CN105388222B - A kind of high performance liquid chromatography determines borneol, camphor, the method for isoborneol content simultaneously - Google Patents
A kind of high performance liquid chromatography determines borneol, camphor, the method for isoborneol content simultaneously Download PDFInfo
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- CN105388222B CN105388222B CN201510670954.8A CN201510670954A CN105388222B CN 105388222 B CN105388222 B CN 105388222B CN 201510670954 A CN201510670954 A CN 201510670954A CN 105388222 B CN105388222 B CN 105388222B
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Abstract
Borneol, camphor, the method for isoborneol content in a kind of high effective liquid chromatography for measuring sample, this method are borneol, camphor, isoborneol content in the chromatographic peak area determination sample using high performance liquid chromatography, including：(1) chromatographic condition is determined；(2) prepared by reference substance solution；(3) preparation of need testing solution；(4) determine；(5) result is calculated：According to calculation formula Cx=CR×AX×WR/AR×WX× 100% calculates borneol, camphor, isoborneol content.Detection speed of the present invention is fast, accurate detection data, favorable reproducibility, sensitivity is high, specificity is strong, operating method is easy, detection reagent and instrument application are extensive, the experiment such as recovery test is investigated and is loaded by precision test, replica test, linear relationship to verify method, as a result this method is shown accurately and reliably, it is convenient and swift, it is with low cost；Detection range is big；Sensitivity, accuracy are high；Chemical analysis field will be widely used in.
Description
Technical field
Invention is related to borneol, more particularly to high performance liquid chromatography determine borneol in the sample such as L-Borneol and borneol,
Camphor, isoborneol content method.
Background technology
It is well known that borneol has tri- kinds of optical isomers of d, l, dl.《Chinese Pharmacopoeia》Version one has been recorded naturally within 2010
Borneol (d-Bomeol), borneol (borneolum syntheticum) and L-Borneol (L-Borneol), L-Borneol therein are new for feverfew Blumea balsamifera
The extracted crystallization being processed into of fresh leaf, its main active is L-Borneol；Borneol is Spore density kapur
Resin and volatile oil processed goods extract the translucent crystalline solid like plum blossom valve bulk, sheet obtained, are near the pure right side
Borneol is revolved, is also also chemically synthesized；Natural borneol for natural canella camphor tree fresh branch, leaf it is extracted be processed into,
For white crystalline powder or flaky crystal, main component is d-Bomeol.《Chinese Pharmacopoeia》Record L-Borneol, borneol, natural ice
Piece：" pungent, hardship, is slightly cold.The thoughts of returning home, spleen, lung channel.Its effect：Inducing resuscitation of having one's ideas straightened out, clearing away heat to and alleviating pain.Fainted for pyreticosis coma, convulsion, in
Wind phlegm is fainted, bstruction of the circulation of vital energy is fainted cruelly, middle evil is gone into a coma, hot eyes, aphtha, abscess of throat, discharging ear sore ".L-Borneol, borneol clinical practice are extremely wide
It is general, it is the conventional Chinese medicine of inducing resuscitation of having one's ideas straightened out especially in the prevention and treatment of cardiovascular and cerebrovascular disease.
Significant active component in natural borneol, borneol and L-Borneol is borneol, and its content is normally used as above-mentioned three kinds
Medicinal material major quality controlling index, the main method of current Borneo amount assay includes gas chromatography, derivatization efficient liquid phase
Chromatography, thin-layered chromatography, infra-red sepectrometry, UV-VIS spectrophotometry etc..Because it has a volatility, and not compared with
Strong UV absorption, its content assaying method uses GC methods or GC-MS methods mostly.At present,《Chinese Pharmacopoeia》2010 editions are included day
Borneol, camphor, isoborneol content assaying method are gas chromatography (GC) in right borneol, borneol and L-Borneol, for its quality control
System.But GC analysis costs are higher, high is required to sample stability, and the pre-column derivatization for the liquid phase measurement reported at present increases
Add the pre-treatment time of sample, and Composition distribution is detected according to the refractive index of material, is a kind of universal
Detector, can detect the component that UV absorption can not be responded, applied widely, and its requirement to sample is relatively low, divide
Analyse cost low, the detection available for industrialized production.In Chinese patent database also without be related to efficient liquid phase determine borneol, camphor tree
The patent of brain, the content of isoborneol, therefore set up high performance liquid chromatography and show poor method Accurate Determining borneol, camphor, isoborneol content
Method have very important significance.
The content of the invention
The present invention is intended to provide simplicity, good separating effect, precision and a kind of high high performance liquid chromatography of accuracy are determined
Borneol, camphor, the method for isoborneol content, to reach above-mentioned purpose, this method employs following technical scheme：
(1) chromatographic condition is determined
Chromatographic column：C18 posts；
Detector：Composition distribution；
Column temperature：30 DEG C~40 DEG C；
Mobile phase：The phosphoric acid solution of methanol -0.4% or acetonitrile-phosphate aqueous solution
Flow velocity：0.6~1mLmin-1；
Sample size：10~20 μ L.
(2) preparation of reference substance solution
Weigh borneol, camphor, isoborneol reference substance it is appropriate, it is accurately weighed, plus 1mgmL is made in methanol-1, it is used as control
Product solution；
(3) preparation of need testing solution
Precision weighs appropriate test sample 10mg, plus methanol dissolving prepares 1mgmL-1, produce need testing solution；
(4) determine
Reference substance solution and need testing solution are subjected to high performance liquid chromatography detection respectively, it is wherein to be checked in need testing solution
The separating degree surveyed between peak is more than 1.5, reference substance solution chromatographic peak and need testing solution chromatographic peak is respectively obtained, according to reference substance
The qualitative of test sample component is carried out with the appearance time of test sample, is calculated to obtain component to be detected by external standard method
Content；
(5) calculation formula
Borneol, camphor, isoborneol content are calculated according to following external standard method calculation formula：
Cx=CR×AX×WR/AR×WX× 100%
In formula：CXFor the content of test sample,
CRFor the content of reference substance,
AXFor the peak area of test sample,
ARFor the peak area of reference substance,
WRFor the sample weighting amount of reference substance, unit：Mg,
WxFor the sample weighting amount of test sample, unit：mg.
It is methanol or acetonitrile in the mobile phase of the above method；The fraction of phosphoric acid is 0.4%.
The present invention detects borneol, camphor, isoborneol content simultaneously using high performance liquid chromatography, different from existing gas phase
Chromatographic process, compared with prior art, with detection speed is fast, accurate detection data, favorable reproducibility, sensitivity is high, specificity
By force, the features such as operating method simplicity, detection reagent and extensive instrument application, by precision test, replica test, linear pass
System investigates and is loaded the experiment such as recovery test and method is verified, as a result shows this method accurately and reliably, convenient and swift, cost
It is cheap；Detection range is big；Sensitivity, accuracy are high；It will be widely used in chemical analysis field.
Brief description of the drawings
Fig. 1 is borneol, camphor, the mixed mark high-efficient liquid phase chromatogram of isoborneol；Fig. 2 Chinese mugwort powder test sample high-efficient liquid phase chromatograms；
The mixed mark high-efficient liquid phase chromatogram of Fig. 3 borneol, camphor, isoborneol；Fig. 4 L-Borneol high-efficient liquid phase chromatograms；Fig. 5 reference substance efficient liquid phases
Chromatogram；Fig. 6 borneols (borneolum syntheticum) high-efficient liquid phase chromatogram.
Embodiment
Following case study on implementation will be helpful to the understanding of the present invention, but these case study on implementation to the present invention only for being said
Bright, the present invention is not limited to these contents.
Embodiment 1
Borneol, camphor, isoborneol content detection in Chinese mugwort powder
(1) chromatographic condition：Chromatographic column：Yi Lite C18 posts；Detector：Composition distribution；Column temperature：40℃；Mobile phase：First
Alcohol：Phosphate aqueous solution is 65：35, flow velocity：1mL·min-1。
(2) preparation of standard solution：The preparation of reference substance solution：Borneol, camphor, the reference substance of isoborneol is weighed to fit
Amount, it is accurately weighed, plus 1mgmL is made in methanol-1, it is used as reference substance solution；
(3) preparation of need testing solution：Precision weighs appropriate test sample 10mg, plus methanol dissolving prepares 1mgmL-1, i.e.,
Obtain need testing solution；
(4) record chromatogram and determine：Reference substance solution and need testing solution are subjected to high performance liquid chromatography detection respectively,
Separating degree wherein in need testing solution between peak to be detected is more than 1.5, respectively obtains reference substance solution chromatographic peak and test sample
Solution chromatographic peak, carries out the qualitative of test sample component according to the appearance time of reference substance and test sample, is carried out by external standard method
Calculate to obtain the content of component to be detected.According to calculation formulaThe content calculated
1 is shown in Table, reference substance high-efficient liquid phase chromatogram is shown in accompanying drawing 1, test sample high-efficient liquid phase chromatogram is shown in accompanying drawing 2.
Borneol, camphor, isoborneol assay result in the Chinese mugwort powder of table 1
Show that stability of the present invention is good by examples detailed above, precision is high, it is adaptable to borneol, camphor, isoborneol it is efficient
The assay of liquid chromatography.
The foundation of the inventive method
1.1 specificity
This method specificity is strong.
1.2 the range of linearity
Precision measure mixed reference substance solution 1,2,3,4,5,8mL be respectively placed in 10mL measuring bottles, with methanol constant volume to carve
Degree, shakes up, is configured to reference substance solution.Sample introduction is analyzed and records chromatogram on HPLC.Respectively with camphor, borneol, isoborneol
Concentration be abscissa, using peak area as ordinate, carry out regression analysis.Camphor, borneol, the regression equation difference of isoborneol
For：
Y=77727X+2635.7 (R2=0.9990)；
Y=79415X+2756.0 (R2=0.9991)；
Y=79284X+4796.6 (R2=0.9993).
The range of linearity is respectively 0.205~1.535,0.207~1.554,0.214~1.607mgmL-1。
1.3 precision test
Precision measures mixed reference substance solution 1.0mL, puts in 10mL measuring bottles, with methanol constant volume to scale, shakes up, and produces pair
According to product solution.Repeat sample introduction to analyze 6 times, record chromatogram.Measuring camphor, borneol, the RSD of the peak area of isoborneol is respectively
0.90%th, 1.07% and 0.68%, show that sample introduction precision is fine.
1.4 replica test
Precision weighs Chinese mugwort powder 6 parts of 10mg samples of sample respectively, and 10mL chromatograms methanol dissolves, in measure under above-mentioned chromatographic condition,
As a result show not detect isoborneol in the sample, camphor, borneol average content be 27.37%, 61.83%, RSD be 1.67%,
1.81%.Show that method repeatability is good, the results are shown in Table 2.
Borneol, camphor, isoborneol content replica test (n=6) in the sample of table 2
1.5 sample solution study on the stability
Precision weighs Chinese mugwort powder sample 10.46mg, is dissolved with methanol and is settled to 10mL, is carried out respectively at 0,2,4,8,12,24h
Determine, camphor, borneol average content 29.55%, 61.43%, RSD are 0.98%, 0.95%, isoborneol content is 0.Show molten
Liquid is stable in 24h, the results are shown in Table 3.
The sample solution study on the stability of table 3
1.6 average recoveries are tested
Precision weighs Chinese mugwort powder sample 20.20mg, 10mL methanol dissolving, shakes up, and draws 5mL in 100mL volumetric flasks, methanol constant volume
To scale, 5mL is drawn respectively in 6 test tubes, two parts are separately added into mixed reference substance solution 1mL (0.100mg containing borneol, camphor tree
Brain 0.069mg, isoborneol 0.071mg)；Two parts are separately added into mixed reference substance solution 1mL (0.200mg containing borneol, camphor
0.138mg, isoborneol 0.143mg)；Two parts are separately added into mixed reference substance solution 1mL (0.310mg containing borneol, camphor
0.207mg, isoborneol 0.218mg).Sample introduction is determined, and borneol, camphor, the average recovery rate of isoborneol and RSD values are tried to achieve in calculating,
It the results are shown in Table 4.
Borneol, camphor, isoborneol average recovery experiment (n=6) in the sample of table 4
Embodiment 2
Borneol, camphor, isoborneol content detection in L-Borneol
(1) chromatographic condition：Chromatographic column：Yi Lite C18 posts, detector：Composition distribution, column temperature：35℃；Mobile phase：Second
Nitrile：0.4% phosphoric acid solution is 55:45；Flow velocity：0.8mL·min-1。
(2) preparation of standard solution：The preparation of reference substance solution：Borneol, camphor, the reference substance of isoborneol is weighed to fit
Amount, it is accurately weighed, plus 1mgmL is made in methanol-1, it is used as reference substance solution；
(3) preparation of need testing solution：Precision weighs appropriate test sample 10mg, plus methanol dissolving prepares 1mgmL-1, i.e.,
Obtain need testing solution；
(4) record chromatogram and determine：Reference substance solution and need testing solution are subjected to high performance liquid chromatography detection respectively,
Separating degree wherein in need testing solution between peak to be detected is more than 1.5, respectively obtains reference substance solution chromatographic peak and test sample
Solution chromatographic peak, carries out the qualitative of test sample component according to the appearance time of reference substance and test sample, is carried out by external standard method
Calculate to obtain the content of component to be detected.5 are the results are shown in Table, borneol content is up to 95.47%, camphor, different dragon wherein in L-Borneol
Brain content is essentially 0, and reference substance high-efficient liquid phase chromatogram is shown in accompanying drawing 3, and the test sample high-efficient liquid phase chromatogram is shown in accompanying drawing 4.
Borneo amount assay (n=5) in the L-Borneol of table 5
Embodiment 3
Borneol, camphor, isoborneol content detection in borneol (borneolum syntheticum)
(1) chromatographic condition：Chromatographic column：Yi Lite C18 posts；Detector：Composition distribution；Column temperature：40℃；Mobile phase：Second
Nitrile：0.4% phosphoric acid solution is 55:45；Flow velocity：1mL·min-1。
(2) preparation of standard solution：The preparation of reference substance solution：Borneol, camphor, the reference substance of isoborneol is weighed to fit
Amount, it is accurately weighed, plus 1mgmL is made in methanol-1, it is used as reference substance solution；
(3) preparation of need testing solution：Precision weighs appropriate test sample 5mg, plus methanol dissolves and is settled to 50mL, prepares
Obtain 0.1mgmL-1, produce need testing solution；
(4) determine：Reference substance solution and need testing solution are subjected to high performance liquid chromatography detection respectively, wherein test sample is molten
Separating degree in liquid between peak to be detected is more than 1.5, respectively obtains reference substance solution chromatographic peak and need testing solution chromatographic peak, root
The qualitative of test sample component is carried out according to the appearance time of reference substance and test sample, is calculated to be checked to obtain by external standard method
Survey the content of component.6 are the results are shown in Table, reference substance high-efficient liquid phase chromatogram is shown in accompanying drawing 5, the test sample high-efficient liquid phase chromatogram is shown in attached
Fig. 6.
Borneol, camphor, isoborneol content in the borneol of table 6 (borneolum syntheticum)
Claims (1)
1. one kind is determined using borneol, camphor, isoborneol content method in high effective liquid chromatography for measuring L-Borneol and borneol sample,
Including：
(1) chromatographic condition is determined
Chromatographic column：C18 posts；
Detector：Composition distribution；
Column temperature：30 DEG C~40 DEG C；
Mobile phase：The phosphoric acid solution of methanol -0.4% or acetonitrile-phosphate aqueous solution
Flow velocity：0.6~1 mLmin-1；
Sample size：10~20 μ L；
(2) preparation of reference substance solution
Weigh borneol, camphor, isoborneol reference substance it is appropriate, it is accurately weighed, plus 1 mgmL is made in methanol-1, it is used as reference substance
Solution；
(3) preparation of need testing solution
Precision weighs the appropriate mg of test sample 10, plus methanol dissolving prepares 1 mgmL-1, produce need testing solution；
(4) determine
Reference substance solution and need testing solution are subjected to high performance liquid chromatography detection respectively, peak to be detected wherein in need testing solution
Between separating degree be more than 1.5, respectively obtain reference substance solution chromatographic peak and need testing solution chromatographic peak, according to reference substance with confession
The appearance time of test product carries out the qualitative of test sample component, is calculated to obtain containing for component to be detected by external standard method
Amount；
(5) calculation formula
Borneol, camphor, isoborneol content are calculated according to following external standard method calculation formula：
Cx=CR×AX×WR/AR×WX×100%
In formula：CXFor the content of test sample,
CRFor the content of reference substance,
AXFor the peak area of test sample,
ARFor the peak area of reference substance,
WRFor the sample weighting amount of reference substance, unit：Mg,
WxFor the sample weighting amount of test sample, unit：mg；
Organic phase is methanol or acetonitrile in the mobile phase, and the fraction of phosphoric acid is 0.4%；Methanol and 0.4% phosphoric acid matter in mobile phase
Amount is than being 65: 35, and acetonitrile and 0.4% phosphoric acid solution mass ratio are 55: 45.
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| CN102091283A (en) * | 2009-12-11 | 2011-06-15 | 无锡济民可信山禾药业股份有限公司 | Method for detecting Xingnaojing injection |
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| CN102091283A (en) * | 2009-12-11 | 2011-06-15 | 无锡济民可信山禾药业股份有限公司 | Method for detecting Xingnaojing injection |
Non-Patent Citations (4)
| Title |
|---|
| Comparative study on camphor enantiomers behavior under the conditions of gas–liquid chromatography and reversed-phase high-performance liquid chromatography systems modified with a- and b-cyclodextrins;Monika Asztemborska等;《Journal of Chromatography A》;20000331;第874卷(第1期);第73-80页 * |
| HPLC-RID 法同时测定复方痱子粉中 4 种成分的含量;彭敏;《现代中药研究与实践》;20140831;第28卷(第4期);第60页第1-2节 * |
| HPLC-RID 法测定紫冬油中冰片含量;汤艳群等;《医药前沿》;20150430;第5卷(第10期);第93页第2节 * |
| 气相色谱法同时测定醒脑静注射液中麝香酮、龙脑、樟脑、异龙脑的含量;方颖等;《中国实验方剂学杂志》;20120430;第18卷(第8期);第96-99页 * |
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