CN108593827A - A kind of method of Determination of sorbic in detection levocarnitine oral solution - Google Patents
A kind of method of Determination of sorbic in detection levocarnitine oral solution Download PDFInfo
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Abstract
The invention belongs to Pharmaceutical Analysis fields, are related to a kind of method detecting Determination of sorbic in levocarnitine oral solution, include the following steps (1) chromatographic condition：Octadecylsilane chemically bonded silica chromatographic column；UV detector wavelength：257nm；Mobile phase：The mixed liquor of phosphate buffer and methanol；(2) test solution is prepared：(3) reference substance solution is prepared：(4) assay method：Precision measures reference substance solution and injects liquid chromatograph, repeats 36 times, records chromatogram, and the relative standard deviation of sorbic acid peak area answers≤2.0%.Precision measure test solution inject liquid chromatograph, record chromatogram, by external standard method with calculated by peak area to get；The advantages that this method has detection cycle short, and reproducibility, accuracy, stability, durability can meet the requirements, and at low cost.
Description
Technical field
The present invention relates to the methods of Determination of sorbic in a kind of detection levocarnitine oral solution in Pharmaceutical Analysis field.
Background technology
(English is entitled for sorbic acid：Sorbicacid) also known as 2,4- hexadienes acid, 2- acrylic acrylic acid.Sorbic acid is
A kind of antisepsis antistaling agent of highly effective and safe, is widely used in the industries such as food, beverage, tobacco, pesticide, cosmetics, it can also be used to
Resin, fragrance and rubber industry.It is very big in the application amount of western developed country but also wideless in the application range of China.
Sorbic acid is a kind of unsaturated fatty acid, and as the aliphatic acid natural with other, sorbic acid participates in metabolic mistake in human body
Journey, and by human consumption and absorption, generate carbon dioxide and water.From the aspects of safety, sorbic acid is a kind of internationally recognized
The preservative of safety (GRAS), safety are very high.FAO (Food and Agriculture Organization of the United Nation), the World Health Organization, U.S. FDA are all to its safety
Give affirmative.Sorbic acid can effectively inhibit mould, the activity of saccharomycete and aerobic bacteria, moreover it is possible to prevent clostridium botulinum,
The harmful microbes such as staphylococcus, salmonella are grown and breeding, but beneficial to anaerobic Bacillus and lactobacillus acidophilus etc.
Microorganism is nearly unavailable, inhibits the effect of development more stronger than bactericidal effect, when to reach the preservation for effectively extending food
Between, and keep the flavor of original food.Its anti-corrosion effect is 5-10 times of similar product sodium benzoate, and toxic side effect is than benzene first
Acid, vitamin C and salt also want low, and toxicity only has a quarter of benzoic acid, the half of salt.Sorbic acid will not produce human body
Raw carcinogenic and teratogenesis.It is the trend of development of food industry that sorbic acid, which replaces sodium benzoate,.But it if adds in food
Sorbic acid is exceeded serious, and long term consumer is taken, and can inhibit bone growth to a certain extent, endangers the health of kidney, liver.Mesh
Before, it has no the pertinent literature detected about Determination of sorbic in levocarnitine oral solution and patent, therefore, develops a kind of
The method of Determination of sorbic is new issue urgently to be resolved hurrily in detection levocarnitine oral solution.
Invention content
The purpose of the present invention is to provide a kind of detection method of Determination of sorbic in levocarnitine oral solution, this method
It is detected using high performance liquid chromatography, by the optimization of chromatographic condition, the selection of composition proportion and mobile phase in mobile phase
Selection, establish it is a kind of detection levocarnitine oral solution in Determination of sorbic method, pass through this method detect sorb
Acid, detection cycle is short, and reproducibility, accuracy, stability, durability can meet the requirements, and has many advantages, such as at low cost.
The object of the present invention is achieved like this：A kind of method of Determination of sorbic in detection levocarnitine oral solution,
The detection method includes the following steps：
(1) chromatographic condition：
Octadecylsilane chemically bonded silica chromatographic column；
UV detector wavelength：257nm；
Mobile phase：The mixed liquor of phosphate buffer and methanol；
(2) test solution is prepared：
(3) reference substance solution is prepared：
(4) assay method：
Precision measures reference substance solution and injects liquid chromatograph, repeats 3-6 times, records chromatogram, sorbic acid peak area
Relative standard deviation answers≤2.0%.Precision measure test solution inject liquid chromatograph, record chromatogram, by external standard method with
Calculated by peak area to get；
(5) calculation formula
In formula：
AIt is right：Reference substance solution main peak area；
ASample：Test solution main peak area；
WIt is right：The weight of reference substance, mg；
P：The purity of reference substance, %；
In the mobile phase, the volume ratio of phosphate buffer and methanol is 65-75：25-35；In the mobile phase, phosphorus
The volume ratio of phthalate buffer and methanol is 70：30；The preparation method of the phosphate buffer is：Phosphatase 11 1.5ml is taken, is added
Water is diluted to 1900ml, with the sodium hydroxide solution tune pH to 2.0-3.0 of 1mol/L, sodium heptanesulfonate 1.1g, shaking is added to make molten
Solution；The octadecylsilane chemically bonded silica chromatographic column is selected from Agilent TC-C18 chromatographic columns, Agilent zorbax SB-
C18 chromatographic columns, Waters symmetry shield RP18 chromatographic columns, Diamonsil C18 chromatographic columns, Agilent
Eclipse XDB-C18 chromatographic columns, Agilent HC-C18 chromatographic columns, Agilent Extend C18 chromatographic columns, Ecosil
C18 chromatographic columns, II C18 chromatographic columns of SHISEIDO MG, SHISEIDO MG C18 chromatographic columns, SHISEIDO ODS C18 chromatographies
Column, Waters XTERRAMS C18 chromatographic columns, 18 chromatographic columns of Waters XTERRARP, Waters Spherisorb ODS2
One kind in chromatographic column；The column temperature of the chromatographic column is 20-40 DEG C, and the flow velocity of the mobile phase is 0.5-2.0ml/min；It is described
The specification of chromatographic column be 250 × 4.6mm, 5 μm, 150 × 4.6mm, 5 μm, 150 × 4.6mm, 3.5 μm, 250 × 4.6mm, 3.5 μm
In one kind；The method for preparing test solution is：Precision measures levocarnitine oral solution 2ml, sets 100ml volumetric flasks
In, it is diluted with water to scale and shakes up, as test solution；It is described prepare reference substance solution method be：Precision weighs sorb
Sour reference substance 15mg, sets in 100ml measuring bottles, appropriate amount of ethanol is added to make dissolving, be diluted with water to scale, shake up；It is accurate again to measure 2ml
It sets in 10ml volumetric flasks, is diluted with water to scale, shake up, as a contrast product solution；The test solution and reference substance solution
Sample size be 5-100 μ l, preferred sample size be 5-20 μ l；The accurate reference substance solution that measures injects liquid chromatograph,
It is repeated 5 times.
The present invention is characterized by its detection method, and principle is that this method is detected using HPLC, and chromatographic column is adopted
With the chromatographic column that octadecylsilane chemically bonded silica is filler, wavelength is chosen most using ultraviolet-uisible spectrophotometer scanning
Big absorbing wavelength, by the optimization of chromatographic condition, the selection of composition proportion, establishes one kind in the selection of mobile phase and mobile phase
The method for detecting Determination of sorbic in levocarnitine oral solution；Methodology validation the result shows that, blank auxiliary (levocarnitine mouth
Take the other compositions in addition to sorbic acid in solution process prescription) it is noiseless to the inspection of sorbic acid, specificity is strong, reproducibility knot
Fruit RSD is 0.7%, the accuracy testing result rate of recovery is 100.5%, RSD 0.3%, stability of solution result RSD are
0.1%, durability result RSD is 0.2%, can be met the requirements.
The detection method of Determination of sorbic compared with prior art, has operation letter in a kind of levocarnitine oral solution
The advantages such as single, convenient, accurate and testing cost is low.It will be widely used in Pharmaceutical Analysis field.
Description of the drawings
The following describes the present invention in detail with reference to the accompanying drawings and embodiments.
Fig. 1 is the blank solution HPLC collection of illustrative plates of the present invention.
Fig. 2 is the test 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.
In figure, 1 is sorbic acid, and 2 be levocarnitine.
Specific implementation mode
With reference to embodiment, the present invention is described further, and embodiment helps to more fully understand the present invention, but
The present invention is not limited only to following embodiments.
Embodiment one
1. chromatographic condition
Chromatographic column：5 μm of (2) 250 × 4.6mm of Diamonsil C18, column temperature：35 DEG C, flow velocity：1.0ml/min detects wave
It is long：257nm, sample size：20μl.
2. prepared by solution
2.1 mobile phase：The mixed liquor of 70 volume phosphate buffers and 30 volumes methanols.
2.2 phosphate buffer：Phosphatase 11 1.5ml is taken, 1900ml is diluted with water to, with sodium hydroxide solution (1mol/L)
About 100ml tune pH to 2.0-3.0 adds sodium heptanesulfonate 1.1g, shaking to make dissolving.
2.3 test solution：Test sample 2, each accurate measurement 2ml is taken to set in 100ml volumetric flasks, be diluted with water respectively
It is shaken up to scale, as test solution 1 and 2.
2.4 reference substance solution：Precision weighs sorbic acid reference substance 15.58mg, 15.25mg, sets in 100ml measuring bottles, adds second
Alcohol makes dissolving in right amount, is diluted with water to scale, shakes up；The accurate 2ml that measures is set in 10ml volumetric flasks again, is diluted with water to scale,
It shakes up, as a contrast product solution 1 and 2.
3. assay method
Reference substance solution 1 is taken to inject liquid chromatograph, 5 needle of parallel sample introduction, 20 μ l of sample size record spectrogram, calculate sorb
Relative standard deviation RSD≤2.0% of sour peak area；Reference substance solution 2 is taken to inject liquid chromatograph, 2 needle of parallel sample introduction, sample introduction
Measure 20 μ l.Test solution 1 and 2 is taken to inject liquid chromatograph, 20 μ l of sample size record collection of illustrative plates.
The peak area of 1 continuous 5 needle of reference substance solution is：
A1=7995.1 A2=7993.5 A3=7991.5 A4=7993.6
A5=7992.9 ATo 1Average=7993.3 RSD=0.1%
The peak area of reference substance solution 2 is：
A1=7825.5 A2=7840.0 ATo 2Average=7832.8
4. sample measures
It takes test solution to inject liquid chromatograph, records collection of illustrative plates, be respectively repeated once.
The main peak area of test solution 1：A1=7593.3 A2=7584.2 ASample 1Average=7588.8
The main peak area of test solution 2：A1=7594.7 A2=7595.6 ASample 2Average=7595.2
5. result of calculation
In formula：
AIt is right：Reference substance solution main peak area；
ASample：Test solution main peak area；
WIt is right：The weight of reference substance, mg；
P：The purity of reference substance, %.
Test solution 1：
Test solution 2：
The selection of composition proportion in two mobile phase of embodiment
1. chromatographic column：5 μm of (2) 250 × 4.6mm of Diamonsil C18, column temperature：35 DEG C, flow velocity：1.0ml/min, detection
Wavelength：257nm, sample size：20μl.
2. prepared by solution
2.1 mobile phase：The mixed liquor of 72 volume phosphate buffers and 28 volumes methanols.
2.2 phosphate buffer：Phosphatase 11 1.5ml is taken, 1900ml is diluted with water to, with sodium hydroxide solution (1mol/L)
About 100ml tune pH to 2.0-3.0 adds sodium heptanesulfonate 1.1g, shaking to make dissolving.
2.3 test solution：Test sample 2, each accurate measurement 2ml is taken to set in 100ml volumetric flasks, be diluted with water respectively
It is shaken up to scale, as test solution 3 and 4.
2.4 reference substance solution：Precision weighs sorbic acid reference substance 15.28mg, 15.62mg, sets in 100ml measuring bottles, adds second
Alcohol makes dissolving in right amount, is diluted with water to scale, shakes up；The accurate 2ml that measures is set in 10ml volumetric flasks again, is diluted with water to scale,
It shakes up, as a contrast product solution 3 and 4.
3. assay method
Reference substance solution 3 is taken to inject liquid chromatograph, 5 needle of parallel sample introduction, 20 μ l of sample size.Spectrogram is recorded, sorb is calculated
Relative standard deviation RSD≤2.0% of sour peak area；Reference substance solution 4 is taken to inject liquid chromatograph, 2 needle of parallel sample introduction, sample introduction
Measure 20 μ l.Test solution 3 and 4 is taken to inject liquid chromatograph, 20 μ l of sample size record collection of illustrative plates.
The peak area of 3 continuous 5 needle of reference substance solution is：
A1=8004.1 A2=8007.8 A3=8005.2 A4=8006.0
A5=7999.6 ATo 1Average=8004.5 RSD=0.1%
The peak area of reference substance solution 4 is：
A1=8161.1 A2=8163.8 ATo 2Average=8162.4
4. sample measures
It takes test sample liquid to inject liquid chromatograph, records collection of illustrative plates, be respectively repeated once.
The main peak area of test solution 3：A1=7755.0 A2=7759.3 ASample 1Average=7757.2
The main peak area of test solution 4：A1=7751.9 A2=7756.3 ASample 2Average=7754.1
5. result of calculation
In formula：
AIt is right：Reference substance solution main peak area；
ASample：Test solution main peak area；
WIt is right：The weight of reference substance, mg；
P：The purity of reference substance, %.
Test solution 3：
Test solution 4：
The selection of three chromatographic column of embodiment
1. chromatographic column：5 μm of 250 × 4.6mm of Agilent zorbax SB-C18, column temperature：35 DEG C, flow velocity：1.0ml/
Min, Detection wavelength：257nm, sample size：20μl.
2. prepared by solution
2.1 mobile phase：The mixed liquor of 70 volume phosphate buffers and 30 volumes methanols.
2.2 phosphate buffer：Phosphatase 11 1.5ml is taken, 1900ml is diluted with water to, with sodium hydroxide solution (1mol/L)
About 100ml tune pH to 2.0-3.0 adds sodium heptanesulfonate 1.1g, shaking to make dissolving.
2.3 test solution：Test sample 2, each accurate measurement 2ml is taken to set in 100ml volumetric flasks, be diluted with water respectively
It is shaken up to scale, as test solution 5 and 6.
2.4 reference substance solution：Precision weighs sorbic acid reference substance 15.05mg, 14.48mg, sets in 100ml measuring bottles, adds second
Alcohol makes dissolving in right amount, is diluted with water to scale, shakes up；The accurate 2ml that measures is set in 10ml volumetric flasks again, is diluted with water to scale,
It shakes up, as a contrast product solution 5 and 6.
3. assay method
Reference substance solution 5 is taken to inject liquid chromatograph, 5 needle of parallel sample introduction, 20 μ l of sample size.Spectrogram is recorded, sorb is calculated
Relative standard deviation RSD≤2.0% of sour peak area；Reference substance solution 6 is taken to inject liquid chromatograph, 2 needle of parallel sample introduction, sample introduction
Measure 20 μ l.Test solution 5 and 6 is taken to inject liquid chromatograph, 20 μ l of sample size record collection of illustrative plates.
The peak area of 5 continuous 5 needle of reference substance solution is：
A1=7892.6 A2=7894.3 A3=7894.3 A4=7890.5
A5=7897.0 ATo 1Average=7893.7 RSD=0.1%
The peak area of reference substance solution 6 is：
A1=7536.4 A2=7534.1 ATo 2Average=7535.2
4. sample measures
It takes test sample liquid to inject liquid chromatograph, records collection of illustrative plates, be respectively repeated once.
The main peak area of test solution 5：A1=7751.9 A2=7758.8 ASample 1Average=7755.4
The main peak area of test solution 6：A1=7755.3 A2=7763.0 ASample 2Average=7759.2
5. result of calculation
In formula：
AIt is right：Reference substance solution main peak area；
ASample：Test solution main peak area；
WIt is right：The weight of reference substance, mg；
P：The purity of reference substance, %.
Test solution 5：
Test solution 6：
Note：All test samples are what Northeast Pharmaceutical Group Shenyang First Pharmaceutical Co., Ltd. produced in present specification
Levocarnitine oral solution, specification：10ml：1g, authentication code：Chinese medicines quasi-word H19990372；Lot number used：170508.
For the octadecylsilane chemically bonded silica that the application uses for the chromatographic column of filler, source is as follows：
Claims (8)
1. a kind of method of Determination of sorbic in detection levocarnitine oral solution, it is characterised in that：The detection method includes such as
Lower step：
(1) chromatographic condition：
Octadecylsilane chemically bonded silica chromatographic column；
UV detector wavelength：257nm；
Mobile phase：The mixed liquor of phosphate buffer and methanol；
(2) test solution is prepared：
(3) reference substance solution is prepared：
(4) assay method：
Precision measure reference substance solution inject liquid chromatograph, repeat 3-6 time, record chromatogram, sorbic acid peak area it is opposite
Standard deviation answers≤2.0%；Precision measures test solution and injects liquid chromatograph, chromatogram is recorded, by external standard method with peak face
Product calculate to get；
(5) calculation formula
In formula：
AIt is right：Reference substance solution main peak area；
ASample：Test solution main peak area；
WIt is right：The weight of reference substance, mg；
P：The purity of reference substance, %.
2. the method for Determination of sorbic, feature exist in a kind of detection levocarnitine oral solution according to claim 1
In in the mobile phase, the volume ratio of phosphate buffer and methanol is 65-75：25-35.
3. the method for Determination of sorbic, feature exist in a kind of detection levocarnitine oral solution according to claim 2
In in the mobile phase, the volume ratio of phosphate buffer and methanol is 70：30.
4. the method for Determination of sorbic, feature exist in a kind of detection levocarnitine oral solution according to claim 1
In the preparation method of the phosphate buffer is：Phosphatase 11 1.5ml is taken, 1900ml is diluted with water to, with the hydrogen-oxygen of 1mol/L
Change sodium solution tune pH to 2.0-3.0, sodium heptanesulfonate 1.1g, shaking is added to make dissolving.
5. the method for Determination of sorbic, feature exist in a kind of detection levocarnitine oral solution according to claim 1
In the octadecylsilane chemically bonded silica chromatographic column is selected from Agilent TC-C18 chromatographic columns, Agilent zorbax SB-
C18 chromatographic columns, Waters symmetry shield RP18 chromatographic columns, Diamonsil C18 chromatographic columns, Agilent
Eclipse XDB-C18 chromatographic columns, Agilent HC-C18 chromatographic columns, Agilent Extend C18 chromatographic columns, Ecosil
C18 chromatographic columns, II C18 chromatographic columns of SHISEIDO MG, SHISEIDO MG C18 chromatographic columns, SHISEIDO ODS C18 chromatographies
Column, Waters XTERRA MS C18 chromatographic columns, 18 chromatographic columns of Waters XTERRA RP, Waters Spherisorb
One kind in ODS2 chromatographic columns.
6. the method for Determination of sorbic, feature exist in a kind of detection levocarnitine oral solution according to claim 1
In the column temperature of the chromatographic column is 20-40 DEG C, and the flow velocity of the mobile phase is 0.5-2.0ml/min；The specification of the chromatographic column
For 250 × 4.6mm, 5 μm, 150 × 4.6mm, 5 μm, 150 × 4.6mm, 3.5 μm, 250 × 4.6mm, one kind in 3.5 μm.
7. the method for Determination of sorbic, feature exist in a kind of detection levocarnitine oral solution according to claim 1
In the method for preparing test solution is：Precision measures levocarnitine oral solution 2ml, sets in 100ml volumetric flasks, uses
Water is diluted to scale and shakes up, as test solution；It is described prepare reference substance solution method be：Precision weighs sorbic acid control
Product 15mg sets in 100ml measuring bottles, appropriate amount of ethanol is added to make dissolving, be diluted with water to scale, shakes up；The accurate 2ml that measures sets 10ml again
In volumetric flask, it is diluted with water to scale, is shaken up, as a contrast product solution.
8. the method for Determination of sorbic, feature exist in a kind of detection levocarnitine oral solution according to claim 1
In the sample size of the test solution and reference substance solution is 5-100 μ l, and preferred sample size is 5-20 μ l；The precision
It measures reference substance solution and injects liquid chromatograph, be repeated 5 times.
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| CN112305114A (en) * | 2020-10-29 | 2021-02-02 | 诺峰药业(成都)有限公司 | High performance liquid chromatography method for determining sorbic acid content in difluprednate eye emulsion |
| 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 |
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| CN112305114A (en) * | 2020-10-29 | 2021-02-02 | 诺峰药业(成都)有限公司 | High performance liquid chromatography method for determining sorbic acid content in difluprednate eye emulsion |
| 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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