A kind of detection method of Chinese materia medica preparation Low Back Pain ball
Technical field
The present invention relates to the quality determining methods of Chinese materia medica preparation, the in particular to quality determining method of Low Back Pain ball.
Background technique
Pain in the loins is common in a variety of diseases such as kidney trouble, rheumatism, lumbar muscle strain and wound, and Chinese medicine thinks mostly due to impression
Cold-dampness, the damp and hot, qi depression to blood stasis of impression, deficiency of the kidney is physically weak causes, so Chinese patent drugs for treatment pain in the loins, it is necessary to dialectical according to cause of disease difference
It selects.
Low Back Pain ball is made of southern rattan, psoralea corylifolia, teasel root, pink reineckea herb, radix achyranthis bidentatae, Chinese yam, and Fang Zhongnan rattan temperature leads to evacuate pathogenic wind, and detumescence stops
Bitterly strong waist and knee, tonifying kidney and strengthening yang is relieving cough and asthma, promoting blood circulation and stopping pain;Promoting the circulation of qi that psoralea corylifolia is supporing yang;Pink reineckea herb regulating blood condition removing toxic substances, evacuate pathogenic wind synthetism;It is continuous
Disconnected and radix achyranthis bidentatae nourishing liver and kidney, strengthening the bones and muscles, dispelling stasis of blood and stimulating the menstrual flow;Chinese yam tonifying spleen nourishing the stomach, engender liquid and benefic lung, kidney tonifying arresting seminal emission;All medicine compatibilities, gather altogether
The benefits of replenishing kidney, strengthening waist, the promoting flow of qi and blood circulation, removing blood stasis and acesodyne.It is mainly used for qi depression to blood stasis caused by treating traumatic injury and lumbar muscle strain etc.
The pain in the loins of card.
Existing Low Back Pain ball quality standard determines psoralen, Isopsoralen, asperosaponin VI, oleanolic acid
Property identify, and have psoralen, Isopsoralen assay, but existing detection method is complicated for operation, precision is not high, practical
Mistake, the controllability existing defects of control of product quality specificity are easy to appear in operating process, it is difficult to Accurate Determining Low Back Pain master
Want composition.
The present invention after study, increases the microscopical characters item of finished product preparation, respectively to Chinese yam in prescription, stir-baked SEMEN PSORALEAE with salt solution,
Teasel root and southern rattan have carried out the identification of microscopic features, it is ensured that containing the main component of major function in preparation, play curative effect.Also mention
Having gone out can be with Accurate Determining Low Back Pain ball to the related substance accurate discrimination method of progress and accurate content assaying method, this method
Quality, it is exclusive it is controllable, stablize, be easy, feasible.
Summary of the invention
The purpose of the present invention is to provide a kind of detection methods of Chinese materia medica preparation Low Back Pain ball
Low Back Pain ball of the present invention, formula composition and the preparation method is as follows:
Preparation method: the above Six-element is ground into fine powder, is sieved, and mixes.Every 100g powder adds 140~150g of refined honey that young mistress is made
Ball to get.
Quality determining method of the present invention, including discrimination method and content assaying method.
Method of the invention shares 4, and any of them one measuring method as Low Back Pain ball can be used, can also group
It closes used as the measuring method to Low Back Pain ball, is preferably applied in combination.
Detection method of the present invention, including discrimination method and content assaying method, wherein discrimination method includes micro-
Sem observation method and thin-layered chromatography,
Microscopic observation, this method include to Chinese yam in prescription, stir-baked SEMEN PSORALEAE with salt solution, teasel root and southern rattan microscopic features mirror
Fixed, discrimination method is as follows:
Take Low Back Pain ball, set microscopically observation: amylum body triangular shape is oval or square is round, and 24~40 μm of diameter, omphalion is short
Gap-like or herringbone shape are Chinese yam;Class polygonal is seen on kind skin palisade cells light brown or rufous, surface, and wall is slightly thick, and cell is containing red
Brown object is stir-baked SEMEN PSORALEAE with salt solution;45 μm of calcium oxalate cluster crystal diameter medicine, is present in the parenchyma cell of light brown yellow shrinkage, constant
It is arranged in rows, is teasel root;Lithocyte is faint yellow, and class ovum, class be rectangular or rectangle, and diameter is about to 60 μm, and wall is thicker, and pit is thin
Close, cavity is larger, for southern rattan.
Thin-layered chromatography, this method include the identification to psoralen, Isopsoralen, asperosaponin, oleanolic acid,
Discrimination method is as follows:
(1) Low Back Pain ball is taken, ethyl acetate is added, is heated to reflux, is filtered, filtrate is evaporated, and residue adds ethyl acetate to make to dissolve, and makees
For test solution;Psoralea corylifolia control medicinal material separately is taken, adds ethyl acetate, is made in the same way of control material solution;Psoralen is taken again
Reference substance, Isopsoralen reference substance, add ethyl acetate that mixed solution is made, as reference substance solution;It is tried according to thin-layered chromatography
It tests, draws above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, using normal hexane-ethyl acetate as solvent, taken
Out, it dries, sprays with alcoholic caustic potash, set and inspected under ultraviolet lamp;In sample chromatogram, with reference medicine chromatography and right
According on the corresponding position of product chromatography, the fluorescence spot of same colored is shown;
(2) Low Back Pain ball is taken, methanol is added, is heated to reflux, is filtered, filtrate is steamed near dry, adds water low-grade fever to make to dissolve, be saturated with water
N-butanol shake extract, merge n-butanol extracting liquid, washed with ammonia solution, n-butanol liquid is evaporated, and it is molten that residue adds water low-grade fever to make
Solution, is added on polyamide column, with ethanol elution, collects eluent, is evaporated, residue adds methanol to make to dissolve, as test solution;
Asperosaponin VI reference substance is taken, adds methanol that solution is made, as reference substance solution;It tests, draws above-mentioned according to thin-layered chromatography
Two kinds of solution are put respectively on same silica gel g thin-layer plate, using chloroform-methanol-water as solvent, are unfolded, take out, dry in the air
On dry, with ethanol solution of sulfuric acid, it is clear to be heated to spot development for spray, inspects under exposure;In sample chromatogram, with reference substance
On the corresponding position of chromatography, the spot of same color is shown;
(3) Low Back Pain ball is taken, ethanol solution hydrochloride is added, is heated to reflux, is filtered, solution, which is concentrated into, closely to be done, and adds water heating to make molten
Solution stands, takes supernatant, is shaken and is extracted with chloroform, merges chloroform liquid, is evaporated, and residue adds methanol to make to dissolve in right amount,
Add neutral alumina, mixes thoroughly, it is dry, is fitted into chromatographic column, successively with water, methanol elution, discards eluent, then with three chloromethanes
Alkane-methanol elution, collects eluent, is evaporated, residue adds methanol to make to dissolve, as test solution;Radix achyranthis bidentatae control medicinal material separately is taken,
Add ethanol solution hydrochloride, is made in the same way of control medicinal material solution;Oleanolic acid reference substance is taken again, adds methanol that solution is made, as right
According to product solution;It is tested according to thin-layered chromatography, draws test solution, control medicinal material and reference substance solution, put respectively in same silicon
On glue G lamellae, using thiacyclohexane-chloroform-acetic ether-methanoic acid as solvent, takes out, dry, spray with vanillin-sulfuric acid
Solution, it is clear to be heated to spot development, inspects in the sunlight;In sample chromatogram, with reference medicine chromatography and reference substance color
It composes on corresponding position, shows the spot of identical color.
Preferably, identified using thin-layered chromatography, the method is as follows:
(1) Low Back Pain ball 3g is taken, is shredded, ethyl acetate 30ml is added, is heated to reflux 30 minutes, is filtered, filtrate is evaporated, and residue adds
Ethyl acetate 2ml makes to dissolve, as test solution;Psoralea corylifolia control medicinal material 0.3g separately is taken, adds ethyl acetate 10ml, same to legal system
Material solution is shone in pairs;Psoralen reference substance, Isopsoralen reference substance are taken again, are added ethyl acetate that every 1ml is made and are contained
The mixed solution of 0.2mg, as reference substance solution;It is tested according to thin-layered chromatography, draws above-mentioned each 2~5 μ l of three kinds of solution, respectively
Point is on same silica gel g thin-layer plate, with normal hexane-ethyl acetate (15:4) for solvent, is unfolded 2 times, takes out, dry, spray with
10% alcoholic caustic potash is set and is inspected under ultraviolet lamp (365nm);In sample chromatogram, with reference medicine chromatography and right
According on the corresponding position of product chromatography, the fluorescence spot of same colored is shown;
(2) Low Back Pain ball 6g is taken, is shredded, methanol 40ml is added, though heat reflux 30 minutes, filtration, filtrate is steamed near dry, and adds water
25ml low-grade fever makes to dissolve, and the n-butanol shaking being saturated with water is extracted 2 times, each 30ml, merges n-butanol extracting liquid, uses ammonia solution
60ml washing, n-butanol liquid is evaporated, and residue adds water 15ml low-grade fever to make to dissolve, be added in polyamide column (60-100 mesh, 3g internal diameter 2cm,
With water prewashing) on, it is eluted with 80% ethyl alcohol 60ml, collects eluent, be evaporated, residue adds methanol 3ml to make to dissolve, as examination
Product solution;Asperosaponin VI reference substance is taken, adds methanol that solution of every 1ml containing 1mg is made, as reference substance solution;According to thin layer
Chromatography experiment draws each 2~5 μ l of above two solution, is put respectively on same silica gel g thin-layer plate, with chloroform-first
10 DEG C of alcohol-water (13:7:2) lower layer's solution arranged below is solvent, is unfolded, and takes out, dries, and is sprayed with 10% sulfuric acid second
Alcoholic solution, it is clear to be heated to spot development at 105 DEG C, inspects under exposure;In sample chromatogram, corresponding to reference substance chromatography
Position on, show same color spot;
(3) this product 18g is taken, is shredded, adds ethanol solution hydrochloride (1 → 10) 80ml, is heated to reflux 1 hour, is filtered, solution is dense
It is reduced to and closely does, add water 40ml heating to make to dissolve, stand, take supernatant, shake extraction 3 times, each 30ml, merging with chloroform
Chloroform liquid, is evaporated, and residue adds methanol to make to dissolve in right amount, adds neutral alumina (100-200 mesh) 3g, mixes thoroughly, dry, is packed into
In chromatographic column (internal diameter 0.9cm), successively with each 30ml elution of water, 20% methanol, eluent is discarded, then with chloroform-first
Alcohol (20:1) 40ml elution, collects eluent, is evaporated, residue adds methanol 1ml to make to dissolve, as test solution;Separately take radix achyranthis bidentatae
Control medicinal material 0.5g, adding ethanol solution hydrochloride, ((1 → 10) 40ml, is made in the same way of control medicinal material solution;Oleanolic acid is taken to compare again
Product add methanol that solution of every 1ml containing 0.2mg is made, as reference substance solution;It is tested according to thin-layered chromatography, it is molten to draw test sample
10~20 μ l of liquid, control medicinal material and each 10 μ l of reference substance solution are put respectively on same silica gel g thin-layer plate, with thiacyclohexane-trichlorine
Methane-acetic ether-methanoic acid (20:5:8:0.1) is solvent, takes out, dries, and is sprayed with 5% vanillin-sulfuric acid solution, 105
It is clear DEG C to be heated to spot development, inspects in the sunlight;In sample chromatogram, with reference medicine chromatography and reference substance chromatography phase
On the position answered, the spot of identical color is shown.
Content assaying method of the present invention is to carry out assay to the effective ingredient in Low Back Pain ball.
Wherein the effective ingredient is in terms of the total amount of psoralen (C11H6O3) and Isopsoralen (C11H6O3).
Content assaying method of the invention uses high-efficient liquid phase analysis method.
Content assaying method of the present invention, comprising the following steps:
(1) preparation of reference substance solution: taking psoralen reference substance, Isopsoralen reference substance appropriate, accurately weighed, adds
Methanol be made mixed solution to get;
(2) preparation of test solution: taking small honey pill or takes big honeyed bolus under weight differential item, shreds, and mixes, and sets tool plug
In conical flask, methanol, weighed weight is added, ultrasonic treatment lets cool, then weighed weight, the weight of less loss is supplied with methanol, is shaken
It is even, filtration, take subsequent filtrate to get;
(3) measuring method: it is accurate respectively to draw reference substance solution and each 10 μ l of test solution, liquid chromatograph is injected, is surveyed
It is fixed to get;
Chromatographic condition: C18 chromatographic column is used, using octadecylsilane chemically bonded silica as filler;It is flowing with acetonitrile-water
Phase.
Preferably, content assaying method of the present invention, comprising the following steps:
(1) preparation of reference substance solution: taking psoralen reference substance, Isopsoralen reference substance appropriate, accurately weighed, adds
Methanol be made every 1ml contain the mixed solution of 10 μ l to get;
(2) preparation of test solution: taking small honey pill or takes big honeyed bolus under weight differential item, shreds, and mixes, takes about
0.6g, it is accurately weighed, it sets in stuffed conical flask, methanol 50ml is added in precision, and weighed weight is ultrasonically treated (power 500W, frequency
40kHz) 1 hour, let cool, then weighed weight, the weight of less loss supplied with methanol, is shaken up, filter, take subsequent filtrate to get;
(3) measuring method: it is accurate respectively to draw reference substance solution and each 10 μ l of test solution, liquid chromatograph is injected, is surveyed
It is fixed to get;
Chromatographic condition: DIKMA, C18 chromatographic column, using octadecylsilane chemically bonded silica as filler are used;With acetonitrile-water
(30:70) is mobile phase;The present invention is isocratic elution, column temperature: flow velocity 1.0ml/min 25 DEG C, Detection wavelength 246nm, is managed
3000 should be not less than by calculating by plate number by psoralen peak.
Reference substance solution determination data table
Test solution determination data table
Calculation formula:
Content average value=2.76mg/g relative deviation (%)=1.3%
Control item: this product saliferous psoralen is with the total amount of psoralen (C11H6O3) and Isopsoralen (C11H6O3)
Meter, the big every ball of honeyed bolus must not be less than 10.26mg;The every 1g of small honey pill must not be less than 1.14mg.
Beneficial effects of the present invention: the side of the content of effective component in the liquid chromatogram measuring Low Back Pain ball that the present invention establishes
Method meets the requirement of methodology validation, and easy to operate, high sensitivity, and measurement is accurate, and strong applicability can be used in the production
The quality of product controls.Meanwhile the blank that the product quantitatively controls has been filled up, so that being able to carry out more effective quality to the product
Analysis more comprehensively reflects the quality condition of product, guarantees the quality stability of the product.Meanwhile the present invention is identified by thin layer
Method identifies Low Back Pain ball Chinese medicine, improves the quality and safety of drug.
Specific embodiment
The present invention is further illustrated by the following examples, but not as limitation of the present invention.
The identification of Chinese yam, stir-baked SEMEN PSORALEAE with salt solution, teasel root, Nan Teng (Hance Pepper Stem and Leaf) in 1 Low Back Pain ball of embodiment
(1) take Low Back Pain ball, set microscopically observation: amylum body triangular shape is oval or square is round, and 24~40 μm of diameter, omphalion
Short gap-like or herringbone shape (Chinese yam).Class polygonal is seen on kind skin palisade cells light brown or rufous, surface, and wall is slightly thick, and cell contains
Rufous object (stir-baked SEMEN PSORALEAE with salt solution).45 μm of calcium oxalate cluster crystal diameter medicine, is present in the parenchyma cell of light brown yellow shrinkage, constant
Be arranged in rows (teasel root).Lithocyte is faint yellow, and class ovum, class be rectangular or rectangle, and diameter is about to 60 μm, and wall is thicker, and pit is thin
Close, cavity is larger (southern rattan (Hance Pepper Stem and Leaf))
(2) Low Back Pain ball 3g is taken, is shredded, ethyl acetate 30ml is added, is heated to reflux 30 minutes, is filtered, filtrate is evaporated, and residue adds
Ethyl acetate 2ml makes to dissolve, as test solution.Psoralea corylifolia control medicinal material 0.3g separately is taken, adds ethyl acetate 10ml, same to legal system
Material solution is shone in pairs.Psoralen reference substance, Isopsoralen reference substance are taken again, are added ethyl acetate that every 1ml is made and are contained
The mixed solution of 0.2mg, as reference substance solution.It is tested according to thin-layered chromatography, draws above-mentioned each 2~5 μ l of three kinds of solution, respectively
Point is on same silica gel g thin-layer plate, with normal hexane-ethyl acetate (15:4) for solvent, is unfolded 2 times, takes out, dry, spray with
10% alcoholic caustic potash is set and is inspected under ultraviolet lamp (365nm).In sample chromatogram, with reference medicine chromatography and right
According on the corresponding position of product chromatography, the fluorescence spot of same colored is shown.
(3) Low Back Pain ball 6g is taken, is shredded, methanol 40ml is added, though heat reflux 30 minutes, filtration, filtrate is steamed near dry, and adds water
25ml low-grade fever makes to dissolve, and the n-butanol shaking being saturated with water is extracted 2 times, each 30ml, merges n-butanol extracting liquid, uses ammonia solution
60ml washing, n-butanol liquid is evaporated, and residue adds water 15ml low-grade fever to make to dissolve, be added in polyamide column (60-100 mesh, 3g internal diameter 2cm,
With water prewashing) on, it is eluted with 80% ethyl alcohol 60ml, collects eluent, be evaporated, residue adds methanol 3ml to make to dissolve, as examination
Product solution.Asperosaponin VI reference substance is taken, adds methanol that solution of every 1ml containing 1mg is made, as reference substance solution.According to thin layer
Chromatography experiment draws each 2~5 μ l of above two solution, is put respectively on same silica gel g thin-layer plate, with chloroform-first
10 DEG C of alcohol-water (13:7:2) lower layer's solution arranged below is solvent, is unfolded, and takes out, dries, and is sprayed with 10% sulfuric acid second
Alcoholic solution, it is clear to be heated to spot development at 105 DEG C, inspects under exposure.In sample chromatogram, corresponding to reference substance chromatography
Position on, show same color spot.
(4) this product 18g is taken, is shredded, adds ethanol solution hydrochloride (1 → 10) 80ml, is heated to reflux 1 hour, is filtered, solution is dense
It is reduced to and closely does, add water 40ml heating to make to dissolve, stand, take supernatant, shake extraction 3 times, each 30ml, merging with chloroform
Chloroform liquid, is evaporated, and residue adds methanol to make to dissolve in right amount, adds neutral alumina (100-200 mesh) 3g, mixes thoroughly, dry, is packed into
In chromatographic column (internal diameter 0.9cm), successively with each 30ml elution of water, 20% methanol, eluent is discarded, then with chloroform-first
Alcohol (20:1) 40ml elution, collects eluent, is evaporated, residue adds methanol 1ml to make to dissolve, as test solution.Separately take radix achyranthis bidentatae
Control medicinal material 0.5g, adding ethanol solution hydrochloride, ((1 → 10) 40ml, is made in the same way of control medicinal material solution.Oleanolic acid is taken to compare again
Product add methanol that solution of every 1ml containing 0.2mg is made, as reference substance solution.It is tested according to thin-layered chromatography, it is molten to draw test sample
10~20 μ l of liquid, control medicinal material and each 10 μ l of reference substance solution are put respectively on same silica gel g thin-layer plate, with thiacyclohexane-trichlorine
Methane-acetic ether-methanoic acid (20:5:8:0.1) is solvent, takes out, dries, and is sprayed with 5% vanillin-sulfuric acid solution, 105
It is clear DEG C to be heated to spot development, inspects in the sunlight.In sample chromatogram, with reference medicine chromatography and reference substance chromatography phase
On the position answered, the spot of identical color is shown.
Embodiment 2
The assay of psoralen and isopsorapen in Low Back Pain ball
Chromatographic condition: DIKMA, C18 chromatographic column, using octadecylsilane chemically bonded silica as filler are used;With acetonitrile-water
(30:70) is mobile phase;Flow velocity is 1.0ml/min, Detection wavelength 246nm.Number of theoretical plate should not by the calculating of psoralen peak
Lower than 3000.
The preparation of reference substance solution: taking psoralen reference substance, Isopsoralen reference substance appropriate, accurately weighed, adds first
Alcohol be made every 1ml contain the mixed solution of 10 μ l to get.
The preparation of test solution: taking small honey pill or takes big honeyed bolus under weight differential item, shreds, and mixes, takes about 0.6g,
It is accurately weighed, it sets in stuffed conical flask, methanol 50ml is added in precision, and weighed weight is ultrasonically treated (power 500W, frequency
40kHz) 1 hour, let cool, then weighed weight, the weight of less loss supplied with methanol, is shaken up, filter, take subsequent filtrate to get.Measurement
Method: it is accurate respectively to draw reference substance solution and each 10 μ l of test solution, inject liquid chromatograph, measurement to get.This product saliferous
For psoralea corylifolia in terms of the total amount of psoralen and isopsorapen, the big every ball of honeyed bolus is no less than 10.26mg, and the every 1g of small honey pill is no less than
1.14mg。
The screening process of test example 1, content assaying method of the present invention
1. the selection of extraction conditions: being dissolved in methanol, ethyl alcohol, benzene, chloroform, acetone according to psoralen, Isopsoralen
Ultrasonic extraction is selected in property, experiment.Respectively with methanol ultrasonic 0.5 hour, 1 hour, 1,5 hour, 2 hours, the results showed that ultrasound
Time is that 1 hour effect is relatively good.In an experiment, respectively using methanol, water, ether as solvent, ultrasonic treatment 1 hour, as a result
Show to measure that effect is best with methyl alcohol process sample, and the impurity that methanol extracts is few, can more preferably protect chromatographic column, therefore we
Method is using methanol as Extraction solvent.
The selection of 1 ultrasonic time of table
The selection of 2 Extraction solvent of table
2. chromatographic condition is investigated
The selection of 2.1 mobile phases: experiment is first investigated with four kinds of not year-on-year mobile phases of acetonitrile and water, ratio point
Not are as follows: 20:55,25:60,30:70,35:75, the results showed that, in the mobile phase of four kinds of different ratios, acetonitrile: water (30:70) point
Best from effect, sample separating degree meets regulation.
2.2 chromatographic conditions and system suitability test: using octadecylsilane chemically bonded silica as filler;With acetonitrile-water
(30:70) is mobile phase;Flow velocity is 1.0ml/min;Column temperature: 25 DEG C, Detection wavelength 246nm.Number of theoretical plate presses psoralen
Peak calculating should be not less than 3000. psoralen (C11H6O3) and Isopsoralen (C11H6O3) reference substance (Zhong Jian institute, Chengdu Puffy
Moral provides, for assay);Acetonitrile is chromatographically pure, other reagents are that analysis is pure.
The preparation of 2.3 reference substance solutions: taking psoralen reference substance, Isopsoralen reference substance appropriate, accurately weighed, adds
Methanol be made every 1ml contain the mixed solution of 10 μ l to get.
The preparation of 2.4 test solutions: taking small honey pill or takes big honeyed bolus under weight differential item, shreds, and mixes, takes about
0.6g, it is accurately weighed, it sets in stuffed conical flask, methanol 50ml is added in precision, and weighed weight is ultrasonically treated (power 500W, frequency
40kHz) 1 hour, let cool, then weighed weight, the weight of less loss supplied with methanol, is shaken up, filter, take subsequent filtrate to get.
3, linear relationship investigation table, table 3
4, precision test
Precision draws psoralen, Isopsoralen reference substance solution, continuous sample introduction 5 times, measures peak area, as a result table 4:
5, stability test
Sample is taken, presses psoralen under [assay] project, the method for Isopsoralen prepares test solution, difference
It is measured in accordance with the law in matching to postpone the 0th, 2,4,6,8 hour, the results are shown in Table 5.
5 stability test investigation table of table
6. reappearance test
Take same sample respectively, 5 parts of stable precision, by high performance liquid chromatography continuous sample introduction 5 times, the results showed that this law weight
Existing property is good, the results are shown in Table 6
7. sample-adding recovery test: precision weighs five parts of sample of knowledge content, is separately added into a certain amount of psoralea corylifolia
Element, Isopsoralen reference substance the results are shown in Table by the method measurement of psoralen, Isopsoralen under [assay] item
7:
Table 7, sample-adding recovery test result
Psoralen average recovery rate is 97.26%, RSD:1.3%, and Isopsoralen average recovery rate is 95.75%,
RSD:0.9%, sample recovery rate test result show that this law rate of recovery is good.
8. the assay of ten batches of samples: by the method for psoralen, Isopsoralen under [assay] item to ten batches
Sample has carried out assay, the results are shown in Table 8:
The assay result of 8, ten batches of samples of table
According to said determination as a result, preferred this product of the present invention contains stir-baked SEMEN PSORALEAE with salt solution with psoralen and isopsorapen
Total amount meter, the big every ball of honeyed bolus is no less than 10.26mg, and the every 1g of small honey pill is no less than 1.14mg.