CN102721754A - A method for determining phthalate content in traditional Chinese medicine by adopting a liquid chromatography-mass spectrometry method - Google Patents
A method for determining phthalate content in traditional Chinese medicine by adopting a liquid chromatography-mass spectrometry method Download PDFInfo
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- 238000012360 testing method Methods 0.000 claims description 15
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- NFOQRIXSEYVCJP-UHFFFAOYSA-N 2-propoxycarbonylbenzoic acid Chemical compound CCCOC(=O)C1=CC=CC=C1C(O)=O NFOQRIXSEYVCJP-UHFFFAOYSA-N 0.000 claims description 9
- QWDBCIAVABMJPP-UHFFFAOYSA-N Diisopropyl phthalate Chemical compound CC(C)OC(=O)C1=CC=CC=C1C(=O)OC(C)C QWDBCIAVABMJPP-UHFFFAOYSA-N 0.000 claims description 9
- DWNAQMUDCDVSLT-UHFFFAOYSA-N diphenyl phthalate Chemical compound C=1C=CC=C(C(=O)OC=2C=CC=CC=2)C=1C(=O)OC1=CC=CC=C1 DWNAQMUDCDVSLT-UHFFFAOYSA-N 0.000 claims description 9
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Abstract
The invention discloses a method for determining phthalate content in traditional Chinese medicine by adopting a liquid chromatography-mass spectrometry method, characterized by comprising the following steps: (1) preprocessing a sample to be measured, and (2) determining the content of one kind or multiple kinds of phthalate in the sample after the preprocessing step by adopting the liquid chromatography-mass spectrometry method. The determining method is advantaged by high sensitivity, strong specificity, excellent accuracy degree, and suitability for determining multi-component residues of phthalate in traditional Chinese medicine such as lanceolata, white peony root, red peony root, radix rehmanniae, and the like.
Description
Technical field
The present invention relates to the assay method of phthalic acid ester content in the Chinese medicine, be specifically related to adopt LC/MS to measure the method for phthalic acid ester content in the Chinese medicines such as Radix Codonopsis, the root of herbaceous peony, the radix paeoniae rubrathe, glutinous rehmannia.
Background technology
Phthalic ester (PAEs); It is the plastifier of one type of widespread use; Can play emollescence, it is widely used in toy, packaging material for food, medical blood bag and sebific duct, vinyl flooring and wallpaper, detersive, lubricating oil, the personal-care supplies hundreds of products such as (like nail polish, hair spray, perfumed soap and shampoos).Research shows; Phthalic ester is being brought into play the effect of similar female hormone in human body and animal body, can disturb endocrine, and man semen amount and sperm quantity are reduced; The sperm motility ability is low; Sperm morphology is unusual, and serious meeting causes carcinoma of testis, is " arch-criminal " who causes man's reproductive problems.This material in the cosmetics can get in the body through women's respiratory system and skin; If too much use, can increase the probability that the women suffers from breast cancer, the reproductive system that also can jeopardize the boy baby of their following fertility is (referring to Shen Xiahong etc.; Phthalate embryonic germ toxicity profile [J]; Chinese public health, in September, 2010,26 (9): 1215).
Harm in view of the phthalate composition; National governments greatly pay close attention to the distribution and the detection of phthalic ester in food and medicine wrappage, toy, the cosmetics; Particularly developed country such as the European Union and the U.S. has set the strict regulation of limiting the quantity of to this.The guidance standard about phthalic ester (2005/84/EC) of European Union's promulgation begins to carry out on January 16th, 2007.On February 6th, 2009; U.S. consumer goods council issue directive document; Requirement was from February 10th, 2009, and the content of diisooctyl phthalate in toy for children or the child care products (DEHP), dibutyl phthalate (DBP), butyl benzyl phthalate 6 kinds of phthalic esters such as (BBP) must not surpass 0.1%.The phthalate of China in media such as food, cosmetics, toy, environment detects existing report, but the limit standard of phthalate composition in the product-free still.
Chinese crude drug may be contaminated in the plantation process.Therefore, be badly in need of the efficient analysis assay method of phthalate composition in the exploitation medicine.
Summary of the invention
The present invention provides a kind of method that adopts liquid chromatography-mass spectrography to measure phthalic acid ester content in the Chinese medicine.This method can be used for the residual examination of phthalic ester in the Chinese medicine, and the standard of phthalic ester provides technical service in the medicine in order to draft.
The present invention realizes through following technical scheme:
A kind of method that is used for measuring the Chinese medicine phthalic acid ester content is provided, it is characterized in that, this method comprises the steps:
(1) Chinese medicine sample to be measured is carried out pre-service;
(2) pass through the content that LC/MS is measured one or more phthalic esters in pretreated sample.
According to of the present invention one preferred embodiment, said phthalic ester comprises phthalic acid mono-methyl, repefral (DMP), phthalic acid mono, diethyl phthalate (DEP), diisopropyl phthalate, n-propyl phthalate, diphenyl phthalate, BBP(Butyl Benzyl Phthalate (BBP), diisobutyl phthalate (DIBP), dicyclohexyl phthalate (DCHP), diisooctyl phthalate (DEHP).
According to of the present invention one preferred embodiment, said Chinese medicine can comprise Radix Codonopsis, the root of herbaceous peony, the radix paeoniae rubrathe and glutinous rehmannia etc.
Preferred embodiment said The pretreatment comprises with first kind of solvent supersonic or backflow or Suo Shi to be extracted according to one of the present invention, and rotary evaporation is to doing then, again with the step of second kind of solvent redissolution.Preferably, said first kind of solvent can be selected from normal hexane, sherwood oil (60～90 ℃) or chloroform, preferred especially normal hexane; Said second kind of solvent can be selected from methyl alcohol, chloroform methanol solution (preferred volume ratio 3: 1) or normal hexane, special particular methanol.
According to of the present invention one preferred embodiment, the condition of said ultrasonic Extraction is: under the ultrasonic frequency of 32～40kHz, 25～45 ℃ of following water-bath ultrasonic Extraction 30～60min of temperature are preferably under the 40kHz frequency, at 30 ℃ of following water-bath ultrasonic Extraction 30min; The condition of refluxing extraction is: 80～100 ℃ of refluxing extraction 1h～2h, preferably at 90 ℃ of refluxing extraction 1h; The condition that Suo Shi extracts is: 80～100 ℃ of Suo Shi extract 4～8h, preferably extract 6h at 90 ℃.Term " Suo Shi extraction " refers to and adopts apparatus,Soxhlet's (250ml flat bottom flask, 80ml are extracted tube out), utilizes solvent refluxing and siphon principle, and solid matter is continuously extracted by neat solvent.
Especially preferred embodiment The pretreatment comprises: precision takes by weighing the Chinese medicine sample powder according to one of the present invention, and the accurate normal hexane that adds is ultrasonic, filters, and precision is measured the subsequent filtrate rotary evaporation to doing, and the accurate methyl alcohol that adds makes dissolving, filters.
According to of the present invention one preferred embodiment, the chromatographic condition of LC/MS is in the said step (2): adopt C 18 posts, moving phase by A mutually with the B phase composition, A is aqueous formic acid mutually, B is acetonitrile or methyl alcohol mutually, the employing gradient elution.
Preferably, the particle diameter of said C18 post is 1.7～5 μ m, and column internal diameter 2.1～4.6mm, column length are 5～25cm, more preferably Agilent ZORBAX SB-C18 post.The concentration of A phase is 0%～0.1%, preferred 0.05% (volume); A: the B volume ratio is 50～2%: 50～98%, and flow velocity is 0.3ml/min.
According to of the present invention one preferred embodiment; The mass spectrum condition of LC/MS is in the said step (2): adopt electric spray ion source; Adopt positive ion mode to carry out data acquisition; 2.5～4.0 kilovolts of capillary voltage, 10～30 volts of taper hole voltages, more preferably capillary voltage is 3.0 kilovolts, 15 volts or 20 volts of taper hole voltages.
The inventive method also comprises the standard items drawing standard curve with various phthalic esters, and the content of phthalic ester in the calculation sample thus.
The inventive method is for be applied to residual detection and the analysis of phthalic ester multicomponent in the Chinese medicine first.The inventive method is highly sensitive, specificity is strong, accuracy is good, can be used for the residual mensuration of multicomponent of phthalic ester in Radix Codonopsis, the root of herbaceous peony, the radix paeoniae rubrathe, glutinous rehmannia or the similar medicinal material.
The inventive method is carried out the sample introduction analysis through liquid chromatography-mass spectrography to the Chinese medicine sample, detects the residual quantity of multiple phthalic ester in the Chinese medicine, and analysis cost is low, the time is short, and stronger practicality is arranged.This method has great importance to the residual detection of phthalic ester multicomponent in the Chinese medicine, and gordian techniquies such as new drug are had good directive significance, can be applicable to many aspects such as new drug development Quality Control.This method can effectively be applied to the formulation of national standard, the research and development of enterprise's new drug, the lifting of enterprise's existing product quality control method.
Description of drawings
Fig. 1 is the chromatogram of the hybrid standard article solution of positive ion mode;
Fig. 2 is the chromatogram of Radix Codonopsis sample solution of the embodiment 1 of positive ion mode.
Fig. 3 is the chromatogram of root of herbaceous peony sample solution of the embodiment 2 of positive ion mode.
Fig. 4 is the chromatogram of radix paeoniae rubrathe sample solution of the embodiment 3 of positive ion mode.
Fig. 5 is the chromatogram of glutinous rehmannia sample solution of the embodiment 4 of positive ion mode.
Fig. 6 is the chromatogram of Radix Codonopsis sample solution of the embodiment 5 of positive ion mode.
Fig. 7 is the chromatogram of root of herbaceous peony sample solution of the embodiment 6 of positive ion mode.
Embodiment
1, materials and methods
1.1 key instrument and reagent
The single quadrupole mass spectrometer (U.S. Waters company) of Waters ZQ, ACQUITY Ultra Performance LC liquid chromatograph (U.S. Waters company), KQ-250DB type numerical control Ultrasound Instrument, ultrapure water machine (U.S. Millipore company).
Phthalic acid mono-methyl, repefral (DMP), phthalic acid mono, diethyl phthalate (DEP), diisopropyl phthalate, n-propyl phthalate, diphenyl phthalate, BBP(Butyl Benzyl Phthalate (BBP), diisobutyl phthalate (DIBP), dicyclohexyl phthalate (DCHP), diisooctyl phthalate (DEHP) standard items (lark prestige Science and Technology Ltd.); Acetonitrile, methyl alcohol, formic acid are chromatographically pure, and normal hexane is analyzed pure.
Radix Codonopsis crude drug: medicinal material market, Wen County, Gansu, lot number: ds-080712.
1.2 experimental technique
1.2.1 chromatographic condition
Agilent ZORBAX SB-C
18Column (3.0 * 150mm, 3.5 μ m); With 0.05% formic acid is the mobile phase A phase, is the Mobile phase B phase with 100% acetonitrile, flow velocity 0.3ml/min; According to the form below carries out gradient elution:
Table 1-1 eluent gradient
1.2.2 mass spectrum condition
Ion gun is ESI source, electro-spray ionization source; Mass spectrum parameters such as capillary voltage, taper hole voltage are seen table 1-2.
Table 1-2 mass spectrum parameter list
1.2.3 the preparation of standard solution
It is an amount of that precision takes by weighing phthalic acid mono-methyl, repefral (DMP), phthalic acid mono, diethyl phthalate (DEP), diisopropyl phthalate, n-propyl phthalate, diphenyl phthalate, BBP(Butyl Benzyl Phthalate (BBP), diisobutyl phthalate (DIBP), dicyclohexyl phthalate (DCHP), diisooctyl phthalate (DEHP) standard items; Add the solution that methyl alcohol is mixed with 1mg/mL, as standard reserving solution.
The above-mentioned storing solution of accurate respectively absorption is an amount of, is diluted to the series standard solution of the said concentration of following table with blank matrix solution.
Table 1-3 series standard solution concentration table
1.2.4 the preparation of need testing solution
Precision takes by weighing Radix Codonopsis sample powder (crossing 40 mesh sieves) 3g, puts in the tool plug Erlenmeyer flask the accurate normal hexane 90ml that adds; 30 ℃ of water-baths ultrasonic (250W, 40kHz) extraction 30min, filtrations; Precision is measured subsequent filtrate 50ml, puts to revolve and steams that 40 ℃ of rotary evaporations are settled to 5ml to doing with dissolve with methanol in the bottle; Filter, promptly get.(annotate: experimentation avoids contacting plastic products, in order to avoid pollute).
2, result
2.1 measure
Above-mentioned each the standard solution sample introduction 5 μ l of accurate respectively absorption analyze the drawing standard curve according to above-mentioned condition sample introduction.The accurate in addition need testing solution 5 μ l that draw analyze according to above-mentioned condition sample introduction, read corresponding amount from typical curve, calculate, and promptly get.
Computing formula:
In the formula:
X (ng/g)---the content of composition to be measured in the sample;
A (ng/mL)---the concentration of composition to be measured in the need testing solution;
M (g)---the quality of sample.
The result shows that detect above-mentioned diisooctyl phthalate (DEHP) in the Radix Codonopsis sample, content is 678ng/g.
2.2 linear relationship
Accurate each the 5 μ l of above-mentioned series standard solution that draw, each component chromatographic peak area to be measured is write down in the sample introduction analysis, is horizontal ordinate (X) with sample introduction concentration, and peak area is ordinate (Y), carries out regretional analysis, and result's (seeing table 1-4) shows that each component lines sexual intercourse is good.
Table 1-4 regression equation and related coefficient
2.3 detectability
With signal to noise ratio (S/N ratio) is 3: 1 computing method, the detectability of standard solution (seeing table 1-5).
Table 1-5 detectability
2.4 precision test
Get matrix standard solution 6, continuous sample introduction 3 times, the record peak area, the result shows that the RSD value of 3 sample introduction peak areas of above-mentioned 11 kinds of compositions is in the 1.2%-5.1% scope, precision is good.
1, materials and methods
1.1 key instrument and reagent
The single quadrupole mass spectrometer (U.S. Waters company) of Waters ZQ, ACQUITY Ultra Performance LC liquid chromatograph (U.S. Waters company), reflux, ultrapure water machine (U.S. Millipore company).
Phthalic acid mono-methyl, repefral (DMP), phthalic acid mono, diethyl phthalate (DEP), diisopropyl phthalate, n-propyl phthalate, diphenyl phthalate, BBP(Butyl Benzyl Phthalate (BBP), diisobutyl phthalate (DIBP), dicyclohexyl phthalate (DCHP), diisooctyl phthalate (DEHP) standard items (lark prestige Science and Technology Ltd.); Acetonitrile, methyl alcohol, formic acid are chromatographically pure, and normal hexane is analyzed pure.
Root of herbaceous peony medicine materical crude slice is available from medicinal material market, Bozhou
1.2 experimental technique
1.2.1 chromatographic condition
Waters BEH C
18Column (2.1 * 100mm, 1.7 μ m); With 0.1% formic acid is the mobile phase A phase, is the Mobile phase B phase with 100% acetonitrile, flow velocity 0.45ml/min; According to the form below carries out gradient elution:
Table 2-1 eluent gradient
1.2.2 mass spectrum condition
Ion gun is ESI source, electro-spray ionization source; Mass spectrum parameters such as capillary voltage, taper hole voltage are seen table 2-2.
Table 2-2 mass spectrum parameter list
1.2.3 the preparation of need testing solution
Precision takes by weighing root of herbaceous peony sample powder (crossing 40 mesh sieves) 3g, puts in the round-bottomed flask, and the accurate normal hexane 90ml that adds, 1h is extracted in 90 ℃ of hot refluxs; Filter, precision is measured subsequent filtrate 50ml, puts to revolve and steams in the bottle 40 ℃ of rotary evaporations to doing; Be settled to 5ml with dissolve with methanol, filter, promptly get.(annotate: experimentation avoids contacting plastic products, in order to avoid pollute).
2, result
2.1 measure
The accurate need testing solution 5 μ l that draw analyze according to above-mentioned condition sample introduction, read corresponding amount from typical curve, calculate, and promptly get.
The result shows that detect above-mentioned diisooctyl phthalate (DEHP) in the root of herbaceous peony sample, content is 1212ng/g.
1, materials and methods
1.1 key instrument and reagent
The single quadrupole mass spectrometer (U.S. Waters company) of Waters ZQ; ACQUITY Ultra Performance LC liquid chromatograph (U.S. Waters company); KQ-250DB numerical control Ultrasound Instrument (Kunshan ultrasonic instrument company limited), ultrapure water machine (U.S. Millipore company).
Phthalic acid mono-methyl, repefral (DMP), phthalic acid mono, diethyl phthalate (DEP), diisopropyl phthalate, n-propyl phthalate, diphenyl phthalate, BBP(Butyl Benzyl Phthalate (BBP), diisobutyl phthalate (DIBP), dicyclohexyl phthalate (DCHP), diisooctyl phthalate (DEHP) standard items (lark prestige Science and Technology Ltd.); Acetonitrile, methyl alcohol, formic acid are chromatographically pure, and sherwood oil (60～90 ℃), chloroform are that analysis is pure.
Radix paeoniae rubrathe medicine materical crude slice is available from medicinal material market, Bozhou
1.2 experimental technique
1.2.1 chromatographic condition
Agilent ZORBAX SB C
18Column (4.6 * 250mm, 5 μ m); With 0.5% formic acid is the mobile phase A phase, is the Mobile phase B phase with 100% acetonitrile, flow velocity 1.0ml/min; According to the form below carries out gradient elution:
Table 3-1 eluent gradient
1.2.2 mass spectrum condition
Ion gun is ESI source, electro-spray ionization source; Mass spectrum parameters such as capillary voltage, taper hole voltage are seen table 3-2.
Table 3-2 mass spectrum parameter list
1.2.3 the preparation of need testing solution
Precision takes by weighing radix paeoniae rubrathe sample powder (crossing 40 mesh sieves) 3g, puts in the tool plug Erlenmeyer flask accurate sherwood oil (60～90 ℃) 90ml that adds; 45 ℃ of water-baths ultrasonic (250W, 36kHz) extraction 1h, filtrations; Precision is measured subsequent filtrate 50ml, puts to revolve and steams in the bottle 40 ℃ of rotary evaporations to doing, and dissolves with chloroform methanol solution (3: 1) to be settled to 5ml; Filter, promptly get.(annotate: experimentation avoids contacting plastic products, in order to avoid pollute).
2, result
2.1 measure
The accurate need testing solution 5 μ l that draw analyze according to above-mentioned condition sample introduction, read corresponding amount from typical curve, calculate, and promptly get.
The result shows that detect above-mentioned diisooctyl phthalate (DEHP) in the radix paeoniae rubrathe sample, content is 925ng/g.
1, materials and methods
1.1 key instrument and reagent
The single quadrupole mass spectrometer (U.S. Waters company) of Waters ZQ, ACQUITY Ultra Performance LC liquid chromatograph (U.S. Waters company), apparatus,Soxhlet's (250ml flask, 80ml are extracted tube out), ultrapure water machine (U.S. Millipore company).
Phthalic acid mono-methyl, repefral (DMP), phthalic acid mono, diethyl phthalate (DEP), diisopropyl phthalate, n-propyl phthalate, diphenyl phthalate, BBP(Butyl Benzyl Phthalate (BBP), diisobutyl phthalate (DIBP), dicyclohexyl phthalate (DCHP), diisooctyl phthalate (DEHP) standard items (lark prestige Science and Technology Ltd.); Acetonitrile, methyl alcohol, formic acid are chromatographically pure, and normal hexane, chloroform are that analysis is pure.
Radix rehmanniae is available from Bai Quan medicinal material market, Jinan.
1.2 experimental technique
1.2.1 chromatographic condition
Agilent ZORBAX SB-C
18Column (2.1 * 50mm, 1.8 μ m); With water is the mobile phase A phase, is the Mobile phase B phase with 100% methyl alcohol, flow velocity 0.25ml/min; According to the form below carries out gradient elution:
Table 4-1 eluent gradient
1.2.2 mass spectrum condition
Ion gun is ESI source, electro-spray ionization source; Mass spectrum parameters such as capillary voltage, taper hole voltage are seen table 4-2.
Table 4-2 mass spectrum parameter list
1.2.3 the preparation of need testing solution
Precision takes by weighing glutinous rehmannia sample powder (crossing 40 mesh sieves) 3g, puts in the tool plug Erlenmeyer flask the accurate normal hexane 90ml that adds; 100 ℃ of Suo Shi extract 4h, filter, and precision is measured subsequent filtrate 50ml; Put to revolve and steam in the bottle 40 ℃ of rotary evaporations to doing; Be settled to 5ml with chloroform methanol solution (1: 10) dissolving, filter, promptly get.(annotate: experimentation avoids contacting plastic products, in order to avoid pollute).
2, result
2.1 measure
The accurate need testing solution 5 μ l that draw analyze according to above-mentioned condition sample introduction, read corresponding amount from typical curve, calculate, and promptly get.
The result shows that detect above-mentioned diisooctyl phthalate (DEHP) in the glutinous rehmannia sample, content is 607ng/g.
1, materials and methods
1.1 key instrument and reagent
The single quadrupole mass spectrometer (U.S. Waters company) of Waters ZQ, ACQUITY Ultra Performance LC liquid chromatograph (U.S. Waters company), reflux (250ml flask), ultrapure water machine (U.S. Millipore company).
Phthalic acid mono-methyl, repefral (DMP), phthalic acid mono, diethyl phthalate (DEP), diisopropyl phthalate, n-propyl phthalate, diphenyl phthalate, BBP(Butyl Benzyl Phthalate (BBP), diisobutyl phthalate (DIBP), dicyclohexyl phthalate (DCHP), diisooctyl phthalate (DEHP) standard items (lark prestige Science and Technology Ltd.); Acetonitrile, methyl alcohol, formic acid are chromatographically pure, and normal hexane is analyzed pure.
The Radix Codonopsis medicine materical crude slice, available from Wen County, Gansu, lot number: ds-080712.
1.2 experimental technique
1.2.1 chromatographic condition
Agilent ZORBAX SB-C
18Column (3.0 * 150mm, 3.5 μ m); With 0.05% formic acid is the mobile phase A phase, is the Mobile phase B phase with 100% acetonitrile, flow velocity 0.3ml/min; According to the form below carries out gradient elution:
Table 5-1 eluent gradient
1.2.2 mass spectrum condition
Ion gun is ESI source, electro-spray ionization source; Mass spectrum parameters such as capillary voltage, taper hole voltage are seen table 5-2.
Table 5-2 mass spectrum parameter list
1.2.3 the preparation of need testing solution
Precision takes by weighing Radix Codonopsis sample powder (crossing 40 mesh sieves) 3g, puts in the round-bottomed flask, and the accurate normal hexane 90ml that adds, 2h is extracted in 80 ℃ of hot refluxs; Filter, precision is measured subsequent filtrate 50ml, puts to revolve and steams in the bottle 40 ℃ of rotary evaporations to doing; Be settled to 5ml with n-hexane dissolution, filter, promptly get.(annotate: experimentation avoids contacting plastic products, in order to avoid pollute).
2, result
2.1 measure
The accurate need testing solution 5 μ l that draw analyze according to above-mentioned condition sample introduction, read corresponding amount from typical curve, calculate, and promptly get.
The result shows that detect above-mentioned diisooctyl phthalate (DEHP) in the Radix Codonopsis sample, content is 664ng/g.
1, materials and methods
1.1 key instrument and reagent
The single quadrupole mass spectrometer (U.S. Waters company) of Waters ZQ, ACQUITY Ultra Performance LC liquid chromatograph (U.S. Waters company), apparatus,Soxhlet's (250ml flask, 80ml are extracted tube out), ultrapure water machine (U.S. Millipore company).
Phthalic acid mono-methyl, repefral (DMP), phthalic acid mono, diethyl phthalate (DEP), diisopropyl phthalate, n-propyl phthalate, diphenyl phthalate, BBP(Butyl Benzyl Phthalate (BBP), diisobutyl phthalate (DIBP), dicyclohexyl phthalate (DCHP), diisooctyl phthalate (DEHP) standard items (lark prestige Science and Technology Ltd.); Acetonitrile, methyl alcohol, formic acid are chromatographically pure, and sherwood oil (60～90 ℃) is analyzed pure.
Root of herbaceous peony medicine materical crude slice is available from medicinal material market, Bozhou
1.2 experimental technique
1.2.1 chromatographic condition
Agilent ZORBAX SB-C
18Column (3.0 * 150mm, 3.5 μ m); With 0.1% formic acid is the mobile phase A phase, is the Mobile phase B phase with 100% acetonitrile, flow velocity 0.3ml/min; According to the form below carries out gradient elution:
Table 2-1 eluent gradient
1.2.2 mass spectrum condition
Ion gun is ESI source, electro-spray ionization source; Mass spectrum parameters such as capillary voltage, taper hole voltage are seen table 6-2.
Table 6-2 mass spectrum parameter list
1.2.3 the preparation of need testing solution
Precision takes by weighing root of herbaceous peony sample powder (crossing 40 mesh sieves) 3g, puts in the round-bottomed flask, and accurate sherwood oil (60～90 ℃) 90ml that adds, 85 ℃ of Suo Shi extract 6h; Filter, precision is measured subsequent filtrate 50ml, puts to revolve and steams in the bottle 40 ℃ of rotary evaporations to doing; Be settled to 5ml with dissolve with methanol, filter, promptly get.(annotate: experimentation avoids contacting plastic products, in order to avoid pollute).
2, result
2.1 measure
The accurate need testing solution 5 μ l that draw analyze according to above-mentioned condition sample introduction, read corresponding amount from typical curve, calculate, and promptly get.
The result shows that detect above-mentioned diisooctyl phthalate (DEHP) in the root of herbaceous peony sample, content is 1242ng/g.
Claims (15)
1. be used for measuring the method for Chinese medicine phthalic acid ester content, it is characterized in that, this method comprises the steps:
(1) testing sample is carried out pre-service;
(2) pass through the content that LC/MS is measured one or more phthalic esters in pretreated sample.
2. method according to claim 1; It is characterized in that said phthalic ester is selected from phthalic acid mono-methyl, repefral, phthalic acid mono, diethyl phthalate, diisopropyl phthalate, n-propyl phthalate, diphenyl phthalate, BBP(Butyl Benzyl Phthalate, diisobutyl phthalate, dicyclohexyl phthalate or diisooctyl phthalate.
3. method according to claim 1 is characterized in that said Chinese medicine is selected from Radix Codonopsis, the root of herbaceous peony, the radix paeoniae rubrathe or glutinous rehmannia.
4. method according to claim 1 is characterized in that, The pretreatment comprises that rotary evaporation is to doing the step of redissolving with second kind of solvent more then with first kind of solvent supersonic or backflow or Suo Shi extraction in the said step (1).
5. method according to claim 4 is characterized in that, said first kind of solvent is selected from normal hexane, sherwood oil or chloroform, preferred normal hexane.
6. method according to claim 4 is characterized in that, said second kind of solvent is selected from methyl alcohol, chloroform methanol solution or normal hexane, particular methanol.
7. method according to claim 4 is characterized in that, the condition of said ultrasonic Extraction is: under the ultrasonic frequency of 32～40kHz, and 25～45 ℃ of following water-bath ultrasonic Extraction 30～60min of temperature; The condition of said refluxing extraction is: 80～100 ℃ of refluxing extraction 1h～2h; The condition that said Suo Shi extracts is: 80～100 ℃ of Suo Shi extract 4～8h.
8. method according to claim 1 is characterized in that, the chromatographic condition of liquid chromatography in the said step (2)-polyphone mass spectroscopy is: adopt the C18 post; Moving phase is by A phase and B phase composition; A is aqueous formic acid mutually, and B is methyl alcohol or acetonitrile mutually, adopts gradient elution.
9. method according to claim 1; It is characterized in that; The mass spectrum condition of LC/MS is in the said step (2): adopt electric spray ion source, adopt positive ion mode to carry out data acquisition, 2.5～4.0 kilovolts of capillary voltage, 10～30 volts of taper hole voltages.
10. according to the said method of claim 9, it is characterized in that capillary voltage is 3.0 kilovolts, taper hole voltage is 15 volts or 20 volts.
11. method according to claim 8 is characterized in that, the particle diameter of said C18 post is 1.7～5.0 μ m, and column internal diameter 2.1～4.6mm, column length are 5～25cm.
12. method according to claim 8 is characterized in that, said C18 post is an Agilent ZORBAX SB-C18 post.
13. method according to claim 8 is characterized in that, wherein the concentration of A phase is 0.0%～0.5%, preferred 0.05%.
14. method according to claim 8 is characterized in that, wherein A: the B volume ratio is 50～2%: 50～98%.
15. method according to claim 8 is characterized in that, wherein the flow velocity of moving phase is 0.2～1ml/min, preferred 0.3ml/min.
Priority Applications (2)
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| CN201110076663.8A CN102721754B (en) | 2011-03-29 | 2011-03-29 | A method for determining phthalate content in traditional Chinese medicine by adopting a liquid chromatography-mass spectrometry method |
| HK13103499.5A HK1176121A1 (en) | 2011-03-29 | 2013-03-20 | A liquid phase chromatography mass spectrometry method for determining the amount of phthalates in chinese medicine |
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| CN201110076663.8A CN102721754B (en) | 2011-03-29 | 2011-03-29 | A method for determining phthalate content in traditional Chinese medicine by adopting a liquid chromatography-mass spectrometry method |
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| CN103226144A (en) * | 2013-03-19 | 2013-07-31 | 北京普赞生物技术有限公司 | Making method of rapid detection kit of dibutyl phthalate, and detection method of rapid detection kit of dibutyl phthalate |
| CN103399104A (en) * | 2013-07-31 | 2013-11-20 | 中国农业科学院茶叶研究所 | Pretreatment kit for detecting phthalic acid ester in tea and treatment method of pretreatment kit |
| CN103760274A (en) * | 2014-02-18 | 2014-04-30 | 云南健牛生物科技有限公司 | Method for detecting phthalic acid esters in food |
| CN103995075A (en) * | 2014-05-08 | 2014-08-20 | 广州甘蔗糖业研究所 | High performance liquid chromatography-tandem mass spectrometry detection method of plasticizer in sugar |
| CN104560358A (en) * | 2014-12-27 | 2015-04-29 | 华南农业大学 | Removal and extraction method for low phthalic acid ester compounds in schisandra chinensis oil |
| CN106404934A (en) * | 2016-08-25 | 2017-02-15 | 宁波检验检疫科学技术研究院 | Method for detecting phthalic acid di-n-propyl ester in washing product |
| CN106680394A (en) * | 2017-01-05 | 2017-05-17 | 上海迪安医学检验所有限公司 | Kit for determining content of 14 environmental hormones in urine in liquid chromatography tandem mass spectrometry |
| CN107727775A (en) * | 2017-11-28 | 2018-02-23 | 福建省国鼎检测技术有限公司 | The detection method of phthalic acid mono-n-butylester in a kind of textile |
| CN109521134A (en) * | 2018-11-24 | 2019-03-26 | 上海化工研究院有限公司 | The deuterium-labeled own ester standard substance of phthalic acid two (2- ethyl) measures and application |
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- 2011-03-29 CN CN201110076663.8A patent/CN102721754B/en active Active
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103226144A (en) * | 2013-03-19 | 2013-07-31 | 北京普赞生物技术有限公司 | Making method of rapid detection kit of dibutyl phthalate, and detection method of rapid detection kit of dibutyl phthalate |
| CN103399104A (en) * | 2013-07-31 | 2013-11-20 | 中国农业科学院茶叶研究所 | Pretreatment kit for detecting phthalic acid ester in tea and treatment method of pretreatment kit |
| CN103760274A (en) * | 2014-02-18 | 2014-04-30 | 云南健牛生物科技有限公司 | Method for detecting phthalic acid esters in food |
| CN103995075A (en) * | 2014-05-08 | 2014-08-20 | 广州甘蔗糖业研究所 | High performance liquid chromatography-tandem mass spectrometry detection method of plasticizer in sugar |
| CN103995075B (en) * | 2014-05-08 | 2016-03-30 | 广州甘蔗糖业研究所 | The using high performance liquid chromatography tandem mass spectrum detection method of plasticiser in a kind of sugar |
| CN104560358A (en) * | 2014-12-27 | 2015-04-29 | 华南农业大学 | Removal and extraction method for low phthalic acid ester compounds in schisandra chinensis oil |
| CN106404934A (en) * | 2016-08-25 | 2017-02-15 | 宁波检验检疫科学技术研究院 | Method for detecting phthalic acid di-n-propyl ester in washing product |
| CN106680394A (en) * | 2017-01-05 | 2017-05-17 | 上海迪安医学检验所有限公司 | Kit for determining content of 14 environmental hormones in urine in liquid chromatography tandem mass spectrometry |
| CN106680394B (en) * | 2017-01-05 | 2019-03-08 | 上海迪安医学检验所有限公司 | Liquid Chromatography-Tandem Mass Spectrometry measures the kit of 14 kinds of Environmental Hormone contents in urine |
| CN107727775A (en) * | 2017-11-28 | 2018-02-23 | 福建省国鼎检测技术有限公司 | The detection method of phthalic acid mono-n-butylester in a kind of textile |
| CN109521134A (en) * | 2018-11-24 | 2019-03-26 | 上海化工研究院有限公司 | The deuterium-labeled own ester standard substance of phthalic acid two (2- ethyl) measures and application |
Also Published As
| Publication number | Publication date |
|---|---|
| HK1176121A1 (en) | 2013-07-19 |
| CN102721754B (en) | 2015-03-25 |
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