CN104958771A - Large-scale produced fermented pollen sterilization method - Google Patents
Large-scale produced fermented pollen sterilization method Download PDFInfo
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Abstract
The invention relates to a large-scale produced fermented pollen sterilization method, in particular to an irradiation sterilization method, i.e., a method of killing microorganisms in fermented pollen through ionizing radiation by using gamma rays irradiated by a radiation source or electron beams generated by an electron accelerator. The method preferably selects 60-gamma rays for radiation sterilization, wherein the radiation dose is 1.0kGy-20.0kGy, preferably 6.0kGy-10.0kGy. After the fermented pollen is sterilized, the number of bacteria satisfies the national legal standard on the quality of drugs or food, and the effective components and sensory index of the fermented pollen are substantially unchanged. The fermented pollen is a product obtained after a period of fermentation at different temperatures by adding one or a plurality of pollens into certain solution (including water) or preparation, and contains one or a plurality of microorganisms such as bacteria, mold and Escherichia coli; after irradiation sterilization, the number of bacteria in products produced under conditions regulated in Good Manufacturing Practice (GMP) also satisfies the legal standard on the quality of drugs or food.
Description
Technical field
The present invention relates to a kind of fermented pollen sterilizing methods of large-scale production; refer to irradiation sterilization method; namely the electron beam utilizing the gamma-rays of radioactive source radiation or electron accelerator to occur carries out ionizing radiation and reaches and kill method of microorganism in fermented pollen, belongs to biomedicine field.
Background technology
The initiation material of some medicine or food have employed fermented pollen, such as, just has in the national drug quality standard of in December, 2014 state food pharmaceuticals administration general bureau (CFDA) the Shenqi Huafen sheet (authentication code: the accurate word B20020762 of traditional Chinese medicines) ratified [WS-5741(B-0741)-2014Z] " with the Pollen Brassicae campestris of hydromel fermentation, Pollen Maydis, Pollen Helianthi and Pollen Typhae ".
Shenqi Huafen sheet is produced in the approval of 1994 01 month 15 Department of Public Health of Nikkei Henan Province, and authentication code is that No. (1993) L-1510064, the accurate word of medicine is defended in Henan; Within 1996, Department of Public Health of Nikkei in February 24 Henan Province is approved as health care medicine, and authentication code is that Henan is defended medicine and is good for No. 0108th, word (1996); November 01 in 2002, Nikkei National Drug Administration re-registered as medicine, authentication code is the accurate word B20020762 of traditional Chinese medicines, drug standard (trying) is WS-5741(B-0741) its quality standard of in December ,-2002,2014 is approved as drug standard WS-5741(B-0741 by state food pharmaceuticals administration general bureau)-2014Z.National drug quality standard [WS-5741(B-0741)-2014Z] display: " Radix Codonopsis, the Radix Astragali, Pollen Typhae, Pollen Brassicae campestris; Pollen Maydis and Pollen Helianthi Six-element, Radix Codonopsis and the Radix Astragali decoct with water three times, collecting decoction; filter, concentrating under reduced pressure, is ground into fine powder after being evaporated to dry cream; Get Pollen Brassicae campestris, Pollen Maydis, Pollen Helianthi and Pollen Typhae to mix, with hydromel, fermentation, plays material, dries, pulverizes to obtain yeast powder; Add Radix Codonopsis, Extract of Huangqi, granulate, dry, tabletting, coating, to obtain final product.”
Adopt the object that Pollen Brassicae campestris, Pollen Maydis, Pollen Helianthi and Pollen Typhae are fermented to be promote pollen broken wall in Shenqi Huafen sheet (the accurate word B20020762 of traditional Chinese medicines) technique, increase the absorption of effective ingredient, subtract the effect of untoward reaction.Research display, after Shenqi Huafen sheet (the accurate word B20020762 of traditional Chinese medicines) fermentation process pollen, rate of breaking pollen wall brings up to more than 97% by less than 10%, effective ingredient improves more than 2 times, curative effect improves 27%, irritated side effect decreases 93%(2014 year Science and Technology in Henan progress prize, certificate number: 2014-J-315-D01/02).The bright three teach problem treated animal experiments of Henan College Of Traditional Chinese Medicine Seedling show, have employed Shenqi Huafen sheet (the accurate word B20020762 of traditional Chinese medicines) prepared by fermented pollen, the cardiovascular and cerebrovascular disease curative effect that treatment anoxia causes improves 27%[Seedling bright three, Jiang Meiqiong, Zhang Xuexia. Shenqi Huafen sheet is on the impact letter of rat motor fatigue model. Chinese experimental pharmacology of Chinese medical formulae magazine 2014; 20 (5): 1221-1226].Nanfang Medical Univ Deng Hong pearl teach problem group proves, have employed prepared by fermented pollen Shenqi Huafen sheet (traditional Chinese medicines accurate word B20020762) to deficiency of both the splenic and pulmonary QI disease [Yi Yankui evident in efficacy, Deng Hongzhu, Dong Changrong, Zhao Yong, Huang Kaizheng, Wang Wei, Lin Jianyun. Shenqi Huafen preparation recipe is inquired into the utilization of deficiency of both the splenic and pulmonary QI disease. Inner Mongol Chinese medicine 2014; (1): 7-10].Key lab of the plateau medicine Ministry of Education of Third Military Medical University is under national science and technology supporting plan (No.2009BAl85803) and army's 11th Five-Year brainstorm subject (No. 08G093) are supported, the Shenqi Huafen sheet (the accurate word B20020762 of traditional Chinese medicines) proving have employed fermented pollen has increases blood oxygen, purify the blood, thrombus, the effects such as vessel softening, lower plateau proteinuria efficacy and be better than Rhodiola rosea capsules, three recoveries capsule, GUIFU DIHUANG WAN and FUFANG DANSHEN DIWAN, play the multiple chronic disease caused by treatment cardio-cerebral-pulmo hypoxic-ischemic, altitude sickness and plateau pika disease [Wang Fuling, Chen Qidong, limit bar Drolma, rise million posts, Guo Hongbin, Li Xiaoping, Zhou Qiquan. Shenqi Huafen sheet intervenes plateau albuminuria observation of curative effect. Chinese tcm emergency 2010, 19 (12): 2038-2040, Shi Zifu, Zhou Qiquan, Xiang Lu, Ma Sanding, Yan Chengjun, Luo Han. the Randomised placebo-controlled trial of 3 kinds of compound Chinese medicinal preparation treatment plateau pika diseases. Journal of Chinese Integrative Medicine 2011, 9 (1): 1-7, Lou Xiaomin, Zhang Xuefeng, Zhou Qiquan, Wang Shengyu. Shenqi Huafen sheet is to the observation of curative effect of deacclimatization between the chronic intermittent hypoxia exposure period of plateau. modern preventive medicine 2013, 40 (7): 1227-1229].
But if do not carry out sterilization treatment to fermented pollen and the medicine produced thereof or food, so the micro organism quantity of fermented pollen and the medicine produced or food very likely exceeds the medicine of national regulation or the limitation standard in microbe of food.Such as, the limitation standard in microbe of the preparation of the oral administration of current national regulation sees the limitation standard in microbe (table 1) of " Chinese Pharmacopoeia version in 2010 " (one) annex XIII C microbial limit test.If do not carry out sterilization treatment, Shenqi Huafen sheet (the accurate word B20020762 of traditional Chinese medicines) microbiological indicator prepared by fermented pollen is difficult to reach above-mentioned requirements.We show in research, and in Shenqi Huafen sheet (traditional Chinese medicines accurate word B20020762), the microorganism of the fermented pollen of non-sterilizing is extremely many, and wherein bacterial population not count enable, the not count enable of yeast and mold number, escherichia coli does not detect.
Table 1 oral Preparation limitation standard in microbe
| Bacterial population | Yeast and mold number | Escherichia coli | |
| Not containing the preparation of the former medicated powder of medical material | < 1000/g | < 100/g | Must not detect |
| Containing the preparation of the former medicated powder of medical material | < 10000/g | < 100/g | Must not detect |
| Containing the preparation of the fermented ingredient such as Semen Sojae Preparatum, Massa Medicata Fermentata | < 100000/g | < 500/g | Must not detect |
According to the requirement of national regulation limitation standard in microbe (table 1), fermented pollen is regarded as containing the fermented ingredient such as Semen Sojae Preparatum, Massa Medicata Fermentata, so Shenqi Huafen sheet (the accurate word B20020762 of traditional Chinese medicines) limitation standard in microbe is bacterial population <100000/g, yeast and mold number <500/g, escherichia coli must not detect; Shenqi Huafen sheet (traditional Chinese medicines accurate word B20020762) is regarded as not containing the preparation of the former medicated powder of medical material, its limitation standard in microbe is bacterial population <1000/g, yeast and mold number <100/g, escherichia coli must not detect.Indicate " preparation containing the fermented ingredient such as Semen Sojae Preparatum, Massa Medicata Fermentata " in " Chinese Pharmacopoeia version in 2010 " (one) annex XIII C microbial limit test, this refers to that crude drug contains containing the fermented ingredient such as Semen Sojae Preparatum, Massa Medicata Fermentata.Whether " preparation containing the fermented ingredient such as Semen Sojae Preparatum, Massa Medicata Fermentata " not to have in clear and definite Shenqi Huafen sheet (the accurate word B20020762 of traditional Chinese medicines) technique with yeast powder prepared by four kinds of pollen is fermented ingredient, therefore, can only by Shenqi Huafen sheet (traditional Chinese medicines accurate word B20020762) as the preparation not containing the former medicated powder of medical material, improve limitation standard in microbe (bacterial population <1000/g, yeast and mold number <100/g, escherichia coli must not detect), just have no objection in theory.
Radix Codonopsis in Shenqi Huafen sheet (the accurate word B20020762 of traditional Chinese medicines) technique, the Radix Astragali decocts with water extraction, is aseptic in theory; If after four kinds of pollen fermentations, increasing sterilization steps, is also aseptic in theory.Therefore, fermented pollen sterilizing must be increased in Shenqi Huafen sheet (the accurate word B20020762 of traditional Chinese medicines) technique, its limitation standard in microbe is easy to the target reaching " bacterial population <1000/g; yeast and mold number <100/g, escherichia coli must not detect ".
Fermented pollen is solid matter, sterilizing methods can not with filter, illumination sterilizing can only carry out surface disinfection, gaseous sterilization may with flavone compound generation chemical reaction in fermented pollen.Although many sterilizing methods, comprise moist heat sterilization, dry heat sterilization, microwave sterilization method can effectively kill microorganism in fermented pollen, reach the legal medicine of the country of national regulation or food quality standard, but likely there is significant change in fermented pollen organoleptic indicator.Therefore, this research is mainly invented a kind of easy, and after making the fermented pollen sterilizing of large-scale production, bacterial number reaches the legal medicine of country or food quality standard, and fermented pollen effective ingredient and organoleptic indicator do not change substantially.
Summary of the invention
A kind of fermented pollen sterilizing methods of large-scale production has been invented in this research; after it is characterized in that the fermented pollen sterilizing of large-scale production, bacterial number reaches the legal medicine of country or food quality standard, and fermented pollen effective ingredient and organoleptic indicator do not change substantially.
This fermented pollen sterilizing methods, is characterized in that irradiation sterilization method, and the electron beam namely utilizing the gamma-rays of radioactive source radiation or electron accelerator to occur carries out ionizing radiation and reaches and kill method of microorganism in fermented pollen.
This irradiation sterilization method, it is characterized in that preferred Co 60 (
60co)-gamma Rays sterilizing, radiation dose 1.0kGy ~ 20.0kGy, preferred radiation dose is 6.0kGy ~ 10.0kGy.
Fermented pollen is solid matter, is therefore not suitable for filtration sterilization process.
Gas sterilization is the method for the gas killing microorganisms with chemosterilant formation, and be applicable in gas stable, the article sterilizing that content liquid is less, conventional chemosterilant has oxirane, gaseous hydrogen peroxide, formaldehyde, ozone etc.During gaseous sterilization process, in fermented pollen, flavone compound is not easy to stablize, and chemosterilant easily and these flavone compound generation chemical reactions.
Research shows, fermented pollen uses moist heat sterilization and dry heat sterilization method, and the too high meeting of temperature significantly changes fermented pollen effective ingredient and organoleptic indicator.
Microwave sterilization method also changes into high temperature and carrys out sterilizing, also can significantly change fermented pollen effective ingredient and organoleptic indicator.
Research also shows, and illumination (comprising ultraviolet) sterilizing methods can only kill the microorganism on surface, unbalanced to microorganism killing in fermented pollen, is difficult to reach the legal medicine of country or food quality standard.
Fermented pollen of the present invention is after one or more pollen add certain solution (comprising water) or preparation, the product at different temperatures after a period of time fermentation, and this product contains one or more microorganisms such as antibacterial, mycete, escherichia coli; These solution (comprising water) or preparation, be the product that one or more raw materials (comprising water) are made, some product adds microorganism.
The product that the fermented pollen of sterilizing is produced under " medicine (or food) production management specification " (GMP) condition, its bacterial number meets legal medicine or food quality standard; The drug standard that country is legal, refers to the medicine sanitary standard that existing country promulgates, as the requirement of limitation standard in microbe in " Pharmacopoeia of People's Republic of China version in 2010 " that Chinese Pharmacopoeia Commission edits; The food quality standard that country is legal, refer to the food hygienic standard that existing country promulgates, as national quality technical supervision management general bureau has promulgated that " standard GB/T 14891.2-94 irradiation pollen sanitary standard " that health ministry is drafted and calendar year 2001 have promulgated the regulation of " standard GB/T 18526.6.2-2001 Irradiating pollen process for sterilizing " that health ministry is drafted for 1994.
Fermented pollen effective ingredient refers to protein, fat, sugar, flavone compound, and fermented pollen organoleptic indicator refers to color and luster, state, abnormal smells from the patient, flavour etc.
Study for the fermentation Pollen Brassicae campestris of large-scale production in Shenqi Huafen sheet (the accurate word B20020762 of traditional Chinese medicines) technique, Pollen Maydis, Pollen Helianthi and Pollen Typhae below, set forth the above results.
The sterilizing research of fermented pollen
According to national relevant regulations, as " Pharmacopoeia of People's Republic of China version in 2010 " (one) adnexa XVI sterilization, radiation sterilization process, moist heat sterilization method, dry heat sterilization method, gaseous sterilization process and filtration sterilization method should be comprised.In addition, the microwave sterilization method and illumination sterilizing methods that do not have to record also is added in " Pharmacopoeia of People's Republic of China version in 2010 " (one) adnexa XVI sterilization in this research.
filtration sterilization method
Be utilize microorganism not by the principle of fine and close tool hole filter material, remove method of microorganism in gas or liquid.Because fermented pollen is solid, filtration sterilization method can not be applied.
gaseous sterilization process
Be the method for the gas killing microorganisms with chemosterilant formation, conventional chemosterilant has oxirane, gaseous hydrogen peroxide, formaldehyde, ozone etc.Gaseous sterilization is applicable in gas stable, the article sterilizing that content liquid is less.Because before fermentation, four kinds of pollen amount of water reach about 20%, and water content is more, and therefore, this fermented pollen is not suitable for gaseous sterilization process.
Flavone compound is the main active skull cap components of pollen, play an important role, but the chemosterilant of gaseous sterilization easily causes flavone compound generation chemical reaction Sum decomposition in pollen effect.
dry heat sterilization method
Be utilize heated dry air to reach killing microorganisms or eliminate the method for heat source substance, sterilising conditions is generally 160 ~ 170 DEG C × more than 120min, 170 ~ 180 DEG C × more than 60min or 250 DEG C × more than 45min.Result of study shows, when only using 160 DEG C × 30min or 170 DEG C × 20min dry heat sterilization, fermented pollen character (color) there occurs obvious change (Fig. 1 and Fig. 2).160 ~ 170 DEG C × 120min or 170 ~ 180 DEG C × 60min dry heat sterilization sterilization effect is good, but it causes fermented pollen main component flavone compound (in anhydrous rutin) to reduce to 564.37 mg/100g(minimizings 43.0% by 989.63mg/100g) and 521.64mg/100g(minimizing 47.3%) (table 2).
Table 2 dry heat sterilization is on the impact of fermented pollen main component and sterilization effect
Can be drawn by above Fig. 1, Fig. 2 and table 2, though dry heat sterilization can effective killing microorganisms, but destroy the character of pollen, result in the degraded of main component flavone compound highly significant, therefore, fermented pollen is unwell to dry heat sterilization.
moist heat sterilization method
Be the method utilizing the means such as high-pressure saturated steam, superheated water spray to make protein in microbial cells, nucleic acid generation degeneration and killing microorganisms, usually adopt the condition of 121 DEG C × 15min, 121 DEG C × 30min or 116 DEG C × 40min.With 121 DEG C × 15min or 116 DEG C × 40min moist heat sterilization, fermented pollen character there occurs significant change (Fig. 3 and Fig. 4).121 DEG C × 15min or 116 DEG C × 40min moist heat sterilization sterilization effect is good, but it causes fermented pollen main component flavone compound (in anhydrous rutin) to reduce to 745.39mg/100g(minimizing 24.7% by 989.63mg/100g) and 751.25mg/100g(minimizing 24.1%) (table 3).
The main component of table 3 moist heat sterilization on fermented pollen and the impact of sterilization effect
From above result, though the effective killing microorganisms of moist heat sterilization energy, but destroy the character of pollen, result in the remarkable degraded of main component flavone compound, therefore, this fermented pollen is not suitable for moist hear heat test.
microwave sterilization method
It is the method adopting microwave (frequency is 300MHz ~ 300000MHz) to irradiate heat energy killing microorganisms and the spore produced.Altofrequency Electromagnetic Field produces molecular polarization phenomenon in material, and in material, water absorbs microwave consumingly polar molecule is rotated, intermolecular friction and heat-dissipating, and the temperature rising of material is with killing microorganisms.Although, temperature required by microwave sterilizating is more xeothermic is low, inside and outside heat simultaneously, heat time heating time is short, thus its to item spoilage comparatively other heat sterilizer be light, but research shows, when fermentation Pollen Brassicae campestris, Pollen Maydis, Pollen Helianthi and Pollen Typhae adopt microwave sterilizating, intensification is rapid, and temperature is higher, also can significantly change fermented pollen effective ingredient and organoleptic indicator, result is as moist heat sterilization and dry heat sterilization method.
illumination sterilizing methods
Refer to illumination, the most often refer to the method that ultraviolet (energy) irradiates killing microorganisms.Illumination, particularly ultraviolet can not only make nucleic acid-protein degeneration, and oxygen in air can be made to produce drop ozone, thus reach common bactericidal action.But illumination sterilizing (comprising ultraviolet disinfection) method belongs to surface sterilizing, be applicable to the sterilizing of irradiating object surface sterilizing, sterilizing room air and distilled water; Be not suitable for the sterilizing of medicinal liquid and the deep sterilizing of solid material.Because light (comprising ultraviolet) is with straightline propagation, can by different surface reflections or absorption, penetration power is faint, simple glass Absorbable rod light (comprising ultraviolet), and the medicine be loaded in container can not use illumination sterilizing.Therefore, fermentation Pollen Brassicae campestris, Pollen Maydis, Pollen Helianthi and Pollen Typhae are difficult to illumination (comprising ultraviolet) sterilizing methods.
radiation sterilization process
That the electron beam utilizing the gamma-rays of radioactive source radiation or electron accelerator to occur carries out ionizing radiation and reaches the method for killing microorganisms, the most frequently used is Co 60 (
60co) radiation sterilization.All available this law sterilizing such as crude drug, intermediate, finished product that not raying destroys.
Pollen radiating sterilizing also sees " standard GB/T 18526.6.2-2001 Irradiating pollen process for sterilizing " and in " standard GB/T 14891.2-94 irradiation pollen sanitary standard ".
Although fermented pollen adopts the radiation sterilization effect of different radiation dose, relevant with pollen kind, attenuation degree, fermented pollen amount etc., the microbial limit of sterilizing after fermentation pollen should reach the legal medicine of country or food quality standard.Research shows, this fermented pollen be suitable for Co 60 (
60co) radiation sterilization, radiation dose is 1.0 kGy ~ 20.0 kGy, is optimized for 6.0 kGy ~ 10.0 kGy's.The radiation sterilization of the fermentation Pollen Brassicae campestris of this large-scale production, Pollen Maydis, Pollen Helianthi and Pollen Typhae mixture, the radiation dose of optimization is 6.0 kGy ~ 10.0 kGy.
For checking fermented pollen
60the stability of Co radiation sterilization, fermented pollen, through sterilizing, after 45 DEG C of oven dry, is put into stability test case with after the sealing of solid medicinal Low Density Polyethylene bag, is carried out accelerating and long-term stable experiment.
Long-term stable experiment: temperature 25 oC ± 2oC, places 12 months under relative humidity 60% ± 10% condition, carries out long-time stability investigation.Duration of test was respectively examined or check once at 0,1,3,6,12 month respectively.Result shows, and fermented pollen main component is comparatively stable, and fermented pollen bacterial regrowth does not change (referring to table 4, table 5, table 6, table 7 and table 8).
Table 4
60the long-term stable experiment (0 month) of Co irradiation after fermentation pollen
Table 5
60the long-term stable experiment (January) of Co irradiation after fermentation pollen
Table 6
60the long-term stable experiment (March) of Co irradiation after fermentation pollen
Table 7
60the long-term stable experiment (June) of Co irradiation after fermentation pollen
Table 8
60the long-term stable experiment (December) of Co irradiation after fermentation pollen
Accelerated stability test: temperature 40oC ± 2oC, places 6 months under relative humidity 75% ± 5% condition, carries out accelerated stability investigation.Duration of test was respectively examined or check once at 0,1,2,3,6 month respectively.Accelerated stability test carries out 3 months, result shows, fermented pollen main component is comparatively stable, and fermented pollen bacterial regrowth does not change (referring to table 9, table 10, table 11 and table 12), and wherein accelerated stability test 0 month result and long-term stable experiment 0 month result (table 4) is consistent.
Table 9
60the accelerated stability test (January) of Co irradiation after fermentation pollen
Table 10
60the accelerated stability test (February) of Co irradiation after fermentation pollen
Table 11
60the accelerated stability test (March) of Co irradiation after fermentation pollen
Table 12
60the accelerated stability test (June) of Co irradiation after fermentation pollen
Can tentatively draw thus, fermented pollen warp
60after Co radiation sterilization, microbial limit and main component keep basicly stable.Therefore, it is feasible for increasing Co 60 sterilizing in Shenqi Huafen blade technolgy.
Irradiation sterilization method radiation dose is studied
This research, with reference to sterilizing methods in " standard GB/T 18526.6.2-2001 Irradiating pollen process for sterilizing ", adopts
60co irradiation sterilization method is as the sterilization process of fermented pollen, and test rating is with reference to relevant regulations in " Chinese Pharmacopoeia version () in 2010 " annex XIII C microbial limit test, " standard GB/T 14891.2-94 irradiation pollen sanitary standard " and " standard GB/T/T30359-2013 bee pollen ".
Get Pollen Brassicae campestris, Pollen Maydis, Pollen Helianthi and Pollen Typhae to mix, warm water stirs evenly, and evenly admixes in pollen with freshly prepared 20% hydromel, fermentation, plays material, obtains fermented pollen.
60co radiation source: Inst. of Isotope, Henan Provincial Academy of Sciences.
Take out Partial fermentation pollen about to check as before sterilizing, remaining fermented pollen is divided into 7 parts, with after the sealing of Low Density Polyethylene bag, put into fiber drum and pack, speed is sent
60co irradiation sterilization.According to " standard GB/T 18526.6.2-2001 Irradiating pollen process for sterilizing ", the minimum effective dose of irradiation pollen is 1 kGy, and maximum tolerated dose is 20 kGy, and therefore, this research is selected
60co irradiation dose is 0 kGY, 1 kGy, 6 kGy, 8 kGy, 10 kGy, 15 kGy and 20 kGy, and fermented pollen sterilization effect refers to table 13.
According to " Chinese Pharmacopoeia version () in 2010 " annex XIII C limit test of microbe, " standard GB/T 14891.2-94 irradiation pollen sanitary standard " and " National Standard of the People's Republic of China GB/T30359-2013 bee pollen ", due to this product be fermentation after pollen, belong to middle product, so this product microbial standard is selected standard with person the strictest in three.
Table 13 is different
60co irradiation dose irradiates the sterilization effect to fermented pollen
As can be seen from table 13 result, with being greater than after 1 kGy irradiation dose carries out irradiation, the antibacterial of in fermented pollen more than 90%, mycete and escherichia coli can be killed.But, after carrying out irradiation with 1 kGy irradiation dose, annex XIII C microbial limit test standard that antibacterial and mold count result just meet " Chinese Pharmacopoeia version () in 2010 ", but highest standard in not meeting " standard GB/T 14891.2-94 irradiation pollen sanitary standard " and " National Standard of the People's Republic of China GB/T30359-2013 bee pollen ", the regulation namely in " National Standard of the People's Republic of China GB/T30359-2013 bee pollen "; With being greater than after 6.0 kGy ~ 10.0 kGy irradiation doses carry out irradiation, can significantly kill antibacterial in fermented pollen, mycete and escherichia coli, the regulation of highest standard in result all meets " Chinese Pharmacopoeia version () in 2010 " annex XIII C microbial limit test, " standard GB/T 14891.2-94 irradiation pollen sanitary standard " and " National Standard of the People's Republic of China GB/T30359-2013 bee pollen "; After carrying out irradiation with 15.0 kGy and 20.0 kGy irradiation doses, can significantly kill antibacterial in fermented pollen, mycete and escherichia coli, but irradiation dose is excessive, significantly beyond irradiation dose maximum standard 10.0 kGy of national regulation.
Therefore, according to this result of study preferably 6.0 kGy ~ 10.0 kGy be best irradiation dose.
Variation of Main Nutritional Components in irradiation sterilization after fermentation pollen
According to the standard of " National Standard of the People's Republic of China GB/T30359-2013 bee pollen ", this research is right
60in Co predose after fermentation pollen, main nutrient composition (protein, total sugar and flavone compound) has carried out detecting (table 14).
Protein content determination
Method for measuring according to " Chinese Pharmacopoeia version () in 2010 " annex IX L N2 method and standard GB/T 5009.5-2010 Protein in Food performs.
Total sugar (in reducing sugar) assay
Perform according to the method for measuring of sucrose in standard GB/T 5009.8-2008 food.
principle
Sample is through after deproteinize, and wherein sucrose is converted into reducing sugar through hydrochloric acid hydrolysis, then is cane sugar content by the difference of reducing sugar before and after reducing sugar test hydrolysis.
reagent
A. 6N hydrochloric acid: measure 50ml hydrochloric acid, be diluted with water to 100ml.
B. C.I. 13020. indicator solution: 0.1% dissolve with ethanol solution.
C. 20% sodium hydroxide solution.
instrument
800 type ultraviolet-uisible spectrophotometers.
operational approach
In sample 10g, adding distil water 150ml, fully stirs evenly, and filters.Filtrate adds chloroform-n-butyl alcohol (volume ratio is 4:1) mixed liquor 150ml, and fully concussion 30 minutes, centrifugal, separately aqueous phase and chloroform phase.
Draw 2 parts of 50ml aqueous phases as sample treatment liquid, be placed in 1000ml volumetric flask.1 part adds 6N hydrochloric acid 5ml, heats 15 minutes in 68 ~ 70 DEG C of water-baths, and the red indicator solution of methylate 2 after cooling, is neutralized to neutrality with 20% sodium hydroxide solution, adds water to scale, mixing.Another 1 part of direct adding distil water is diluted to 100ml, measures reducing sugar respectively.
calculate
X=（R
2 - R
1）×0.95
In formula: cane sugar content in X-sample, %;
R
2content of reducing sugar after-hydrolysis process, %;
R
1-without hydrolysis process content of reducing sugar, %;
0.95-reducing sugar (with glucose meter) is scaled the coefficient of sucrose.
Flavone compound (in anhydrous rutin) assay
Method according to standard GB/T/T30359-2013 Flavonoids in Bee Pollens performs.
reagent
A. except as otherwise noted, analytical reagent and distilled water or deionized water is at least used in analysis.
B. dehydrated alcohol.
C. 70% (V/V) ethanol: purified water 30ml adds dehydrated alcohol 70ml, to obtain final product.
D. 5% sodium nitrite solution: take 5.0g sodium nitrite, be settled to 100ml by purified water.
E. 10% aluminum nitrate solution: take 17.6g aluminum nitrate, be settled to 100ml by purified water.
F. sodium hydroxide test solution: take 4.3g sodium hydroxide, be settled to 100ml by purified water.
G. rutin standard solution: control substance of Rutin stock solution: precision takes control substance of Rutin 50 mg, puts in 50ml measuring bottle, and add 70% appropriate amount of ethanol, jolting makes dissolving, and is diluted to scale.
H. control substance of Rutin uses solution: accurate absorption 10ml control substance of Rutin stock solution, put in 50ml measuring bottle, add 70% appropriate amount of ethanol, jolting makes dissolving, and is diluted to scale.
instrument and equipment
800 type ultraviolet-uisible spectrophotometers.
the preparation of sample
Get fermented pollen sample mill comminuted powder 1.0g ~ 3.0g, accurately weighedly (be accurate to 0.0001 g), add 70% ethanol 40ml, reflux 2 hours, cooling, filters, puts in 50ml measuring bottle, with 70% washing with alcohol residue, washing liquid is incorporated in measuring bottle, is settled to scale, shakes up with 70% ethanol.The accurate 2ml that draws puts in 10 ml measuring bottles, with 70% ethanol dilution to scale, shakes up.
analytical procedure
A. the preparation of standard curve: precision measure reference substance use solution 0,1,2,3,4,5,6ml, put in 25ml measuring bottle respectively, respectively add water to 6.0ml, add 5% sodium nitrite solution and each 1ml of 10% aluminum nitrate solution, shake up, place 6 minutes, hydro-oxidation sodium test solution 10ml, add water to scale, shake up, place 15 minutes.According to spectrophotometry, measuring absorbance at 510nm wavelength place, take trap as vertical coordinate, concentration Wei Transverse coordinate, drawing standard curve.
B. sample tests: the sample solution 2ml that accurate absorption prepares, puts in 25 ml measuring bottles, adds water to 6.0ml, add 5% sodium nitrite solution and each 1ml of 10% aluminum nitrate solution, shake up, place 6 minutes, hydro-oxidation sodium test solution 10ml, add water to scale, shake up, place 15 minutes.According to spectrophotometry, measure absorbance at 510nm wavelength place, bring light absorption value into regression equation, calculate, to obtain final product.
calculate
Flavones content is calculated as follows, and result of calculation is accurate or remain into 2 significant digits.
X = m/(w×d×1000) ×100%
In formula: X-flavones content, %;
M-by rutin quality in the article colorimetric liquid that standard curve calculates, unit is milligram (mg);
The quality of w-sample, unit is gram (g);
D-dilution ratio
tolerance
The relative error that same operator twice parallel determinations allows should be not more than 3%.
Table 14 is different
60co irradiation dose irradiates the impact on main nutrient composition in fermented pollen
As can be seen from Table 14,1kGy, 6kGy, 8kGy and 10 kGy irradiation doses are adopted
60after Co irradiates, pollen protein, saccharide, flavonoid content all do not have significant change.
Therefore, this research considers according to table 13 and table 14 result, selects 6.0 kGy ~ 10.0 kGy irradiation doses as fermented pollen
60co irradiation sterilization method is comparatively suitable.
Literature research result
fermented pollen increases the reasonability of radiation sterilization
Radiation sterilization appears at the United States Patent (USP) collection of nineteen thirty the earliest, and its mechanism is after microorganism is subject to irradiation, produces a large amount of active particle, has an effect with protein, DNA etc., cause cell death.
Japan in 1972 the available radiation of approval suppresses potato sprout, and radiation sterilization is used for spongaion and produces by the U.S. general strong (Upjohn) company at first.FAO (Food and Agriculture Organization of the United Nation) (FAO), International Atomic Energy Agency (IAEA) and the World Health Organization (WHO) repeatedly hold International Radiation Food Science symposium, evaluate irradiated food safety.In October, 1980, more than the 30 year safety research result in various countries is concluded by above-mentioned three " the irradiated food joint specialist committee " that tissue forms greatly, confirm " for storing object; any food is subject to the irradiation (gamma-rays and 10 below MeV electron beams) of grand mean dosage 10 below kGy; no longer need the detection carrying out toxicology aspect, does not also have special dietary and microorganism problem ".Nineteen eighty-three, the Codex Committee on Food (CAC) of FAO and WHO has formally issued " the general regulation of irradiated food ".1999, European Union promulgated the skeleton instruction 1999/2/EC of " food of ion exposure process " and has performed instruction 1999/3/EC, allows irradiation medicinal herbs, spice and plant sauce.
1986, health ministry put into effect " irradiated food administration of health regulation (provisional) ", within 1996, has promulgated " irradiated food administration of health way ", within 1997, issue "
60co irradiation Chinese medicine sterilizing dose standard ", clearly " the method is only for the sterilizing for domestic circulation Chinese medicine ", in annex, allow the medical material of irradiation sterilization to have 203 kinds, wherein Chinese patent medicine 70, allow the medical material kind of low dosage (3kGy) irradiation to have 5, do not allow the medical material kind of irradiation to have 2 (containing the medical materials of gentiopicrin as Radix Gentianae Macrophyllae, Radix Gentianae and goods thereof) ".
Radiation sterilization has been included in " Pharmacopoeia of People's Republic of China version in 2005 " (one) annex XVI sterilization, indicating conventional radiation sterilization absorbed dose is 25 kGy, requires " verifying that the dosage used does not affect by the safety of sterilizing article, effectiveness and stability ".Also radiation sterilization has been recorded in the aseptic medicine annex of 2010 version " Good manufacturing practice " (GMP), basically identical to the regulation of radiation sterilization in this regulation and European Union GMP.
National quality technical supervision management general bureau has promulgated " standard GB/T 14891.2-94 irradiation pollen sanitary standard " that health ministry is drafted for 1994, calendar year 2001 has promulgated and " standard GB/T 18526.6.2-2001 Irradiating pollen process for sterilizing " that health ministry is drafted has provided technical standard to pollen radiating sterilizing.
Therefore, fermented pollen increases irradiation sterilization, legal principle is rational.
the effectiveness of irradiation sterilization
1998, the use such as Zhong Hailuo
60co-gamma-ray irradiation 7 kinds of native Sichuan crude drugs, after display 8kGy irradiation, Chinese crude drug principle active component, tissue morphology and sense organ condition are constant, have no side effect.2000, the reports such as Owczarczyk, after the radiation of Radix Et Rhizoma Rhei etc. 10 kind of plant medicine, medicinal ingredient and active not change.Calendar year 2001, straight space and Xue Zhimou research report, 20 ~ 50 kGy
60co irradiation is to vitamin A, B
1, B
2, B
6, D, E and K impact is little.2002, Lei Xi etc. were with heavy dose of
60co-gamma-ray irradiation Radix Angelicae Sinensis and Radix Codonopsis, do not change its composition and content.2002, Koseki etc. adopted 0,10,20,30 kGy's
60co-gamma-rays, to Herba Ocimi (Herba Ocimi Pilosi), arithoke, Herba Rosmarini Officinalis irradiation, has no the significant change of flavonoid, quintessence oil, tannin, phenols, carotene.2003, the employings such as Huang Zhiyong
60co irradiation bee pollen, prove with 8.0kGy doses of sterilization best results, escherichia coli sterilizing rate >90%, mycete sterilizing rate reaches 99.6%, little on protein, saccharide and amino acid whose impact in bee pollen, but vitamin and catalase loss about 30%.2006, the reports such as Zhu Jun, 1kGy, 3kGy, 5kGy and 7kGy
60co radiation is good to soybean protein sterilization effect, and does not affect physical and chemical index, functional and aminoacid.
With regard to research, the effectiveness of natural drug (comprising fermented pollen) radiation sterilization is unquestionable.
the production safety of irradiation sterilization
The regulation of current domestic radiation sterilization only have " irradiated food administration of health way ", "
60c0 irradiation Chinese medicine sterilizing dose standard " and " Pharmacopoeia of People's Republic of China version in 2010 ", there is mutual connection problem, easily cause the misgivings of safety.By contrast, " Pharmacopoeia of People's Republic of China version in 2010 " regulation is comparatively perfect.
The research of Chinese medicine, natural drug irradiation sterilization carries out the contrast after predose based on quality standard, and result is insufficient.First, the effective ingredient of most Chinese medicine and natural drug is not yet clear, infers that its mass change is obviously not objective with composition several in quality standard; Secondly, Chinese medicine and natural drug are difficult to an effective ingredient, an impurity to determine its effectiveness and safety.
People generally worry " radiation sterilization exists production safety problem to natural drug (comprising pollen) ".In December in 2006 7 days State Food and Drug Administration's file [No. (2006) 598, state's food medicine prison peace] " notice about regaining Lishizhen Medicine Group Co., Ltd.'s " drug's GMP certificate " ", just there is the defect of the said firm's " about 60Coradiation but do not have relative recording in bath manufacturing records, radiation dose cannot be provided ".
Therefore, fermented pollen increases secondary is safe according to sterilizing.
Accompanying drawing explanation
The change of Fig. 1 160 DEG C × 30min dry heat sterilization fermented pollen character.
The change of Fig. 2 170 DEG C × 20min dry heat sterilization fermented pollen character.
The Character change of Fig. 3 moist heat sterilization (121 DEG C × 15min) fermented pollen.
The Character change of Fig. 4 moist heat sterilization (116 DEG C × 40min) fermented pollen.
Claims (10)
1. a fermented pollen sterilizing methods for large-scale production, after it is characterized in that the fermented pollen sterilizing of large-scale production, bacterial number reaches the legal medicine of country or food quality standard, and fermented pollen effective ingredient and organoleptic indicator do not change substantially.
2. the fermented pollen sterilizing methods of claim 1, is characterized in that irradiation sterilization method, and the electron beam namely utilizing the gamma-rays of radioactive source radiation or electron accelerator to occur carries out ionizing radiation and reaches and kill method of microorganism in fermented pollen.
3. the irradiation sterilization method of claim 2, it is characterized in that preferred Co 60 (
60co)-gamma Rays sterilizing, radiation dose 1.0kGy ~ 20.0kGy, preferred radiation dose is 6.0kGy ~ 10.0kGy.
4. the fermented pollen sterilizing methods of the large-scale production of claim 1; it is characterized in that being not suitable for filtration sterilization, illumination sterilizing and gaseous sterilization process; moist heat sterilization, dry heat sterilization and microwave sterilization method significantly can change fermented pollen effective ingredient and organoleptic indicator, and difficulty can reach the legal medicine of country or food quality standard.
5. fermented pollen, after it is characterized in that one or more pollen add certain solution (comprising water) or preparation, the product at different temperatures after a period of time fermentation, this product contains one or more microorganisms such as antibacterial, mycete, escherichia coli.
6. the solution (comprising water) of claim 5 or preparation, it is characterized in that the product made with one or more raw materials (comprising water), some product adds microorganism.
7., after the fermented pollen sterilizing of claim 1, it is characterized in that the product that the fermented pollen of sterilizing is produced under " medicine (or food) production management specification " (GMP) condition, its bacterial number meet the legal medicine of country or food quality standard.
8. the drug standard that the country of claim 1 and 7 is legal, its feature is exactly the medicine sanitary standard that existing country promulgates, as the requirement of limitation standard in microbe in " Pharmacopoeia of People's Republic of China version in 2010 " that Chinese Pharmacopoeia Commission edits.
9. the food quality standard that the country of claim 1 and 7 is legal, its feature is exactly the food hygienic standard that existing country promulgates, as national quality technical supervision management general bureau has promulgated that " standard GB/T 14891.2-94 irradiation pollen sanitary standard " that health ministry is drafted and calendar year 2001 have promulgated the regulation of " standard GB/T 18526.6.2-2001 Irradiating pollen process for sterilizing " that health ministry is drafted for 1994.
10. the fermented pollen effective ingredient of claim 1 and organoleptic indicator, it is characterized in that fermented pollen effective ingredient refers to protein, fat, sugar, flavone compound, fermented pollen organoleptic indicator refers to color and luster, state, abnormal smells from the patient, flavour etc.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107875410A (en) * | 2017-10-27 | 2018-04-06 | 湖南省核农学与航天育种研究所 | A kind of sterilizing methods of ultra micro traditional Chinese medicine powder |
| CN110367430A (en) * | 2019-07-18 | 2019-10-25 | 中国辐射防护研究院 | One kind preventing the super calibration method of Polygala tenuifolia aflatoxins |
| CN111514320A (en) * | 2020-03-23 | 2020-08-11 | 贵阳学院 | Method for killing ulcer disease pathogenic bacteria PSA in kiwi fruit pollen |
| CN112294982A (en) * | 2020-09-21 | 2021-02-02 | 中国科学院武汉植物园 | Cobalt radiation sterilization method for kiwi fruit pollen with canker pathogen |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20050024872A (en) * | 2003-09-05 | 2005-03-11 | 주식회사 에스앤텍 | Fermented pollen extracts with antimicrobial activity |
-
2015
- 2015-07-01 CN CN201510376663.8A patent/CN104958771A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20050024872A (en) * | 2003-09-05 | 2005-03-11 | 주식회사 에스앤텍 | Fermented pollen extracts with antimicrobial activity |
Non-Patent Citations (5)
| Title |
|---|
| 中华人民共和国卫生部: "《GB 14891.2-94 辐照花粉卫生标准》", 31 August 1994 * |
| 中华人民共和国国家质量监督检验检疫总局: "《GB/T 18526.6.2-2001 花粉辐照杀菌工艺》", 31 March 2002 * |
| 国家药典委员会: "《中华人民共和国药典 2010年版（一部）》", 31 January 2010 * |
| 张学峰等: "《蜂产品加工》", 31 August 2002 * |
| 骆和生等: "《免疫中药学 中药免疫药理与临床》", 30 April 1999 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107875410A (en) * | 2017-10-27 | 2018-04-06 | 湖南省核农学与航天育种研究所 | A kind of sterilizing methods of ultra micro traditional Chinese medicine powder |
| CN110367430A (en) * | 2019-07-18 | 2019-10-25 | 中国辐射防护研究院 | One kind preventing the super calibration method of Polygala tenuifolia aflatoxins |
| CN111514320A (en) * | 2020-03-23 | 2020-08-11 | 贵阳学院 | Method for killing ulcer disease pathogenic bacteria PSA in kiwi fruit pollen |
| CN111514320B (en) * | 2020-03-23 | 2021-08-10 | 贵阳学院 | Method for killing ulcer disease pathogenic bacteria PSA in kiwi fruit pollen |
| CN112294982A (en) * | 2020-09-21 | 2021-02-02 | 中国科学院武汉植物园 | Cobalt radiation sterilization method for kiwi fruit pollen with canker pathogen |
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