Five, embodiment:
The present invention is not subjected to the restriction of following embodiment, can determine concrete embodiment according to technical scheme of the present invention and actual conditions.
Below in conjunction with embodiment and accompanying drawing the present invention is further described:
Embodiment, shown in accompanying drawing 7 and 8, this optical fiber in site online drug dissolution/release test instrument comprises light source 19, is no less than one y-type optical fiber chemical sensor probe 20, digestion instrument 18 and detecting device 21, the light source input end of y-type optical fiber chemical sensor probe 20 is connected on the light source 19, the test side of y-type optical fiber chemical sensor probe 20 places in the digestion instrument 18, and the light source output terminal of y-type optical fiber chemical sensor probe 20 is connected on the input end of detecting device 21.
Wherein: light source can adopt deuterium lamp or xenon lamp or mercury lamp or xenon/mercury lamp or laser instrument, detecting device can adopt charge-coupled detector(CCD) CCD, cmos image sensor CMOS, electric charge injector CID or diode array DAD detecting device, and concrete selection comes to determine as required.As shown in Figure 8, detecting device can adopt single channel or multichannel detector, and y-type optical fiber chemical sensor probe can adopt single or many y-type optical fiber chemical sensor probes.
Fibre optic chemical sensor probe among the present invention can adopt one of following fibre optic chemical sensor probe:
As shown in Figure 1, one of this fibre optic chemical sensor probe comprises jacket layer 1, optical fiber 2 and sensitive membrane 3, the test side 4 of optical fiber is equipped with sensitive membrane 3, the length of its optical fiber 2 is that 25cm to 250cm, diameter are 10.0 μ m to 2500 μ m, can determine concrete length and diameter according to actual needs, it is applicable to makes mini optical fibre original position drug dissolution/release test instrument.
As shown in Figure 2, two of this fibre optic chemical sensor probe comprises sleeve pipe 5, optical fiber 2 and reflective mirror 6, the test side 4 of optical fiber is installed in the inner end 7 of sleeve pipe 5, outer end 8 at sleeve pipe 5 is equipped with reflective mirror 6, one injection port 11 is arranged at the middle part 10 of sleeve pipe 5, the length of its optical fiber 2 is that 25cm to 250cm, diameter are 0.1cm to 2.50cm, determines concrete length and diameter too according to actual needs; As shown in Figure 2, according to the mensuration of different solid pharmaceutical preparations being carried out dissolution rate or release, produce the fibre optic chemical sensor probe of different light paths, be specially adapted to mensuration, be applicable to the fibre optic chemical sensor probe of making ad hoc type particular solid pharmaceutical preparation (tablet, capsule etc.) dissolution rate or release.
Shown in accompanying drawing 2 and 3, three of this fibre optic chemical sensor probe is with two difference of above-mentioned fibre optic chemical sensor probe: two injection port 11 is arranged at the middle part 10 of sleeve pipe 5.
Shown in accompanying drawing 2,3,4 and 5, four of this fibre optic chemical sensor probe is with two difference of above-mentioned fibre optic chemical sensor probe: the injection port 11 that a big lateral opening is arranged at the middle part 10 of sleeve pipe 5.
Shown in accompanying drawing 5 and 6, five of this fibre optic chemical sensor probe is with four difference of above-mentioned fibre optic chemical sensor probe: the outer end 8 at sleeve pipe 5 is equipped with adjusting light path bar 12 by engage thread or inserted mode, at the inner end 13 of regulating light path bar 12 reflective mirror 6 is installed, the injection port 11 of a big side opening is arranged at the middle part 10 of sleeve pipe 5.By regulating light path bar its light paths of 12 scalable (being the distance between reflective mirror and the optical fiber test side), thereby be applicable to mensuration to different solid pharmaceutical preparations (tablet, capsule etc.) dissolution rate or release, thereby it can be used as universal fibre optic chemical sensor probe, can adopt ultraviolet/visible/near infrared absorption, reflection and phosphorescence/fluorescence/light sources such as Raman spectrum radiation.
To detect performance in order further improving, as shown in Figure 6, sensitive membrane 3 can be installed in the inboard of reflective mirror 6; In order to be applicable to the detection of ultraviolet light, can be coated with ultraviolet reflectance film 14 in reflective mirror 6 outsides; In order to be applicable to more weak light source, optical fiber focus lamp 15 can be installed in the test side 4 of optical fiber.In order to increase adaptability, optical fiber can adopt y-type optical fiber or branch type optical fiber or not branch type optical fiber or single-mode fiber or multimode optical fiber.
The most preferred embodiment of the present invention that above technical characterictic has constituted, it has stronger adaptability and best implementation result, can increase and decrease non-essential technical characterictic according to actual needs.
Sensitive membrane among the present invention can adopt following three class disclosed methods to obtain respectively.
1. " molecular probe+polymkeric substance+plastifier " solvent evaporation method is made even sensitive membrane; Its disclosed document is:
[1] Seitz, WR., etc. based on the fibre optic chemical sensor of immobilizing indicator.Analytical chemistry, CRC Crit Rev, 1988; 19:135-139. (Seitz, WR., et al, Chemical Sensors based on immobilizedindicators and fiber optic, Anal.Chem., CRC Crit Rev, 1988; 19:135-139.)
[2] W.Rudolt Seitz. fibre optic chemical sensor polymkeric substance indicator substrate.Biosensor technology. (W.Rudolt Seitz.Polymeric Indicator Substrates for Fiber Optic Chemical Sensors.Biosensor Technology.)
[3] Kuit Seiler, Kemin Wang, Matthias Kuratli and Wilhelm Simon; A kind of ethanol selectivity optical fiber sensing membrane based on the reversible chemical identifying.The analytical chemistry journal, 1991; 244:151-1601. (KuitSeiler, Kemin Wang, Matthias Kuratli and Wilhelm Simon; Development of anethanol-selective optode membrane based on a reversible chemical recognitionprocess.Anal.Chim.Acta, 1991; 244:151-1601.)
[4] Otto S.Wolfbeis. chemical sensitisation indicator dye.Fibre optic chemical sensor, 1997,4:53-107. (Otto S.Wolfbeis.Chemical sensing using indicator dyes.Optical fiber sensor.1997,4:53-107.)
2. " sol-gel " legal system sensitive membrane (mesh is arranged, get rid of solid content disturb) in the sensitive membrane surface distributed, its disclosed document is:
[5] Rainer Klein, the ammoniacal sensor that Edgar Voges. optics is integrated. Dutch analytical chemistry magazine, (1994) 349:394-398. (Rainer Klein, Edgar Voges.Integrated-optic ammonia sensor.Fresenius J Anal.Chem. (1994) 349:394-398.)
[6] David J.Blyth, Sarah J.Poynter and David A.Russell is with the fixing calcium biology sensor of photosensitive protein of sol-gal process.The analyst, 1996,12, vol.121 (1975-1978). (David J.Blyth, Sarah J.Poynter and David A.Russell, Calcium Biosensing with a Sol-gelImmobilized Photo protein.Analyst, December 1996, vol.121 (1975-1978) .)
[7] R.C.Hughes, S.V.Patel, M.W.Jenkins, T.J.Boyle, T.J.Gardner and C.J.Brinker is based on collosol and gel porous membrane chemical catalysis sensor.(R.C.Hughes,S.V.Patel,M.W.Jenkins,T.J.Boyle,T.J.Gardner?and?C.J.Brinker,Thin?film?Porous?MembranesBased?on?Sol-gel?Chemistry?for?Catalytic?Sensors.)
[8] the collosol and gel membrane technology prepares sulfate anion chemistry microsensor (Sulfate Anion-SensingChemical Microsensors Prepared by the Sol-gel Membrane Technology.)
[9] O.Lev, M.Tsionsky, L. Rabinovich, the collosol and gel sensor of et al. organic decoration.Analytical chemistry, 1995, Vol.67 (1): 22A-30A. (O.Lev, M.Tsionsky, L.Rabinovich, et al.Organically modified Sol-Gel Sensors.Analytical Chemistry, 1995, Vol.67 (1): 22A-30A.)
[10] Dave B.C., Dunn B., Valentine J.S.et al. sol-gel embedding legal system is equipped with biology sensor.Analytical chemistry, 1994,66 (22): 1120A-1127A. (Dave B.C., Dunn B., Valentine J.S.et al.Sol-Gel encapsulation methods for biosensors.Anal.Chem.[J], 1994,66 (22): 1120A-1127A.)
[11] Kwang E.Chung, Esther H.Lan, Michael S.Davidson. glass embedding myoglobin is measured oxygen in water.Analytical chemistry, 1995 (67): 1505-1509. (Kwang E.Chung, Esther H.Lan, MichaelS.Davidson.Measurement of dissolved oxygen in water using glass-encapsulatedMyoglobin.Anal.Chem.1995 (67): 1505-1509.)
[12] Upvan Narang, Paras n.Prasad, and Frank V.Bright. is based on the glucose biological sensor of collosol and gel derivatization.Analytical chemistry, 1994 (66): 3139-3144. (Upvan Narang, Paras n.Prasad, and Frank V.Bright.Glucose biosensor based on Sol-Gel-derived platform.Anal.Chem.1994 (66): 3139-3144.)
[13] Faida A.El-Essi, Ali Z.Abu Zuhri, Suleiman I.Al-Khalil, et al. is with the fixing hydrogen peroxide that produces of horseradish peroxidase spectrographic determination enzymatic of collosol and gel.Talanta,1997(44)2051-2058.(Faida?A.El-Essi,Ali?Z.Abu?Zuhri,Suleiman?I.Al-Khalil,et?al.Spectrophometric?determination?of?enzymatically?generated?hydrogen?peroxideusing?Sol-gel?immobilized?horseradish?peroxidase.Talanta,1997(44)2051-2058.)
[14] L.M.Ellerby, C.R.Nishida, S.A.Yamanka, the transparent porous silica glass of et.al. Prepared by Sol Gel Method embedding albumen.Science [J], 1992,225:1113-1115. (L.M.Ellerby, C.R.Nishida, S.A.Yamanka, et.al.Encapsulation of protein in transparent porous silicate glassesprepared by the sol-gel method.Science[J], 1992,225:1113-1115.)
3. the covalent bonding legal system is equipped with sensitive membrane, and its disclosed document is:
[15] the optical fiber fluorescence sensing method of evaluation antibody technique for fixing.The analytical chemistry journal, 229 (1990) 169-176. (Evaluation of antibody immobilization techniques for fiber optic-basedfluoroimmunosensing.Analytica Chimica Acta, 229 (1990) 169-176.)
[16] optical fiber biosensor of immobilized enzyme.Biosensor technique.(Fiber?Optic-Based?BiosensorsUtilizing?Immobilized?Enzyms.Biosensor?Technology.)
[17] Susan L.R.Barker, Yunde Zhao and Michael A.Marletta; Solubility guanine cyclase heme based on dye marker is applied to the highly sensitive of cell and high selectivity optical fiber nitrogen monoxide biology sensor.Analytical chemistry, 1999,71,2071-2075. (Susan L.R.Barker, Yunde Zhao and Michael A.Marletta; Cellular applications of a sensitive and selective fiber-optic NitricOxide biosensor based on a dye-labeled heme domain of soluble guanylate cyclase.Anal.Chem.1999,71,2071-2075.)
[18] Julia Cordek, Xinwen Wang and Weihong Tan; Fibre-optical probe directly fixedly the glutamate dehydrogenasa be used for hypersensitive and measure glutamate.Analytical chemistry, 1999,71,1529-1533. (Julia Cordek, XinwenWang and Weihong Tan; Direct immobilization of Glutamate dehydrogenase on opticalfiber probes for ultrasensitive glutamate detection.Anal.Chem.1999,71,1529-1533.)
[19] Zhongping Chen, D.L.Kaplan and H.Gao; Molecule integrated multi-layer enzyme: be used to develop a kind of based on chemiluminescent optical fiber biosensor.Materials Science and Engineering, C4 (1996) 155-159. (Zhongping Chen, D.L.Kaplan and H.Gao; Molecular assembly of multiplayer enzyme:toward thedevelopment of a chemiluminescence-based fiber optic biosensor.Material Scienceand Engineering.C4 (1996) 155-159.)
[20] Frank Kleinjung, Krank F.Bier and Axel Warsinke; The optical fibre bio gas sensor is used for selectivity and measures nanomole DNA oligomer.The analytical chemistry journal, 350 (1997) 51-58. (Frank Kleinjung, Krank F.Bier and Axel Warsinke; Fibre-optic genosensor for specific determinationof femtomolar DNA oligomer.Analytical Chimica Acta 350 (1997) 51-58.)
[21] M.A.Karymov, A.A.Kruchinin and Yu.A.Tarantov; The direct fixed dna in fibre-optic waveguide surface is used to measure the biology sensor of molecule.Sensor and actuator B 29 (1995) 324-327. (M.A.Karymov, A.A.Kruchinin and Yu.A.Tarantov; Fixation of DNA directly on optical waveguidesurfaces for molecular probe biosensor development.Sensors and actuators B 29 (1995) 324-327.)
More than the sensitive membrane of three kinds of methods preparation be applicable to the monitoring of various chemical constitutions in liquid or the gas respectively.
The principle of work of fibre optic chemical sensor probe among the present invention:, be applicable to the process monitoring on the throne of mensuration " ultraviolet/visible/near infrared absorption, reflection and phosphorescence/fluorescence/raman radiation spectrum " respectively according to different analytic targets.
One, the measurement of ultraviolet/visible/near infrared absorption or reflectance spectrum radiation: optical signals optical fiber feeds back to detecting device or spectrophotometer, and output signal meets following mathematic(al) representation:
a?or?K:E
1%、E
1‰?or?ε/λ
max、1cm。C:g/100ml、g/L?or?mol/L
Two, the actinometry of fluorescent/phosphorescent and Raman spectrum: spectral signal feeds back to ultraviolet/visible/near infrared spectrometer or fluorospectrophotometer or detecting device by optical fiber, and signal output meets following mathematic(al) representation:
I(F、P?or?R)=Φ
0×I
0×ε×L×C
I (F): fluorescence intensity, I (P): phosphorescence intensity, I (R): Raman light intensity, Φ: constant term, I
0: incident intensity, ε: absorptivity, L: light path, C: analyte concentration.
Spectral signal feeds back to detecting device or fluorescent/phosphorescent/Raman spectrometer by optical fiber, obtains fluorescence, phosphorescence or Raman spectrum.
Three, the measurement of the polynary quencher spectral radiance of fluorescence: in optical fiber end device sensitive membrane, accurate adjustable distance is arranged, form microcuvette.Analyte absorbs radiation (the first inner filtration IFE from light source in this space
1), weaken the exciting light that arrives sensitive membrane (reagent phase), the film emitted fluorescence is weakened.When the fluorescence spectrum of the ultraviolet-visible UV-VIS spectrum of analyte and sensitive membrane overlaps, also will absorb fluorescence (second inner filtration, the IFE of sensitive membrane
2), resonance energy takes place simultaneously shift (FTE) or dynamics quencher.Mathematic(al) representation:
First is the dynamics quencher; Second is static quenching; The 3rd is the resonance energy transfer; The 4th is first inner filtration; The 5th is second inner filtration; F
0Be respectively quencher Q with F and add fluorescence intensity preceding and that the adding back is measured, [Q] is broad sense quencher concentration, τ
0Be fluorescence decay life-span, K
QBe dynamics quenching constant, K
SThe static quenching constant, E
ExQuencher is to the molar absorptivity of exciting light, E
ExQuencher is to radiative molar absorptivity, and b is the effective light path of light by the agent of slightly going out.Following formula calculates through multivariate linear model, obtains two and simplifies mathematic(al) representation:
F
0/ F=Kpq[Q]+C or log (F
0/ F)=Kpq[Q]+C
F
0Fluorescence intensity when/F exists for blank and analyte, Kpq claims apparent quenching constant, characterizes the sensitivity of response.C is an intercept.Because multifactorial compatible and addition has improved mensuration sensitivity.Chemical reaction does not take place based on the physical optics principle in response in the mensuration process, therefore have good reversibility and long-life.
The present invention is when using, preferably by the detected data of computer acquisition detecting device, by to the spectral characteristic in object 200~1100nm to be measured interval in real time, original position, on-line analysis, by multiple mathematical model, set up the corresponding relation of drug concentration and spectrum.Realization just can be finished drug dissolution/release test overall process easily to the on-line automaticization operation of analytic target real-time data acquisition and processing, spectrum demonstration, stripping curve Real time dynamic display, data storage, data management, parameter extraction and reporting printing.
Application example of the present invention is as follows:
Application examples 1: the drug dissolution test of fast release formulation Clozapine sheet.Adopt Self-control method with dissolution rate test solvent preparation Clozapine series standard solution, instrument of the present invention shows acquisition serial absorption spectrum (accompanying drawing 9).With 285nm is to measure wavelength.Select No. 5 probes of №, 900ml is a solvent with hydrochloric acid solution (9 → 1000), changes blue laws, 50 rev/mins, measure 30 minutes stripping curves (accompanying drawing 10), and do control experiment by official method simultaneously.Obtain data consistent.But rapid delivery of pharmaceuticals drug concentration variation in the very short time of beginning is very fast as can be seen from real-time monitoring curve, and sample analysis can't obtain preceding 2 minutes more information.
Application examples 2: disintegrating tablet piroxicam tablet drug dissolution test.The piroxicam reference substance is with dissolution rate test solvent configuration series standard solution, and instrument of the present invention shows acquisition serial absorption spectrum (accompanying drawing 11).Piroxicam has absorption maximum at the 332nm place, as measuring wavelength.Select No. 2 probes of №, 900ml is a solvent with hydrochloric acid solution (9 → 1000), changes blue laws, 50 rev/mins, measure 40 minutes stripping curves (accompanying drawing 12), and do control experiment by official method simultaneously.Obtain data consistent.But this law is compared with official method, not only reduces operate miss and the more information amount is provided.
Applicating example 3: not disintegrating tablet vitamin B2 sheet drug dissolution test.The vitamin B2 reference substance is with dissolution rate test solvent configuration series standard solution, and instrument of the present invention shows the serial absorption spectrum of acquisition, at the 264nm place absorption maximum is arranged, and good linear is arranged, as measuring wavelength (accompanying drawing 13).Select No. 2 probes of №, the oar method is measured 25 minutes.Obtain the real-time stripping curve of vitamin B2 (accompanying drawing 14), do control experiment by official method simultaneously.This law and official method obtain data consistent.