• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    Device for fluorimetric determination of tertiary amines. A device for the determination by fluorescence of tertiary amines by means of reagents is described, for this the device is equipped with a plate of transparent and colorless material, another plate of the same material and of the same dimensions on which a groove or channel is practiced. to circulate the fluid of the problem sample. Likewise, a valve is available to regulate the gas inlet in the channel of the device together with a pump to introduce the sample in the channel of the device. A capillary tube is used to conduct the sample from the pump to the entrance of the device channel, and an LED responsible for causing the emission of fluorescence together with an optical filter. All this complemented with a fluorescence detector and an optical pound to drive the light emitted to it. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2684868A1
    申请号:ES201730530
    申请日:2017-03-31
    公开日:2018-10-04
    发明作者:Gabriel GARCÍA HERBOSA；Tomás TORROBA PÉREZ；José Vicente CUEVAS VICARIO；Victor GARCÍA CALVO；José GARCÍA CALVO
    申请人:Universidad de Burgos；
    IPC主号:
    专利说明:

    The present invention relates to a device for fluorescence determination of tertiary amines using the. The device is based on the use of microfluids moved with a syringe pump, lighting system and fluorescence conduction by means of an optical fiber to a detector external to the device.
    STATE OF THE TECHNIQUE
    The detection by means of portable devices of potentially dangerous substances, such as toxic or flammable gases, or somehow interesting is one of the 15 main contributions to building an increasingly safe society. One of the most interesting aspects of portable detection systems is the ability to provide reliable data in short periods of time. Normally it is desired to obtain the quantification data of the substance of interest instantaneously. For example, the detection of levels of ca, nitrogen oxides in gases
    20 combustion residuals in heating boilers or in vehicles are obtained immediately by applying immersion probes in the fluid. However, in the detection and determination of, for example, the amount of ethyl alcohol present in the drivers' breath requires that the person analyzed provide the sample to be analyzed.
    25 Drugs and their abusive use are a serious health problem being the cause of millions of serious illnesses or injuries, and many of the main social problems, such as driving under their effects, violence, stress and child abuse. In addition, the addiction derived from this consumption is a chronic disease,
    30 characterized by the compulsive search for drugs and their use despite the negative consequences associated with it.
    Among the most addictive drugs is heroin, an opioid derived from morphine, and administered intravenously is two to four times more potent than this. Both this drug, as the rest of compounds belonging to the opioid family, and more generally, tertiary amines, have very
    Various applications such as drugs, pesticides and surfactants, and therefore the development of sensitive analytical methods for their determination, especially in aqueous solution, is a desirable aspect.
    5 Portable devices currently marketed to detect and quantifyimmediate way the presence of drugs in biological media (DrugTest 5000, CocaChip, etc.) are based on enzymatic reactions that are detected byelectronic absorption spectroscopy or UV-VIS. The costs for each analysis areconsiderably high. In addition, the results of the analyzes are considered
    10 only indicators when establishing possible sanctions. Unlike the devices currently marketed, the invention presented here is based on the detection of the fluorescent emission that appears when mixing a reagent kit under the conditions required by the device. It is well known that fluorescence is a much more sensitive technique than that of visible ultraviolet absorption.
    DESCRIPTION OF THE INVENTION
    A portable device for the determination of tertiary amines based on the fluorescence emitted by an deaerated sample is described in the present invention.
    20 contains the analyte and the selected reagents, which can be halogenated xanthene pigments. Tertiary amines are drugs such as opiates and / or their derivatives in such a way that the device of the invention is used for the detection and / or quantification of heroin in samples.
    Therefore, a device for the detection and quantification of tertiary amino acids that allows to know the concentration of tertiary amine present in the test sample is presented through the fluorescent emission induced by illumination.
    30 Additionally, the device operates in an aqueous medium and uses stable materials, making it valid for the in situ determination of substances such as heroin and opiates, in urine and saliva samples.
    Therefore, a device for the detection and / or quantification of fluorimetric tertiary amines is described herein, characterized in that it comprises the following components:
    to) A smooth plate of transparent and colorless material.
    b) Another plate of the same material and of the same dimensions on which it
    Practice a groove or channel to circulate the fluid in the test sample.
    C) A valve that allows regulating the entry of gas into the channel of the device.
    d) A syringe pump to introduce the sample into the channel of the device.
    and) Join your flexible metal capillary bo to drive the sample from the pump
    syringe to the entrance of the device channel.
    F) A first LED emitting at 495 nm (green or blue) responsible for causing the
    excitation responsible for fluorescence emission.
    g) An optical filter that only allows the passage of wavelengths less than 500
    nm.
    h) A fluorescence detector.
    i) Fiber optic to drive the emitted light to the detector.
    j) Areceiverinhethatlocate the problem sample during the process of
    illumination.
    k) A second LED lightwhite to illuminate the sample during the time
    necessary.
    one) A tube to discard the sample that has run through the device channel
    during the measurement process.
    With the device provided by the present invention it is possible to detect mainly tertiary amines.
    In a preferred embodiment, the transparent and colorless plate material is, for example, glass, polycarbonate, methacrylate, polyesters, resins, polyamides, polyethylenes. In an even more preferred embodiment, the transparent and colorless plate material is polycarbonate.
    In another possible preferred embodiment, the two plates are smooth and assembled with a separator on which the channel through which the fluid circulates has been etched.
    In another more preferred embodiment, the valve that regulates the entry of gas into the channel of the device may have manual or automatic control.
    In another even more preferred embodiment, a syringe pump or any similar part of the market that is used for the same purpose of introducing the sample at a certain rate is used.
    5 In another still more preferred embodiment, a capillary, tube or carrying system is usedshows it to the circuit.
    The device of the invention can be equipped with a LED or any other lighting system that achieves the excitation of the sample so that an emission 10 of this occurs; while an optical filter can be used to prevent the wavelengths at which the sample is emitted from reaching it.
    The device can be complemented with a light emission detector to collect the light signal from the sample to be analyzed.
    In another preferred embodiment, any type of optical fiber is used to carry the signal to the detector.
    In another preferred embodiment, any chamber, cuvette, vessel, or channel 20 designed for irradiation of the sample is used.
    In another preferred embodiment, a led or lighting system is used that is capable of irradiating the sample and producing the chemical reaction.
    In another preferred embodiment, a tube or any valid system is used to evacuate the sample from the device after the analysis is completed.
    In the present invention, opiates are understood as both natural alkaloids from opium, such as morphine, codeine and thebaine, among others; as to its 30 semi-synthetic derivatives such as heroin and oxycodone, among others; and to completely synthetic opioids, such as pethidine and methadone, among others.
    Throughout the description and claims the word "comprises" and its variants are not intended to exclude other technical characteristics, components or steps. For those skilled in the art, other objects, advantages and characteristics of the invention will be derived partly from the description and partly from the practice of the
    invention. The following examples and figures are provided by way of illustration, and
    They are not intended to be limiting of the present invention.
    BRIEF DESCRIPTION OF THE FIGURES
    Figure 1. Shows a diagram in which the different components of the device of the invention are appreciated.
    Figure 2. Shows a graph showing the linear dependence observed in the
    10 calibration of the device with different concentrations of fluorescein in a range of 0.1 ~ M to 10 ~ M.
    Figure 3. It shows a graph showing the variation of the emission intensity values obtained in the calibration of the device as a function of the heroin concentration.
    PREFERRED REALIZATION
    In a preferred embodiment of the object of the invention there is a device (1)
    20 for fluorimetric determination of tertiary amines shown in Figure 1, which comprises at least a first plate (2) of colorless and transparent material in light in visible spectrum, and a second plate (3) also of colorless and transparent material in light in visible spectrum of approximately the same size as the first plate (2) and partially comprising a channel (4) intended to drive
    A sample fluid, both plates (2,3) being separated by a sheet (5) of inert material disposed between the two plates (2,3) where said sheet (5) partially comprises channel (4); in such a way that the channel (4) can be defined between the sheet (5) and the second plate (3). In a more preferred embodiment, sheet (5) is not available and channel (4) is defined only
    30 on the second plate (3).
    Said channel (4) is associated with a valve (6) that can be manual or a digitally controlled solenoid valve and is intended to control at least the gas inlet in the channel (4) which is also responsible for receiving a sample 35 which is introduced therein by means of a pump (7), preferably of syringes,
    connected to the channel (4) by means of a preferably flexible capillary tube (8) that conducts the sample from the pump (7) to an inlet of the channel (4).
    To carry out the object of the invention, a first emitter (9) of
    5 light radiation intended to cause the excitation responsible for the emission offluorescence, this can be an LED type emitter that emits light radiation witha wavelength of approximately 495 nm, which is associated withan optical filter (10) intended to allow the passage of wavelengths less than 500nm, so that a fluorescence detector (11) associated with an optical fiber (12) for
    10 to conduct the radiation emitted by the first emitter (9) to the detector (11) carry out the fluorescence detection produced by the reaction of the sample when it is irradiated by the first emitter (9) since the sample is held in a container (15), a chamber or a cuvette or the like, intended to accommodate the sample while it receives the radiation from the first emitter (9). The detector (1 1) can be found
    15 arranged externally to the device (1) or being part of it giving rise to possible different configurations according to requirements.
    To be able to visualize the sample and its route, there is a second emitter (13) intended to illuminate the sample, which is of the LED type and emits white light and which
    20 finds the side of the container (15).
    Finally, and once the determination of the presence of tertiary amines and their eventual quantification is made, a drain (14) is available, such as a pipe connected to the channel (4), intended to discard the sample once it has been
    25 run through the channel (4).
    Next, the invention will be illustrated by tests carried out by the inventors, which show the effectiveness of the device (1) of the invention.
    Unless otherwise indicated, the different materials, concentrations, and the like specified herein are illustrative only and should not be construed as limiting the scope of the invention.
    The basic operation of the device (1) is in a configuration for the detection and quantification of tertiary amines that allows to know the concentration of tertiary amine present in the test sample through the emission
    Fluorescent induced by lighting with the LEO, device (1) presenting the valve (6) which in this basic configuration is controlled manually.
    The device (1) has a sample inlet to inject the sample to analyze the5 which is connected by the capillary tube (8) in this case metallic to the pump (7)in this case syringe (see figure 1).
    The injected sample, driven by gas, travels through the channel (4) of the microfluidic device (1) until a certain point is reached at which the sample is excited
    10 by means of the light coming from an LED acting as the first emitter (9), which crosses the optical filter (10) of all wavelengths greater than 500 nm to proceed to collect the emission by means of the optical fiber (12) connected to the detector (11) fluorescence.
    15 Once the emission intensity data has been collected, the sample is pushed again by opening the valve (6) until the fluid reaches and fills the cuvette that acts as a container (15) and then the drive stops of the fluid by closing the valve (6). The container (15) is illuminated by the second emitter (13) that emits white light, since in a possible embodiment
    20 preferred both are located together. The lighting is stopped and the sample fluid is suctioned by means of the syringe pump (7) until it is positioned again on the said point at which the sample is excited by the light from an LED. The emission intensity data is collected again by means of the optical fiber (12) connected to the fluorescence detector (11), and
    25 the valve (6) is subsequently opened and the fluid is pushed to the outlet to empty the device circuit (1).
    The circuit can be washed with distilled water injected and driven through the entire route as many times as deemed appropriate.
    Calibration of the device (1) with fluorescein.
    The necessary calibration before measuring samples such as urine, blood, serum, blood plasma, sweat, saliva, or hair, and is carried out by preparing various solutions with different concentrations of fluorescein that are successively introduced into the device (1) and the emission data is recorded
    for each of the concentrations. Linearity is observed in the relationship between concentration and emission intensity (Figure 2).
    The graph resulting from these measurements conforms to the following equation:
    5 Ir = AC + B
    where Ir is the intensity observed, A is the slope of the line and e is the concentration of fluorescein.
    Device calibration (1) for measuring heroin concentrations.
    Samples containing heroin were prepared with a concentration range from 1O. 7M to 5x106M. Next, the sample is taken with a syringe and
    injected into the device (1) by the syringe pump and the sequence indicated above in the description of the invention is followed.
    In particular, Figure 3 shows the results obtained for calibration with heroin.
    The resulting graph of these measurements fits the following equation: 20 Ir = A (l -1O-8e)
    where Ir is the variation in intensity before and after illuminating the sample at a determined wavelength, and e is the concentration of heroin in the sample.
    权利要求:
    Claims (9)
    [1]
    1. Device (1) for fluorimetric determination of tertiary amines, characterized in that it comprises:
    • at least one first plate (2) of colorless and transparent material to light in visible spectrum,
    • a second plate (3) of colorless and transparent material in spectrum light
    visible that at least partially comprises a channel (4) intended to conduct a sample,
    • a valve (6) associated with the channel (4) and intended to control the gas inlet in the channel (4),
    • a pump (7) intended to introduce the sample into the channel (4),
    • a flexible capillary tube (8) intended to drive the sample from the pump (7) 15 to a channel inlet (4),
    • a first emitter (9) of light radiation intended to cause the excitation responsible for the fluorescence emission.
    • an optical filter (10) associated with the first emitter (9) and intended to allow the passage of wavelengths less than 500 nm,
    20 • a fluorescence detector (1 1) associated with an optical fiber (12) to drive the radiation emitted by the first emitter (9) to the detector (1 1),
    • a container (15) intended to accommodate the sample while it receives the radiation from the first emitter (9),
    • a second emitter (13) intended to illuminate the sample, and
    25 • a drain (14) connected to the channel (4) and intended to discard the sample once it has run through the channel (4).
    [2]
    2. Device according to claim 1 characterized in that it additionally comprises a sheet (5) of inert material disposed between the two plates (2,3)
    30 which comprises at least partially channel (4) such that the channel (4) is defined between the sheet (5) and the second plate (3).
    [3]
    3. Device according to claim 1 or 2 characterized in that the first emitter (9)
    It is configured to emit light radiation with a wavelength of 495 35 nm.
    [4]
    4. Device according to claim 1 or 2 characterized in that the second emitter
    (13) is configured to emit white light.
    [5]
    5. Device according to claim 1 or 2 characterized in that the valve (6) is a5 digitally controlled solenoid valve.
    [6]
    6. Device according to claim 1 or 2 characterized in that the pump (7) is a syringe pump.
    Device according to claim 1 or 2, characterized in that the emitters (9,13) are LEO type.
    [8]
    8. Device according to claim 1 or 2 characterized in that the container (15) is a chamber or a bucket or the like.
    [9]
    9. Device according to claim 1 or 2 characterized in that the drain (14) is a pipe.
    [10]
    10. Device according to any of the preceding claims where the sample
    20 is selected from: urine, blood, serum, blood plasma, sweat, saliva, and hair.
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    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    US4008397A|1975-04-24|1977-02-15|Hoffmann-La Roche Inc.|Fluorometer flow cell|
    US4172227A|1978-07-21|1979-10-23|Becton, Dickinson And Company|Flow microfluorometer|
    CN2432579Y|2000-06-28|2001-05-30|中国科学院合肥智能机械研究所|Fluorescence analyzer for heroine|
    WO2005111586A1|2004-05-13|2005-11-24|As Laser Diagnostic Instruments|A portable device and method for on-site detection and quantification of drugs|
    WO2012098241A2|2011-01-20|2012-07-26|City University|Fibre-optic sensor|
    法律状态:
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    优先权:
    申请号 | 申请日 | 专利标题
    ES201730530A|ES2684868B1|2017-03-31|2017-03-31|DEVICE FOR FLUORIMETRIC DETERMINATION OF TERTIARY AMINES|ES201730530A| ES2684868B1|2017-03-31|2017-03-31|DEVICE FOR FLUORIMETRIC DETERMINATION OF TERTIARY AMINES|
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