CN105823830B - One surveys the methods for commenting tanshin polyphenolic acid B and schizandrin content in method measurement Yixinfumai particle more - Google Patents
One surveys the methods for commenting tanshin polyphenolic acid B and schizandrin content in method measurement Yixinfumai particle more Download PDFInfo
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- CN105823830B CN105823830B CN201510013254.1A CN201510013254A CN105823830B CN 105823830 B CN105823830 B CN 105823830B CN 201510013254 A CN201510013254 A CN 201510013254A CN 105823830 B CN105823830 B CN 105823830B
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- YEFOAORQXAOVJQ-RZFZLAGVSA-N schisandrol a Chemical compound C1[C@H](C)[C@@](C)(O)CC2=CC(OC)=C(OC)C(OC)=C2C2=C1C=C(OC)C(OC)=C2OC YEFOAORQXAOVJQ-RZFZLAGVSA-N 0.000 title claims abstract description 196
- 229930193195 schizandrin Natural products 0.000 title claims abstract description 98
- YEFOAORQXAOVJQ-UHFFFAOYSA-N wuweizischun A Natural products C1C(C)C(C)(O)CC2=CC(OC)=C(OC)C(OC)=C2C2=C1C=C(OC)C(OC)=C2OC YEFOAORQXAOVJQ-UHFFFAOYSA-N 0.000 title claims abstract description 98
- 239000002253 acid Substances 0.000 title claims abstract description 97
- 239000002245 particle Substances 0.000 title claims abstract description 82
- 238000000034 method Methods 0.000 title claims abstract description 62
- 238000005259 measurement Methods 0.000 title claims abstract description 16
- 239000013558 reference substance Substances 0.000 claims abstract description 59
- 239000012085 test solution Substances 0.000 claims abstract description 38
- 239000000243 solution Substances 0.000 claims abstract description 34
- 238000010828 elution Methods 0.000 claims abstract description 23
- 238000012937 correction Methods 0.000 claims abstract description 21
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 144
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 54
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 34
- 238000002360 preparation method Methods 0.000 claims description 31
- 238000001514 detection method Methods 0.000 claims description 22
- 239000012071 phase Substances 0.000 claims description 22
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 17
- 235000019253 formic acid Nutrition 0.000 claims description 17
- 239000012467 final product Substances 0.000 claims description 16
- 239000000706 filtrate Substances 0.000 claims description 14
- 238000004090 dissolution Methods 0.000 claims description 11
- 238000002604 ultrasonography Methods 0.000 claims description 11
- 239000000463 material Substances 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 7
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 7
- 240000006079 Schisandra chinensis Species 0.000 claims description 6
- 235000008422 Schisandra chinensis Nutrition 0.000 claims description 6
- 235000019640 taste Nutrition 0.000 claims description 5
- 241000208340 Araliaceae Species 0.000 claims description 4
- 239000009636 Huang Qi Substances 0.000 claims description 4
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 claims description 4
- 235000003140 Panax quinquefolius Nutrition 0.000 claims description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- 235000008434 ginseng Nutrition 0.000 claims description 4
- 238000004587 chromatography analysis Methods 0.000 claims description 3
- 239000007791 liquid phase Substances 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 229920001353 Dextrin Polymers 0.000 claims description 2
- 239000004375 Dextrin Substances 0.000 claims description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 2
- 239000006071 cream Substances 0.000 claims description 2
- 235000019425 dextrin Nutrition 0.000 claims description 2
- 239000008101 lactose Substances 0.000 claims description 2
- 238000004811 liquid chromatography Methods 0.000 claims description 2
- 238000012856 packing Methods 0.000 claims description 2
- 229930188195 rebaudioside Natural products 0.000 claims description 2
- 239000006228 supernatant Substances 0.000 claims description 2
- 239000011260 aqueous acid Substances 0.000 claims 1
- 239000008187 granular material Substances 0.000 claims 1
- 238000012360 testing method Methods 0.000 abstract description 29
- 239000007788 liquid Substances 0.000 abstract description 14
- 238000003908 quality control method Methods 0.000 abstract description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 28
- 239000000047 product Substances 0.000 description 14
- 239000000945 filler Substances 0.000 description 12
- 239000003814 drug Substances 0.000 description 10
- 238000000605 extraction Methods 0.000 description 8
- 239000004615 ingredient Substances 0.000 description 8
- 238000011084 recovery Methods 0.000 description 8
- 239000000741 silica gel Substances 0.000 description 8
- 229910002027 silica gel Inorganic materials 0.000 description 8
- 239000002904 solvent Substances 0.000 description 8
- 238000003556 assay Methods 0.000 description 7
- 239000007787 solid Substances 0.000 description 7
- 238000011835 investigation Methods 0.000 description 6
- 239000008188 pellet Substances 0.000 description 6
- 150000001343 alkyl silanes Chemical group 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 5
- 239000012982 microporous membrane Substances 0.000 description 5
- 238000000825 ultraviolet detection Methods 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 description 4
- 239000000377 silicon dioxide Substances 0.000 description 4
- 230000003252 repetitive effect Effects 0.000 description 3
- 238000013112 stability test Methods 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 210000001367 artery Anatomy 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- RZVAJINKPMORJF-UHFFFAOYSA-N Acetaminophen Chemical compound CC(=O)NC1=CC=C(O)C=C1 RZVAJINKPMORJF-UHFFFAOYSA-N 0.000 description 1
- 206010002383 Angina Pectoris Diseases 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 230000001427 coherent effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000011868 grain product Nutrition 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 150000007965 phenolic acids Chemical class 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 238000013441 quality evaluation Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 229930192961 schisandrol Natural products 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
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- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention relates to one kind one to survey the methods for commenting tanshin polyphenolic acid B and schizandrin content in method measurement Yixinfumai particle more.Specially prepare Yixinfumai particle test solution and schizandrin reference substance solution, it is injected separately into high performance liquid chromatograph, gradient elution is used under specific chromatographic condition of the invention, measure the content of schizandrin, it is surveyed by one and comments method more, the relative correction factor between tanshin polyphenolic acid B and schizandrin is utilized to calculate the content of tanshin polyphenolic acid B in Yixinfumai particle.The survey quality control levels for commenting method to further improve the product that the present invention establishes more, while saving testing cost.
Description
Technical field
The present invention relates to a kind of analyzing detecting methods of drug, and in particular to a kind of survey using one comments method measurement invigorating heart multiple more
The method of tanshin polyphenolic acid B and schizandrin content in arteries and veins particle.
Background technique
Yixinfumai particle is the drug that Tianjin Tasly (Liaoning) Pharmaceutical Co., Ltd. produces, and major function is
Supplementing qi and nourishing yin, multiple arteries and veins of invigorating blood circulation；For deficiency of both qi and yin, painstaking effort internal resistance, chest impediment and cardialgia is uncomfortable in chest not relax, palpitaition knotted pulse.With load medicine
Measure the characteristics such as high, rapid-action, free from extraneous odour, easy to carry.Said preparation is by sun-dried ginseng, Radix Astragali, Radix Ophiopogonis, Schisandra chinensis, Radix Salviae Miltiorrhizae, Rhizoma Chuanxiong
The compound preparation of 6 taste medicinal materials composition, for the product, existing detection method only includes under character, 2 colour developings identifications and check item
Coherent detection item, do not carry out the qualitative or quantitative detection of medicinal material, cannot reflect the quality condition of product comprehensively.
Due to the complexity of plant chemical ingredient, multi objective assay has become plant origin Control of drug quality
Common recognition.Several ingredients in Chinese medicine are measured simultaneously with traditional measuring method, not only face that reference substance is rare, and round of visits is long, also
Energy consumption, time-consuming.One survey comments method by existing intrinsic function relationship and proportionate relationship method between effective component of chinese medicine, measures one
Ingredient (reference substance is easy to get, inexpensively, responder) realizes multiple ingredients (reference substance be difficult to obtain or difficult supplier) Simultaneous Determination.No
It addresses only Chinese medicine multicomponent quantitatively and reference substance present in the control of multi objective quality lacks problem, while realizing that multi objective is same
Walk quality control reaction traditional Chinese medicine quality.
The present invention is surveyed using one comments method to establish tanshin polyphenolic acid B and the five tastes in product for ministerial drug Schisandra chinensis and adjutant Radix Salviae Miltiorrhizae more
The method of sub- alcohol first assay has filled up the blank of Yixinfumai particle quantitative control, so that being able to carry out more to the product
Effective quality analysis, more comprehensively reflect product quality condition, guarantee the quality stability of the product, at the same save detection at
This.
Summary of the invention
Of the present invention one surveys the sides for commenting tanshin polyphenolic acid B and schizandrin content in method measurement Yixinfumai particle more
Method, method includes the following steps:
Step 1: the preparation of schizandrin reference substance solution；
Step 2: the preparation of Yixinfumai particle test solution；
Step 3: schizandrin reference substance solution and Yixinfumai particle test solution are injected separately into efficient liquid phase
Chromatograph measures the content of schizandrin in Yixinfumai particle test solution；
Step 4: surveying the relative correction factor f and pellet phenol for commenting method, utilizing between tanshin polyphenolic acid B and schizandrin by one more
The content of tanshin polyphenolic acid B in sour B calculated by peak area Yixinfumai particle.
Wherein, step 1, schizandrin reference substance solution the preparation method is as follows:
Take schizandrin reference substance appropriate, it is accurately weighed, add 70% methanol that every 1ml 0.030- containing schizandrin is made
0.045mg to get.
Wherein, step 2, Yixinfumai particle test solution the preparation method is as follows:
Take Yixinfumai particle finely ground in right amount, it is accurately weighed, add 5-20 times of 70% methanol for measuring w/v, ultrasound is molten
Solution, lets cool to room temperature, the weight of less loss is supplied with 70% methanol, is shaken up, and is centrifuged, and filtering takes subsequent filtrate to obtain the final product.
Preferably, test solution the preparation method is as follows:
Take Yixinfumai particle finely ground in right amount, it is accurately weighed, add 70% methanol of 10 times of amount w/vs, ultrasonic 20-
40min dissolution, lets cool to room temperature, the weight of less loss is supplied with 70% methanol, is shaken up, and is centrifuged, and crosses 0.45 μm of miillpore filter, takes continuous
Filtrate to obtain the final product.
Most preferably, in the preparation method of test solution, preferably ultrasound 30min dissolution.
Wherein, in step 3, the high performance liquid chromatography, chromatographic condition is as follows:
Using C18 chromatographic column；Mobile phase: A%: B%=0.1% aqueous formic acid: acetonitrile, condition of gradient elution such as table 1
It is shown；Column temperature: 25 DEG C -35 DEG C；Flow velocity: 0.8-1.5ml/min；Detection wavelength: 220-280nm；Sample volume: 5-20 μ l.
1 condition of gradient elution of table
Preferably, chromatographic condition is as follows:
C18 chromatographic column；Mobile phase: A%: B%=0.1% aqueous formic acid: acetonitrile；Condition of gradient elution is shown in Table 2；Column
Temperature: 30 DEG C；Flow velocity: 1.0ml/min；Detection wavelength: 250nm；Sample volume: 10 μ l.
2 gradient elution table of table
Wherein, step 4, the relative correction factor between tanshin polyphenolic acid B and schizandrin is to calculate as follows
It arrives:
1. preparing tanshin polyphenolic acid B and schizandrin reference substance solution:
Take tanshin polyphenolic acid B reference substance appropriate, it is accurately weighed, add 70% methanol that every 1ml 0.0655- containing tanshin polyphenolic acid B is respectively prepared
0.5244mg to get；
Take schizandrin reference substance appropriate, it is accurately weighed, add 70% methanol that every 1ml is respectively prepared containing schizandrin
0.01827-0.07308mg to get；
2. prepare Yixinfumai particle test solution: where the preparation method of Yixinfumai particle test solution with
It is identical in step 2, specifically:
Take Yixinfumai particle finely ground in right amount, it is accurately weighed, add 70% methanol of 10 times of amount w/vs, ultrasonic 20-
40min dissolution, lets cool to room temperature, the weight of less loss is supplied with 70% methanol, is shaken up, and is centrifuged, and crosses 0.45 μm of miillpore filter, takes continuous
Filtrate to obtain the final product.
Most preferably, in the preparation method of test solution, preferably ultrasound 30min dissolution.
3. tanshin polyphenolic acid B and schizandrin reference substance solution and Yixinfumai particle test solution are injected separately into height
Effect liquid phase chromatogram instrument measures the content of tanshin polyphenolic acid B and schizandrin in Yixinfumai particle test solution, wherein efficiently
The chromatographic condition of liquid chromatogram is identical with step 3, specific as follows:
Using C18 chromatographic column；Mobile phase: A%: B%=0.1% aqueous formic acid: acetonitrile；The following institute of condition of gradient elution
Show；Column temperature: 25 DEG C -35 DEG C；Flow velocity: 0.8-1.5ml/min；Detection wavelength: 220-280nm；Sample volume: 5-20 μ l；
Preferably, chromatographic condition is as follows:
C18 chromatographic column；Mobile phase: A%: B%=0.1% aqueous formic acid: acetonitrile；Condition of gradient elution is shown in Table 2；Column
Temperature: 30 DEG C；Flow velocity: 1.0ml/min；Detection wavelength: 250nm；Sample volume: 10 μ l.
2 gradient elution table of table
4. utilizing formula:Calculate the relative correction between tanshin polyphenolic acid B and schizandrin
Factor f, wherein ASchizandrinFor schizandrin reference substance peak area, CSchizandrinFor schizandrin reference substance concentration；ATanshin polyphenolic acid BFor
Tanshin polyphenolic acid B reference substance peak area, CTanshin polyphenolic acid BFor tanshin polyphenolic acid B reference substance concentration.
Yixinfumai particle of the present invention is that the original of Tianjin Tasly (Liaoning) Pharmaceutical Co., Ltd. grinds medicine
Product, prescription are as follows: sun-dried ginseng 75-225 parts by weight, Radix Astragali 75-225 parts by weight, Radix Ophiopogonis 75-225 parts by weight, Schisandra chinensis 50-
150 parts by weight, Radix Salviae Miltiorrhizae 100-300 parts by weight, Rhizoma Chuanxiong 50-150 parts by weight and appropriate amount of auxiliary materials.
Preferably, Yixinfumai particle prescription of the invention is as follows: 150 parts by weight of sun-dried ginseng, 150 parts by weight of Radix Astragali, wheat
150 parts by weight of winter, 100 parts by weight of Schisandra chinensis, 200 parts by weight of Radix Salviae Miltiorrhizae, 100 parts by weight of Rhizoma Chuanxiong and appropriate amount of auxiliary materials.
Yixinfumai particle of the invention, preparation method are as follows: the above 6 taste medicinal material adds water to cook 2 times, adds for the first time
10 times of water amounts, decoct 2 hours, and second plus 8 times of water amounts decoct 1 hour, merge decoction liquor, stand 12 hours, draw supernatant
Liquid is concentrated into the clear cream of relative density 1.20-1.30 (80 DEG C of surveys), 1% rebaudioside is added, is uniformly mixed, qinghuo reagent 1
Part, be added 1 part of dextrin, dry, pulverize, cross 60 meshes, be added 2 parts of lactose, mix, be made 600g, packing to get.
Currently preferred experimental method, chromatographic condition and solvent extraction process are obtained by screening, screening process
It is as follows:
1, the selection of test article treating method
(1) investigation of Extraction solvent
In conjunction with Yixinfumai grain products process recipes, different Extraction solvents is chosen, investigates test article treating method.
1. sample is finely ground, 1.0g is taken, it is accurately weighed, it sets in 25ml conical flask, adds water 10ml, weighed weight, ultrasound
30min dissolves, and takes out, lets cool to room temperature, the weight of less loss is supplied with water, shake up (centrifugation), crosses 0.45 μm of miillpore filter, takes
Subsequent filtrate to obtain the final product.As a result as shown in Figure 1.
2. sample is finely ground, 1.0g is taken, it is accurately weighed, it sets in 25ml conical flask, adds 30% methanol 10ml, weighed weight surpasses
Sound 30min dissolution, takes out, lets cool to room temperature, the weight of less loss is supplied with 30% methanol, shake up (centrifugation), crosses 0.45 μm of micropore
Filter membrane takes subsequent filtrate to obtain the final product.As a result as shown in Figure 2.
3. sample is finely ground, 1.0g is taken, it is accurately weighed, it sets in 25ml conical flask, adds 50% methanol 10ml, weighed weight surpasses
Sound 30min dissolution, takes out, lets cool to room temperature, the weight of less loss is supplied with 50% methanol, shake up (centrifugation), crosses 0.45 μm of micropore
Filter membrane takes subsequent filtrate to obtain the final product.As a result as shown in Figure 3.
4. sample is finely ground, 1.0g is taken, it is accurately weighed, it sets in 25ml conical flask, adds 70% methanol 10ml, weighed weight surpasses
Sound 30min dissolution, takes out, lets cool to room temperature, the weight of less loss is supplied with 70% methanol, shake up (centrifugation), crosses 0.45 μm of micropore
Filter membrane takes subsequent filtrate to obtain the final product.As a result as shown in Figure 4.
5. sample is finely ground, 1.0g is taken, it is accurately weighed, it sets in 25ml conical flask, adds 100% methanol 10ml, weighed weight surpasses
Sound 30min dissolution, takes out, lets cool to room temperature, the weight of less loss is supplied with 100% methanol, shake up (centrifugation), crosses 0.45 μ micropore
Filter membrane takes subsequent filtrate to obtain the final product.As a result as shown in Figure 5.
The investigation of the above processing method is as the result is shown: 1., 2., 3., 5. peak area is smaller for test article treating method, cannot
Meet the requirement of accurate quantitative analysis, and the peak area of test article treating method 4. is higher, therefore primarily determine 70% methanol solution conduct
Extraction solvent.
(2) investigation of test sample processing extracting method condition
The careful investigation of condition is 4. extracted based on above test article treating method, respectively to liquid-solid ratio, ultrasound when
Between investigated, as a result such as table 3 and profiling results: liquid-solid ratio 5:1 is shown in Fig. 6；Liquid-solid ratio 10:1 is shown in Fig. 7；Liquid-solid ratio 20:1
See Fig. 8.
The investigation result of 3 test sample of table processing extracting method condition
2, the selection of chromatographic condition
(1) selection of condition of gradient elution
Tanshin polyphenolic acid B and schisandrol are separated using chromatographic column Agilent ZORBAX SB-C18 (4.6 × 250mm, 5 μm)
Condition of gradient elution is carried out investigation adjustment, as a result the gradient strip shown in table 2 under the premise of meeting separating degree requirement by first
Under part, the separating degree of tanshin polyphenolic acid B and schizandrin is met the requirements, when analysis a length of 65min.
(2) selection of different sample volumes
Different sample volumes are investigated to be tested, when as a result sample volume is respectively 10 μ l, 15 μ l, 20 μ l, tanshin polyphenolic acid B with it is latter
Separating degree is followed successively by 1.67,1.65,1.62 (integration method is trough to trough) between chromatographic peak, as a result such as Fig. 9, Figure 10 and figure
Shown in 11.Therefore, it is seen that increase sample volume reduce the separating degree between tanshin polyphenolic acid B and latter chromatographic peak, sample volume number
On the separating degree of schizandrin and other impurities peak without influence, therefore select 10 μ l sample volumes.
(3) selection of different column temperatures
The case where investigating 35 DEG C, 30 DEG C and 25 DEG C respectively, as a result as shown in Figure 12, Figure 13 and Figure 14.Column temperature as the result is shown
When being 35 DEG C, the chromatographic peak of tanshin polyphenolic acid B and the front, which has, to intersect, and influences its separating degree；Column temperature be 25 DEG C when, tanshin polyphenolic acid B and its
Chromatographic peak separating degree is good below, but its chromatographic peak has certain broadening, and appearance time delay simultaneously influences subsequent chromatographic peak simultaneously
Appearance time and peak width；When column temperature is 30 DEG C, tanshin polyphenolic acid B and chromatographic peak separating degree behind are good, therefore in above-mentioned chromatography ladder
Suggest that column temperature uses 30 DEG C when spending elution program.
3, in Yixinfumai particle tanshin polyphenolic acid B and schizandrin assay methodological study
(1) preparation of standard curve
Take tanshin polyphenolic acid B reference substance appropriate, it is accurately weighed, add 70% methanol be respectively prepared every 1ml containing tanshin polyphenolic acid B 0.06555,
0.1311, the serial solution of 0.2622,0.41952,0.5244mg, it is accurate respectively to measure 10 μ l, liquid chromatograph is injected, by step
Chromatographic condition analysis in rapid 3, measures peak area, and with reference substance sample introduction concentration (mg/ml) for abscissa, peak area value is vertical sits
Mark, acquires regression equation: tanshin polyphenolic acid B Y=10000000X+34833, r=0.9998.The result shows that tanshin polyphenolic acid B is 0.06555
It is linear good within the scope of~0.5244mg, it the results are shown in Table 4.
4 tanshin polyphenolic acid B standard curve of table
Take schizandrin reference substance appropriate, it is accurately weighed, add 70% methanol that every 1ml is respectively prepared containing schizandrin
0.01827, the serial solution of 0.027405,0.03654,0.05481,0.07308mg, it is accurate respectively to measure 10 μ l, inject liquid
Chromatography is analyzed by the chromatographic condition in step 3, measures peak area, with reference substance sample introduction concentration (mg/ml) for abscissa,
Peak area value is ordinate, acquires regression equation: schizandrin Y=20000000X+9823.2, r=0.9998.As a result table
Bright schizandrin is linear good within the scope of 0.01827~0.07308mg, the results are shown in Table 5, linear map is as shown in Figure 15.
5 schizandrin standard curve of table
(2) sample introduction precision test
Tanshin polyphenolic acid B, schizandrin reference substance are taken, prepares reference substance solution, chromatographic condition are as follows: with 18 according to step 1
Alkyl silane bonded silica gel is filler, Agilent ZORBAX SB-C18 (4.6 × 250mm, 5 μm) chromatographic column；UV detection
Device；Mobile phase: A%: B%=0.1% aqueous formic acid: acetonitrile；Condition of gradient elution is shown in Table 2.Column temperature: 30 DEG C；Flow velocity:
1.0ml/min；Detection wavelength: 250nm；Sample volume: 10 μ l.Continuous sample introduction 6 times, measure tanshin polyphenolic acid B, schizandrin peak face
Product, measuring peak area average value is respectively 2322178,719510, and the RSD of peak area is respectively 0.82%, 0.39%, is conformed to
It asks, the results are shown in Table 6, map is as shown in figure 16.
6 sample introduction precision test of table
(3) repetitive test
Yixinfumai particulate samples are taken, prepare the test solution of basic, normal, high 3 kinds of concentration, concentration range is 50%~
150%, totally 9 parts, prepare test solution, chromatographic condition according to step 2 are as follows: with octadecylsilane chemically bonded silica be filling
Agent, Agilent ZORBAX SB-C18 (4.6 × 250mm, 5 μm) chromatographic column；UV detector；Mobile phase: A%: B%=0.1%
Aqueous formic acid: acetonitrile；Condition of gradient elution is shown in Table 2.Column temperature: 30 DEG C；Flow velocity: 1.0ml/min；Detection wavelength: 250nm；Into
Sample amount: 10 μ l.Measure the content of tanshin polyphenolic acid B, schizandrin in every part of sample.Tanshin polyphenolic acid B, schizandrin in results sample
Average content is 1.8572mg/g, 0.3916mg/g, and RSD is respectively 1.7%, 2.8%, and repeatability is good, is as a result seen
Table 7, map is as shown in figure 17.
7 repetitive test of table
(4) stability test
Yixinfumai particulate samples are taken, accurately weighed 1.0g prepares test solution, chromatographic condition according to step 2 are as follows: with
Octadecylsilane chemically bonded silica is filler, Agilent ZORBAX SB-C18 (4.6 × 250mm, 5 μm) chromatographic column；UV inspection
Survey device；Mobile phase: A%: B%=0.1% aqueous formic acid: acetonitrile；Condition of gradient elution is shown in Table 2.Column temperature: 30 DEG C；Flow velocity:
1.0ml/min；Detection wavelength: 250nm；Sample volume: 10 μ l.Respectively at 0,2,4,6,8,12 and 24 hour, measure red in sample
Phenolic acid B, schizandrin peak area, the RSD for measuring peak area value is respectively 0.50%, 0.63%, the results showed that test sample is molten
Liquid measures stabilization in 24 hours, the results are shown in Table 8, map is as shown in figure 18.
8 stability test of table
(5) recovery test
Yixinfumai particulate samples are taken, the 1/2 of test sample sample weighting amount is taken, accurately weighed 9 parts, it is (molten to be separately added into reference substance
Liquid) in right amount, according to step 2 prepare 3 various concentrations containing corresponding reference substance test solution (with contained by test solution to
The concentration for surveying ingredient is 100%, and 3 various concentrations are 70%~130%), each concentration prepares 3 parts of test solutions, chromatography
Condition are as follows: using octadecylsilane chemically bonded silica as filler, Agilent ZORBAX SB-C18 (4.6 × 250mm, 5 μm) color
Compose column；UV detector；Mobile phase: A%: B%=0.1% aqueous formic acid: acetonitrile；Condition of gradient elution is shown in Table 2.Column temperature: 30
℃；Flow velocity: 1.0ml/min；Detection wavelength: 250nm；Sample volume: 10 μ l.It is analyzed by above-mentioned chromatographic condition, calculates the rate of recovery, knot
Fruit tanshin polyphenolic acid B, schizandrin average recovery rate be 101.1%, RSD is respectively 1.45%, the results are shown in Table 9 and table 10, map
See that Figure 19 and Figure 20, recovery test meet the requirements.
Tanshin polyphenolic acid B recovery test in 9 Yixinfumai particle of table
Schizandrin recovery test in 10 Yixinfumai particle of table
(6) serviceability test
Yixinfumai particulate samples are taken, prepare test solution according to step 2, use Agilent ZORBAX C-18 respectively
(4.6 × 250mm, 5 μm)；OmniBond Hubble C-18 (5 μm of 4.6 × 250mm) chromatographic column, chromatographic condition are as follows: with 18
Alkyl silane bonded silica gel is filler, Agilent ZORBAX SB-C18 (4.6 × 250mm, 5 μm) chromatographic column；UV detection
Device；Mobile phase: A%: B%=0.1% aqueous formic acid: acetonitrile；Condition of gradient elution is shown in Table 2.Column temperature: 30 DEG C；Flow velocity:
1.0ml/min；Detection wavelength: 250nm；Sample volume: 10 μ l.Tanshin polyphenolic acid B, schizandrin content in sample are measured, is as a result seen
Table 11, chromatogram is shown in Figure 21 and Figure 22.
Influence (n=2) of the different chromatographic columns of table 11 to assay
Measurement result shows that different brands chromatographic column is on the result of assay without influence.
(7) determination of number of theoretical plate
The separation situation and theoretical cam curve for comprehensively considering the different chromatographic columns of 2 kinds of Yixinfumai particle, will as a result, be shown in Table 11
The number of theoretical plate of tanshin polyphenolic acid B and schizandrin must not be defined as lower than 10000.
(8) determination of correction factor
Consider that the amount of index components and the response of detector are directly proportional, in multi objective quality evaluation in a certain range
When, internal reference object is made with typical case's effective component a certain in medicinal material, establishes the relative correction factor between internal reference object and other ingredients to be measured.
In the method, foundation measures tanshin polyphenolic acid B and schizandrin in Yixinfumai particle simultaneously with schizandrin reference substance and contains
The method of amount.
F is correction factor, and formula is as follows:
As internal reference object reference substance peak area, Cs internal reference object reference substance concentration；Ai ingredient reference substance peak area to be measured, Ci are to be measured
Ingredient reference substance concentration
It refers in this method i.e.:
Table 12
12 parts of samples are calculated by correction factor (f), compare the RAD value that two methods calculate content of danshinolic acid B, specific number
According to being shown in Table 13.
Table 13
As can be seen from the above table, the RAD value of the content of two methods measurement is respectively less than 3%.
The invention has the benefit that
1, the survey that the present invention the establishes methods for commenting tanshin polyphenolic acid B in method measurement Yixinfumai particle, Schisandra chinensis content, symbol more
The requirement of methodology validation, and easy to operate, high sensitivity are closed, measurement is accurate, and strong applicability can be used in the matter to the product
Amount control.
2, tanshin polyphenolic acid B and five in a kind of reference substance measurement Yixinfumai particle of schizandrin is used only in method of the invention
Taste alcohol first content provides more perfect detection method for the quality control of said preparation, while saving testing cost.
3, method of the invention has filled up the blank of Yixinfumai particle quantitative control, so that being able to carry out more to the product
Effective quality analysis more comprehensively reflects the quality condition of product, guarantees the quality stability of the product.
Detailed description of the invention
Fig. 1 Extraction solvent: water
Fig. 2 Extraction solvent: 30% methanol
Fig. 3 Extraction solvent: 50% methanol
Fig. 4 Extraction solvent: 70% methanol
Fig. 5 Extraction solvent: 100% methanol
Fig. 6 liquid-solid ratio 5:1
Fig. 7 liquid-solid ratio 10:1
Fig. 8 liquid-solid ratio 20:1
10 μ l sample volume of Fig. 9
15 μ l sample volume of Figure 10
20 μ l sample volume of Figure 11
35 DEG C of Figure 12 column temperature
30 DEG C of Figure 13 column temperature
25 DEG C of Figure 14 column temperature
The linear map of Figure 15
Figure 16 sample introduction precision map
Figure 17 repetitive test map
Figure 18 stability test map
Figure 19 tanshin polyphenolic acid B rate of recovery map
Figure 20 schizandrin rate of recovery map
Figure 21 OmniBond Hubble C-18 chromatographic column spectrogram
Figure 22 Agilent ZORBAX C-18 chromatographic column spectrogram
Specific embodiment
Instrument selected by the present invention and material are as follows:
1, high performance liquid chromatograph: U.S. Waters HPLC-2695-2996, Empower work station.
2, chromatographic column
Agilent ZORBAX SB-C18 1. (4.6 × 250mm, 5 μm) chromatographic column: it is suitable for low pH value chromatographic column, it is fixed
Mutually using octadecylsilane chemically bonded silica as filler, Anjelen Sci. & Tech. Inc.
OmniBond HPLC Column Hubble 2. C18 (4.6 × 250mm, 5 μm) chromatographic column: the wide pH value of versatility
Chromatographic column, stationary phase are up to 99.999% high-purity silica gel as filler using purity, Beijing ohm Buddhist nun Science and Technology Ltd..
3, reagent: acetonitrile (chromatographically pure, Tianjin Concord Technology Co., Ltd.).
Methanol (analyzes pure, Tianjin Concord Technology Co., Ltd.).
Formic acid (analyzes pure, Tianjin Chemical Reagents Factory No.1).
4, reference substance: tanshin polyphenolic acid B (Nat'l Pharmaceutical & Biological Products Control Institute, lot number: 111562-201313, assay
With)；Schizandrin (Nat'l Pharmaceutical & Biological Products Control Institute, lot number: 110857-201211, assay use)
5, sample: Yixinfumai particle (Tianjin Tasly (Liaoning) Pharmaceutical Co., Ltd., number: test 1, test
2, test 3, the preparation method of the middle Yixinfumai particle provided is made through the invention)
Embodiment 1
Taking number is the Yixinfumai particle for testing 1, measures the content of its tanshin polyphenolic acid B and schizandrin.
Step 1: the preparation of schizandrin reference substance solution
Take schizandrin reference substance appropriate, it is accurately weighed, add 70% methanol that every 1ml is made containing schizandrin
The solution of 0.036mg to get.
Step 2: the preparation of Yixinfumai particle test solution
It takes Yixinfumai particle finely ground in right amount, takes about 1.0g, it is accurately weighed, add 70% methanol ultrasound 30min of 10ml molten
Solution is taken out, and places to room temperature, the weight of less loss is supplied with 70% methanol, is shaken up, and is centrifuged, and crosses 0.45 μm of filtering with microporous membrane, takes
Filtrate to obtain the final product.
Step 3: schizandrin reference substance solution and test solution being injected separately into high performance liquid chromatograph, with 18
Alkyl silane bonded silica gel is filler, Agilent ZORBAX SB-C18 (4.6 × 250mm, 5 μm) chromatographic column；UV detection
Device；Mobile phase: A%: B%=0.1% aqueous formic acid: acetonitrile；Condition of gradient elution is shown in Table 2.Column temperature: 30 DEG C；Flow velocity:
1.0ml/min；Detection wavelength: 250nm；Sample volume: 10 μ l.The content for measuring schizandrin in Yixinfumai particle is
0.3116mg/g。
Step 4: surveying the relative correction factor f and pellet phenol for commenting method, utilizing between tanshin polyphenolic acid B and schizandrin by one more
The content of tanshin polyphenolic acid B is 2.3325mg mg/g in sour B calculated by peak area Yixinfumai particle.
It is actually detected to the content progress of tanshin polyphenolic acid B in Yixinfumai particle under chromatographic condition described in step 3, it measures every
The content of tanshin polyphenolic acid B is 2.4471mg/g in gram Yixinfumai particle, with the content of danshinolic acid B phase according to correction factor measurement
Than the RAD value that two methods measure content of danshinolic acid B is 2.6%.
Embodiment 2
Taking number is the Yixinfumai particle for testing 2, measures the content of its tanshin polyphenolic acid B and schizandrin.
Step 1: the preparation of schizandrin reference substance solution
Take schizandrin reference substance appropriate, it is accurately weighed, add 70% methanol that every 1ml is made containing schizandrin
The solution of 0.030mg to get.
Step 2: the preparation of Yixinfumai particle test solution
It takes Yixinfumai particle finely ground in right amount, takes about 1.0g, it is accurately weighed, add 70% methanol ultrasound 20min of 5ml to dissolve,
It takes out, places to room temperature, the weight of less loss is supplied with 70% methanol, is shaken up, be centrifuged, cross 0.45 μm of filtering with microporous membrane, take filter
Liquid to obtain the final product.
Step 3: schizandrin reference substance solution and test solution being injected separately into high performance liquid chromatograph, with high-purity
Silica gel is filler, OmniBond HPLC Column Hubble C18 (4.6 × 250mm, 5 μm) chromatographic column；UV detector；
Mobile phase: A%: B%=0.1% aqueous formic acid: acetonitrile；Condition of gradient elution is shown in Table 2.Column temperature: 25 DEG C；Flow velocity: 1.5ml/
min；Detection wavelength: 220nm；Sample volume: 5 μ l.The content for measuring schizandrin in Yixinfumai particle is 0.4344mg/g.
Step 4: surveying the relative correction factor f and pellet phenol for commenting method, utilizing between tanshin polyphenolic acid B and schizandrin by one more
The content of tanshin polyphenolic acid B is 1.7023mg/g in sour B calculated by peak area Yixinfumai particle.
Actually detected measure often is carried out to the content of tanshin polyphenolic acid B in Yixinfumai particle under chromatographic condition described in step 3
The content of tanshin polyphenolic acid B is 1.7924mg/g in gram Yixinfumai particle, with the content of danshinolic acid B phase according to correction factor measurement
Than the RAD value that two methods measure content of danshinolic acid B is 2.6%.
Embodiment 3
Taking number is the Yixinfumai particle for testing 3, measures the content of its tanshin polyphenolic acid B and schizandrin.
Step 1: the preparation of schizandrin reference substance solution
Take schizandrin reference substance appropriate, it is accurately weighed, add 70% methanol that every 1ml is made containing schizandrin
The solution of 0.040mg to get.
Step 2: the preparation of test solution
It takes Yixinfumai particle finely ground in right amount, takes about 1.0g, it is accurately weighed, add 70% methanol ultrasound 40min of 20ml molten
Solution is taken out, and places to room temperature, the weight of less loss is supplied with 70% methanol, is shaken up, and is centrifuged, and crosses 0.45 μm of filtering with microporous membrane, takes
Filtrate to obtain the final product.
Step 3: schizandrin reference substance solution and test solution being injected separately into high performance liquid chromatograph, with 18
Alkyl silane bonded silica gel is filler, Agilent ZORBAX SB-C18 (4.6 × 250mm, 5 μm) chromatographic column；UV detection
Device；Mobile phase: A%: B%=0.1% aqueous formic acid: acetonitrile；Condition of gradient elution is shown in Table 2.Column temperature: 35 DEG C；Flow velocity:
0.8ml/min；Detection wavelength: 280nm；Sample volume: 20 μ l.The content for measuring schizandrin in Yixinfumai particle is
0.3968mg/g。
Step 4: surveying the relative correction factor f and pellet phenol for commenting method, utilizing between tanshin polyphenolic acid B and schizandrin by one more
The content of tanshin polyphenolic acid B is 1.8510mg/g in sour B calculated by peak area Yixinfumai particle.
It is actually detected to the content progress of tanshin polyphenolic acid B in Yixinfumai particle under chromatographic condition described in step 3, it measures every
The content of tanshin polyphenolic acid B is 1.7955mg/g in gram Yixinfumai particle, with the content of danshinolic acid B phase according to correction factor measurement
Than the RAD value that two methods measure content of danshinolic acid B is 1.6%.
Embodiment 4
Taking number is the Yixinfumai particle for testing 3, measures the content of its tanshin polyphenolic acid B and schizandrin.
Step 1: the preparation of schizandrin reference substance solution
Take schizandrin reference substance appropriate, it is accurately weighed, add 70% methanol that every 1ml is made containing schizandrin
The solution of 0.045mg to get.
Step 2: the preparation of Yixinfumai particle test solution
It takes Yixinfumai particle finely ground in right amount, takes about 1.0g, it is accurately weighed, add 70% methanol ultrasound 30min of 15ml molten
Solution is taken out, and places to room temperature, the weight of less loss is supplied with 70% methanol, is shaken up, and is centrifuged, and crosses 0.45 μm of filtering with microporous membrane, takes
Filtrate to obtain the final product.
Step 3: schizandrin reference substance solution and test solution being injected separately into high performance liquid chromatograph, with high-purity
Silica gel is filler, OmniBond HPLC Column Hubble C18 (4.6 × 250mm, 5 μm) chromatographic column；UV detector；
Mobile phase: A%: B%=0.1% aqueous formic acid: acetonitrile；Condition of gradient elution is shown in Table 2.Column temperature: 32 DEG C；Flow velocity: 1.0ml/
min；Detection wavelength: 250nm；Sample volume: 10 μ l.The content for measuring schizandrin in Yixinfumai particle is 0.3825mg/
g。
Step 4: surveying the relative correction factor f and pellet phenol for commenting method, utilizing between tanshin polyphenolic acid B and schizandrin by one more
The content of tanshin polyphenolic acid B is 1.7682mg/g in sour B calculated by peak area Yixinfumai particle.
It is actually detected to the content progress of tanshin polyphenolic acid B in Yixinfumai particle under chromatographic condition described in step 3, it measures every
The content of tanshin polyphenolic acid B is 1.7334mg/g in gram Yixinfumai particle, with the content of danshinolic acid B phase according to correction factor measurement
Than the RAD value that two methods measure content of danshinolic acid B is 1.0%.
Embodiment 5
Taking number is the Yixinfumai particle for testing 3, measures the content of its tanshin polyphenolic acid B and schizandrin.
Step 1: the preparation of schizandrin reference substance solution
Take schizandrin reference substance appropriate, it is accurately weighed, add 70% methanol that every 1ml is made containing schizandrin
The solution of 0.036mg to get.
Step 2: the preparation of test solution
It takes Yixinfumai particle finely ground in right amount, takes about 1.0g, it is accurately weighed, add 10ml70% methanol ultrasound 30min to dissolve,
It takes out, places to room temperature, the weight of less loss is supplied with 70% methanol, is shaken up, be centrifuged, cross 0.45 μm of filtering with microporous membrane, take filter
Liquid to obtain the final product.
Step 3: schizandrin reference substance solution and test solution being injected separately into high performance liquid chromatograph, with 18
Alkyl silane bonded silica gel is filler, Agilent ZORBAX SB-C18 (4.6 × 250mm, 5 μm) chromatographic column；UV detection
Device；Mobile phase: A%: B%=0.1% aqueous formic acid: acetonitrile；Condition of gradient elution is shown in Table 2.Column temperature: 28 DEG C；Flow velocity:
1.0ml/min；Detection wavelength: 250nm；Sample volume: 10 μ l.The content for measuring schizandrin in Yixinfumai particle is
0.3846mg/g。
Step 4: surveying the relative correction factor f and pellet phenol for commenting method, utilizing between tanshin polyphenolic acid B and schizandrin by one more
The content of tanshin polyphenolic acid B is 1.7811mg/g in sour B calculated by peak area Yixinfumai particle.
It is actually detected to the content progress of tanshin polyphenolic acid B in Yixinfumai particle under chromatographic condition described in step 3, it measures every
The content of tanshin polyphenolic acid B is 1.7721mg/g in gram Yixinfumai particle, with the content of danshinolic acid B phase according to correction factor measurement
Than the RAD value that two methods measure content of danshinolic acid B is 0.26%.
Claims (7)
1. one kind one surveys the methods for commenting tanshin polyphenolic acid B and schizandrin content in method measurement Yixinfumai particle more, specifically include
Following steps:
Step 1: the preparation of schizandrin reference substance solution；
Step 2: the preparation of Yixinfumai particle test solution；
Step 3: schizandrin reference substance solution and Yixinfumai particle test solution are injected separately into high performance liquid chromatography
Instrument measures the content of schizandrin in Yixinfumai particle test solution；
Step 4: being surveyed by one and comment method more, utilize the relative correction factor f and tanshin polyphenolic acid B between tanshin polyphenolic acid B and schizandrin
The content of tanshin polyphenolic acid B in calculated by peak area Yixinfumai particle；
Wherein, step 1 schizandrin reference substance solution the preparation method is as follows: take schizandrin reference substance appropriate, it is accurate
It is weighed, add 70% methanol be made every 1ml 0.030-0.045mg containing schizandrin to get；
Wherein, step 2 Yixinfumai particle test solution the preparation method is as follows: take Yixinfumai particle finely ground in right amount, essence
It is close weighed, add 5-20 times of 70% methanol for measuring w/v, ultrasonic dissolution is let cool to room temperature, supplies less loss with 70% methanol
Weight, shake up, be centrifuged, filtering, take subsequent filtrate to obtain the final product；
Wherein, step 3 high performance liquid chromatography chromatographic condition is as follows: using C18 chromatographic column；Mobile phase: A%: B%=0.1% first
Aqueous acid: acetonitrile；Condition of gradient elution is as follows；Column temperature: 25 DEG C -35 DEG C；Flow velocity: 1.0ml/min；Detection wavelength:
220-280nm；Sample volume: 5-20 μ l,
2. the method for claim 1, wherein step 2 Yixinfumai particle test solution the preparation method is as follows: taking
Yixinfumai particle is finely ground in right amount, accurately weighed, adds 70% methanol of 10 times of amount w/vs, and ultrasonic 20-40min dissolves,
It lets cool to room temperature, the weight of less loss is supplied with 70% methanol, is shaken up, be centrifuged, cross 0.45 μm of miillpore filter, take subsequent filtrate to obtain the final product.
3. the method for claim 1, wherein in the preparation method of step 2 Yixinfumai particle test solution, ultrasound
30min dissolution.
4. the method for claim 1, wherein step 3 high performance liquid chromatography chromatographic condition is as follows: C18 chromatographic column；Flowing
Phase: A%: B%=0.1% aqueous formic acid: acetonitrile；Condition of gradient elution is as follows；Column temperature: 30 DEG C；Flow velocity: 1.0ml/
min；Detection wavelength: 250nm；Sample volume: 10 μ l,
5. the method for claim 1, wherein step 4, the relative correction factor between tanshin polyphenolic acid B and schizandrin
It is calculated as follows:
1. preparing tanshin polyphenolic acid B and schizandrin reference substance solution: take tanshin polyphenolic acid B reference substance appropriate, it is accurately weighed, add 70% first
Alcohol be respectively prepared every 1ml 0.0655-0.5244mg containing tanshin polyphenolic acid B to get；Take schizandrin reference substance appropriate, precision claims
It is fixed, add 70% methanol be respectively prepared every 1ml 0.01827-0.07308mg containing schizandrin to get；
2. preparing Yixinfumai particle test solution: take Yixinfumai particle finely ground in right amount, it is accurately weighed, add 10 times of amount weight
70% methanol of volume ratio, ultrasonic 20-40min dissolution, lets cool to room temperature, the weight of less loss is supplied with 70% methanol, is shaken up, from
The heart crosses 0.45 μm of miillpore filter, takes subsequent filtrate to obtain the final product；
3. tanshin polyphenolic acid B and schizandrin reference substance solution and Yixinfumai particle test solution are injected separately into efficient liquid
Chromatography measures the content of tanshin polyphenolic acid B and schizandrin in Yixinfumai particle test solution, wherein efficient liquid phase
Chromatography chromatographic condition is as follows: using C18 chromatographic column；Mobile phase: A%: B%=0.1% aqueous formic acid: acetonitrile；Gradient elution
Condition is as follows；Column temperature: 25 DEG C -35 DEG C；Flow velocity: 1.0ml/min；Detection wavelength: 220-280nm；Sample volume: 5-20 μ l；
4. utilizing formula:Calculate the relative correction factor between tanshin polyphenolic acid B and schizandrin
F, wherein ASchizandrinFor schizandrin reference substance peak area, CSchizandrinFor schizandrin reference substance concentration；ATanshin polyphenolic acid BFor red phenol
Sour B reference substance peak area, CTanshin polyphenolic acid BFor tanshin polyphenolic acid B reference substance concentration.
6. method a method as claimed in any one of claims 1 to 5, wherein the Yixinfumai particle prescription is as follows: sun-dried ginseng 75-225 weight
Measure part, Radix Astragali 75-225 parts by weight, Radix Ophiopogonis 75-225 parts by weight, Schisandra chinensis 50-150 parts by weight, Radix Salviae Miltiorrhizae 100-300 parts by weight,
Rhizoma Chuanxiong 50-150 parts by weight and appropriate amount of auxiliary materials.
7. method as claimed in claim 6, wherein the Yixinfumai preparation method of granules is as follows: the above 6 taste medicinal material adds water
It decocts 2 times, for the first time plus 10 times of water are measured, decoction 2 hours, second plus 8 times of water amounts, decoction 1 hour, and merging decoction liquor is stood
12 hours, Aspirate supernatant, being concentrated into relative density at 80 DEG C was the clear cream of 1.20-1.30, and 1% rebaudioside is added,
It is uniformly mixed, 1 part of qinghuo reagent, is added 1 part of dextrin, dry, pulverize, cross 60 meshes, be added 2 parts of lactose, mix, 600g is made,
Packing to get.
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| CN114965779A (en) * | 2022-05-30 | 2022-08-30 | 华润三九医药股份有限公司 | Method for determining content of lignans in schisandra chinensis by multi-evaluation method and application |
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