CN107941741A - The method that methylenum careuleum burst size in Plasma Inactivation bag is detected using near infrared spectrum - Google Patents
The method that methylenum careuleum burst size in Plasma Inactivation bag is detected using near infrared spectrum Download PDFInfo
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- CN107941741A CN107941741A CN201710993811.XA CN201710993811A CN107941741A CN 107941741 A CN107941741 A CN 107941741A CN 201710993811 A CN201710993811 A CN 201710993811A CN 107941741 A CN107941741 A CN 107941741A
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- 230000002779 inactivation Effects 0.000 title claims abstract description 74
- 238000002329 infrared spectrum Methods 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title claims abstract description 37
- 238000001514 detection method Methods 0.000 claims abstract description 53
- 239000000758 substrate Substances 0.000 claims abstract description 20
- 230000005540 biological transmission Effects 0.000 claims abstract description 19
- 238000004458 analytical method Methods 0.000 claims abstract description 11
- 239000012086 standard solution Substances 0.000 claims abstract description 8
- 230000003595 spectral effect Effects 0.000 claims description 16
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 15
- 239000010453 quartz Substances 0.000 claims description 14
- 238000002835 absorbance Methods 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 6
- 238000012360 testing method Methods 0.000 claims description 6
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- 239000005304 optical glass Substances 0.000 claims description 2
- 238000013102 re-test Methods 0.000 claims 1
- 238000000491 multivariate analysis Methods 0.000 abstract description 8
- 210000002381 plasma Anatomy 0.000 description 82
- 239000000523 sample Substances 0.000 description 33
- 238000001228 spectrum Methods 0.000 description 29
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- 238000009659 non-destructive testing Methods 0.000 description 4
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- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
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- 238000012795 verification Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 241001062009 Indigofera Species 0.000 description 1
- 238000004497 NIR spectroscopy Methods 0.000 description 1
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- 239000012153 distilled water Substances 0.000 description 1
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- 235000013399 edible fruits Nutrition 0.000 description 1
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N21/359—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light using near infrared light
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- Spectroscopy & Molecular Physics (AREA)
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- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
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- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
The present invention relates to a kind of near-infrared spectrometer, including a transmission platform, transmission platform includes a substrate, two pieces of risers being parallel to each other are equipped with vertically on substrate, respectively offer corresponding transmissive window on two pieces of risers, transmissive window is covered with transparent optics window, an objective table is equipped between two pieces of risers, determinand is between two optics windows, and at least one riser can be slided along the surface of substrate so that determinand is clamped between two optics windows.The invention further relates to a kind of method that methylenum careuleum burst size in Plasma Inactivation bag is detected using near infrared spectrum, including：Light path between two optics windows is fixed as 2.0 5.0mm；Plasma Inactivation bag is placed on objective table, near infrared spectrum detection is carried out to multiple plasma standard solution, the multivariate analysis model of methylenum careuleum burst size is established using PLS methods；Near infrared spectrum detection is carried out to unknown sample, utilizes the burst size of analysis model analysis methylenum careuleum.
Description
Technical field
The present invention relates to methylene in field of biological detection, more particularly to a kind of detection Plasma Inactivation bag using near infrared spectrum
The method of blue burst size.
Background technology
Transfusion infection virus in order to prevent, blood used in clinic starch widely used methylenum careuleum luminescence method and go out to plasma viral
Living, this is that China is uniquely approved single part blood constituent Viral inactivation techniques in Clinical practice.Inactivation process is in Plasma Inactivation
Bag in carry out, and in Plasma Inactivation bag inactivator methylenum careuleum release, be that aseptically blood plasma flows through methylenum careuleum memory
When by gradually dissolve and with blood plasma together enter inactivation bag, which is the release process of methylenum careuleum, its burst size is to determine
Determine the key parameters of inactivation of virus degree.However, the burst size is related with many factors, including flow velocity, environment temperature, methylenum careuleum
Moisture absorption degree and memory in the material of immobilized methylenum careuleum, production technology etc..If methylenum careuleum releases in Plasma Inactivation bag
High-volume excessive, then methylene blue in blood plasma residual quantity is excessive, is easy to cause cumulative toxicity；If burst size is too low, and cannot be guaranteed it
Inactivation of virus effect.Mass data shows that virus inactivator methylenum careuleum must assure that in a very narrow burst size scope, China
It is defined as 0.9-1.3 μm of ol/L.In order to as far as possible easily and fast, delicately detect burst size, occurred largely being based on
The method for sampling detection, including solid phase extractions-spectrophotometry, enhanced sensitivity fluorescent spectrometry, HPLC, chemoluminescence method, resonance are auspicious
Sharp scattering spectrometry.Sampling detection, which needs to extract sample from Plasma Inactivation bag using draw-out device, to be detected, due to extraction
Operation will cause to damage to Plasma Inactivation bag, and this detection method that damages easily causes blood plasma secondary pollution in sampling, including
Bacterium, chemical pollutant and particulate matter, cause the blood plasma in Plasma Inactivation bag to be no longer available for human body, cause blood plasma
Waste.Also this is based on, the blood plasma detected through burst size cannot also be used further to clinic.Therefore, it is applied to for each bag
Clinical blood plasma, it is impossible to carry out such based on the inactivation quality control for damaging detection.In fact, blood in Plasma Inactivation bag
The reliable in quality of slurry or not it is only a statistical result.Non-Destructive Testing to methylenum careuleum burst size in inactivation bag, is to realize
The key issue of each bag of Plasma Inactivation quality control.
At present, the lossless quick detection of sample is usually carried out using near infrared spectroscopy, with the matter suitable for production process
Amount control.Wherein, the Non-Destructive Testing that diffusing reflection near infrared spectrum carries out sample is should be most commonly used, but this method has
Certain limitation：This method applies in general to solid, powder, body of paste first, is then difficult to be applicable in for fluid sample, due to
Only generation is extremely weak after illumination injection liquid diffuses, and most of or even all transmitted lights, therefore, it is difficult to unrestrained according to it
Reflect the quantitative detection for certain material that near infrared spectrum is carried out in liquid.
The near infrared spectrum detection method of fast quantification is carried out for the existing material in liquid, it usually needs from tested
Sampling is detected in container, mainly including two ways.The first, the fluid sample of acquirement is placed in cuvette, then
The cell rack for being put into instrument is detected；Second, by the liquid of the tested container of liquid fiber probe insertion with slit
Among, tested liquid is flowed directly among the slit as the fibre-optical probe of cuvette, and to realize sampling operation, and optical fiber will be believed
Number carry out spectrometer is transmitted again, to realize quick detection.Any mode among either two kinds, is to damage detection side
The defects of method, this method, has hereinbefore been described in detail.And plasma bags are directly detected using existing infrared spectrometer
Very big problem can be run into, first, more bubble is contained in Plasma Inactivation bag, if these bubbles are in detection zone and can to survey
Measure result and produce very big error；Secondly, when Plasma Inactivation bag directly detects, since bag body is softer, its is in irregular shape, spectrum
The light path of detection is difficult to fix, and the content for calculating determinand is difficult to using quantitative equation.Certainly, the blood plasma inactivated in bag contains greatly
The large biological molecule of amount, C-H bond therein can produce substantial amounts of near infrared light spectrum signal, also by interference detection results, to detection
Method brings certain difficulty.
The content of the invention
In order to solve the above technical problems, the object of the present invention is to provide one kind to utilize near infrared spectrum detection Plasma Inactivation bag
The method of middle methylenum careuleum burst size, the method overcome the prior art and is difficult to accurately carry out near-infrared to sample in irregular shape
The defects of quantitative, realize lossless, quick, the accurate analysis of the methylenum careuleum burst size in Plasma Inactivation bag.
On the one hand, the present invention provides a kind of near-infrared spectrometer, including light source, monochromator, detector and meter
Calculation machine processing information system, further includes a transmission platform, and transmission platform includes a substrate, and being equipped with two pieces vertically on substrate mutually puts down
Capable starting stave and the second riser, corresponding transmissive window, transmissive window covering are respectively offered on starting stave and the second riser
There is transparent optics window, and optics window is located on the opposite side of two pieces of risers, is set between starting stave and the second riser
There is an objective table, the soft bag of determinand is filled for carrying, and the objective table is located at the lower section of optical window piece lower edge, fills to be measured
Between two optics windows, at least one riser can move horizontally the soft bag of thing along the surface of the substrate so that fill
The light path that the soft bag of determinand is clamped between two optics windows and between two optics windows can adjust.
Further, objective table is located at least 2cm below optical window piece lower edge.
Further, starting stave is fixedly connected with substrate, and objective table is fixedly connected with starting stave, is opened on the second riser
Bar hole equipped with face objective table, bar hole are adapted with the shape of objective table, and objective table can be slidably inserted into bar hole.
Further, guide rail is fixedly connected with substrate, the second riser can slide along guide rail level and can be locked in guide rail
Any position, so as to fix distance (i.e. light path) between two optics windows.
Further, a positioning table is fixedly connected with one side of second riser away from starting stave, the second riser passes through
Positioning table is slidably connected with guide rail, and positioning table is threaded with a screw rod, and the axial direction of screw rod and the second riser are perpendicular, on guide rail
A fixed baffle is fixedly connected with, fixed baffle opens up a through hole, and screw rod is arranged in through hole.
Further, the material of optics window is quartz or optical glass.
Further, transmissive window is rectangular.
Further, the area of optics window is more than the area of transmissive window.
On the other hand, discharged present invention also offers one kind using methylenum careuleum near infrared spectrum detection Plasma Inactivation bag
The method of amount, using above-mentioned near-infrared spectrometer, comprises the following steps：
(1) the distance between starting stave and the second riser are adjusted, is fixed as the light path between two optics windows
2.0-5.0mm；Preferably, light path 3.5mm；
(2) by Plasma Inactivation bag be placed on the objective table of near-infrared spectrometer and be located at two optics windows it
Between, and slightly firmly extruded in the top of bag, Plasma Inactivation bag is closely contacted with two optics windows, and Plasma Inactivation bag
The plane for being extruded rear formation rule and its surface is pressed into plane, known methylenum careuleum content is wherein filled in Plasma Inactivation bag
Plasma standard solution；
(3) in wave number 4000-10000cm-1Under conditions of to plasma standard solution carry out near infrared spectrum detection, repeat survey
Multiple samples are tried, according to the near infrared spectrum testing result of multiple samples, the multivariable of methylenum careuleum burst size is established using PLS methods
Analysis model；
(4) the Plasma Inactivation bag to be measured containing methylenum careuleum is placed on the objective table, and slightly exerted oneself in the top of bag
Extruding, makes Plasma Inactivation bag closely be contacted with two optics windows, and Plasma Inactivation bag is extruded the plane of rear formation rule；
(5) in wave number 4000-10000cm-1Under conditions of carry out near infrared spectrum detection, utilize the methylenum careuleum burst size
Multivariate analysis model determine the burst size of methylenum careuleum in Plasma Inactivation bag.
Further, in step (2), the concentration of methylenum careuleum is 0.312 μm of ol/L-2.959 μ in plasma standard solution
mol/L。
Further, in step (3), the number of plasma standard solution is 52.
Further, in step (3) and step (5), the computerized near infrared scan resolution ratio of near infrared spectrum detection is
8cm-1, scanning times are 32 times, sweep speed 0.3165.
Further, in step (3) and step (5), the scanning aperture of near infrared spectrum detection is 22, gain 1.
Further, in step (3) and step (5), after near infrared spectrum detection, 3999.7-4265.8cm is selected-1Area
Between absorbance average as drift value, drift elimination is carried out to the near infrared spectrum of detection.
Further, in step (3) and step (5), after near infrared spectrum detection, 5350-6600cm is selected-1With
7200-10001cm-1The absorbance in two sections is as rich informative spectral.Preferably, after near infrared detection, 5827.9- is selected
6464.3cm-1And 7467.1-9009.9cm-1The absorbance in two sections is as rich informative spectral.
Further, in step (3), it is 11 to select number of principal components, by height distribution 11 samples of random selection of concentration
This is forecast set, and using remaining 41 samples as calibration set, the multi-variables analysis mould of methylenum careuleum burst size is established using PLS methods
Type.
Further, the formula of the multivariate analysis model of methylenum careuleum burst size is：C=k1A1+k2A2+···+knAn,
Wherein, in c representative samples methylenum careuleum concentration, A1、A2···AnThe richness selected in detection spectrum respectively from the sample
The corresponding wavelength in informative spectral section is λ1、λ2···λnThe absorbance at place, k1、k2···knReturned for corresponding PLS and be
Number.
Further, before step (2), institute will be placed on filled with air and original Plasma Inactivation bag by further including
State the step of being used as spectral background after progress near infrared spectrum detection on the objective table of near-infrared spectrometer.
According to the above aspect of the present invention, the present invention has at least the following advantages：
The structure setting of the transmission platform of the near-infrared spectrometer of the present invention can realize the lossless inspection of Plasma Inactivation bag
Light path is fixed during survey, and can effectively solve the problem that the problem of bubble in Plasma Inactivation bag is present among light path, is made quantitative
As a result it is accurate, reliable.It is rule by the Plasma Inactivation bag extruding between two pieces of optics windows due to the squeezing action of optics window
Plane, simultaneously as the density of bubble is relatively low, under optics window extruding force, the gas in Plasma Inactivation bag therebetween
Bubble has been rushed in the Plasma Inactivation bag beyond optics window crimping section naturally, so that bubble does not appear in detection zone
Domain.
The inventive method achieves under non-sampling mode, the Asia in Plasma Inactivation bag can be achieved outside Plasma Inactivation bag
Lossless, quick, the accurate analysis of first indigo plant burst size.Model calculates concentration
98.16% (calibration set) and 98.93% (forecast set), the average recovery rate of senior middle school's low concentration group sample is 103.6%-
110.8%.
Described above is only the general introduction of technical solution of the present invention, in order to better understand the technological means of the present invention,
And can be practiced according to the content of specification, below with presently preferred embodiments of the present invention and coordinate attached drawing describe in detail as after.
Brief description of the drawings
Fig. 1 is the dimensional structure diagram of present invention transmission platform；
Fig. 2 is the leading flank structure diagram of present invention transmission platform；
Fig. 3 is the left surface structure diagram of present invention transmission platform；
Fig. 4 is the enlarged structure schematic diagram of Fig. 3 centre circles A；
Fig. 5 is the overlooking the structure diagram of present invention transmission platform；
Fig. 6 is the atlas of near infrared spectra after baseline drift elimination in embodiment 2；
Fig. 7 is the residual error of the PLS models of the cross-verification based on leave-one-out；
Fig. 8 is the multivariate analysis model figure of methylenum careuleum burst size；
Description of reference numerals：
The second risers of 1-；2- quartz plates；3- Plasma Inactivation bags；4- transmissive windows；5- substrates；6- starting staves；7- objective tables；
8- positioning tables；9- screw rods；10- guide rails；11- fixed baffles.
Embodiment
With reference to the accompanying drawings and examples, the embodiment of the present invention is described in further detail.Implement below
Example is used to illustrate the present invention, but is not limited to the scope of the present invention.
Embodiment 1
Referring to Fig. 1-5, a kind of near-infrared spectrometer of the invention, including light source, monochromator, detector and computer
Processing information system, further includes a transmission platform, and transmission platform includes a substrate 5, and two pieces are connected with substrate 5 and is mutually put down
Capable 6 and second riser 1 of starting stave, 6 and second riser 1 of starting stave are set perpendicular to substrate 5.Respectively opened up on two pieces of risers
There is corresponding rectangular transmission window 4,4 upper cover of transmissive window is equipped with square suprasil piece 2, and quartz plate 2 is located at two pieces of riser phases
To side on, the area of quartz plate 2 is more than the area of transmissive window 4.Starting stave 6 is fixedly connected with substrate 5, starting stave 6
On be also fixedly connected with objective table 7, objective table 7 is located at least 2cm below 2 lower edge of quartz plate, and objective table 7 is filled for placement and treated
Survey the soft bag of thing.The upper surface of objective table 7 is parallel with substrate 5, and the bar hole of face objective table 7 is offered on the second riser 1,
Bar hole is adapted with the shape of objective table 7, so that objective table 7 is slidably inserted into bar hole.It is fixedly connected on substrate 5
There are two guide rails 10, guide rail 10 is set perpendicular to the second riser 1, and the second riser 1 can be slided along guide rail 10 is horizontal.Second riser 1
A positioning table 8 is fixedly connected with one side away from starting stave 6, positioning table 8 is threaded with a screw rod 9, the axial direction of screw rod 9
Perpendicular with the second riser 1, positioning table 8 is slidably connected with guide rail 10, and a fixed baffle 11 is fixedly connected with guide rail 10, fixed
Baffle 11 opens up a through hole, and screw rod 9 is arranged in through hole.
When carrying out sample detection, such as the sample detection in Plasma Inactivation bag 3, first adjusting screw rod make it drive second to erect
Plate 1 is moved along guide rail 10, it is slid into the distance of needs, glide direction is vertical with the plane where riser, then adjusts spiral shell
Bar 9 makes the second riser 1 fix the light path in this position, that is, securing transmission detection.And second riser 1 in sliding process, carry
Thing platform 7 is slidably inserted into the bar hole set thereon, to facilitate adjusting light path and ensure under different light paths, quartz plate
2 can clamp Plasma Inactivation bag 3.Plasma Inactivation bag 3 is placed on objective table 7, and is slightly firmly extruded in the top of bag,
Plasma Inactivation bag is set closely to be contacted with two optics windows so that Plasma Inactivation bag 3 is clamped between two quartz plates 2.
When quartz plate 2 extrudes Plasma Inactivation bag 3 so that the shape of 3 formation rule of Plasma Inactivation bag between two blocks of quartz plates 2, thus
Exact light path (distance indicated such as the graticule " 5 " in Fig. 4 is to detect light path) is secured, and in Plasma Inactivation bag 3
Bubble is migrated to plasma bags part more than edge on quartz plate 2 so that is not had bubble between quartz plate 2, is prevented it from being tied to detection
Fruit impacts.
Embodiment 2
In following embodiments, the material used includes：The non-inactivation human plasma that Suzhou City Red Cross center blood station provides, makes
With the filter filtrating leukocytes in disposable viral inactivating corollary apparatus, the blood plasma (i.e. blank plasma) without methylenum careuleum is obtained.
Disposable blood device for taking and transfusing (transfering bag, specification comprising Plasma Inactivation bag：35ml, product batch number：170428, Si Chuannan
Ge Er bio tech ltd).Methylenum careuleum hydrochloride (analyzes pure, the content based on dry product>97%, Sigma-Aldrich
Company), experimental water is tri-distilled water.Near infrared spectrometer (model：NEXUS, Thermo companies of the U.S.), its transmit platform according to
Embodiment 1 is transformed.
The present embodiment is the method for building up of the multivariate analysis model of methylenum careuleum burst size, specific as follows：
(1) methylenum careuleum storing solution is configured first：Weigh methylenum careuleum 1.3953g (analysis is pure, loss on drying rate for 10.6%),
It is placed in the volumetric flask of 500mL, with PBS solution constant volume.Precision measures 1.00ml methylene blue solutions, then is settled to PBS solution
1000ml, up to 7.8 μm of ol/L methylenum careuleum storing solutions.Lucifuge, be sealed.
(2) it is reconfigured at sample to be tested：First with methylenum careuleum storing solution and PBS buffer (pH=7.4), use is progressively dilute
Firsts and seconds dilution is prepared in interpretation of the law.Afterwards, with storing solution, dilution and blood plasma, accurate compound concentration is 0.312-2.959
The methylenum careuleum plasma solutions totally 52 of μm ol/L.Finally, adopted in defeated transfering bag and filled to each blood with primary sterilization syringe
Methylenum careuleum plasma solutions, up to sample to be tested.
(3) the near infrared spectrum detection of sample
Outer surface with alcohol wipe test plasma inactivation bag and the quartz plate on transmission platform, as described in Example 1
The distance between 6 and second riser 1 of starting stave is adjusted, the light path between two optics windows is fixed as 3.5mm.Afterwards,
Plasma Inactivation bag is placed on transmission platform according to the method for embodiment 1, the position that Plasma Inactivation bag is put every time is tried one's best or so
Both sides are alignd, and measure sample spectrum.Instrument parameter is：Wave number 4000-10000cm-1, light path 3.5mm, resolution ratio 8cm-1, scanning
Number 32, sweep speed 0.3165, aperture 22, gain 1.Before detecting sample on the day of, with filled with air, untapped blood plasma
Bag is inactivated as spectral background.
(4) by the NIR transmittance spectroscopy of sample in the plasma bags for detecting acquisition with 3999.7-4265.8cm-1The light in section
Compose and carry out baseline drift calibration for standard.Select 5827.9-6464.3cm-1And 7467.1-9009.9cm-1For rich information section,
Using the spectrum of average centralization as model variable, it is 11 to select number of principal components, by height distribution 11 samples of random selection of concentration
This is forecast set, and using remaining 41 samples as calibration set, the multi-variables analysis mould of methylenum careuleum burst size is established in PLS methods
Type.
For the lower wave number region of near infrared spectrum, one section of spectrum baseline will usually occur.It should be horizontal line, no
With concentration sample the section spectrum should it is completely overlapped, without statistical discrepancy, and trap average is 0.In Fig. 6,
3999.7-4265.8cm-1Section is the spectrum baseline of each sample.However, in original spectrum, the baseline hair of various concentrations sample
Drift is given birth to, drift value is about 7, and this drift value unrelated with concentration will have an impact the accuracy of methylenum careuleum burst size.
With the detection spectrum of each sample in 3999.7-4265.8cm-1The absorbance average in section drifts about it as drift value
Eliminate, up to the spectrum after the elimination drift of all samples.
From the spectrum (Fig. 6) after baseline drift elimination it can be found that the plasma sample of methylenum careuleum goes out in whole spectrum range
5350-6600cm is showed-1And 7200-10001cm-1The obvious absorption peaks in two sections, are absworption peak spectral regions.For the former,
It is made of two acromions of height, in 5350-5828cm-1Lower wave number region, not only trap is relatively low for spectrum, essentially without peak
Horizontal line, and there is awave burr, the spectral signal of this kind of low signal-to-noise ratio will influence the precision of Quantitative Analysis Model.
And high wavenumber range 6464-6600cm-1Spectrum, for the end spectrum of the absworption peak, trap is relatively low, and signal is weaker, equally
Also low signal-to-noise ratio is shown as.Therefore, the 5827.9-6464.3cm of the absworption peak spectral regions is selected-1Spectrum be rich signal spectrum.
Similarly, in 7200-10001cm-1Absworption peak spectral regions, select 7467.1-9009.9cm-1Spectrum be rich signal spectrum.
With 5827.9-6464.3cm-1And 7467.1-9009.9cm-1Spectrum be rich informative spectral, with average centralization
Spectrum be model variable, with calibration set sample press leave-one-out cross-verification, PLS when calculating different number of principal components
The residual error (Fig. 7) of model prediction methylenum careuleum concentration.We are it can be found that with the increase of number of principal components, the concentration residual error of prediction
With the trend being gradually reduced, show that the accuracy of model gradually increases.After number of principal components >=11, what prediction residual reduced
Degree substantially diminishes, and it is relatively low to the contribution rate of model accuracy to illustrate that number of principal components continues increase.Certainly, number of principal components is excessive
It can cause the over-fitting of model, therefore, it is number of principal components to select 11.
Identification variable using the rich informative spectral in two sections of selection as methylenum careuleum burst size, based on foregoing definite master
Component number, through spectrum average centralization, the multivariate analysis model of burst size is established using PLS methods, wherein dense by methylenum careuleum
The height distribution of degree, 11 samples of random selection are forecast set, remaining 41 are forecast set, as a result referring to Fig. 8, in figure ▲
Calibration set sample is represented, ● represent forecast set sample.It can be found that in multivariate analysis model, the correction of calibration samples is dense
Degree can be corresponded preferably with actual concentration, and in whole detectable concentration section, it is (diagonal that they are distributed in actual concentration
Line) near, its coefficient R corr=98.16%.Using the model of foundation, to the methylenum careuleum in independent forecast set sample
Burst size is predicted (i.e. pattern detection), also it is not difficult to find that the prediction concentrations of these samples still show good pair
It should be related to, they can be also distributed near actual concentration, its coefficient R pred is up to 98.93%.It is indicated above that this
The model that invention is established has good accuracy.
In order to illustrate the accuracy of model, the sample of design senior middle school low concentration group, carries out recovery test, the results are shown in Table 1.
As seen from Table 1, no matter concentration level, each concentration group has the good rate of recovery, and the rate of recovery does not have in 103.6%-110.8%
There is concentration level correlation, there is randomness, illustrate the lossless detection method established accurately, reliably.Certainly, lowest rate of return is low
To 96%, and highest is then up to 119%, this for lossless, quick determination method, particularly biological specimen a detection,
It is entirely an acceptable accuracy.
The accuracy of the non-damaging detection methylene blue in blood plasma burst size of 1 near infrared spectrum of table
Result above shows that the present invention utilizes the good penetration of near infrared light, is established using NIR transmittance spectroscopy
Blood adopts the Nondestructive Testing Model and method of blood-plasma virus killing agent methylenum careuleum burst size in defeated transfering bag.Will outside plasma bags into
The NIR transmittance spectroscopy of sample is as identification variable in the plasma bags that row Non-Destructive Testing obtains, with 3999.7-4265.8cm-1Area
Between spectrum for standard carry out baseline drift calibration, select 5827.9-6464.3cm-1And 7467.1-9009.9cm-1Section
Spectrum is rich informative spectral, and using the spectrum of average centralization as model variable, it is 11 to select number of principal components, randomly chooses 11 samples
This is forecast set, and using remaining 41 samples as calibration set, the multi-variables analysis mould of methylenum careuleum burst size is established in PLS methods
Type.The formula of the model is as follows：
C=k1A1+k2A2+···+knAn, wherein, the concentration of methylenum careuleum, A in c representative samples1、A2···AnRespectively
It is λ for the corresponding wavelength in rich informative spectral section selected in the detection spectrum from the sample1、λ2···λnThe extinction at place
Degree, k1、k2···knFor corresponding PLS regression coefficients.
It is respectively 98.16% (calibration set) and 98.93% (prediction to the related coefficient of each concentration of specimens that model, which calculates concentration,
Collection), the average recovery rate of senior middle school's low concentration group sample is in 103.6%-110.8%.Thus illustrate, the near-infrared that the present invention establishes
Transmission spectrum method can realize that lossless, quick, accurate analyze adopts the burst size of methylenum careuleum in defeated transfering bag outside plasma bags.Indication
The monitoring to adopting blood-plasma virus killing quality in defeated transfering bag can be realized under conditions of non-secondary pollution by.
Embodiment 3
The multivariate analysis model for present embodiments providing a kind of methylenum careuleum burst size using above-mentioned foundation carries out blood plasma
The detection method of middle methylenum careuleum burst size, it is specific as follows：
Outer surface with alcohol wipe test plasma inactivation bag and the quartz plate on transmission platform, according to the side of embodiment 1
The distance between method adjustment 6 and second riser 1 of starting stave, makes the light path between two optics windows be fixed as 3.5mm.Then
The Plasma Inactivation bag (the Plasma Inactivation bag for releasing methylenum careuleum) of unknown methylenum careuleum content is placed on objective table, Plasma Inactivation
The position put every time of bag is tried one's best the right and left alignment, measures sample spectrum.Instrument parameter is：Wave number 4000-10000cm-1, light path
3.5mm, resolution ratio 8cm-1, scanning times 32, sweep speed 0.3165, aperture 22, gain 1.Before detecting sample on the day of, with filling
There are air, untapped Plasma Inactivation bag as spectral background.Then by measured NIR transmittance spectroscopy with 3999.7-
4265.8cm-1The spectrum in section carries out baseline drift calibration for standard.Select 5827.9-6464.3cm-1And 7467.1-
9009.9cm-1For rich information section, the formula for the model that the absorbance substitution embodiment 2 at each wavelength in the section is established
In, calculate the methylenum careuleum content in Plasma Inactivation bag.
The above is only the preferred embodiment of the present invention, is not intended to limit the invention, it is noted that for this skill
For the those of ordinary skill in art field, without departing from the technical principles of the invention, can also make it is some improvement and
Modification, these improvements and modifications also should be regarded as protection scope of the present invention.
Claims (10)
1. a kind of near-infrared spectrometer, including light source, monochromator, detector and computer treatmenting information system, its feature
It is：The infrared spectrometer further includes a transmission platform, and the transmission platform includes a substrate, is equipped with vertically on the substrate
Corresponding transmission is respectively offered on two pieces of starting staves being parallel to each other and the second riser, the starting stave and the second riser
Window, the transmissive window is covered with transparent optics window, and the optics window is located on the opposite side of two pieces of risers, described
An objective table is equipped between starting stave and the second riser, the soft bag of determinand is filled for carrying, the objective table is located at light
The lower section of window lower edge is learned, for the soft bag for filling determinand between two optics windows, at least one riser can be along institute
The surface for stating substrate moves horizontally, and the soft bag for filling determinand is clamped between two optics windows and two optical windows
Light path between piece can adjust.
2. near-infrared spectrometer according to claim 1, it is characterised in that：The starting stave is consolidated with the substrate
Fixed connection, the objective table are fixedly connected with the starting stave, and objective table described in face is offered on second riser
Bar hole, the bar hole are adapted with the shape of the objective table, and the objective table can be slidably inserted into the bar hole.
3. near-infrared spectrometer according to claim 1, it is characterised in that：It is fixedly connected with and leads on the substrate
Rail, second riser can slide along guide rail level and can be locked in any position of guide rail.
4. near-infrared spectrometer according to claim 1, it is characterised in that：The material of the optics window is quartz
Or optical glass.
A kind of 5. method that methylenum careuleum burst size in Plasma Inactivation bag is detected using near infrared spectrum, it is characterised in that using power
Profit requires the near-infrared spectrometer any one of 1-4, comprises the following steps：
(1) the distance between described starting stave and the second riser are adjusted, is fixed as the light path between two optics windows
2.0-5.0mm；
(2) by Plasma Inactivation bag be placed on the objective table of the near-infrared spectrometer and be located at two optics windows it
Between, Plasma Inactivation bag is closely contacted with two optics windows, contain wherein filling known methylenum careuleum in the Plasma Inactivation bag
The plasma standard solution of amount；
(3) in wave number 4000-10000cm-1Under conditions of near infrared spectrum detection is carried out to plasma standard solution, retest is more
A sample, according to the near infrared spectrum testing result of multiple samples, the multi-variables analysis of methylenum careuleum burst size is established using PLS methods
Model；
(4) the Plasma Inactivation bag to be measured containing methylenum careuleum is placed on the objective table, make Plasma Inactivation bag closely with two
Optics window contacts；
(5) in wave number 4000-10000cm-1Under conditions of carry out near infrared spectrum detection, utilize the more of the methylenum careuleum burst size
Variable analysis model determines the burst size of methylenum careuleum in Plasma Inactivation bag.
6. according to claim 5 using the method for methylenum careuleum burst size near infrared spectrum detection Plasma Inactivation bag, it is special
Sign is：In step (2), the concentration of methylenum careuleum is 0.312 μm of ol/L-2.959 μm of ol/L in the plasma standard solution.
7. according to claim 5 using the method for methylenum careuleum burst size near infrared spectrum detection Plasma Inactivation bag, it is special
Sign is：In step (3) and step (5), the computerized near infrared scan resolution ratio of the near infrared spectrum detection is 8cm-1, scanning
Number is 32 times, sweep speed 0.3165.
8. according to claim 5 using the method for methylenum careuleum burst size near infrared spectrum detection Plasma Inactivation bag, it is special
Sign is：In step (3), after near infrared spectrum detection, 3999.7-4265.8cm is selected-1The absorbance average conduct in section
Drift value, drift elimination is carried out to infrared spectrum testing result.
9. according to claim 5 using the method for methylenum careuleum burst size near infrared spectrum detection Plasma Inactivation bag, it is special
Sign is：In step (3) and step (5), after near infrared spectrum detection, 5350-6600cm is selected-1And 7200-10001cm-1
The absorbance in two sections is as rich informative spectral.
10. according to claim 5 using the method for methylenum careuleum burst size near infrared spectrum detection Plasma Inactivation bag, it is special
Sign is：Before step (2), the near infrared light will be placed on filled with air and original Plasma Inactivation bag by further including
The step of spectral background being used as on the objective table of spectrometer after progress near infrared spectrum detection.
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