CN107064377A - A kind of method for detecting levocarnitine content in compound vitamin C and L levocarnitine chewable tablets - Google Patents
A kind of method for detecting levocarnitine content in compound vitamin C and L levocarnitine chewable tablets Download PDFInfo
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- CN107064377A CN107064377A CN201611204973.2A CN201611204973A CN107064377A CN 107064377 A CN107064377 A CN 107064377A CN 201611204973 A CN201611204973 A CN 201611204973A CN 107064377 A CN107064377 A CN 107064377A
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- levocarnitine
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
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Abstract
A kind of method for being applied to detect levocarnitine content in compound vitamin C and L levocarnitine chewable tablets in analysis field, the detection method comprises the following steps：Chromatographic condition：Chromatographic column：Using octadecylsilane chemically bonded silica as filler, 5 μm of particle diameter；Column temperature：30‑40℃；Flow velocity：0.8‑1.2ml/min；Detection wavelength：225nm；Sample size：10‑50μl；Mobile phase：Phosphate buffer and methanol；Levocarnitine content is detected by this method, detection cycle is short, and reappearance, the degree of accuracy, stability can meet the requirements, and reduce testing cost.
Description
Technical field
The present invention relates to left card in a kind of detection compound vitamin C in Pharmaceutical Analysis field and L- levocarnitine chewable tablets
The method of Buddhist nun spit of fland content.
Background technology
(English is entitled for levocarnitine：), also known as l-cn Carnitine.Early in, the Gulewitsch of Russia in 1905
Just carnitine is found that with Krimberg from beef stain meter.Nineteen twenty-seven, Tomita and Sendju determine chemical constitution.Zuo Ka
Ni Ting is the composition of food, is widely present in nature, content highest in chevon, about 2.1g/kg, and vegetalitas
Content is few even without, it is considered to be the nutrient of biostearin in food.Levocarnitine major function is to promote lipid generation
Thank, ischemic, anoxic for muscle cell tissue, levocarnitine is its main energy sources, left Ni Kating is applied to chronic renal
Decline chronic hemodialysis patient because Secondary cases carnitine lack produce a series of syndromes, clinical manifestation be cardiomyopathy, skeletal myopathy,
Muscle spasmus etc. in arrhythmia cordis, hyperlipidemia, and low blood pressure and dialysis.The levocarnitine detection reported both at home and abroad has a variety of sides
Method, conventional at present has enzyme assay, radioisotope method, optical method and high performance liquid chromatography.Tried needed for enzyme assay
Mostly costly, operating procedure is complicated for agent, takes longer.Isotope method not only operates loaded down with trivial details, measure precision poor, and
There is radioisotope pollution environment and the problem of operator damages.Although optical method cost is low, poor specificity, sensitivity
It is low, it is difficult to meet analysis method requirement.And in compound preparation, pharmaceutical preparation composition is complicated, principal component content is relatively low, auxiliary material group
Divide the detection of interference principal component chromatographic peak, cause content analysis difficult.At present, have no on levocarnitine content in compound preparation
The pertinent literature and patent of detection, therefore, the method for developing levocarnitine content in a kind of compound preparation is urgently to be resolved hurrily
New problem.
The content of the invention
It is an object of the invention to provide levocarnitine content in a kind of compound vitamin C and L- levocarnitine chewable tablets
Detection method, this method is detected using high performance liquid chromatography (concentration, Gradient Elution Method), passes through the excellent of chromatographic condition
Change, the selection of composition proportion and the selection of mobile phase in mobile phase when concentration, gradient elution, establish a kind of detection compound preparation
The method of middle levocarnitine content, the levocarnitine detected by this method, detection cycle is short, reappearance, the degree of accuracy, stability
It can meet the requirements, and reduce testing cost.
The object of the present invention is achieved like this：Left card in one kind detection compound vitamin C and L- levocarnitine chewable tablets
The method of Buddhist nun spit of fland content, the detection method comprises the following steps：
(1) chromatographic condition：
Chromatographic column：C18 posts, specification 250 × 4.6mm, 5 μm
Column temperature：30-40℃
Flow velocity：0.8-1.2ml/min
UV-detector wavelength：225nm
Sample size：10-50μl
Mobile phase：Phosphate buffer and methanol
(2) preparation of phosphate buffer：
Phosphatase 11 1.5ml is taken, 1900ml is diluted with water to, pH is adjusted to 2.4 with sodium hydroxide solution (1mol/L) about 100ml,
Plus sodium heptanesulfonate 1.1g, shaking makes dissolving；
(3) preparation of need testing solution：
Take this product appropriate, accurately weighed, finely ground, precision weighs fine powder in right amount, equivalent to levocarnitine 20mg, in 100ml
In measuring bottle, add water appropriate, shaking makes dissolving, is diluted with water to scale, shakes up, filter, take subsequent filtrate as need testing solution；
(4) preparation of reference substance solution：
Precision weighs levocarnitine reference substance 20mg, puts in 100ml measuring bottles, is dissolved in water and is diluted to scale, shakes up, and makees
For reference substance solution；
(5) assay method：
Precision measures reference substance solution 10-50 μ l injection liquid chromatographs, is repeated 5 times, records chromatogram, levocarnitine
Number of theoretical plate must not be less than 1200, and the relative standard deviation of levocarnitine peak area answers≤2.0%；Precision measures need testing solution
10-50 μ l inject liquid chromatograph, record chromatogram, by external standard method with calculated by peak area, produce；
(6) calculation formula
In formula：
AIt is right：Reference substance solution main peak area；
ASample：Need testing solution main peak area；
WIt is right：The weight of reference substance, mg；
WFor：The weight of test sample, mg；
WAverage piece weight：The average piece weight of test sample, mg；
WLabelled amount：The sign content of test sample, mg；
P：The purity of reference substance, %.
Mobile phase phosphate buffer and methanol use concentration, gradient elution method in the assay method, are specially：Wash
Disengage 0-10 minutes, phosphate buffer 98%, methanol 2% after beginning；Elution start it is latter 10-11 minutes, phosphate buffer by
98% to 80%, methanol is by 2% to 20%；Elution starts latter 11-31 minutes, phosphate buffer 80%, methanol 20%；Elution
31-32 minutes after beginning, phosphate buffer is by 80% to 98%, and methanol is by 20% to 2%；Elution start it is latter 32-62 minutes,
Phosphate buffer 98%, methanol 2%；The C18 posts are selected from Agilent TC-C18 posts.
The present invention is characterized by its detection method, and this method is detected using HPLC (concentration, Gradient Elution Method),
By the optimization of chromatographic condition, the selection of composition proportion and the selection of mobile phase in mobile phase, are established when concentration, gradient elution
A kind of method of levocarnitine content in detection compound preparation.
The detection method and prior art phase of levocarnitine content in a kind of compound vitamin C and L- levocarnitine chewable tablets
Than detecting the content of levocarnitine by this method, the detection cycle per sample needle was shorten to 1 hour by 2 hours, and methodology is tested
Card result show, reappearance result RSD be the 0.42%, degree of accuracy testing result rate of recovery be 100.0%, RSD be 0.15%, it is molten
Liquid stability result RSD is 0.43%, can be met the requirements, and reduces testing cost, will be widely used in Pharmaceutical Analysis
In field.
Brief description of the drawings
Below in conjunction with the accompanying drawings and embodiment the present invention is described in detail.
Fig. 1 is the blank solution HPLC collection of illustrative plates of the present invention.
Fig. 2 is the need testing solution HPLC collection of illustrative plates of the present invention.
Fig. 3 is the reference substance solution HPLC collection of illustrative plates of the present invention.
The need testing solution HPLC collection of illustrative plates of Fig. 4 original methods.
Embodiment
Following instance will be helpful to the understanding to the present invention, but these examples to the present invention only for being illustrated, this hair
It is bright to be not limited to these contents.
Embodiment one
1. chromatographic condition
Chromatographic column：5 μm of 250 × 4.6mm of Agilent (Agilent) TC-C18, column temperature：30-40 DEG C, flow velocity：0.8-
1.2ml/min, Detection wavelength：225nm, sample size：10-50μl.
2. it is prepared by solution
2.1 mobile phase：Phosphate buffer and methanol
2.2 phosphate buffer：Phosphatase 11 1.5ml is taken, 1900ml is diluted with water to, with sodium hydroxide solution (1mol/L)
About 100ml adjusts pH to 2.4, plus sodium heptanesulfonate 1.1g, and shaking makes dissolving.
2.3 need testing solution：Take this product appropriate, it is accurately weighed, it is finely ground, precision weigh fine powder 84.96mg, 83.38mg,
83.64mg, puts in 100ml measuring bottles respectively, adds water appropriate, shaking makes dissolving, is diluted with water to scale, shakes up, filters, take continuous filter
Liquid is used as need testing solution 1,2 and 3.
2.4 reference substance solution：Levocarnitine reference substance 20.58mg, 21.48mg are weighed, is put respectively in 100ml measuring bottles, plus
Water dissolves and is diluted to scale, shakes up, and is used as reference substance solution 1 and 2.
3. assay method
Using mobile phase phosphate buffer and methanol concentration, Gradient Elution Method；
Elution starts latter 0-10 minutes, phosphate buffer 98%, methanol 2%；Elution starts latter 10-11 minutes, phosphoric acid
Salt buffer 98% → 80%, methanol 2% → 20%；Elution starts latter 11-31 minutes, phosphate buffer 80%, methanol
20%；Elution starts latter 31-32 minutes, phosphate buffer 80% → 98%, methanol 20% → 2%；Elution starts rear 32-62
Minute, phosphate buffer 98%, methanol 2%.
Take blank auxiliary to prepare blank solution in right amount, inject liquid chromatograph；
Reference substance solution 1 is taken to inject liquid chromatograph, the parallel pin of sample introduction 5, sample size 10-20 μ l.Spectrogram is recorded, calculates left
Relative standard deviation RSD≤2.0% of Carnitine peak area；Need testing solution is taken to inject liquid chromatograph, sample size 10-50 μ
L,
Record collection of illustrative plates.
The peak area of continuous 5 pin of reference substance solution 1 is：
A1=33519 A2=33551 A3=33429 A4=33609
A5=33609 ATo 1Average=33514 RSD=0.21%
The peak area of reference substance solution 2 is：
A1=35162 A2=35151 ATo 2Average=35156
4. sample is determined
Take test liquid to inject liquid chromatograph, record collection of illustrative plates, be respectively repeated once.
The main peak area of test sample 1：A1=23575 A2=23492 ASample 1Average=23534
The main peak area of test sample 2：A1=23316 A2=23236 ASample 2Average=23276
The main peak area of test sample 3：A1=23087 A2=23174 ASample 3Average=23130
5. result of calculation
In formula：
AIt is right：Reference substance solution main peak area；
ASample：Need testing solution main peak area；
WIt is right：The weight of reference substance, mg；
WFor：The weight of test sample, mg；
WAverage piece weight：The average piece weight of test sample, mg；
WLabelled amount：The sign content of test sample, mg；
P：The purity of reference substance, %.
Test sample 1：
Test sample 2：
Test sample 3：
When the concentration of embodiment two, gradient elution in mobile phase composition proportion selection
Other conditions be the same as Example one, using mobile phase phosphate buffer and methanol concentration, Gradient Elution Method；
Elution starts latter 0-10 minutes, phosphate buffer 85%, methanol 15%；Elution starts latter 10-11 minutes, phosphoric acid
Salt buffer 85% → 70%, methanol 15% → 30%；Elution starts latter 11-31 minutes, phosphate buffer 70%, methanol
30%；Elution starts latter 31-32 minutes, phosphate buffer 70% → 85%, methanol 30% → 15%；Elution starts rear 32-
62 minutes, phosphate buffer 85%, methanol 15%.
The chromatographic peak of test sample is disturbed by auxiliary material, it is impossible to detected.
The selection of the mobile phase of embodiment three
Other conditions be the same as Example one, mobile phase is used as using phosphate buffer and acetonitrile.Phosphate buffer：Take phosphorus
Sour 11.5ml, is diluted with water to 1900ml, and pH to 2.4, plus sodium heptanesulfonate are adjusted with sodium hydroxide solution (1mol/L) about 100ml
1.1g, shaking makes dissolving.
The chromatographic peak of test sample is disturbed by auxiliary material, it is impossible to detected.
Claims (3)
1. a kind of method for detecting levocarnitine content in compound vitamin C and L- levocarnitine chewable tablets, it is characterised in that：Should
Detection method comprises the following steps：
(1) chromatographic condition：
Chromatographic column：C18 posts, specification 250 × 4.6mm, 5 μm
Column temperature：30-40℃
Flow velocity：0.8-1.2ml/min
UV-detector wavelength：225nm
Sample size：10-50μl
Mobile phase：Phosphate buffer and methanol
(2) preparation of phosphate buffer：
Phosphatase 11 1.5ml is taken, 1900ml is diluted with water to, pH to 2.4, plus heptan are adjusted with sodium hydroxide solution (1mol/L) about 100ml
Alkyl sulfonic acid sodium 1.1g, shaking makes dissolving；
(3) preparation of need testing solution：
Take this product appropriate, accurately weighed, finely ground, precision weighs fine powder in right amount, equivalent to levocarnitine 20mg, in 100ml measuring bottles
In, add water appropriate, shaking makes dissolving, is diluted with water to scale, shakes up, filter, take subsequent filtrate as need testing solution；
(4) preparation of reference substance solution：
Precision weighs levocarnitine reference substance 20mg, puts in 100ml measuring bottles, is dissolved in water and is diluted to scale, shakes up, as right
According to product solution；
(5) assay method：
Precision measures reference substance solution 10-50 μ l injection liquid chromatographs, is repeated 5 times, records chromatogram, the theory of levocarnitine
Plate number must not be less than 1200, and the relative standard deviation of levocarnitine peak area answers≤2.0%；Precision measures need testing solution 10-
50 μ l inject liquid chromatograph, record chromatogram, by external standard method with calculated by peak area, produce；
(6) calculation formula
In formula：
AIt is right：Reference substance solution main peak area；
ASample：Need testing solution main peak area；
WIt is right：The weight of reference substance, mg；
WFor：The weight of test sample, mg；
WAverage piece weight：The average piece weight of test sample, mg；
WLabelled amount：The sign content of test sample, mg；
P：The purity of reference substance, %.
2. levocarnitine content in a kind of detection compound vitamin C according to claim 1 and L- levocarnitine chewable tablets
Method, it is characterised in that：Mobile phase phosphate buffer and methanol use concentration, gradient elution side in the assay method
Method, be specially：Elution starts latter 0-10 minutes, phosphate buffer 98%, methanol 2%；Elution starts latter 10-11 minutes, phosphorus
Phthalate buffer is by 98% to 80%, and methanol is by 2% to 20%；Elution starts latter 11-31 minutes, phosphate buffer 80%,
Methanol 20%；Elution starts latter 31-32 minutes, and phosphate buffer is by 80% to 98%, and methanol is by 20% to 2%；Elution is opened
32-62 minutes after beginning, phosphate buffer 98%, methanol 2%.
3. levocarnitine content in a kind of detection compound vitamin C according to claim 1 and L- levocarnitine chewable tablets
Method, it is characterised in that：The C18 posts are selected from Agilent TC-C18 posts.
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|---|---|---|---|
| CN201611204973.2A CN107064377A (en) | 2016-12-23 | 2016-12-23 | A kind of method for detecting levocarnitine content in compound vitamin C and L levocarnitine chewable tablets |
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| CN201611204973.2A CN107064377A (en) | 2016-12-23 | 2016-12-23 | A kind of method for detecting levocarnitine content in compound vitamin C and L levocarnitine chewable tablets |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107688071A (en) * | 2017-10-10 | 2018-02-13 | 济南维瑞医药科技开发有限公司 | A kind of method that furfural in vitamin C compound preparation is determined with HPLC methods |
| CN108287205A (en) * | 2018-03-02 | 2018-07-17 | 常州兰陵制药有限公司 | Method in relation to substance in Levocarnitine determined by HPLC and its injection |
| CN117368378A (en) * | 2023-12-01 | 2024-01-09 | 山东齐都药业有限公司 | Method for detecting content of auxiliary materials in levocarnitine oral solution |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103439444A (en) * | 2013-09-10 | 2013-12-11 | 广西大学 | High efficiency liquid chromatography method for detecting carnitine content in fish plasma |
| CN103690975A (en) * | 2013-12-27 | 2014-04-02 | 广州白云山天心制药股份有限公司 | Method of utilizing levocarnitine aqueous solution to produce trace 2(5H)-furanone, and measuring and application of trace 2(5H)-furanone |
-
2016
- 2016-12-23 CN CN201611204973.2A patent/CN107064377A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103439444A (en) * | 2013-09-10 | 2013-12-11 | 广西大学 | High efficiency liquid chromatography method for detecting carnitine content in fish plasma |
| CN103690975A (en) * | 2013-12-27 | 2014-04-02 | 广州白云山天心制药股份有限公司 | Method of utilizing levocarnitine aqueous solution to produce trace 2(5H)-furanone, and measuring and application of trace 2(5H)-furanone |
Non-Patent Citations (5)
| Title |
|---|
| KHOSHKAM,ROGHAIEH; AFSHAR, MINOO: "alidation of a Stability-Indicating RP-HPLC Method for Determination of l-Carnitine in Tablets.", 《INTERNATIONAL SCHOLARLY RESEARCH NOTICES》 * |
| 付彪: "HPLC法测定注射用左卡尼汀的含量", 《黑龙江医药》 * |
| 国家药典委员会: "《药品标准征求意见稿》", 2 April 2015, HTTP://WWW.CHP.ORG.CN/UPLOAD/SITES/CHP/RESOURCE/2014B/2015040208393276118.PDF * |
| 廖彩霞: "高效液相色谱法测定注射用左卡尼汀的含量", 《中国医院药学杂志》 * |
| 林淼: "反相高效液相色谱法测定饲料及饲料添加剂中L-肉碱含量", 《理化检验-化学分册》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107688071A (en) * | 2017-10-10 | 2018-02-13 | 济南维瑞医药科技开发有限公司 | A kind of method that furfural in vitamin C compound preparation is determined with HPLC methods |
| CN108287205A (en) * | 2018-03-02 | 2018-07-17 | 常州兰陵制药有限公司 | Method in relation to substance in Levocarnitine determined by HPLC and its injection |
| CN117368378A (en) * | 2023-12-01 | 2024-01-09 | 山东齐都药业有限公司 | Method for detecting content of auxiliary materials in levocarnitine oral solution |
| CN117368378B (en) * | 2023-12-01 | 2024-03-19 | 山东齐都药业有限公司 | Method for detecting content of auxiliary materials in levocarnitine oral solution |
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Application publication date: 20170818 |