CN104914025A - Method for rapidly detecting spore value in ganoderma spore powder - Google Patents
Method for rapidly detecting spore value in ganoderma spore powder Download PDFInfo
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- CN104914025A CN104914025A CN201510295671.XA CN201510295671A CN104914025A CN 104914025 A CN104914025 A CN 104914025A CN 201510295671 A CN201510295671 A CN 201510295671A CN 104914025 A CN104914025 A CN 104914025A
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Abstract
The invention relates to a determination method for a fungal spore wall breaking condition and particularly relates to a method for rapidly detecting a spore value in ganoderma spore. The method for rapidly detecting the spore value in the ganoderma spore powder, provided by the invention, is characterized by comprising the following steps: taking spore powder B to be detected to prepare a proper suspension solution; after enabling the volume to be constant, uniformly mixing; and observing and counting the quantity of complete spores and the quantity of wall-broken spores in a unit by using a microscope with a C6 objective lens micrometer; and calculating the spore value, namely the percentage value of the quantity of the complete spores in the total sum of the quantity of the complete spores and the quantity of the wall-broken spores. The determination method has the advantages that the area of the complete spores and the wall-broken spores of the uniformly-dispersed spore powder is counted by using the C6 micrometer, so that the aims of simplicity in operation, low price and representative detection results are realized.
Description
Technical field
The present invention relates to a kind of fungal spore powder miospore value detection method, especially to detect fast after breaking trachytectum of glossy ganoderma and without with the spore value in batch non-ganoderma spove powder, to belong to the field of Chinese medicines.
Background technology
Reishi sporule be glossy ganoderma in the growth and maturity phase, to be hit by a bullet that shoot out and small avette reproduction cell and the seed of glossy ganoderma from glossy ganoderma lamella.It has condensed the elite of glossy ganoderma, whole inhereditary material of tool glossy ganoderma and health-care effect.The shell of lucidum spore powder comprises the two-layer sporoderm be made up of very hard chitin and glucosan; Sporoderm quality is hard, has the character such as acid and alkali-resistance, anti-oxidant, withstand voltage heat resistanceheat resistant, the anti-digestion of resistance to enzyme, limits human body absorbing nutritional labeling in Reishi sporule.Sporoderm is broken or after eliminating, the effective constituent wrapped tightly by sporoderm could farthest directly be absorbed by human body stomach.Have data to show, the lucidum spore powder of broken wall is conducive to the stripping of the compositions such as triterpene, Thick many candies, crude fat, and wherein Thick many candies content exceeds 70% than non-ganoderma spove powder.If the directly lucidum spore powder of edible non-broken wall, human body only can utilize the effective constituent of about 12%, and takes the lucidum spore powder after broken wall, and utilization rate of active components can reach more than 95%.Therefore, can the size of the sporoderm-broken rate of ganoderma spove powder be exactly the key point that utilize effective component of ganoderma lucidum spore powder to the full extent.For lucidum spore powders numerous on market, sporoderm-broken rate is often by the important indicator as this product quality of measurement.
At present, the quality standard mainly sporoderm-broken rate of ganoderma spove powder, the mensuration Primary Reference Ministry of Agriculture standard " mensuration of NY/T1677-2008 ganoderma spove powder sporoderm-broken rate " of sporoderm-broken rate and national standard " GB/T29344-2012 lucidum spore powder pick and process technical manual " appendix A " assay method of ganoderma spove powder sporoderm-broken rate ".In these two standards, sporoderm-broken rate measuring principle is all counted by the spore of blood counting chamber to non-broken wall, then calculates the quantity of intact spore contained by unit weight, utilizes formula
Sporoderm-broken rate of spore (%)=(spore complete before spore/broken wall complete after 1-broken wall) × 100%
Calculate " sporoderm-broken rate " of spore.The typical curve between lucidum spore powder quality and spore number need be set up, utilize this typical curve to determine the spore number of non-spore powder with crushed sporoderm in same batch certain mass spore powder with crushed sporoderm to be measured, obtain the sporoderm-broken rate of spore powder with crushed sporoderm.But, this kind of method needs with non-spore powder with crushed sporoderm in batch spore powder with crushed sporoderm to be measured, and in real life consumer buy be ganoderma spove powder, cannot obtain with batch non-ganoderma spove powder, and then cannot detect its sporoderm-broken rate, thus the present invention proposes to detect its spore value.
Except blood counting chamber method, at present the assay method of the sporoderm-broken rate of open report also has water load in conjunction with the method for microtechnic, suspension method in conjunction with physics technology detecting method and chemical fingerprint detection method.But the detecting step of chemical composition analysis method is complicated, instrument is expensive, and testing cost is higher.
Summary of the invention
The object of the invention is to overcome above-mentioned weak point of the prior art, provide one to make the average degree of scatter of spore higher, measure the method for ganoderma spove powder spore value comparatively accurately.The spore value intact spore number be in ganoderma spove powder accounts for the percent value of intact spore number and exosporium-broken spore number summation.Because exosporium-broken spore number cannot count, adopt C6 type eyepiece micrometer, count its area, little lattice internal substance is designated as 1 little lattice more than 1/2, and discontented 1/2 is designated as 0 little lattice, and lucky 1/2 is designated as 0.5 little lattice.Respectively to non-wall-broken ganoderma spore and wall-broken ganoderma spore counting, non-wall-broken ganoderma spore number accounts for non-wall-broken ganoderma spore number and the mean percent ratio of exosporium-broken spore number summation, is spore value.
The present invention is a kind of method of spore value of quick detection Reishi sporule, it is characterized in that:
(1) get the lucidum spore powder B of spore value to be measured, dry to constant weight, and use on a certain amount of chloral hydrate test solution and glass mortar and repeatedly softly grind well, ultrasonic mixing, after be settled to same volume, mix, obtain corresponding suspending liquid;
(2) getting above-mentioned lucidum spore powder suspending liquid point drops on microslide, under the microscope, uses C6 type eyepiece micrometer, reads little lattice number shared by intact spore and broken spore respectively; Little lattice include intact spore or broken spore is designated as 1 little lattice more than 1/2, and discontented 1/2 is designated as 0 little lattice, and lucky 1/2 is designated as 0.5 little lattice; Utilize formula
Spore value (%)=[intact spore number ÷ (intact spore number+exosporium-broken spore number)] × 100%
Calculate spore value; Wherein, spore value refers to that the intact spore number in ganoderma spove powder accounts for the percent value of intact spore number and exosporium-broken spore number summation, and intact spore number refers to the grid shared by intact spore, and exosporium-broken spore number refers to the grid shared by broken spore.And with the spore value of conidia powder A for judgment criteria.
The present invention selects suspension to be chloral hydrate, after conidia powder is uniformly dispersed, quantitative point sample is in microslide, through test comparison, select the dispersion effect of chloral hydrate test solution conidia powder effective compared with zinc sulfate test solution etc., because its density and viscosity are conducive to the physical stability of spore in suspending liquid, but this is not the most important reason of the present invention.
In order to accurate measurement, the spore powder with crushed sporoderm used is required to be the ganoderma spove powder not adding any auxiliary material, and should remove impurity, dries to constant weight for 60 DEG C.
Reassemble to increase the dispersiveness of spore in chloral hydrate and reducing spore, after constant volume, carry out mechanical raking, as ultrasonic mixing.
After breaking trachytectum, the spore count of non-broken wall contained complete in the single visual field is less, in order to accurate measurement, the amount of the spore powder with crushed sporoderm got should suitably increase, if conidia powder amount is too many, on microslide, spore powder with crushed sporoderm is assembled, and can not form individual layer conidia powder under the visual field.
The invention has the advantages that and utilize same lucidium spore powder wall breaking and intact spore number to immobilize, dispersed in conidia powder suspending liquid, thus it is accurate to reach technology, and simple to operate cheap, the object that testing result is representative.
Accompanying drawing explanation
Spore distribution (the non-wall-broken ganoderma spore of note: A. under Fig. 1 microscope in zinc sulfate ethanolic solution; B. wall-broken ganoderma spore).
Non-exosporium-broken spore distribution (the non-wall-broken ganoderma spore of note: A. under Fig. 2 microscope after the process of this patent method; B. wall-broken ganoderma spore).
Exosporium-broken spore distribution (the non-wall-broken ganoderma spore of note: A. under Fig. 3 microscope after the process of this patent method; B. wall-broken ganoderma spore).
C6 eyepiece micrometer miospore distribution (the non-wall-broken ganoderma spore of note: A. under Fig. 4 microscope; B. wall-broken ganoderma spore).
The spore value analysis result of sample in Fig. 5 embodiment 1.
The spore value analysis result of sample in Fig. 6 embodiment 2.
The spore value analysis result of sample in Fig. 7 embodiment 3.
The spore value analysis result of sample in Fig. 8 embodiment 4.
Embodiment
Below further illustrate concrete steps of the present invention and condition:
Embodiment 1:
1, spore suspension is prepared
(1) get 4 parts and (derive from the sample of different manufacturers or same producer different batches respectively, in table 1, sample number into spectrum is P01-P04) lucidum spore powder A(broken wall Shuai≤98% of appropriate known sporoderm-broken rate) and commercially available unknown sporoderm-broken rate conidia powder B(conidia powder B adopt conventional method, namely national standard " GB/T29344-2012 lucidum spore powder pick and process technical manual " appendix A " assay method of ganoderma spove powder sporoderm-broken rate ", records its sporoderm-broken rate >98%; In table 1, sample number into spectrum is P05), dry 5 hours at 60 DEG C, baking oven.
(2) accurate quantification takes conidia powder A and the B(W=0.04g of drying).
(3) above-mentioned conidia powder is put into the glass mortar of diameter 5mm, repeatedly grind well conidia powder gently on a small quantity with chloral hydrate test solution makes its agglomerate disperse as far as possible, and with chloral hydrate constant volume in 5ml volumetric flask, more ultrasonic 20min, spore is fully disperseed in suspending liquid.
2, observe and count
(1) draw the above-mentioned detection spore suspension of 10 μ l with liquid-transfering gun, be placed in the edge of cover glass, liquid is slowly infiltrated, unnecessary liquid thieving paper is absorbed, and after waiting a moment, under 400 power microscopes, observes spore, as shown in Figure 2,3, spore on average disperses.
(2) under the microscope of 10 × 40 multiples, 5 × 5mm on C6 micrometer is read respectively
2intact spore number and exosporium-broken spore number in large lattice, moving stage scale, make 25 visuals field be evenly distributed in whole slide, the mean value in 25 visuals field is spore value.Every part of parallel 6 slides of liquid to be measured, calculate their average, N
aand N
b.
3, data calculate
(1) spore value (%)=[intact spore number ÷ (intact spore number+exosporium-broken spore number)] × 100%
Intact spore number is little lattice number shared by intact spore, and exosporium-broken spore number is little lattice number shared by exosporium-broken spore, and concrete outcome asks for an interview Fig. 5.
The spore value analysis result (see Figure of description) of Fig. 5 sample
Note: sample source is as follows: P01(Sichuan Fu Zheng medicine company Ltd, lot number: 140801); P02(Chengdu Kanghua Pharmaceutical Co., Ltd., lot number: L140701); P03(Chengdu Kanghua Pharmaceutical Co., Ltd., lot number: L131101); P04(Chengdu Kanghua Pharmaceutical Co., Ltd., lot number: L140101); P05(lotus pond Chinese Medicinal Materials Markets).
From the data in Fig. 5, utilize detection method provided by the invention can detect the spore value of sample fast, and result is accurate, with its corresponding sporoderm-broken rate of broken wall situation of its reaction Reishi sporule in error range, simple to operate cheap, there is promotional value.
Embodiment 2:
1, spore suspension is prepared
(1) the non-broken wall of lucidum spore powder C(of appropriate known sporoderm-broken rate is got), dry 5 hours at 60 DEG C, baking oven.
(2) accurate quantification takes the conidia powder C(W=0.04g of drying).
(3) above-mentioned conidia powder is put into the glass mortar of diameter 5mm, repeatedly grind well conidia powder gently on a small quantity with chloral hydrate test solution makes its agglomerate disperse as far as possible, and with chloral hydrate constant volume in 5ml volumetric flask, more ultrasonic 20min, spore is fully disperseed in suspending liquid.
2, observe and count
(1) draw the above-mentioned detection spore suspension of 10 μ l with liquid-transfering gun, be placed in the edge of cover glass, liquid is slowly infiltrated, unnecessary liquid thieving paper is absorbed, and after waiting a moment, under 400 power microscopes, observes spore, as shown in Figure 2,3, spore on average disperses.
(3) under the microscope of 10 × 40 multiples, 5 × 5mm on C6 micrometer is read respectively
2intact spore number and exosporium-broken spore number in large lattice, moving stage scale, make 25 visuals field be evenly distributed in whole slide, the mean value in 25 visuals field is spore value.Every part of parallel 6 slides of liquid to be measured, calculate their average, N
c.
4, data calculate
(1) spore value (%)=[intact spore number ÷ (intact spore number+exosporium-broken spore number)] × 100%
Intact spore number is little lattice number shared by intact spore, and exosporium-broken spore number is little lattice number shared by exosporium-broken spore, and concrete outcome asks for an interview Fig. 6.
The spore value analysis result (see Figure of description) of Fig. 6 sample
Embodiment 3:
1, spore suspension is prepared
(1) the lucidum spore powder D(sporoderm-broken rate 50% of appropriate known sporoderm-broken rate is got), dry 5 hours at 60 DEG C, baking oven.
(2) accurate quantification takes the conidia powder D(W=0.04g of drying).
(3) above-mentioned conidia powder is put into the glass mortar of diameter 5mm, repeatedly grind well conidia powder gently on a small quantity with chloral hydrate test solution makes its agglomerate disperse as far as possible, and with chloral hydrate constant volume in 5ml volumetric flask, more ultrasonic 20min, spore is fully disperseed in suspending liquid.
2, observe and count
(1) draw the above-mentioned detection spore suspension of 10 μ l with liquid-transfering gun, be placed in the edge of cover glass, liquid is slowly infiltrated, unnecessary liquid thieving paper is absorbed, and after waiting a moment, under 400 power microscopes, observes spore, as shown in Figure 2,3, spore on average disperses.
(4) under the microscope of 10 × 40 multiples, 5 × 5mm on C6 micrometer is read respectively
2intact spore number and exosporium-broken spore number in large lattice, moving stage scale, make 25 visuals field be evenly distributed in whole slide, the mean value in 25 visuals field is spore value.Every part of parallel 6 slides of liquid to be measured, calculate their average N
d.
5, data calculate
(1) spore value (%)=[intact spore number ÷ (intact spore number+exosporium-broken spore number)] × 100%
Intact spore number is little lattice number shared by intact spore, and exosporium-broken spore number is little lattice number shared by exosporium-broken spore, and concrete outcome asks for an interview Fig. 7.
The spore value analysis result (see Figure of description) of Fig. 7 sample
Embodiment 4:
1, spore suspension is prepared
(1) the lucidum spore powder E(sporoderm-broken rate 75% of appropriate known sporoderm-broken rate is got), dry 5 hours at 60 DEG C, baking oven.
(2) accurate quantification takes the conidia powder E(W=0.04g of drying).
(3) above-mentioned conidia powder is put into the glass mortar of diameter 5mm, repeatedly grind well conidia powder gently on a small quantity with chloral hydrate test solution makes its agglomerate disperse as far as possible, and with chloral hydrate constant volume in 5ml volumetric flask, more ultrasonic 20min, spore is fully disperseed in suspending liquid.
2, observe and count
(1) draw the above-mentioned detection spore suspension of 10 μ l with liquid-transfering gun, be placed in the edge of cover glass, liquid is slowly infiltrated, unnecessary liquid thieving paper is absorbed, and after waiting a moment, under 400 power microscopes, observes spore, as shown in Figure 2,3, spore on average disperses.
(5) under the microscope of 10 × 40 multiples, 5 × 5mm on C6 micrometer is read respectively
2intact spore number and exosporium-broken spore number in large lattice, moving stage scale, make 25 visuals field be evenly distributed in whole slide, the mean value in 25 visuals field is spore value.Every part of parallel 6 slides of liquid to be measured, calculate their average N
e.
6, data calculate
(1) spore value (%)=[intact spore number ÷ (intact spore number+exosporium-broken spore number)] × 100%
Intact spore number is little lattice number shared by intact spore, and exosporium-broken spore number is little lattice number shared by exosporium-broken spore, and concrete outcome asks for an interview Fig. 8.
The spore value analysis result (see Figure of description) of Fig. 8 sample
Claims (4)
1. detect a method for lucidum spore powder miospore value fast, it is characterized in that:
Get lucidum spore powder B to be measured, dry to constant weight, and repeatedly softly grind well with on a certain amount of chloral hydrate test solution and glass mortar, ultrasonic mixing, after be settled to same volume, mix, obtain corresponding suspending liquid;
Getting above-mentioned lucidum spore powder suspending liquid point drops on microslide, under the microscope, uses C6 type eyepiece micrometer, reads little lattice number shared by intact spore and broken spore respectively; Wherein, little lattice include intact spore or broken spore is designated as 1 little lattice more than 1/2, and discontented 1/2 is designated as 0 little lattice, and lucky 1/2 is designated as 0.5 little lattice; Utilize formula
Spore value (%)=
× 100%
Calculate spore value; Wherein, spore value refers to that the intact spore number in ganoderma spove powder accounts for the percent value of intact spore number and exosporium-broken spore number summation, and intact spore number refers to the grid shared by intact spore, and exosporium-broken spore number refers to the grid shared by broken spore.
2. a method for quick detection lucidum spore powder miospore value according to claim 1, is characterized in that, use density and viscosity all larger chloral hydrate test solution mixing conidia powder, make it not lump, be convenient to observe counting.
3. a method for quick detection Reishi sporule oidium value according to claim 1, is characterized in that, conidia powder B to be measured is first dried before using.
4. a method for quick detection Reishi sporule oidium value according to claim 1, it is characterized in that, described mixing refers to and first repeatedly softly grinds well with glass mortar, ultrasonic mixing after constant volume.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113155251A (en) * | 2021-04-02 | 2021-07-23 | 陕西金慧方中药科技有限公司 | Method for measuring weight of orchid seeds |
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2015
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| CN113155251A (en) * | 2021-04-02 | 2021-07-23 | 陕西金慧方中药科技有限公司 | Method for measuring weight of orchid seeds |
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