CN105203673B - The high-efficiency liquid chromatography method for detecting of Compound Solanum Nigrum sulfathiazole - Google Patents
The high-efficiency liquid chromatography method for detecting of Compound Solanum Nigrum sulfathiazole Download PDFInfo
- Publication number
- CN105203673B CN105203673B CN201510780966.6A CN201510780966A CN105203673B CN 105203673 B CN105203673 B CN 105203673B CN 201510780966 A CN201510780966 A CN 201510780966A CN 105203673 B CN105203673 B CN 105203673B
- Authority
- CN
- China
- Prior art keywords
- mobile phase
- sulfathiazole
- solanum nigrum
- methanol
- 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.)
- Expired - Fee Related
Links
Abstract
The invention discloses a kind of high-efficiency liquid chromatography method for detecting of Compound Solanum Nigrum sulfathiazole, described Compound Solanum Nigrum sulfathiazole is mainly made up of black nightshade, pennywort, pithecellobium clypearia, predominantly detects gallic acid in Compound Solanum Nigrum sulfathiazole, Quercetin, Kaempferide, solasonine, the content of solamargine.The present invention carries out substep detection according to this two class of the alkaloids substance such as the phenolic acid such as gallic acid, Quercetin, Kaempferide and solamargine, solasonine in the sulfathiazole, this detection method detection accuracy and detection rates are high, and use it for the assay of the sulfathiazole, this method ensures the accuracy and advance of the sulfathiazole quality inspection standard, can as the sulfathiazole quality control important indicator.
Description
Technical field
The present invention relates to drug tests.It is more particularly related to which a kind of Compound Solanum Nigrum sulfathiazole is efficient
Liquid chromatography detecting method.
Background technology
Herbal mixture is the principal mode of tcm clinical practice medication, is traditional Chinese medicine and pharmacy characteristic and marrow.Chinese medicine is one by many
The complex system of composition, multifactor composition, its chemical composition diversity and complexity are its curative effect material bases, during foundation meets
The modern mass control system of medical feature, captures quality analysis of traditional Chinese medicine and evaluates problem, improve the existing quality control side of Chinese medicine
Method turns into the positive research topic of people.
Setting up must be based on characteristics of Traditional Chinese Medicine to the quality control system of Chinese medicine.Herbal mixture mass action feature is determined
Chinese medicine is different from Western medicine.Traditional Chinese medicine quality control method is tackled multiple active ingredients and is controlled.Only in this way, the quality set up
Control system can just be really achieved control traditional Chinese medicine quality, ensure Chinese medicine purpose safe and effective for medication.Traditional Chinese medicine quality control should conform to the principle of simplicity
Single single component content, which is determined, to be turned to using advanced technology as means, multicomponent, multi objective assay.
Compound Solanum Nigrum sulfathiazole is mainly made up of black nightshade, pennywort, pithecellobium clypearia, its medicinal active ingredient have gallic acid,
Quercetin, Kaempferide, solasonine, solamargine etc., with clearing heat and detoxicating, cough-relieving apophlegmatic, dispersing swelling and dissipating binds analgesic efficacy, plus
Strong antibacterial, antiviral, antiphlogistic effects, improve immunity, and diuresis antidiarrheal cures mainly body surface, respiratory system, digestive system, urinary system
Infection and the inflammation such as system.
Compound Solanum Nigrum sulfathiazole is as herbal mixture, only from appearance character, simple discriminating and single component assay etc.
Angle carries out quality control to Compound Solanum Nigrum sulfathiazole, it is clear that can not accurately illustrate its inherent quality comprehensively, it is impossible to ensure that it is treated
Effect.Control its effect, it is necessary to according to Chinese medicine multicomponent, Mutiple Targets, multipath action character, its material is integrally controlled
System, effectively characterizes the quality of Chinese medicine on the whole, sets up quality standard system safely, effectively, stable.
Compound Solanum Nigrum sulfathiazole is the medicine of new research and development processing, and Compound Solanum Nigrum sulfathiazole raw material comprising 3 taste Chinese medicines and
There is complicated interaction between traditional Chinese medicine ingredients, cause to set up and fully and effectively reflect Compound Solanum Nigrum sulfathiazole inherent quality
There is larger difficulty with the effective substance method of curative effect, therefore, current this area is present to that can characterize compound dragon comprehensively
The demand of the detection method of this medicine of certain herbaceous plants with big flowers sulfathiazole.
The content of the invention
In order to solve the above problems, it is an object of the invention to provide a kind of detection method of Compound Solanum Nigrum sulfathiazole, the party
Method can compared with the medicine activity component of complete detection Compound Solanum Nigrum sulfathiazole content, so as to characterize and control its inherent quality.
To achieve these goals, the invention provides a kind of high performance liquid chromatography detection side of Compound Solanum Nigrum sulfathiazole
Method, described Compound Solanum Nigrum sulfathiazole is mainly made up of black nightshade, pennywort, pithecellobium clypearia, and the detection method is included using efficient
Gallic acid, Quercetin in the liquid chromatography detection Compound Solanum Nigrum sulfathiazole, Kaempferide, solasonine, Australia eggplant side
Alkali.
Preferably, gallic acid, Quercetin, the Kaempferia galanga in Compound Solanum Nigrum sulfathiazole are detected using described detection method
The content of element comprises the following steps：
1) chromatographic condition：
Chromatographic column：Octadecylsilane chemically bonded silica is used for filler；
Mobile phase：The mixed liquor of methanol and phosphoric acid solution, the volume fraction of phosphoric acid solution is 0.4%, methanol and phosphoric acid water
Volume ratio be 55:45；
Flow velocity：1mL/min；
Column temperature：25℃；
Gradient wavelength detecting：0~15min, Detection wavelength is 270nm；15~30min, Detection wavelength is 362nm；
2) mixing contrast solution is prepared：Precision weighs gallic acid reference substance, Quercetin reference substance, Kaempferol reference substance and fitted
0.5mg/mL containing gallic acid, Quercetin 1mg/mL, Kaempferol 0.5mg/mL reference substance mixed solution is made in amount, plus methanol；
3) need testing solution is prepared：This product powder about 2g is taken, accurately weighed, precision adds methanol 100ml, weighed weight,
Refluxing extraction 1h, lets cool, and supplies less loss weight, and filtering, precision measures filtrate 50mL, plus hydrochloric acid 15mL, water-bath be heated at reflux and
Hydrolysis process 2h, filtering, filtrate is put in 100mL measuring bottles, washed with a little methanol, and washing lotion is incorporated in same measuring bottle, plus methanol is extremely
Scale, shakes up, and is filtered with 0.45nm miillpore filters, takes its filtrate；
4) determine：It is accurate respectively to draw after reference substance solution and each 20 μ L of need testing solution, injection high performance liquid chromatograph
It is measured；
5) calculate：Theoretical cam curve is calculated by Quercetin peak, should be not less than 5000.Peak area is recorded, using external standard method meter
Calculate gallic acid, Quercetin, the content of Kaempferide.
Preferably, solasonine, the solamargine in Compound Solanum Nigrum sulfathiazole are detected using described detection method
Content comprises the following steps：
1) chromatographic condition：
Chromatographic column：Chromatographic column uses octadecylsilane chemically bonded silica for filler；
Mobile phase：Acetonitrile and Na2HPO4The mixed liquor of solution, Na2HPO4For 2mol/L, acetonitrile and Na2HPO4The body of solution
Product is than being 39:61；
Flow velocity：1mL/min；
Detection wavelength：205nm；
Column temperature：25℃；
2) mixing contrast solution is prepared：Precision weighs solasonine reference substance, solamargine reference substance in right amount, plus methanol
1mg/mL containing solasonine, solamargine 1mg/mL reference substance mixed solution is made；
3) need testing solution is prepared：Accurately weighed Compound Solanum Nigrum sulfathiazole coarse powder 2g, precision adds 100mL methanol, accurate
Weighed, refluxing extraction 1h is let cool, and the weight of less loss is supplied with methanol, is shaken up, and filtration, precision measures subsequent filtrate 50mL, is evaporated,
Plus 1% hydrochloric acid solution 50mL dissolving, it is evaporated, is washed with a little methanol, washing lotion is incorporated in 10mL measuring bottles, plus methanol is to scale, shakes
It is even, filtered with 0.45nm miillpore filters, take its filtrate；
4) determine：It is accurate respectively to draw reference substance mixed solution and each 20 μ L of need testing solution, inject high performance liquid chromatography
It is measured after instrument；
5) calculate：Theoretical cam curve is calculated by solasonine peak, should be not less than 5000.Peak area is recorded, using external standard method
Calculate solasonine, the content of solamargine.
Preferably, gallic acid, Quercetin, the Kaempferia galanga in Compound Solanum Nigrum sulfathiazole are detected using described detection method
Element content step 1) described in constant gradient wavelength detecting can also using equiwavelength detection, wavelength is 254nm.
Preferably, gallic acid, Quercetin, the Kaempferia galanga in Compound Solanum Nigrum sulfathiazole are detected using described detection method
Element content step 1) described in mobile phase can also be：Mobile phase A is methanol, and Mobile phase B is 0.4% phosphoric acid solution, is entered
Row gradient elution, the gradient elution program is as follows, and wherein mobile phase ratio is percent by volume：
0~13min, mobile phase A is：2%, Mobile phase B is：88%；
13~14min, mobile phase A is：2%~52%, Mobile phase B is：48%~88%；
14~30min, mobile phase A is：52%, Mobile phase B is：48%；
More than 30min, mobile phase A is：55%, Mobile phase B is：45%.
Preferably, using the solasonine in described detection method simultaneously detection Compound Solanum Nigrum sulfathiazole, Australia eggplant
The step 1 of side alkali content) phosphoric acid solution can select glacial acetic acid, formic acid, trichloroacetic acid in any one acidic materials
To replace.
Preferably, using the solasonine in described detection method simultaneously detection Compound Solanum Nigrum sulfathiazole, Australia eggplant
The step 1 of side alkali content) described in mobile phase can also be：Mobile phase A is acetonitrile, and Mobile phase B is that 1% volume fraction phosphoric acid is molten
Liquid, carries out gradient elution, and the gradient elution program is as follows, and wherein mobile phase ratio is percent by volume：
0~5min, mobile phase A is：22%, Mobile phase B is：78%；
5~20min, mobile phase A is：22%~25%, Mobile phase B is：78%~75%；
20~25min, mobile phase A is：25%, Mobile phase B is：75%；
More than 25min, mobile phase A is：20%, Mobile phase B is：80%.
Preferably, using the solasonine in described detection method simultaneously detection Compound Solanum Nigrum sulfathiazole, Australia eggplant
The step 1 of side alkali content) described in Na2HPO4Ammoniacal liquor, diethylamine, triethylamine, NaH can be selected2PO4And Na3PO4In it is any
A kind of material is replaced.
The present invention according to the phenolic acid such as gallic acid, Quercetin, Kaempferide in the sulfathiazole and solamargine,
This two class of the alkaloids such as solasonine carries out substep detection, and this detection method testing result accuracy is high, but also can carry
High detection rates.And the assay of the sulfathiazole is used it for, this method ensures the sulfathiazole quality inspection standard
Accuracy and advance, can as the sulfathiazole quality control important indicator.Not not having in detection Compound Solanum Nigrum sulfathiazole
When the phenolic acids such as gallate-based, Quercetin, Kaempferide, eluted using the mixed liquor of methanol and phosphoric acid solution.By
It is stronger in the eluting power of methanol, after the phosphoric acid solution for adding 0.4% volume fraction, in detection Compound Solanum Nigrum sulfathiazole
When gallic acid, Quercetin, Kaempferide, mobile phase polarity can be preferably adjusted, the quality of chromatographic peak appearance, example is improved
Such as：Reduce hangover, peak type narrows, absorption peak increases.At the same time it can also be improved methanol eluting power, so as to
To detect the content of the gallic acid in Compound Solanum Nigrum sulfathiazole, Quercetin, Kaempferide exactly.But the phosphoric acid solution of addition
Will bearing in atmosphere in chromatographic column, and ratio can not be too high, the phenomenon salted out otherwise occurs, therefore by first in mixed liquor
Alcohol and volume fraction are deployed into 55 for the volume ratio of 0.4% phosphoric acid solution:It is advisable when 45.According to gallic acid, Quercetin, mountain
The characteristic of a kind of apple element, these medicines are detected in the way of gradient wavelength exactly in different time, different fluctuations.Detect compound
Gradient elution is carried out when gallic acid, Quercetin in black nightshade antiphlogistic, Kaempferide can increase the de- ability of washing of mobile phase, carry
Height is de- to wash efficiency.
When detecting the alkaloids such as solamargine, solasonine, using acetonitrile and Na2HPO4The mixed liquor of solution enters
Row elution.Because the eluting power of acetonitrile is very strong, 2mol/L Na is added2HPO4After solution, in detection solamargine, Australia
When the alkaloids such as continent solanine, mobile phase polarity can be preferably adjusted, the quality of chromatographic peak appearance is improved, such as：Hangover reduction,
Peak type narrows, absorption peak increases.But the Na of addition2HPO4Solution will bearing in atmosphere in chromatographic column, and ratio is not
Can be too high, otherwise there is the phenomenon salted out, therefore by acetonitrile in mixed liquor and 2mol/L
Na2HPO4The volume ratio of solution is deployed into 39:It is advisable when 61.Solasonine, Australia in detection Compound Solanum Nigrum antiphlogistic
Gradient elution is carried out during the solamargine of continent can increase the de- ability of washing of mobile phase, improve to take off and wash efficiency.
Embodiment
With reference to embodiment, the present invention is described in further detail, to make those skilled in the art with reference to explanation
Book word can be implemented according to this.
Embodiment 1
Gallic acid, the Mongolian oak in isocratic elution detection Compound Solanum Nigrum sulfathiazole are carried out using high performance liquid chromatography (HPLC)
Pi Su, Kaempferide, solasonine, solamargine content.
1st, using the gallic acid in high effective liquid chromatography for detecting detection Compound Solanum Nigrum sulfathiazole, Quercetin, mountain
The content of a kind of apple element comprises the following steps：
1) chromatographic condition：
Chromatographic column：Octadecylsilane chemically bonded silica is used for filler；
Mobile phase：The mixed liquor of methanol and phosphoric acid solution, the volume fraction of phosphoric acid solution is 0.4%, methanol and phosphoric acid water
Volume ratio be 55:45；
Flow velocity：1mL/min；
Column temperature：25℃；
Gradient wavelength detecting：0~15min, Detection wavelength is 270nm；15~30min, Detection wavelength is 362nm；
2) mixing contrast solution is prepared：Precision weighs gallic acid reference substance, Quercetin reference substance, Kaempferol reference substance and fitted
0.5mg/mL containing gallic acid, Quercetin 1mg/mL, Kaempferol 0.5mg/mL reference substance mixed solution is made in amount, plus methanol；
3) need testing solution is prepared：This product powder about 2g is taken, accurately weighed, precision adds methanol 100ml, weighed weight,
Refluxing extraction 1h, lets cool, and supplies less loss weight, and filtering, precision measures filtrate 50mL, plus hydrochloric acid 15mL, water-bath be heated at reflux and
Hydrolysis process 2h, filtering, filtrate is put in 100mL measuring bottles, washed with a little methanol, and washing lotion is incorporated in same measuring bottle, plus methanol is extremely
Scale, shakes up, and is filtered with 0.45nm miillpore filters, takes its filtrate；
4) determine：Accurate respectively to draw reference substance solution and each 20 μ L of need testing solution, injection high performance liquid chromatograph enters
Determined after row：
5) calculate：Theoretical cam curve is calculated by Quercetin peak, should be not less than 5000.Peak area is recorded, using external standard method meter
Gallic acid, Quercetin, the content of Kaempferide are calculated, as shown in table 1.
2nd, solasonine, the solamargine in Compound Solanum Nigrum sulfathiazole are detected using high effective liquid chromatography for detecting
Content comprises the following steps：
1) chromatographic condition：
Chromatographic column：Chromatographic column uses octadecylsilane chemically bonded silica for filler；
Mobile phase：Acetonitrile and Na2HPO4The mixed liquor of solution, Na2HPO4For 2mol/L, acetonitrile and Na2HPO4The body of solution
Product is than being 39:61；
Flow velocity：1mL/min；
Detection wavelength：205nm；
Column temperature：25℃；
1) mixing contrast solution is prepared：Precision weighs solasonine reference substance, solamargine reference substance in right amount, plus methanol
1mg/mL containing solasonine, solamargine 1mg/mL reference substance mixed solution is made；
2) need testing solution is prepared：Accurately weighed Compound Solanum Nigrum sulfathiazole coarse powder 2g, precision adds 100mL methanol, accurate
Weighed, refluxing extraction 1h is let cool, and the weight of less loss is supplied with methanol, is shaken up, and filtration, precision measures subsequent filtrate 50mL, is evaporated,
Plus 1% hydrochloric acid solution 50mL dissolving, it is evaporated, is washed with a little methanol, washing lotion is incorporated in 10mL measuring bottles, plus methanol is to scale, shakes
It is even, filtered with 0.45nm miillpore filters, take its filtrate；
3) determine：It is accurate respectively to draw reference substance mixed solution and each 20 μ L of need testing solution, inject high performance liquid chromatography
It is measured after instrument；
4) calculate：Theoretical cam curve is calculated by solasonine peak, should be not less than 5000.Peak area is recorded, using external standard method
Solasonine, the content of solamargine are calculated, as shown in table 1.
Embodiment 2
Carried out using high performance liquid chromatography (HPLC) gallic acid in gradient degree elution detection Compound Solanum Nigrum sulfathiazole,
Quercetin, Kaempferide, solasonine, solamargine content.
1st, using the gallic acid in high effective liquid chromatography for detecting detection Compound Solanum Nigrum sulfathiazole, Quercetin, mountain
The content of a kind of apple element comprises the following steps：
1) chromatographic condition：
Chromatographic column：Octadecylsilane chemically bonded silica is used for filler；
Mobile phase：Mobile phase A is methanol, and Mobile phase B is 0.4% phosphoric acid solution, carries out gradient elution, the gradient elution
Program is as follows, and wherein mobile phase ratio is percent by volume：
0~13min, mobile phase A is：2%, Mobile phase B is：88%；
13~14min, mobile phase A is：2%~52%, Mobile phase B is：48%~88%；
14~30min, mobile phase A is：52%, Mobile phase B is：48%；
More than 30min, mobile phase A is：55%, Mobile phase B is：45%；
Flow velocity：1mL/min；
Column temperature：25℃；
Gradient wavelength detecting：0~15min, Detection wavelength is 270nm；15~30min, Detection wavelength is 362nm；
2) mixing contrast solution is prepared：Precision weighs gallic acid reference substance, Quercetin reference substance, Kaempferol reference substance and fitted
0.5mg/mL containing gallic acid, Quercetin 1mg/mL, Kaempferol 0.5mg/mL reference substance mixed solution is made in amount, plus methanol；
3) need testing solution is prepared：This product powder about 2g is taken, accurately weighed, precision adds methanol 100ml, weighed weight,
Refluxing extraction 1h, lets cool, and supplies less loss weight, and filtering, precision measures filtrate 50mL, plus hydrochloric acid 15mL, water-bath be heated at reflux and
Hydrolysis process 2h, filtering, filtrate is put in 100mL measuring bottles, washed with a little methanol, and washing lotion is incorporated in same measuring bottle, plus methanol is extremely
Scale, shakes up, and is filtered with 0.45nm miillpore filters, takes its filtrate；
4) determine：It is accurate respectively to draw after reference substance solution and each 20 μ L of need testing solution, injection high performance liquid chromatograph
It is measured；
5) calculate：Theoretical cam curve is calculated by Quercetin peak, should be not less than 5000.Peak area is recorded, using external standard method meter
Gallic acid, Quercetin, the content of Kaempferide are calculated, as shown in table 1.
2nd, solasonine, the solamargine in Compound Solanum Nigrum sulfathiazole are detected using high effective liquid chromatography for detecting
Content comprises the following steps：
1) chromatographic condition：
Chromatographic column：Chromatographic column uses octadecylsilane chemically bonded silica for filler；
Mobile phase：Mobile phase A is acetonitrile, and Mobile phase B is 1% volume fraction phosphoric acid solution, carries out gradient elution, the ladder
Degree elution program is as follows, and wherein mobile phase ratio is percent by volume：
0~5min, mobile phase A is：22%, Mobile phase B is：78%；
5~20min, mobile phase A is：22%~25%, Mobile phase B is：78%~75%；
20~25min, mobile phase A is：25%, Mobile phase B is：75%；
More than 25min, mobile phase A is：20%, Mobile phase B is：80%.
Flow velocity：1mL/min；
Detection wavelength：205nm；
Column temperature：25℃；
2) mixing contrast solution is prepared：Precision weighs solasonine reference substance, solamargine reference substance in right amount, plus methanol
1mg/mL containing solasonine, solamargine 1mg/mL reference substance mixed solution is made；
3) need testing solution is prepared：Accurately weighed Compound Solanum Nigrum sulfathiazole coarse powder 2g, precision adds 100mL methanol, accurate
Weighed, refluxing extraction 1h is let cool, and the weight of less loss is supplied with methanol, is shaken up, and filtration, precision measures subsequent filtrate 50mL, is evaporated,
Plus 1% hydrochloric acid solution 50mL dissolving, it is evaporated, is washed with a little methanol, washing lotion is incorporated in 10mL measuring bottles, plus methanol is to scale, shakes
It is even, filtered with 0.45nm miillpore filters, take its filtrate；
4) determine：It is accurate respectively to draw reference substance mixed solution and each 20 μ L of need testing solution, inject high performance liquid chromatography
It is measured after instrument；
5) calculate：Theoretical cam curve is calculated by solasonine peak, should be not less than 5000.Peak area is recorded, using external standard method
Solasonine, the content of solamargine are calculated, as shown in table 1.
Embodiment 3
The nutgall during gradient elution is determined in Compound Solanum Nigrum sulfathiazole is carried out using ultra-performance liquid chromatography (UPLC)
Acid, Quercetin, Kaempferide, solasonine, solamargine content.
Ultra-performance liquid chromatography principle in this experiment with it is essentially identical with high performance liquid chromatography in embodiment 2,
Changed place have it is following some：
(1) using little particle, high-performance particulate stationary phase.HPLC chromatographic column, such as：Common 18 alkyl silica gel bonding
Post, particle diameter is 5um, and UPLC chromatographic columns, reaches 3.5um, or even 1.7um, and such aperture is more conducive to material separation.
(2) use of high pressure pump.Because the chromatographic column particle diameter used reduces, produced pressure is also natural when using
Increase exponentially.Therefore the infusion pump of liquid chromatogram also accordingly changes over the infusion pump of super-pressure.
(3) sensitive detectors of high-speed sampling speed.
(4) using low diffusion, low cross pollution automatic sampler.It is equipped with sample introduction probe and pressure in pin and aids in sample introduction skill
Art；
(5) instrument total system optimization design.Chromatographic work station is equipped with multi software platform, realizes ultra high efficiency liquid phase analysis
The automatic conversion of method and high efficient liquid phase analysis method.
The result of ultra-performance liquid chromatography detection Compound Solanum Nigrum sulfathiazole is as shown in table 1.
Table 1HPLC methods and UPLC methods determine Contents of Main Components in Compound Solanum Nigrum sulfathiazole
Same batch sample assay result data is compared referring to table 1.From the data of table 1, it can be seen that high-efficient liquid phase color
Spectrometry (HPLC) and ultra-performance liquid chromatography (UPLC) and two kinds of elution methods determine main component in Compound Solanum Nigrum sulfathiazole and contained
Measure result basically identical.
6 batches of assay data analyses more than, different batches Compound Solanum Nigrum sulfathiazole mass discrepancy is larger, mainly by
The influence of the factors such as herbal species, the place of production, processing, preparation is caused.Can be by specifying high-quality Chinese medicine black nightshade, pennywort, pithecellobium clypearia
Active constituent content in kind and the place of production etc., standardized processing production process, raising or stable Compound Solanum Nigrum sulfathiazole.Australia eggplant
Alkali, solamargine have the bioactivity such as anticancer, antibacterial, and content is higher in black nightshade medicinal material, can be used as evaluation black nightshade
The index components of quality of medicinal material, meanwhile, gallic acid, Quercetin, the Kaempferide content in pennywort and pithecellobium clypearia are higher, can be with
It is used as the index components for evaluating pennywort and pithecellobium clypearia quality.Could dictate that this product every containing gallic acid, Quercetin, Kaempferide,
Solasonine, the content of solamargine should be no less than 8mg, 2mg, 0.2mg, 1.5mg, 1.5mg respectively
Although embodiment of the present invention is disclosed as above, it is not restricted in specification and embodiment listed
With it can be applied to various suitable the field of the invention completely, can be easily for those skilled in the art
Other modification is realized, therefore under the universal limited without departing substantially from claim and equivalency range, the present invention is not limited
In specific details and shown here as the embodiment with description.
Claims (6)
1. a kind of high-efficiency liquid chromatography method for detecting of Compound Solanum Nigrum sulfathiazole, it is characterised in that described Compound Solanum Nigrum anti-inflammatory
Piece is mainly made up of black nightshade, pennywort, pithecellobium clypearia, and the detection method includes detecting the compound using high performance liquid chromatography
Gallic acid, Quercetin, Kaempferide, solasonine, solamargine in black nightshade sulfathiazole；It is same using described detection method
When detection Compound Solanum Nigrum antiphlogistic in gallic acid, Quercetin, the content of Kaempferide comprise the following steps：
1) chromatographic condition：
Chromatographic column：Octadecylsilane chemically bonded silica is used for filler；
Mobile phase：The mixed liquor of methanol and phosphoric acid solution, the volume fraction of phosphoric acid solution is 0.4%, the body of methanol and phosphoric acid water
Product is than being 55:45；
Flow velocity：1mL/min；
Column temperature：25℃；
Gradient wavelength detecting：0~15min, Detection wavelength is 270nm；15~30min, Detection wavelength is 362nm；
2) mixing contrast solution is prepared：Precision weighs gallic acid reference substance, Quercetin reference substance, Kaempferol reference substance in right amount,
Plus 0.5mg/mL containing gallic acid, Quercetin 1mg/mL, Kaempferol 0.5mg/mL reference substance mixed solution is made in methanol；
3) need testing solution is prepared：This product powder about 2g is taken, accurately weighed, precision adds methanol 100ml, weighed weight, backflow
1h is extracted, lets cool, supplies less loss weight, is filtered, precision measures filtrate 50mL, plus hydrochloric acid 15mL, and water-bath is heated at reflux and hydrolyzed
2h is handled, filtering, filtrate is put in 100mL measuring bottles, washed with a little methanol, and washing lotion is incorporated in same measuring bottle, plus methanol is to scale,
Shake up, filtered with 0.45nm miillpore filters, take its filtrate；
4) determine：It is accurate respectively to draw progress after reference substance solution and each 20 μ L of need testing solution, injection high performance liquid chromatograph
Determine；
5) calculate：Theoretical cam curve is calculated by Quercetin peak, should be not less than 5000, records peak area, is calculated and not had using external standard method
Gallate-based, Quercetin, the content of Kaempferide.
2. the high-efficiency liquid chromatography method for detecting of Compound Solanum Nigrum sulfathiazole according to claim 1, it is characterised in that use institute
The detection method stated is while detect that the solasonine in Compound Solanum Nigrum antiphlogistic, solamargine content comprise the following steps：
1) chromatographic condition：
Chromatographic column：Chromatographic column uses octadecylsilane chemically bonded silica for filler；
Mobile phase：Acetonitrile and Na2HPO4The mixed liquor of solution, Na2HPO4For 2mol/L, acetonitrile and Na2HPO4The volume ratio of solution
For 39:61；
Flow velocity：1mL/min；
Detection wavelength：205nm；
Column temperature：25℃；
2) mixing contrast solution is prepared：Precision weighs solasonine reference substance, solamargine reference substance in right amount, plus methanol is made
1mg/mL containing solasonine, solamargine 1mg/mL reference substance mixed solution；
3) need testing solution is prepared：Accurately weighed Compound Solanum Nigrum sulfathiazole coarse powder 2g, precision adds 100mL methanol, accurately weighed,
Refluxing extraction 1h, is let cool, and the weight of less loss is supplied with methanol, is shaken up, and filtration, precision measures subsequent filtrate 50mL, is evaporated, plus 1%
Hydrochloric acid solution 50mL dissolves, and is evaporated, is washed with a little methanol, washing lotion is incorporated in 10mL measuring bottles, plus methanol is to scale, shakes up, and uses
0.45nm miillpore filters are filtered, and take its filtrate；
4) determine：It is accurate respectively to draw after reference substance mixed solution and each 20 μ L of need testing solution, injection high performance liquid chromatograph
It is measured；
5) calculate：Theoretical cam curve is calculated by solasonine peak, should be not less than 5000；Peak area is recorded, is calculated using external standard method
The content of solasonine, solamargine.
3. the high-efficiency liquid chromatography method for detecting of Compound Solanum Nigrum sulfathiazole according to claim 1, it is characterised in that step 1)
Described mobile phase is replaced with：Mobile phase A is methanol, and Mobile phase B is 0.4% phosphoric acid solution, carries out gradient elution, gradient elution
Program is as follows, and wherein mobile phase ratio is percent by volume：
0~13min, mobile phase A is：2%, Mobile phase B is：88%；
13~14min, mobile phase A is：2%~52%, Mobile phase B is：48%~88%；
14~30min, mobile phase A is：52%, Mobile phase B is：48%；
More than 30min, mobile phase A is：55%, Mobile phase B is：45%.
4. the high-efficiency liquid chromatography method for detecting of Compound Solanum Nigrum sulfathiazole according to claim 1, it is characterised in that step 1)
The phosphoric acid solution is replaced from any one material in glacial acetic acid, formic acid, trichloroacetic acid.
5. the high-efficiency liquid chromatography method for detecting of Compound Solanum Nigrum sulfathiazole according to claim 2, it is characterised in that step 1)
Described mobile phase is replaced with：Mobile phase A is acetonitrile, and Mobile phase B is 1% volume fraction phosphoric acid solution, carries out gradient elution, ladder
Degree elution program is as follows, and wherein mobile phase ratio is percent by volume：
0~5min, mobile phase A is：22%, Mobile phase B is：78%；
5~20min, mobile phase A is：22%~25%, Mobile phase B is：78%~75%；
20~25min, mobile phase A is：25%, Mobile phase B is：75%；
More than 25min, mobile phase A is：20%, Mobile phase B is：80%.
6. the high-efficiency liquid chromatography method for detecting of Compound Solanum Nigrum sulfathiazole according to claim 2, it is characterised in that step 1)
Described Na2HPO4From ammoniacal liquor, diethylamine, triethylamine, NaH2PO4And Na3PO4In any one material replace.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510780966.6A CN105203673B (en) | 2015-11-13 | 2015-11-13 | The high-efficiency liquid chromatography method for detecting of Compound Solanum Nigrum sulfathiazole |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510780966.6A CN105203673B (en) | 2015-11-13 | 2015-11-13 | The high-efficiency liquid chromatography method for detecting of Compound Solanum Nigrum sulfathiazole |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105203673A CN105203673A (en) | 2015-12-30 |
| CN105203673B true CN105203673B (en) | 2017-10-31 |
Family
ID=54951464
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510780966.6A Expired - Fee Related CN105203673B (en) | 2015-11-13 | 2015-11-13 | The high-efficiency liquid chromatography method for detecting of Compound Solanum Nigrum sulfathiazole |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105203673B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105954390A (en) * | 2016-04-25 | 2016-09-21 | 广西壮族自治区梧州食品药品检验所 | Method for detecting content of kaempferide in momordica grosvenori tea by using ASE-HPLC |
| CN108205033A (en) * | 2018-01-04 | 2018-06-26 | 吉林师范大学 | A kind of method of solanine and black nightshade polysaccharide in synchronous extracting and developing, measure black nightshade Chinese olive |
| CN109946397A (en) * | 2019-03-26 | 2019-06-28 | 广州莱泰制药有限公司 | A kind of pithecellobium clypearia method of inspection |
-
2015
- 2015-11-13 CN CN201510780966.6A patent/CN105203673B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN105203673A (en) | 2015-12-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102106918B (en) | Method for simultaneously preparing volatile oil, total flavones and total alkaloids from immature bitter oranges | |
| KR20150001777A (en) | Method for extracting and separating ginkgolides | |
| CN104459003B (en) | The construction method of contracting spring standard preparation finger-print and characteristic spectrum and quality determining method | |
| CN105203673B (en) | The high-efficiency liquid chromatography method for detecting of Compound Solanum Nigrum sulfathiazole | |
| CN105486771A (en) | Fingerprint spectrum detecting method of common goldenrop particle compound preparation | |
| CN105353063B (en) | Compound cold drug stream livering ingredient standard finger-print and construction method, application | |
| CN103645251B (en) | A kind of fingerprint atlas detection method of compound donkey-hide gelatin preparation | |
| CN102058814B (en) | General flavone extractive of four medicaments including grassleaf sweetflag rhizome and preparation method thereof | |
| CN107315058A (en) | A kind of method of total ginkgoic acid in detection ginkgo biloba succi | |
| CN102520103A (en) | Contrast Chinese patent medicine extract of pills of six ingredients with rehmannia, and preparation method and application thereof | |
| CN103585204B (en) | Thesium granule with high stability and preparation method thereof | |
| CN102743598A (en) | var. Denneanum extract and formula particles thereof | |
| CN103575821A (en) | Detection method of 14 chemical components in Tangminling preparation | |
| CN104330497A (en) | Method for determining fluoroquinolone drug residual in beef | |
| CN104833754B (en) | A kind of attached sweet drug detection method | |
| CN103940942B (en) | A kind of detection method of CHANGYANNING preparation | |
| CN112763609B (en) | Research method for screening and extracting process of anti-asthma active ingredients of chamomile | |
| CN106769964B (en) | The method of quality control of Herba Achilleae extract | |
| CN103063792B (en) | Quality test method of phlegm eliminating and cough stopping granules for children | |
| CN103372112B (en) | A kind of Rhizoma Alismatis drop pill and preparation method thereof | |
| CN101618069B (en) | Capsule preparation for treating bruise as well as bleeding and easing pain and preparation method and use thereof | |
| CN101339168A (en) | Thunder god vine medicinal materials triptolide content determination method | |
| CN108020611A (en) | A kind of method of a variety of biflavone contents in detection gingko leaf preparation | |
| CN103267824A (en) | Method for detecting wind-cold cold granules | |
| CN103923138B (en) | A kind of preparation method of Nicotifiorin and application 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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20171031 Termination date: 20211113 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |