CN102692396A - Endotoxin detection system and quantitative detection method thereof - Google Patents
Endotoxin detection system and quantitative detection method thereof Download PDFInfo
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- CN102692396A CN102692396A CN2012101965968A CN201210196596A CN102692396A CN 102692396 A CN102692396 A CN 102692396A CN 2012101965968 A CN2012101965968 A CN 2012101965968A CN 201210196596 A CN201210196596 A CN 201210196596A CN 102692396 A CN102692396 A CN 102692396A
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- 238000001514 detection method Methods 0.000 title claims abstract description 71
- 239000002158 endotoxin Substances 0.000 title claims abstract description 59
- 239000002245 particle Substances 0.000 claims abstract description 16
- 239000007788 liquid Substances 0.000 claims abstract description 11
- 201000009310 astigmatism Diseases 0.000 claims abstract description 6
- 238000012545 processing Methods 0.000 claims abstract description 4
- 238000000034 method Methods 0.000 claims description 14
- 239000000243 solution Substances 0.000 claims description 14
- 238000007689 inspection Methods 0.000 claims description 10
- 238000002347 injection Methods 0.000 claims description 8
- 239000007924 injection Substances 0.000 claims description 8
- 238000000149 argon plasma sintering Methods 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 claims description 7
- 239000000523 sample Substances 0.000 claims description 4
- 238000001228 spectrum Methods 0.000 claims description 3
- 239000005304 optical glass Substances 0.000 claims description 2
- 238000012544 monitoring process Methods 0.000 abstract description 9
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 238000006243 chemical reaction Methods 0.000 abstract description 2
- 239000003814 drug Substances 0.000 abstract description 2
- 230000001960 triggered effect Effects 0.000 abstract description 2
- 239000003795 chemical substances by application Substances 0.000 abstract 1
- 231100000284 endotoxic Toxicity 0.000 description 6
- 230000002346 endotoxic effect Effects 0.000 description 6
- 235000014347 soups Nutrition 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 240000007164 Salvia officinalis Species 0.000 description 4
- 235000005412 red sage Nutrition 0.000 description 4
- 238000004848 nephelometry Methods 0.000 description 3
- 239000012086 standard solution Substances 0.000 description 3
- 239000008215 water for injection Substances 0.000 description 3
- 238000004364 calculation method Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000000108 ultra-filtration Methods 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000239218 Limulus Species 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 229920006008 lipopolysaccharide Polymers 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000002510 pyrogen Substances 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000011155 quantitative monitoring Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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Abstract
The invention discloses an endotoxin detection system and a quantitative detection method thereof and belongs to the field of endotoxin monitoring. By the system, a light path detection system is integrated on the basis of the conventional laser particle size detector, by double-laser source irradiation, output light enters a detection pool through a light path, is received by a multi-point scattered light receiver, is subjected to photoelectric conversion and signal amplifying processing and enters a quantitative arithmetic unit; and the quantitative arithmetic unit fits the relativity between the astigmatism and the endotoxin concentration according to the scattering intensity difference of the endotoxin particles at three or more different angles, the endotoxin concentration is calculated and displayed, and when the endotoxin concentration exceeds a limit value, an early warning system is triggered, so that the online monitoring is realized. By the system, the quantitative detection of the endotoxin can be realized, the system is high in detection sensitivity, short in detection time and high in reliability, an agent is not consumed, and pollution to the liquid medicine is avoided.
Description
Technical field
The present invention relates to a kind of endotoxin detection technique, relate in particular to a kind of endotoxin detection system and quantitative detecting method thereof that detects based on the laser particle diameter, belong to endotoxin monitoring field.
Background technology
Bacterial endotoxin is a lipopolysaccharides, also is called as liposome, is the composition of Gram-negative bacteria mantle, extensively is present in occurring in nature, and this material gets into blood of human body can cause heating, is commonly called as pyrogen reaction.Because this type of material possibly cause the people and produce serious adverse reaction, therefore in drug injection, all need strict control.
At present, water for injection and injection production run also can't be carried out on-line monitoring to endotoxin, and endotoxic detection adopts limulus reagent test or rabbit method to check after also depending on and producing end.The sensitivity that these methods and process not only detect is low, and in case when finding that endotoxin in the product exceeds standard, entire block will be with waste disposal, this not only can bring economic loss to enterprise, also can bring huge clinical practice risk to enterprise simultaneously.Therefore, in order to ensure water for injection and injection quality safety, prevent that the burst in the production run from polluting, the integrality of ultra filtration membrane realized significant to endotoxic on-line monitoring during monitoring was produced.
Laser particle diameter detector is not applied to endotoxic detection as yet at present; We analyze the higher Ma Erwen Nano ZSZEW3600 type laser particle size analyzer of sensitivity; It can detect particle in the solution of endotoxin concns more than 10EU/ml, and this can't satisfy endotoxic detectable concentration in the injection.Laser particle diameter detector is different at the astigmatic signal of different angles according to particle, the astigmatism of the multiple spot more than 2 o'clock signal is carried out granularmetric analysis, but can't detect particle concentration, therefore can't be used for endotoxic detection by quantitative.
Summary of the invention
The present invention is directed to the defective that prior art exists, and propose a kind of detection sensitivity is high, can realize online detection endotoxin detection system and quantitative detecting method thereof.
This endotoxin detection system comprises lasing light emitter, short focus lens, grating, long focus lens, detection cell, scattered light receiver, photoelectric commutator, signal amplifier, signal processor, data acquisition unit, quantitative arithmetical unit, data display equipment and prior-warning device; Wherein: the output light of lasing light emitter is successively through getting into detection cell behind short focus lens, grating and the long focus lens; The light that appears of detection cell receives the laggard photoelectric commutator of going into by the scattered light receiver; The output signal of photoelectric commutator carries out endotoxin concns calculating through getting into quantitative arithmetical unit behind signal amplifier, signal processor and the data acquisition unit successively, and quantitatively the output of arithmetical unit connects data display equipment and prior-warning device.
The quantitative detecting method of this endotoxin detection system is: the scattered light signal intensity of endotoxin particle in three above different angles detection solution; According to light scattering formula I=KCf (D) f (E); Calculate f (D) and f (E) by the scattered light signal of any two angles wherein; With light scattering formula abbreviation is S=fC+g, calculates the endotoxin concns of solution again with the scattered light signal of all the other angles, realizes detection by quantitative;
In the above formula: I is a scattered light signal intensity; K is a proportionality constant; C is an endotoxin concns; F (D) is the particle diameter parametric function; F (E) is the scattering spectrum function; S is an astigmatism, and S is that the ln linearization data processing by I obtains; Related coefficient f=Kf (D) f (E); G is a corrected parameter.
Technique effect:
1, detection does not consume reagent, soup is not had any pollution, economic environmental protection.
2, light path system has been carried out integrated, improved detection sensitivity, can detect the above endotoxin solution of 0.5EU/ml.
3, combined quantitative operational method, realized endotoxic detection by quantitative the multiple spot more than 3 o'clock astigmatism signal.
4, detecting pattern is divided into two kinds of online detection and offline inspection, and applying flexible, practicality is good, detection time is short, can satisfy the on-line monitoring requirement of water for injection, injection and integrity of ultrafiltration membrane.
5, set up early warning system, triggered early warning in the time of can exceeding standard at the endotoxin concns of solution, realized on-line monitoring, reliability is high.
Description of drawings
Fig. 1 is the composition structural representation of system of the present invention, label title among the figure: 1, lasing light emitter; 2, lack focus lens; 3, grating; 4, long focus lens; 5, detection cell; 6, scattered light receiver; 7, photoelectric commutator; 8, signal amplifier; 9, signal processor; 10, data acquisition unit; 11, quantitative arithmetical unit; 12, data display equipment; 13, prior-warning device.
Fig. 2 is an endotoxin quantitative Analysis principle schematic.
Fig. 3 is the detection early warning process flow diagram of system of the present invention.
Embodiment
Be described further in the face of the present invention down.
The structure of endotoxin detection system of the present invention is as shown in Figure 1; Comprise lasing light emitter 1, short focus lens 2, grating 3, long focus lens 4, detection cell 5, scattered light receiver 6, photoelectric commutator 7, signal amplifier 8, signal processor 9, data acquisition unit 10, quantitative arithmetical unit 11, data display equipment 12 and prior-warning device 13; Wherein: the output light of lasing light emitter 1 gets into detection cell 5 through short focus lens 2, grating 3 and long focus lens 4 backs successively; The light that appears of detection cell 5 receives the laggard photoelectric commutator 7 of going into by scattered light receiver 6; The output signal of photoelectric commutator 7 gets into quantitative arithmetical unit 11 through signal amplifier 8, signal processor 9 and data acquisition unit 10 backs successively and carries out endotoxin concns calculating, and quantitatively the output of arithmetical unit 11 connects data display equipment 12 and prior-warning device 13.
The present invention carries out integrated on the basis of conventional laser particle diameter detector to light path system, the scattering light path that is preferably about 90 ° detects.Lasing light emitter 1 is the double light beam laser source, adopts the FLS-2600B lasing light emitter of Canadian EXFO company; Short focus lens 2 adopts the ED lens; Grating 3 adopts fiber coupler type fiber grating; Long focus lens 4 adopts low consumption NV-202m lens.Detection cell 5 is processed by optical glass; Comprise two kinds in online detection cell and offline inspection pond: the two ends up and down of online detection cell are provided with liquid outlet and inlet; Online detection cell is connected in the injection production line pipeline through liquid outlet and inlet and forms the loop, to realize the continuous detecting of soup, on liquid outlet and inlet, is equipped with electromagnetic valve; Also be provided with ultrasonic probe at the inlet place, be used to get rid of bubble and disturb; The offline inspection pond is the detachable detection cell of semitight, and this detection cell top is provided with filling opening, is used for manually changing solution.Scattered light receiver 6 adopts the Picometrix LLC high speed multiple spot wide angle scattered light receiver module of API company, is multi-point and arranges at a distance from logarithm; Photoelectric commutator 7 adopts U.S.'s perseverance to open the HESMC photoelectric commutator of Electronics Co., Ltd.; Signal amplifier 8 is made up of amplifying circuit and filtering circuit, and wherein amplifying circuit is connected into the differential amplifier circuit form by three ICL7650 amplifiers, and wave filter is RC network commonly used; Signal processor 9 adopts the CONVERTER-VLC.602 programmable signal processor of Italian VAL.CO; The A/D converter Max132 of the U.S. letter of data acquisition unit 10 employings company.Quantitatively arithmetical unit 11 is our unit's independent research, and quantitatively the calculating principle of arithmetical unit is as shown in Figure 2, according to light scattering formula I=KbCf (D) f (E) f (Δ λ); In the formula: I is a scattered light signal intensity, and K is a proportionality constant, and b is a sample path length to be measured; C is that (unit: EU/ml), f (D) is the particle diameter parametric function to endotoxin concns, and f (E) is the scattering spectrum function; F (Δ λ) is a function of wavelength, and sample path length to be measured (colorimetric pool) is fixing, and wavelength is fixed; So b and f (Δ λ) are constant, the light scattering formula can be reduced to I=KCf (D) f (E).Endotoxin standard solution detection with variable concentrations can draw the constant K value, and the scattered light signal intensity of system's endotoxin particle in three above different angles detection solution is according to I=KCf (D) f (E); Calculate f (D) and f (E) by the scattered light signal of any two angles wherein; With the light scattering simplified formula is S=fC+g, and in the formula: S is an astigmatism, and S is that the ln linearization data processing by I obtains; Related coefficient f=Kf (D) f (E); G is a corrected parameter, and the scattered light signal with all the other angles calculates endotoxin content in the solution again, realizes detection by quantitative.Data display equipment 12 adopts 128 * 64 dot matrix display terminals of the golden company of initiating, and is used to show detected endotoxin concns value; Prior-warning device 13 adopts integrated achievement electronics CMS7000-500 alarm monitoring software and piezoelectric type early warning system; The detection early warning flow process of system is as shown in Figure 3; In early warning software, can preestablish the endotoxin concns limit value, attention device is reported to the police when endotoxin concns surpasses limit value in the soup that detects.
The use flow process that the present invention is online when detecting is: according to producing needs; Quantitatively preestablishing endotoxin concns limit value and coefficient f and parameter g in the arithmetical unit; Open production line, make solution get into online detection cell, calculate endotoxin concns according to S=fC+g; When this concentration during less than limit value system keep operation, early warning system is reported to the police when this concentration is higher than limit value.
Operation steps during offline inspection of the present invention is: quantitatively preestablishing coefficient f and parameter g in the arithmetical unit; Take out the offline inspection pond; Clean up with pure water, with solution to be measured swing wash 2～3 times after, add solution to be measured; Detection cell is placed native system, calculate endotoxin concns according to S=fC+g.
The endotoxin off-line detection by quantitative instance of red sage roo drip liquid is provided below:
Change the offline inspection pond, get the working stamndard endotoxin, use the inspection water to be mixed with the standard solution of concentration 100EU/ml; Re-use the inspection water and progressively dilute standard solution for series concentration; The series concentration that is mixed with is respectively 0.5,1,5,10,50EU/ml, places detection cell to detect solution, carries out the data regression Calculation through quantitative arithmetical unit; Linear equation is: S=0.056C+0.165, R
2=0.997, result of calculation is seen table 1.
Table 1
(Shanghai Worldbest & Anhui Jinhui Pharmaceutical Co., Ltd. provides, lot number: 05110209), adopt native system detection method and TAL nephelometry to calculate endotoxin content respectively, the result sees table 2 to get the red sage roo drip liquid intermedium.
Table 2
The result shows that endotoxin content that native system records and TAL nephelometry are comparatively approaching.The TAL nephelometry is for Chinese Pharmacopoeia records method at present, and it is comparatively accurate to measure the result, and the result that native system records is close with it, and this has also explained the reliability of native system.
The online detection by quantitative instance of endotoxin of red sage roo drip liquid is provided below:
Get two parts of red sage roo drip liquids, a (Shanghai Worldbest & Anhui Jinhui Pharmaceutical Co., Ltd. provides, lot number: 10011605), another part carries out on-line monitoring for the soup after polluting respectively to two parts of soups for normal soup.Detected parameters is set: f is 0.056, and g is 0.165, and the endotoxin limit value is 1EU/mL, opens production line and detects, and calculates endotoxin concns through regression equation S=0.056C+0.165, and the result sees table 3.
Table 3
The result shows that when contaminated with endotoxins surpassed limit value, native system can be reported to the police, and can satisfy the quantitative monitoring of the contaminated with endotoxins situation in the injection production.
Claims (7)
1. endotoxin detection system; It is characterized in that: comprise lasing light emitter (1), short focus lens (2), grating (3), long focus lens (4), detection cell (5), scattered light receiver (6), photoelectric commutator (7), signal amplifier (8), signal processor (9), data acquisition unit (10), quantitative arithmetical unit (11), data display equipment (12) and prior-warning device (13); Wherein: the output light of lasing light emitter (1) gets into detection cell (5) through short focus lens (2), grating (3) and long focus lens (4) back successively; The light that appears of detection cell (5) receives the laggard photoelectric commutator (7) of going into by scattered light receiver (6); The output signal of photoelectric commutator (7) gets into quantitative arithmetical unit (11) through signal amplifier (8), signal processor (9) and data acquisition unit (10) back successively and carries out endotoxin concns calculating, and quantitatively the output of arithmetical unit (11) connects data display equipment (12) and prior-warning device (13).
2. endotoxin detection system according to claim 1 is characterized in that: said lasing light emitter (1) adopts the double light beam laser source.
3. endotoxin detection system according to claim 1 is characterized in that: said detection cell (5) is processed by optical glass, and detection cell (5) comprises online detection cell and offline inspection pond.
4. endotoxin detection system according to claim 1 is characterized in that: said scattered light receiver (6) adopts the multiple spot wide angle scattered light receiver more than 3.
5. endotoxin detection system according to claim 3; It is characterized in that: said online detection cell is provided with liquid outlet and inlet; Online detection cell is connected in the injection production line pipeline through this liquid outlet and inlet; Be equipped with electromagnetic valve on said liquid outlet and the inlet, also be provided with the ultrasonic probe that is used to get rid of the bubble interference at the inlet place.
6. endotoxin detection system according to claim 3 is characterized in that: said offline inspection pond is the detachable detection cell of semitight, and this detection cell top is provided with filling opening.
7. quantitative detecting method based on the described endotoxin detection system of claim 1 is characterized in that:
This method thes contents are as follows: the scattered light signal intensity of endotoxin particle in three above different angles detection solution; According to light scattering formula I=KCf (D) f (E); Calculate f (D) and f (E) by the scattered light signal of any two angles wherein; With light scattering formula abbreviation is S=fC+g, calculates the endotoxin concns of solution again with the scattered light signal of all the other angles, realizes detection by quantitative;
In the above formula: I is a scattered light signal intensity; K is a proportionality constant; C is an endotoxin concns; F (D) is the particle diameter parametric function; F (E) is the scattering spectrum function; S is an astigmatism, and S is that the ln linearization data processing by I obtains; Related coefficient f=Kf (D) f (E); G is a corrected parameter.
Priority Applications (6)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210196596.8A CN102692396B (en) | 2012-06-14 | 2012-06-14 | Endotoxin detection system and quantitative detection method thereof |
| EP13803561.3A EP2863206A4 (en) | 2012-06-14 | 2013-06-13 | Endotoxin detection systems and detection methods thereof |
| JP2015516431A JP6093854B2 (en) | 2012-06-14 | 2013-06-13 | Detection method based on endotoxin detection system |
| US14/408,016 US9958431B2 (en) | 2012-05-14 | 2013-06-13 | Endotoxin detection systems and detection methods thereof |
| PCT/CN2013/077191 WO2013185621A1 (en) | 2012-06-14 | 2013-06-13 | Endotoxin detection systems and detection methods thereof |
| US15/877,627 US20180224426A1 (en) | 2012-06-14 | 2018-01-23 | Endotoxin detection systems and detection methods thereof |
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| CN201210196596.8A CN102692396B (en) | 2012-06-14 | 2012-06-14 | Endotoxin detection system and quantitative detection method thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103278476A (en) * | 2013-06-08 | 2013-09-04 | 南京拓鉒医药科技有限公司 | Endotoxin detection system and limit detection method thereof |
| WO2013185621A1 (en) * | 2012-06-14 | 2013-12-19 | 南京拓鉒医药科技有限公司 | Endotoxin detection systems and detection methods thereof |
| CN110064446A (en) * | 2013-12-06 | 2019-07-30 | 百克特瑞欧扫描有限责任公司 | The test tube component of optical measurement is carried out for the characteristic to particle in fluid sample |
| CN112504922A (en) * | 2020-10-20 | 2021-03-16 | 华南师范大学 | Online measurement system and method for particle size distribution of atmospheric particulates |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013185621A1 (en) * | 2012-06-14 | 2013-12-19 | 南京拓鉒医药科技有限公司 | Endotoxin detection systems and detection methods thereof |
| CN103278476A (en) * | 2013-06-08 | 2013-09-04 | 南京拓鉒医药科技有限公司 | Endotoxin detection system and limit detection method thereof |
| CN110064446A (en) * | 2013-12-06 | 2019-07-30 | 百克特瑞欧扫描有限责任公司 | The test tube component of optical measurement is carried out for the characteristic to particle in fluid sample |
| CN112504922A (en) * | 2020-10-20 | 2021-03-16 | 华南师范大学 | Online measurement system and method for particle size distribution of atmospheric particulates |
| CN112504922B (en) * | 2020-10-20 | 2022-09-02 | 华南师范大学 | Online measurement system and method for particle size distribution of atmospheric particulates |
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