CN102297860A - Rapid detection method of diclofenac sodium added in antirheumatic Chinese patent medicines or health foods - Google Patents
Rapid detection method of diclofenac sodium added in antirheumatic Chinese patent medicines or health foods Download PDFInfo
- Publication number
- CN102297860A CN102297860A CN2011101328378A CN201110132837A CN102297860A CN 102297860 A CN102297860 A CN 102297860A CN 2011101328378 A CN2011101328378 A CN 2011101328378A CN 201110132837 A CN201110132837 A CN 201110132837A CN 102297860 A CN102297860 A CN 102297860A
- Authority
- CN
- China
- Prior art keywords
- sample
- liquid
- chinese patent
- ethyl acetate
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000003814 drug Substances 0.000 title claims abstract description 33
- 229940079593 drug Drugs 0.000 title claims abstract description 18
- 235000013402 health food Nutrition 0.000 title claims abstract description 14
- 230000003356 anti-rheumatic effect Effects 0.000 title claims abstract description 11
- 239000003435 antirheumatic agent Substances 0.000 title claims abstract description 11
- 238000001514 detection method Methods 0.000 title abstract description 24
- 229960001193 diclofenac sodium Drugs 0.000 title abstract description 5
- JGMJQSFLQWGYMQ-UHFFFAOYSA-M sodium;2,6-dichloro-n-phenylaniline;acetate Chemical compound [Na+].CC([O-])=O.ClC1=CC=CC(Cl)=C1NC1=CC=CC=C1 JGMJQSFLQWGYMQ-UHFFFAOYSA-M 0.000 title abstract description 5
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims abstract description 49
- 238000000034 method Methods 0.000 claims abstract description 48
- 239000007788 liquid Substances 0.000 claims abstract description 46
- 239000000706 filtrate Substances 0.000 claims abstract description 9
- 239000007787 solid Substances 0.000 claims abstract description 8
- 239000002024 ethyl acetate extract Substances 0.000 claims abstract description 5
- 239000002775 capsule Substances 0.000 claims description 16
- UGAPHEBNTGUMBB-UHFFFAOYSA-N acetic acid;ethyl acetate Chemical compound CC(O)=O.CCOC(C)=O UGAPHEBNTGUMBB-UHFFFAOYSA-N 0.000 claims description 4
- 239000012530 fluid Substances 0.000 claims description 3
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 abstract description 26
- 229910017604 nitric acid Inorganic materials 0.000 abstract description 26
- 238000006243 chemical reaction Methods 0.000 abstract description 4
- 239000000203 mixture Substances 0.000 abstract description 4
- 230000008901 benefit Effects 0.000 abstract description 2
- 230000001105 regulatory effect Effects 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 93
- 239000000523 sample Substances 0.000 description 61
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 33
- 239000003795 chemical substances by application Substances 0.000 description 20
- 239000013558 reference substance Substances 0.000 description 19
- 230000000802 nitrating effect Effects 0.000 description 17
- 239000000047 product Substances 0.000 description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- 239000006187 pill Substances 0.000 description 8
- 230000014759 maintenance of location Effects 0.000 description 7
- 238000005303 weighing Methods 0.000 description 7
- 230000000694 effects Effects 0.000 description 6
- 230000036541 health Effects 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 208000025747 Rheumatic disease Diseases 0.000 description 4
- 238000004587 chromatography analysis Methods 0.000 description 4
- 238000004090 dissolution Methods 0.000 description 4
- 239000008297 liquid dosage form Substances 0.000 description 4
- 239000000041 non-steroidal anti-inflammatory agent Substances 0.000 description 4
- 229940021182 non-steroidal anti-inflammatory drug Drugs 0.000 description 4
- 230000000552 rheumatic effect Effects 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 208000006820 Arthralgia Diseases 0.000 description 3
- 208000002193 Pain Diseases 0.000 description 3
- -1 aromatic amine compounds Chemical class 0.000 description 3
- 210000000988 bone and bone Anatomy 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000013642 negative control Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000003826 tablet Substances 0.000 description 3
- 238000004809 thin layer chromatography Methods 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- 206010006002 Bone pain Diseases 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 208000017667 Chronic Disease Diseases 0.000 description 2
- 240000001723 Entada phaseoloides Species 0.000 description 2
- 208000008558 Osteophyte Diseases 0.000 description 2
- 206010033799 Paralysis Diseases 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 230000000202 analgesic effect Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- UDSAIICHUKSCKT-UHFFFAOYSA-N bromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C2=CC=CC=C2S(=O)(=O)O1 UDSAIICHUKSCKT-UHFFFAOYSA-N 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 201000010934 exostosis Diseases 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- 238000006396 nitration reaction Methods 0.000 description 2
- 150000003180 prostaglandins Chemical class 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 229940098465 tincture Drugs 0.000 description 2
- 210000002700 urine Anatomy 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 101800004538 Bradykinin Proteins 0.000 description 1
- 235000003255 Carthamus tinctorius Nutrition 0.000 description 1
- 244000020518 Carthamus tinctorius Species 0.000 description 1
- MDNWOSOZYLHTCG-UHFFFAOYSA-N Dichlorophen Chemical compound OC1=CC=C(Cl)C=C1CC1=CC(Cl)=CC=C1O MDNWOSOZYLHTCG-UHFFFAOYSA-N 0.000 description 1
- ITIONVBQFUNVJV-UHFFFAOYSA-N Etomidoline Chemical compound C12=CC=CC=C2C(=O)N(CC)C1NC(C=C1)=CC=C1OCCN1CCCCC1 ITIONVBQFUNVJV-UHFFFAOYSA-N 0.000 description 1
- 208000012671 Gastrointestinal haemorrhages Diseases 0.000 description 1
- QXZGBUJJYSLZLT-UHFFFAOYSA-N H-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-OH Natural products NC(N)=NCCCC(N)C(=O)N1CCCC1C(=O)N1C(C(=O)NCC(=O)NC(CC=2C=CC=CC=2)C(=O)NC(CO)C(=O)N2C(CCC2)C(=O)NC(CC=2C=CC=CC=2)C(=O)NC(CCCN=C(N)N)C(O)=O)CCC1 QXZGBUJJYSLZLT-UHFFFAOYSA-N 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102100035792 Kininogen-1 Human genes 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000008469 Peptic Ulcer Diseases 0.000 description 1
- 244000018633 Prunus armeniaca Species 0.000 description 1
- 235000009827 Prunus armeniaca Nutrition 0.000 description 1
- 208000008765 Sciatica Diseases 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 238000003811 acetone extraction Methods 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- QXZGBUJJYSLZLT-FDISYFBBSA-N bradykinin Chemical compound NC(=N)NCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(=O)NCC(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CO)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)CCC1 QXZGBUJJYSLZLT-FDISYFBBSA-N 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 239000008298 dragée Substances 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- LJQKCYFTNDAAPC-UHFFFAOYSA-N ethanol;ethyl acetate Chemical compound CCO.CCOC(C)=O LJQKCYFTNDAAPC-UHFFFAOYSA-N 0.000 description 1
- 238000003810 ethyl acetate extraction Methods 0.000 description 1
- 238000010812 external standard method Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 201000006334 interstitial nephritis Diseases 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 239000012982 microporous membrane Substances 0.000 description 1
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 1
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 description 1
- 230000036407 pain Effects 0.000 description 1
- 208000011906 peptic ulcer disease Diseases 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229940006198 sodium phenylacetate Drugs 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000007940 sugar coated tablet Substances 0.000 description 1
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a rapid detection method of diclofenac sodium added in antirheumatic Chinese patent medicines or health foods. The method comprises steps that: (1) a solid sample is added to ethyl acetate and a mass volume concentration is regulated to 0.05 to 0.10g/mL, the mixture is shaken until dissolved, the solution is filtered, and the obtained filtrate is a liquid to be detected; or a liquid sample is added to ethyl acetate with a volume 2.5 to 10 times that of the liquid sample, the mixture is settled, and an ethyl acetate extract is obtained as a liquid to be detected; (2) 1mL of the liquid to be detected is fetched, 1mL of a 65% nitric acid solution is added to the liquid, and the mixture is settled and laminated for 20 to 30 seconds; if a lower layer solution is reddish brown, the sample contains diclofenac sodium. The detection method provided by the invention has advantages of simple operation, high specificity, and rapid color reaction. The method can be used in in-site rapid detection of diclofenac sodium doped in antirheumatic Chinese patent medicines or health foods.
Description
Technical field
The present invention relates to utilize the detection of precipitating action, be specifically related to the detection of C14H10Cl2NNaO2 in the medicine non-biological material.
Background technology
C14H10Cl2NNaO2 (Diclofenac Sodium), chemistry 2-[(2 by name, 6-dichlorophenyl) amino] sodium phenylacetate.The structural formula of antiphen sodium as the formula (1), molecular formula is C
12H
10ClN
2Cl
2NNaO
2, molecular weight is 318.13, Cas number is 15307-79-6.C14H10Cl2NNaO2 is the colourless crystallization powder, and fusing point is 283-285 ℃; Be soluble in ethanol, be slightly soluble in water, insoluble in chloroform.
C14H10Cl2NNaO2 is a kind of NSAID (non-steroidal anti-inflammatory drug), and its main mechanism is by suppressing the synthetic of maincenter prostaglandin, brings into play that it is analgesic, analgesia, antiinflammation.It can suppress leukocytic gathering by suppressing the synthetic of prostaglandin, reduces the formation of bradykinin, suppresses effect performance antiinflammations such as hematoblastic aggegation.
There is potential cardiovascular and hemorrhage of digestive tract risk in the use of C14H10Cl2NNaO2, the patient is after having taken the Chinese patent drug or health food that contains C14H10Cl2NNaO2 under the unwitting situation, might produce serious consequence, endanger healthy, among the patient as long-term oral NSAID (non-steroidal anti-inflammatory drug), peptic ulcer takes place in nearly 10%～25% patient, and severe complications can cause hemorrhage or perforation; It is unusual to cause 10% patient the slightly impaired biochemistry of liver to occur; Can cause urine protein, cast, can occur red, leucocyte etc. in the urine, severe patient can cause interstitial nephritis etc.Therefore, this medicine in use should be strict with indication, abides by the regulation of package insert and use under physician guidance.Though this medicine belongs to non-prescribed medicine at present mostly,, there is certain potential risk if take for a long time.
Rheumatism is commonly called as " not dead cancer ", is at every moment endangering human beings'health, obstinate, and outbreak makes patient's pain repeatedly, makes the doctor have a headache.Authority's investigation shows that about 400,000,000 people of global rheumatisant are class illness the hugest in the medical domain, and rheumatism has become " chronic disease " that endangers health of people constantly, and 2000 so far, and annual rheumatisant's medication amount of money has all surpassed 200 hundred million! In China, have at least 200,000,000 people tormented by this chronic disease, 80% is effectively treated, and controls repeatly and does not heal, and sb.'s illness took a turn for the worse, is forced to the edge of paralysis, according to relevant expert's statistics, 40% rheumatic sufferers appearance paralysis arranged national every year.
At present, treating rheumatismal medicine has chemical medicine, compound Chinese medicinal preparation and pure Chinese medicinal preparation, and wherein, chemical medicine onset is rapid, and result of treatment is obvious, but has taken than serious adverse for a long time.
And treatment rheumatoid disease product (comprising Chinese patent drug and the health food) spinoff that the natural drug Chinese herbal medicine is made is less,
But onset is slower, and result of treatment does not have chemical medicine remarkable.
The C14H10Cl2NNaO2 that is used for the treatment of rheumatoid disease is because cheap, and the certain control rheumatic and the effect of rheumatoid arthritis are arranged.Not only think to treat rheumatoid disease fast but also think the psychology that spinoff is little in order to cater to the patient, C14H10Cl2NNaO2 is added in the medicine and health care health food of treatment rheumatism, rheumatism repeatly in recent years.
At present, detect the main high-efficient liquid phase technique of method and the high performance liquid chromatogram chromaticness logotype method of doping C14H10Cl2NNaO2 in medicine and the health products.But said method need use expensive analytical instrument, to the environmental requirement height, needs fixedly to put, and the mobile big scene of uncomfortable cooperation is detected.Use acetone extraction to obtain sample in the document (), add bromophenol blue in sample, positive reference substance presents blueness, and the negative control product present faint yellow.But the present invention adopts said method to carry out replica test, and it is blue to find that positive does not occur, and occurring blue is not positive, and the shown color of positive is inconsistent, and negative sample also shows the color consistent with positive.
Summary of the invention
The technical matters of solution of the present invention provides a kind of method for quick that adds C14H10Cl2NNaO2 in on-the-spot Chinese patent drug that detects antirheumatic of drug surveilance department or the health food, this method accuracy height of being applicable to.
The present invention is as described below in order to solve the problems of the technologies described above technical scheme:
Add the method for quick of C14H10Cl2NNaO2 in a kind of Chinese patent drug of antirheumatic or the health food, this method is made up of following steps:
(1) getting solid sample adding ethyl acetate configuration quality volumetric concentration is 0.05-0.10g/mL solution, and jolting is molten looses, and filters, and gets filtrate as liquid to be measured; Perhaps get fluid sample, the ethyl acetate that adds 2.5-10 times of volume mixes, and leaves standstill and gets acetic acid ethyl acetate extract as liquid to be measured;
(2) get 1mL liquid to be measured, add the 1mL65% salpeter solution, standing demix after 20～30 seconds, if lower floor's solution presents rufous, then contains C14H10Cl2NNaO2 in the judgement sample.
Measuring samples in the method for detection C14H10Cl2NNaO2 of the present invention can be fluid sample or solid sample, and when measuring samples was solid sample, ethyl acetate was to extract the wherein optimal selection of C14H10Cl2NNaO2, and its principle is as described below:
The principle of the inventive method is the physicochemical property according to C14H10Cl2NNaO2, and is easily molten in ethanol, molten in the water part omitted, insoluble characteristics in methenyl choloride, according to its dissolubility, this method adopts ethyl acetate as extraction solvent, extraction C14H10Cl2NNaO2 is wherein removed other impurity.C14H10Cl2NNaO2 is an aromatic amine compounds, can carry out nitration reaction with nitric acid, after this type of aromatic amine compounds is introduced nitro, generates the rufous material, thereby differentiates.
Chemical equation is: R-H+HO-NO
2=R-NO
2+ H
2O
Method for quick of the present invention detect that the process of C14H10Cl2NNaO2 is in the product to be tested, use the C14H10Cl2NNaO2 in the ethyl acetate extraction testing sample earlier; As detection agent, make the nitrated formation rufous of C14H10Cl2NNaO2 material with salpeter solution.
In the method for quick of the present invention, when sample to be checked is red tablet, its red dressing need be removed, add acetic acid ethyl dissolution after the pulverizing again; When sample to be checked is pill, directly pulverize or shred, add acetic acid ethyl dissolution again; When sample to be checked is capsule, directly get content, add acetic acid ethyl dissolution again; Oral liquid is directly got dose, adds acetic acid ethyl dissolution again.
According to the design philosophy of above-mentioned testing process, in the selection of the reagent of each step, following a large amount of examination and exploration have been carried out.
2, the selection of nitrating agent
C14H10Cl2NNaO2 is an aromatic amine compounds, can carry out nitration reaction with the nitric acid of variable concentrations, and generates the rufous material.The inventive method adopts the nitric acid of nine kinds of variable concentrations to test as nitrating agent, 80% nitric acid, 75%, 70%, 65%, 60% nitric acid, 55% nitric acid, 50% nitric acid, 45% nitric acid, 40% nitric acid, 35% nitric acid, 30% nitric acid and the various variable concentrations nitrating agents of 10% nitric acid are used in contrast, test by the method for drafting respectively, the result shows: use 80%～70% nitric acid to make nitrating agent and develop the color immediately; Use 65% nitric acid to develop the color after about 10～20 seconds as nitrating agent; Use 60% nitric acid to develop the color after about 40 seconds as nitrating agent; Use 55% nitric acid to develop the color after about 60 seconds as nitrating agent; Use 50% nitric acid to develop the color after about 90 seconds as nitrating agent; Use 45% nitric acid to develop the color after about 120 seconds as nitrating agent; Use 40%～35% nitric acid to show slightly faint yellow after 120 seconds as nitrating agent; Longer just apparent faint yellow even colour developing of time when using 30%～10% nitric acid as nitrating agent.See with 80%, 75%, 70% nitric acid from effect to be better than 65% nitric acid, but corrosivity is stronger, reacts more violent as the nitrating agent effect; More consuming time and effect is very unobvious when making nitrating agent with 60% following nitric acid.Select for use 65% nitric acid as nitrating agent, all more suitable on the time and the extent of reaction; So selecting concentration for use is that 65% nitric acid is as nitrating agent.
For the ease of implementing detection method of the present invention, can adopt conventional method that described all ingredients is made the reagent for quickly examining box, also can make the fast detecting suit of the C14H10Cl2NNaO2 that uses on the suitable medicine detection vehicle.
Compared with prior art, the present invention has following beneficial effect: in the detection method of the present invention, the specificity of used extract and detection agent is strong, remove interfering material by utilizing the dissolubility difference of different material to separate in extraction and the reextraction process, make the highly sensitive of chromogenic reaction, assay accuracy height, chromogenic reaction is swift in response simultaneously, phenomenon is obvious, so the present invention is applicable to whether mix C14H10Cl2NNaO2 in field quick detection medicine, Chinese patent drug and the health products.
Description of drawings
Fig. 1 is a C14H10Cl2NNaO2 lowest detection limit.Test tube 1 is a negative control sample bone spur pain-eliminating capsule, test tube 2 adds each 25mg of C14H10Cl2NNaO2 reference substance, test tube 3 adds C14H10Cl2NNaO2 reference substance 20mg, test tube 4 adds C14H10Cl2NNaO2 reference substance 15mg, test tube 5 adds C14H10Cl2NNaO2 reference substance 10mg, and test tube 6 adds C14H10Cl2NNaO2 reference substance 5mg.
Fig. 2 is the testing result of document 1 described method.Test tube 1 is a Co treating rheumatic ostealgia Yiganning capsule, and Central Plains, Henan folk secret development popularization center produces lot number 2009.11.08; Test tube 2 is the fine and soft strong bone capsule for treating arthralgia aggravated by cold of first, and Taiqian County, Henan Province people's livelihood pharmaceutcal corporation, Ltd produces, lot number 2009.11.08; Test tube 3 is efficient capsule for curing rheumatism, and positive magnificent medicine company incorporated company produces lot number 2009.11.08; Test tube 4 is Tongning capsules, and people's livelihood pharmaceutcal corporation, Ltd in Henan produces, lot number 100706; Test tube 5 is rheumatism jinxs, the big pharmacy of gift scholar (Hong Kong); Test tube 6 is FENGSHIDING JIAONANGs, and Tonghua continuous heavy rain pharmaceutcal corporation, Ltd of shaking produces lot number 100901; Test tube 7 is compound Small Meridian-Activating Pills, and the Tongrentang Pharmaceutical Factory, Beijing TongrenTang Co., Ltd produces, lot number 0913425; Test tube 8 is TONGREN DAHUOLUO WANs, and the Tongrentang Pharmaceutical Factory, Beijing TongrenTang Co., Ltd produces, lot number 0013509; Test tube 9 is safe and sound an of rheumatism, and Tongyao Pharmaceutical Group Co., Ltd produces, lot number 100703; Test tube 10 is Entada phaseoloides sheets, and peace pharmaceutcal corporation, Ltd in Guangdong produces lot number 100101.
Fig. 3 is the testing result of the method for the invention.Test tube 1 is a Co treating rheumatic ostealgia Yiganning capsule, and Central Plains, Henan folk secret development popularization center produces lot number 2009.11.08; Test tube 2 is the fine and soft strong bone capsule for treating arthralgia aggravated by cold of first, and Taiqian County, Henan Province people's livelihood pharmaceutcal corporation, Ltd produces, lot number 2009.11.08; Test tube 3 is efficient capsule for curing rheumatism, and positive magnificent medicine company incorporated company produces lot number 2009.11.08; Test tube 4 is Tongning capsules, and people's livelihood pharmaceutcal corporation, Ltd in Henan produces, lot number 100706; Test tube 5 is rheumatism jinxs, the big pharmacy of gift scholar (Hong Kong); Test tube 6 is FENGSHIDING JIAONANGs, and Tonghua continuous heavy rain pharmaceutcal corporation, Ltd of shaking produces lot number 100901; Test tube 7 is compound Small Meridian-Activating Pills, and the Tongrentang Pharmaceutical Factory, Beijing TongrenTang Co., Ltd produces, lot number 0913425; Test tube 8 is TONGREN DAHUOLUO WANs, and the Tongrentang Pharmaceutical Factory, Beijing TongrenTang Co., Ltd produces, lot number 0013509; Test tube 9 is safe and sound an of rheumatism, and Tongyao Pharmaceutical Group Co., Ltd produces, lot number 100703; Test tube 10 is Entada phaseoloides sheets, and peace pharmaceutcal corporation, Ltd in Guangdong produces lot number 100101.
Embodiment
Following example will help those of ordinary skill in the art further to understand the present invention, but not limit the present invention in any form.
The detection of example 1 capsule
Sample to be checked: (indicate Taiqian County, Henan Province people's livelihood pharmaceutcal corporation, Ltd produces the fine and soft strong bone capsule for treating arthralgia aggravated by cold of first, lot number 20091108), high performance liquid chromatography-DAD testing result: in the test sample chromatogram, showing identical chromatographic peak with C14H10Cl2NNaO2 reference substance retention time place, be positive, contain C14H10Cl2NNaO2.
Detect step:
(1) sample thief is 2, and its content (about 0.5g) is added 5mL ethyl acetate, close plug, and about 1 minute of strong up and down jolting is left standstill slightly, filters, and gets filtrate as liquid to be measured;
(2) get liquid 1mL to be measured in the 10ml test tube, slowly add 65% salpeter solution 1ml along test tube wall, standing demix was observed after 20～30 seconds;
Interpretation as a result: lower floor's solution is rufous, contains C14H10Cl2NNaO2 in the interpret sample, and is consistent with high performance liquid chromatography-DAD testing result.
The detection of example 2 pills
Sample to be checked: sciatica ball (indicate apricot woods medicine company Co., Ltd. and supervise production, lot number 090108)
High performance liquid chromatography-DAD testing result: in the test sample chromatogram,, be negative, do not contain C14H10Cl2NNaO2 showing with the C14H10Cl2NNaO2 reference substance retention time colourless spectrum peak that exists together mutually.
The inventive method detects:
(1) get described pill 2 balls (about 0.4g), pulverize, add 5mL ethyl acetate, close plug, about 1 minute of strong up and down jolting is left standstill slightly, filters, and gets filtrate as liquid to be measured;
((2) get liquid 1mL to be measured in the 10ml test tube, slowly add 65% salpeter solution 1ml along test tube wall, and standing demix was observed after 20～30 seconds;
Interpretation as a result: lower floor's solution does not have change color, does not contain C14H10Cl2NNaO2 in the interpret sample, and is consistent with high performance liquid chromatography-DAD testing result.
The detection of example 3 tablets
Sample to be checked: FUFANG FENGSHINING PIAN (indicate Guangdong LuoFoshan Medicine Co., Ltd and produce lot number L09D090).
High performance liquid chromatography-DAD testing result: in the test sample chromatogram,, be negative, do not contain C14H10Cl2NNaO2 not having the chromatographic peak demonstration with place, the identical peak of C14H10Cl2NNaO2 reference substance retention time.
The inventive method detects:
(1) get 1 of described pill, remove sugar-coat (about 0.25g), pulverize, add 5mL ethyl acetate, close plug, about 1 minute of strong up and down jolting is left standstill slightly, filters, and gets filtrate as liquid to be measured;
(2) get liquid 1mL to be measured in the 10ml test tube, slowly add 65% salpeter solution 1ml along test tube wall, standing demix was observed after 20～30 seconds;
Interpretation as a result: lower floor's solution does not have change color, does not contain C14H10Cl2NNaO2 in the interpret sample, and is consistent with high performance liquid chromatography-DAD testing result.
The detection of example 4 tinctures
Sample to be checked: safflower analgesic tincture (indicate Ltd of South of Guangxi pharmacy group and produce lot number 090401).
High performance liquid chromatography-DAD testing result: in the test sample chromatogram, do not have chromatographic peak at place, the identical peak of C14H10Cl2NNaO2 reference substance retention time and show, be negative, do not contain C14H10Cl2NNaO2
The inventive method detects:
(1) gets the about 1mL of sample to be checked, add 5mL ethyl acetate, shake up, as test liquid;
(2) get liquid 1mL to be measured in the 10ml test tube, slowly add 65% salpeter solution 1ml along test tube wall, standing demix was observed after 20～30 seconds;
Interpretation as a result: lower floor's solution colour shoals, and does not contain C14H10Cl2NNaO2 in the interpret sample, and is consistent with high performance liquid chromatography-DAD testing result.
The detection of the liquid dosage form of example 5 other types
Sample to be checked: liquid dosage form (indicate Ltd of South of Guangxi pharmacy group and produce lot number 090401).
High performance liquid chromatography-DAD testing result: in the test sample chromatogram, do not have chromatographic peak at place, the identical peak of C14H10Cl2NNaO2 reference substance retention time and show, be negative, do not contain C14H10Cl2NNaO2
The inventive method detects:
(1) gets the about 0.5mL of sample to be checked, add 5mL ethyl acetate, leave standstill and get acetic acid ethyl acetate extract as liquid to be measured;
(2) get liquid 1mL to be measured in the 10ml test tube, slowly add 65% salpeter solution 1ml along test tube wall, standing demix was observed after 20～30 seconds;
Interpretation as a result: lower floor's solution colour shoals, and does not contain C14H10Cl2NNaO2 in the interpret sample, and is consistent with high performance liquid chromatography-DAD testing result.
The detection of the liquid dosage form of example 6 other types
Sample to be checked: liquid dosage form (indicate Ltd of South of Guangxi pharmacy group and produce lot number 090401).
High performance liquid chromatography-DAD testing result: in the test sample chromatogram, do not have chromatographic peak at place, the identical peak of C14H10Cl2NNaO2 reference substance retention time and show, be negative, do not contain C14H10Cl2NNaO2
The inventive method detects:
(1) gets the about 2mL of sample to be checked, add 5mL ethyl acetate, shake up, leave standstill and get acetic acid ethyl acetate extract as liquid to be measured;
(2) get liquid 1mL to be measured in the 10ml test tube, slowly add 65% salpeter solution 1ml along test tube wall, standing demix was observed after 20～30 seconds;
Interpretation as a result: lower floor's solution colour shoals, and does not contain C14H10Cl2NNaO2 in the interpret sample, and is consistent with high performance liquid chromatography-DAD testing result.
The contrast experiment 1: the determining of lowest detectable limit
Get negative sample respectively " bone spur pain-eliminating capsule " (Tonghua continuous heavy rain pharmaceutcal corporation, Ltd of shaking, lot number 101001) each dose (4) is in 20mL tool plug test tube, add each 25mg of C14H10Cl2NNaO2 reference substance, 20mg, 15mg, 10mg and 5mg respectively from second test tube, add ethyl acetate 5mL respectively, close plug, about 1 minute of strong up and down jolting is left standstill slightly, filter, divide and get filtrate as test liquid; Get the about 1ml of test liquid in the 10ml test tube, slowly add 1mL 65% salpeter solution, observe after about 20～30 seconds, the results are shown in Figure 1 along test tube wall.Among Fig. 1, the solution in first test tube is not developing the color after 180 seconds after adding nitrating agent, and the solution in second to the 5th test tube adds nitrating agent and all shows rufous after 20～30 seconds, a kind ofly shows faint yellow at last, so minimum detectability is 2mg.
The contrast experiment 2: the inventive method detects the effect experiment of commercial goods
1, test sample: collect 30 in (mainly buying by market) sample from market, name of product and product batch number see Table 1.
2, adopt the inventive method that test sample is detected, adopt thin-layer chromatography and high performance liquid chromatography-DAD to detect contrast simultaneously.
(1) thin-layered chromatography: the sample of getting a dose adds ethanol 10mL in tool plug conical flask, and jolting was extracted 1 minute, filters, and filtrate is as need testing solution; It is an amount of that other gets the C14H10Cl2NNaO2 reference substance, adds ethanol and make the solution that every 1mL contains 2.5mg, in contrast product solution.According to thin-layered chromatography (Chinese Pharmacopoeia version appendix in 2005 VI B) test, draw above-mentioned two kinds of each 1uL of solution respectively, put respectively on same gel GF 254 plate, with ethyl acetate-ethanol (15: 5) is developping agent, launches, and takes out, dry, put under the ultraviolet lamp (254nm) and inspect.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, if show the fluorescence spot of same color, then may contain C14H10Cl2NNaO2 in the decidable sample.
(2) high performance liquid chromatography
It is the chromatographic column of packed column that chromatographic condition and system flexibility adopt octadecylsilane chemically bonded silica, with diode array formula detecting device (DAD) is detecting device, with methyl alcohol-0.2% glacial acetic acid solution (3: 2) is moving phase, the detection wavelength is 281nm, 30 ℃ of column temperatures, sample size is 10uL, and the theoretical cam curve of C14H10Cl2NNaO2 is not less than 10000, with the degree of separation of other chromatographic peak greater than 1.5.
The preparation precision of reference substance solution takes by weighing the about 15mg of C14H10Cl2NNaO2 reference substance, puts in the 100mL measuring bottle, adds an amount of methyl alcohol jolting and makes dissolving, adds methyl alcohol to scale, shakes up, in contrast product solution.
The preparation of need testing solution grinds after the weighing if test sample is tablet (answering the desaccharification clothing as sugar coated tablet), takes by weighing the sample of a dose then, put in the 100mL measuring bottle, it is an amount of to add methyl alcohol, sonicated 10 minutes, put and be chilled to room temperature, add methyl alcohol and be diluted to scale, shake up; If test sample is a capsule, take out content after the weighing and grind, take by weighing the sample of a dose then, put in the 100mL measuring bottle, it is an amount of to add methyl alcohol, and sonicated 10 minutes is put and is chilled to room temperature, adds methyl alcohol and is diluted to scale, shakes up; If test sample is a pill, grind after the weighing or shred, take by weighing the sample of a dose then, put in the 100mL measuring bottle, it is an amount of to add methyl alcohol, and sonicated 10 minutes is put and is chilled to room temperature, adds methyl alcohol and is diluted to scale, shakes up; If test sample is an oral liquid, directly measure the sample of a dose, put in the 100mL measuring bottle, add methyl alcohol and be diluted to scale, shake up; If test sample is a tincture, directly measure the sample of about 1mL, put in the 100mL measuring bottle, it is an amount of to add methyl alcohol, and sonicated 10 minutes is put and is chilled to room temperature, adds methyl alcohol and is diluted to scale, shakes up.
Determination method is drawn each 10uL of above-mentioned solution behind filtering with microporous membrane respectively, injects liquid chromatograph, measures, and calculates promptly by external standard method.
The result judges if appearance then can tentatively be proved conclusively and contain C14H10Cl2NNaO2 in the test sample with consistent with the retention time of C14H10Cl2NNaO2 reference substance chromatographic peak in the test sample chromatographic peak.
3, result:
The result is as shown in table 1, and in 30 batches of test samples, the inventive method detects 8 batches of positives, 22 batches of feminine genders.Confirm to find that by high performance liquid chromatography wherein 8 batches is true positives, 22 batches of true negatives, 0 feminine gender of permitting a leave, by above data statistics as can be known, the inventive method sensitivity, accuracy and specificity are as follows:
Sensitivity S e=true positives/(true positives+false negative)=8/ (8+0)=1.00
Accuracy AR=(true positives+true negative)/sum=(8+22)/31=1.00
Specificity Sp=true negative/(true negative+false positive)=23/23=1.00
This shows that the inventive method has very high accuracy.
Table 1
Embodiment 3
The described method of method of the present invention and document (1) compares
1, test sample: collect 30 in (mainly buying by market) sample from market, name of product and product batch number see Table 1.2, adopt the inventive method and document 1 described method that test sample is detected, adopt thin-layer chromatography and high performance liquid chromatography-DAD to detect contrast simultaneously.
Detection method:
The method of the invention, its step is shown in embodiment 1.
Document 1 described method: (1) gets 1 dose of test sample (solid pharmaceutical preparation porphyrize, pill can add a spot of zeyssatite porphyrize, liquid preparation elder generation evaporate to dryness), puts in the tool plug conical flask, adds acetone 10mL, and jolting 5min filters, and filtrate is as need testing solution.
(2) get each 2mL of need testing solution, reference substance solution, negative control solution and positive control solution, put respectively in the test tube, drip 1 of 0.05% bromophenol blue solution, jolting.If it is blue that solution shows, then contain C14H10Cl2NNaO2, faint yellow if solution shows, then do not contain C14H10Cl2NNaO2.
Thin-layer chromatography: with embodiment 2.
High performance liquid chromatography-DAD detects, with embodiment 2.
Result: as shown in table 2.Can learn from table 2, result and the described phenomenon of document (1) of using the described method of document (1) to detect have discrepancy: positive does not occur blue, what blueness occurs is not positive, and the shown color of positive is inconsistent, and negative sample also shows the color consistent with positive.Therefore the described method of document (1) exists detection accuracy low, is suitable for the little shortcoming of medicine scope.The method of the invention testing result is identical with the liquid phase chromatography assay with thin-layered chromatography, and the high advantage of detection accuracy is just arranged.
Table 2
Claims (3)
1. add the method for quick of C14H10Cl2NNaO2 in the Chinese patent drug of an antirheumatic or the health food, this method is made up of following steps:
(1) getting solid sample adding ethyl acetate configuration quality volumetric concentration is 0.05-0.10g/mL solution, and jolting is molten looses, and filters, and gets filtrate as liquid to be measured; Perhaps get fluid sample, the ethyl acetate that adds 2.5-10 times of volume mixes, and leaves standstill and gets acetic acid ethyl acetate extract as liquid to be measured;
(2) get 1mL liquid to be measured, add the 1mL65% salpeter solution, standing demix after 20～30 seconds, if lower floor's solution presents rufous, then contains C14H10Cl2NNaO2 in the judgement sample.
2. add the method for quick of C14H10Cl2NNaO2 in the Chinese patent drug of a kind of antirheumatic according to claim 1 or the health food, it is characterized in that, when described antirheumatic Chinese patent drug and health food were the tablet of dressing, described solid sample was the content of dressing.
3. add the method for quick of C14H10Cl2NNaO2 in the Chinese patent drug of a kind of antirheumatic according to claim 1 or the health food, it is characterized in that, when described antirheumatic Chinese patent drug and health food were capsule, described solid sample was the content of capsule.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101328378A CN102297860A (en) | 2011-05-20 | 2011-05-20 | Rapid detection method of diclofenac sodium added in antirheumatic Chinese patent medicines or health foods |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101328378A CN102297860A (en) | 2011-05-20 | 2011-05-20 | Rapid detection method of diclofenac sodium added in antirheumatic Chinese patent medicines or health foods |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102297860A true CN102297860A (en) | 2011-12-28 |
Family
ID=45358424
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011101328378A Pending CN102297860A (en) | 2011-05-20 | 2011-05-20 | Rapid detection method of diclofenac sodium added in antirheumatic Chinese patent medicines or health foods |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102297860A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103293153A (en) * | 2013-05-20 | 2013-09-11 | 韶关市食品药品检验所 | Rapid screening method for illegal addition of diclofenac sodium in anti-rheumatism Chinese patent medicines or healthcare products |
| CN103616375A (en) * | 2013-11-11 | 2014-03-05 | 江苏省食品药品检验所 | Method and kit for detecting oxicams non-steroidal anti-inflammatory drugs in wind-damp expelling Chinese patent medicines and health-care foods |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101531607A (en) * | 2009-01-20 | 2009-09-16 | 鲁南制药集团股份有限公司 | Improved method for preparing aceclofenac |
| CN101762673A (en) * | 2009-12-09 | 2010-06-30 | 中国人民武装警察部队药品仪器检验所 | Method for detection of illegally added chemical in traditional Chinese medicine preparation |
-
2011
- 2011-05-20 CN CN2011101328378A patent/CN102297860A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101531607A (en) * | 2009-01-20 | 2009-09-16 | 鲁南制药集团股份有限公司 | Improved method for preparing aceclofenac |
| CN101762673A (en) * | 2009-12-09 | 2010-06-30 | 中国人民武装警察部队药品仪器检验所 | Method for detection of illegally added chemical in traditional Chinese medicine preparation |
Non-Patent Citations (3)
| Title |
|---|
| 国家食品药品监督管理局: "《国家药品标准》", 30 April 2005, article "新药转正标准第46册" * |
| 郑凯: "HPLC法测定双氯芬酸钠肠溶片的含量", 《西北医学杂志》, vol. 20, no. 1, 31 December 2005 (2005-12-31) * |
| 闫韶华: "HPLC法测定双氯芬酸钠肠溶片的含量及含量均匀度", 《数理医药学杂志》, vol. 22, no. 6, 31 December 2009 (2009-12-31) * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103293153A (en) * | 2013-05-20 | 2013-09-11 | 韶关市食品药品检验所 | Rapid screening method for illegal addition of diclofenac sodium in anti-rheumatism Chinese patent medicines or healthcare products |
| CN103616375A (en) * | 2013-11-11 | 2014-03-05 | 江苏省食品药品检验所 | Method and kit for detecting oxicams non-steroidal anti-inflammatory drugs in wind-damp expelling Chinese patent medicines and health-care foods |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105372377B (en) | A kind of quality determining method of bulk drug forsythin | |
| CN104730171B (en) | Radix Paeoniae Rubra, Flos Lonicerae multi-target ingredient content assaying method in a kind of compound Chinese medicinal preparation | |
| CN102269751B (en) | Detection method of Liuweinengxiao preparation | |
| CN102879495B (en) | Antidotal capsule of Tibetan medicine compound and quality detection method of preparation of antidotal capsule | |
| CN101829266A (en) | Method for detecting quality of bezoar snake bile bulbus fritilariae liquid | |
| CN105510312B (en) | A kind of method for quick of orlistat | |
| CN102269752B (en) | Detection method for pharmaceutical composition preparation | |
| CN102393391B (en) | Rapid determination method of melatonin adulterated in Chinese patent medicine and health food | |
| CN102297860A (en) | Rapid detection method of diclofenac sodium added in antirheumatic Chinese patent medicines or health foods | |
| CN100577677C (en) | Caulis trachelospermi total lignans extractive, extraction method and medicine use of the active constituent thereof | |
| Fitrianasari et al. | Qualitative Analysis Of Drug Substances in Rheumatic Jamu Samples Using Thin Layer Chromatography | |
| CN104998071A (en) | Compound preparation ofherb of dense flower Bulbophyllum and preparation and detection method for said compound preparation | |
| CN101647993A (en) | Medicament for treating flu and preparation and detection method thereof | |
| CN101606950A (en) | A kind of pediatric paracetamol,atificial cowbezoar and chlorphenamine maleate granule and method of quality control | |
| CN101278976A (en) | Quality control method of bone-knitting medicine for traumatology department | |
| CN104597197B (en) | A kind of detection method of the long-pending defaecation pharmaceutical preparation that disappears | |
| Braimah et al. | Impurity profiling of paracetamol dosage forms used in maiduguri metropolis | |
| El-Shaheny et al. | Analysis of ofloxacin and flavoxate HCl either individually or in combination via a green chromatographic approach with a pharmacokinetic study of ofloxacin in biological samples | |
| CN106404953B (en) | A kind of quality detection method of penicillin skin test freeze-dried powder | |
| CN115575546A (en) | Construction method and content determination method of HPLC (high Performance liquid chromatography) fingerprint spectrum of Wuzi Yanzong soft capsule | |
| CN1923264B (en) | Capsule comprising artemisia capillaries and rhizoma imperatae for treating hepatitis | |
| Ahmad et al. | RP-UHPLC and HPTLC method development and validation for analysis of andrographolide from herbal hepatoprotective formulation | |
| CN107449844B (en) | Method for determining dissolution rate of dimercaptosuccinic acid preparation | |
| CN112666278A (en) | Limit detection method for strychnine in Huatuo reconstruction pills | |
| CN101596229A (en) | The method of quality control of Ramulus Et Folium Pithecellobii Lucidi extract and preparation thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20111228 |