method according to clai 3, and including the step of treating said abnormal individual with measured quantities of said block ng agent. 5. A method according to claim 3, and including the step of treating said abnormal individual so as to promote autogenetic production of said blocking agent in said abnormal individual. 6. A method according to claim 1, wherein said body fluid comprises serum. 7. A method according to claim 1, wherein said body fluid comprises cerebrospinal fluid. 8. A method according to claim 1, wherein said body fluid comprises plasma. 9. A method according to claim 1, wherein said body fluid comprises urine. 10. A method according to claim 1, wherein said body fluid comprises saliva. 11. A method according to claim 1, wherein said body fluid derives from a body tissue. 12. A method according to claim 11, wherein said tissue comprises brain tissue. 13. A method for developing pharmaceuticals for disorders in living organisms which method comprises the steps of analyzing body fluids from normal and afflicted (abnormal) individuals to generate analysis patterns representative of individuals' electrochemically active small- molecule inventory, comparing said patterns to determine differences in said patterns representative of (a) the presence or absence of specific electroche ically active compounds in said patterns, or (b) differences in the concentration of said compounds in said patterns; and introducing into a pharmaceutical carrier one or a mixture of chemical compounds known to act at sites within metabolic pathways of said organisms to (a) block or poten iate those metabolic pathways identified as abnormal by the pattern, or (b) supply or retard those unique compounds identified as abnormal in said patte rns . 14. A method according to claim 13, and including the steps of passing said body fluids sequentially through a liquid chromatograph c column for achieving time-space separation of the materials eluting from the column and an electrochemical detection apparatus whereby to generate electrochemical patterns of said electrochemically acti e compounds. 15. A method according to claim 13, wherein said normal fluid contains a blocking agent for said abnormality, and including the step of examining said chemical analysis patterns to identify said blocking agent . 16. A method according to claim 13, wherein said body fluid comprises serum. 17. A method according to claim 13, wherein sa d body fluid comprises cerebrospinal fluid. 18. A method according to claim 1, wherein said body fluid comprises plasma. 19. A method according to claim 13, wherein said body fluid comprises urine. 20. A method according to claim 13, wherein said body fluid comprises saliva. 21. A method according to claim 13, wherein said body fluid derives from a body tissue. 22. A method according to claim 21, wherein said tissue comprises brain tissue. 23. A method for identifying pharmacological materials for treating li ing organisms, wh ch method comprises the steps of analyzing body fluids from normal and afflicted (abnormal) individuals to generate chemical analysis patterns representative of the electrochemically active small molecule inventory contained in said normal and abnormal body fluids; comparing sa d patterns and identifying differences in said patterns representative of (a) the presence or absence of specific compounds in said patterns, or (b) differences in the concentration of said compounds n the metabolic pathways of said individuals; and selecting from chemical compounds known to act at sites within said metabolic pathways one or a mixture of compounds to (a) block or potentiate those metabolic pathways identified as abnormal by the pattern, or (b) supply or retard those unique compounds identified as abnormal in said patterns. 24. A method according to claim 23, and including the steps of passing said body fluids sequentially through a liquid ch ro atographi c column for achieving time-space separation of the materials eluting from the column and an electrochemical detect on apparatus whereby to generate electrochemical patterns of said electrochemically active compounds. 25. A method according to claim 23, wherein said body fluid comprises a fluid selected from the group consisting of serum, cerebrospinal fluid, plasma, urine and saliva. 26. A method according to claim 23, wherein said body fluid derives from a body tissue. 27. A method according to clai 26, wherein said tissue comprises brain tissue. 28. A method of identifying an abo mality in a body fluid of an individual which comprises the steps of determining the presence and relative concentration of electrochemically active- compounds contained in said body fluid by passing a sample containing said body fluid sequentially through a liquid chromatographic column for achieving time-space separation of materials eluting from the column and an electrochemical detection apparatus for electrochemically testing a sample containing electrochemically active materials in solution, the detection apparatus compr sing a plurality of coulometric cells arranged in series, maintaining the testing electrodes of said coulometric cells at different potentials, said cells operating at progress vely varying potentials along the path of flow of the eluant through the cells, and recording the results of coulometric measurements from sa d cells so as to separate said measurements by measuring potential as well as by time of elution, and comparing the said- relative concentrations thus obtained with known concentrations of normal or abnormal body fluids. 29. A method for identifying an abnormality in a body fluid of an individual which comprises the steps of determining the presence and relative concentration of elec rochemically active compounds contained in said body fluid by passing a sample containing said body fluid sequentially through a liquid chromatograph c column by step gradients for ac ieving ti e spaced separa on of the materials eluting from the column and an electrochemical detection apparatus for electrochemically testing the eluant from said column, the detection apparatus comprising a plurality of coulometric cells arranged n series, maintaining the coulometric electrodes of said cells at different potentials, said cells operating at progressi ely varying potentials along the path of flow of the eluant through the cells, recording the results of coulometric measurements from sa d cells so as to separate said measurements by measuring potential as well by time of elution, and comparing said measurements thus obtained with measurements of normal and abnormal body fluids to provide dentification and relevant concentrations of constituents coeluting from the column. 30. A method of identifying abnormalities in the small molecule inventory of an individual comprising the steps in sequence of: determining the presence and relative concentration of electroche ically active constituents in the metabolic pathways containing such abnormalities by passing a sample containing said body fluid sequentially through a liquid chromatograph column for achieving time-space separation of the constituents eluting from the column and an electrochemical detection apparatus comprising a plural ty of coulometric cells arranged in series; maintaining the coulometric electrodes of said cells at different potentials, said cells operating at progressi ely varying potentials along the path of flow of the eluant through the cells, said plurality of cells being sufficient in number to separate by measuring poten ials those relevant constituents coeluted from the chromatographic column at any instant of time and recording the results of such coulometric measurements so as to separate said measurements by measuring potential as well as by time of elution and comparing the said relevent concentrations thus obtained with known concentrations of normal or abnormal body fluids and thereby identify changes of rates through the various metabolic pathways of said relevant constituents as well as unknown metabolites and abnormal precursors for normal metabolites. 31. A method of determining the relative concentration of principal constituents in the metabolic pathways of a-n ani al by passing a sample containing body fluid from said animal sequentially through a liqu d chromatograph column for achieving time-space separation . of the constituents eluting from the column and an electrochemical detection apparatus comprising a plurality of coulometric cells arranged in series; maintaining the coulometric electrodes of said cells at different potentials, said cells operating at progressively varying potentials along the path of flow of the eluant through the cells, said plurality of cells being sufficient in number to separate by measuring potentials those relevant constituents coeluted from the chromatograp ic column at any instant of ti e, and recording the results of such coulometric measurements so as to separate said measurements by measuring potent al as well as by ime of elution. 32. A method for treat ng a disorder in a living organism which method comprises the steps of analyzing body fluids from normal and afflicted (abnormal) individuals to generate analysis patterns representative of constituents of the metabolic pathways of said individuals; comparing said patterns to deter ine differences in said patterns representative of (a) the presence or absence of specific compounds in said patterns, or (b) differences in the concentrations of said compounds in said patterns; introducing into a pharmaceutically acceptable carrier therefor one o r a mixture of chemical compounds known to act at sites within said metabolic pathways to (a) block or potentiate those metabolic pathways identified as abnormal by the pattern, or (b) supply or retard those unique compounds identified as abnormal in said patterns; and administering to said living organism said identified compounds in said pharmaceutically acceptable carrier so as to (a) block or potentiate those metabolic pathways identified as abnormal by the pattern, or (b) supply or retard those unique compounds identified as abnormal in said patterns. 33. A method of treating a disease state in an animal, comprising the steps in sequence of identifying abnormali ies in a body fluid of said animal by determining the relative concentration of the principal constituents in the metabolic pathways containing such abnormal ties by passing a sample containing said body fluid sequentially through a liquid chromatograph column for achieving time-space separation of the constituents eluting from the column and an electrochemical detection apparatus comprising a plurality of coulometric cells arranged in series; maintaining the coulometric electrodes of said cells at different potentials, said cells operating at progressively varying potentials along the path of flow of the eluant through the cells, said plurality of cells being sufficient in number to separate by measuring potentials those relevant constituents coeluted from the chromatographic column at any instant of time and recording the results of such coulometric measurements so as to separate said measurements by measuring potential as well as by time of elution and comparing the said relevent concentrations thus obtained with known concentrations of normal or abnormal body fluids and thereby identifying changes of rates through the various metabolic pathways of said relevant constituents as well as unknown metabolites and abnormal precursors for normal metabolites, and identify those compounds responsible for sai changes of rates, and administering to said animal said identified compounds so as to provoke a change of rate through said metabolic pathways of said individual substantially to approach that found in said normal fluids. 34. A pharmaceutical produced according to the method of claim 1. 35. A pharmaceutical produced according to the method of cla m 13.

Inventors: MATSON WAYNE R, MATSON, WAYNE, R.
Applicants: ESA INC, ESA, INC.
Application year: 1985
Main classification: G01N27
IPC codes: A61B10, G01N27, G01N30, G01N33