阅读说明：本技术 一种盐酸左布比卡因注射液光学异构体的检验方法 (A kind of method of inspection of Levobupivacaine HCL injection optical isomer ) 是由 吕海亮 余丹 刘亚军 王晓翠 关东 谢斌 陈新民 兰柳琴 于 2019-08-30 设计创作，主要内容包括：本发明属于药物分析技术领域,涉及一种盐酸左布比卡因注射液光学异构体的检验方法,该方法利用方便快捷的高效液相色谱法,对盐酸左布比卡因注射液光学异构体进行检测,可用于监测盐酸左布比卡因注射液的质量,本发明所述的检测方法对盐酸左布比卡因注射液光学异构体的色谱峰分离度高、具有较高的系统适用性,同时在专属性、精密度、准确度方面都符合标准,具有专属性强、系统适用性良好、精密度良好、准确度良好、线性关系良好、耐用性良好等特点,同时本发明的测定方法操作简单,所用时间比较短,成本低,耗时短。(The invention belongs to Pharmaceutical Analysis technical fields, it is related to a kind of method of inspection of Levobupivacaine HCL injection optical isomer, this method utilizes conveniently high performance liquid chromatography, Levobupivacaine HCL injection optical isomer is detected, it can be used for monitoring the quality of Levobupivacaine HCL injection, detection method of the present invention is high to the chromatographic peak separating degree of Levobupivacaine HCL injection optical isomer, system suitability with higher, simultaneously in specificity, precision, it is all complied with standard in terms of accuracy, it is strong with specificity, system suitability is good, precision is good, accuracy is good, linear relationship is good, the features such as good tolerance, measuring method of the invention simultaneously is easy to operate, time used is shorter, it is at low cost, it is time-consuming short.)

1. a kind of method of inspection of Levobupivacaine HCL injection optical isomer, comprising the following steps:

(1) solution is prepared, it is molten to prepare blank solution, system suitability solution, test solution and contrast solution, the blank respectively Liquid is dilution；

(2) measuring method: after the system stabilizes, respectively by blank solution, system suitability solution, contrast solution, test solution Inject liquid chromatograph, record chromatogram, chromatographic condition is as follows: chromatographic column: amylose bonded silica gel is filler, flow velocity 1.0 ± 0.1mL/min, column temperature: 25 ± 5 DEG C, sample volume: 10-20 μ l, Detection wavelength: 215nm, mobile phase are diethylamine: acetonitrile System.

2. according to the method described in claim 1, it is characterized by: the dilution is water；

The preparation steps of the system suitability solution are as follows: weigh appropriate hydrochloric acid Bupivacaine reference substance, be placed in volumetric flask, add Dilution dissolves and is diluted to scale, shakes up；

The test solution: pipetting appropriate hydrochloric acid chirocaine injection, be placed in volumetric flask, adds diluted to quarter Degree, shakes up；

The contrast solution: appropriate test solution is pipetted, is placed in volumetric flask, is added dilution to dissolve and be diluted to scale, shake up；

The mobile phase are as follows:

Mobile phase A: volumetric concentration is 0.1% diethylamine solution, i.e., volumetric concentration is 0.1%DEA；

Mobile phase B: acetonitrile；

The diethylamine is HPLC grades；

The acetonitrile is HPLC grades；

The water is HPLC grades；

The chromatographic column be Chiral INA250 × 4.6mm, 5 μm.

3. method according to claim 1 or 2, which is characterized in that the mobile phase is that volumetric concentration is 0.1%DEA- second Nitrile=40:60.

Technical field

The present invention relates to a kind of methods of inspection of Levobupivacaine HCL injection optical isomer, belong to Pharmaceutical Analysis skill Art field.

Background technique

Levobupivacaine HCL, English name: Levobupivacaine Hydrochloride, chemical name are as follows: S- (-)- 1- butyl-N- (2,6- 3,5-dimethylphenyl) -2- piperidine formyl amine hydrochlorate, molecular formula are as follows: C₁₈H₂₈N₂OHCl, chemical structural formula It is as follows:, Levobupivacaine HCL is amides The laevoisomer of local anesthetic Bupivacaine, is mainly used for surgery Epidural Block, and chirocaine has lower Nerve and Cardiovascular Toxicity, but the myocardiac inhibition effect of its dextroisomer is strong, therefore controls in Levobupivacaine HCL Dextroisomer is extremely important to the quality control of drug, and the dextroisomer structural formula II of Bupivacaine is as follows:。

Detection method in " Chinese Pharmacopoeia " first enlarged edition of version in 2015 about Levobupivacaine HCL optical isomer In, with 0.02mol/L phosphate buffer solution, (potassium dihydrogen phosphate 2.72g adds water 800ml to make to dissolve, with 0.1mol/L hydroxide It is mobile phase that sodium solution, which adjusts pH value to 7.0)-isopropanol (90:10), and α 1- acid glycoprotein bonded silica gel filler is chromatographic column, Detection wavelength is 215nm, with the optical isomer in this method detection Levobupivacaine HCL, the right Bupivacaine of institute's sample Fail to reach baseline separation with chirocaine, the separating degree not met between right Bupivacaine and chirocaine chromatographic peak is answered >=1.5 requirement.

Our company independent research a kind of method of inspection of Levobupivacaine HCL injection optical isomer, this method disclose A kind of method of inspection of Levobupivacaine HCL injection optical isomer, the right Bupivacaine of this method institute sample and left Bupivacaine separating degree is greater than 1.5, and separating degree is good, while in system suitability, repeatability, stability, precision and accurate Degree aspect meets the guideline of Chinese Pharmacopoeia method validation.

Summary of the invention

The object of the present invention is to provide a kind of method of inspection of Levobupivacaine HCL injection optical isomer, this method It is convenient, efficient, accurate, Chinese Pharmacopoeia method validation is complied fully in terms of system suitability, repeatability, specificity, accuracy Guideline, can be used for Levobupivacaine HCL injection quality control.

To achieve the above object, the invention provides the following technical scheme:

A kind of method of inspection of Levobupivacaine HCL injection optical isomer, comprising the following steps:

(1) solution is prepared, it is molten to prepare blank solution, system suitability solution, test solution and contrast solution, the blank respectively Liquid is dilution.

(2) measuring method: after the system stabilizes, respectively by blank solution, system suitability solution, contrast solution, test Solution injects liquid chromatograph, records chromatogram, and chromatographic condition is as follows: chromatographic column: amylose bonded silica gel is filler, stream 1.0 ± 0.1mL/min of speed, column temperature: 25 ± 5 DEG C, sample volume: 10-20 μ l, Detection wavelength: 215nm, mobile phase are diethylamine: second Nitrile system.

Preferably, the dilution is water；

The preparation steps of the system suitability solution are as follows: weigh appropriate hydrochloric acid Bupivacaine reference substance, be placed in volumetric flask, add Dilution dissolves and is diluted to scale, shakes up；

The test solution: pipetting appropriate hydrochloric acid chirocaine injection, be placed in volumetric flask, adds diluted to quarter Degree, shakes up；

The contrast solution: appropriate test solution is pipetted, is placed in volumetric flask, is added dilution to dissolve and be diluted to scale, shake up；

The mobile phase are as follows:

Mobile phase A: volumetric concentration is 0.1% diethylamine solution；

Mobile phase B: acetonitrile；

The diethylamine is HPLC grades；

The acetonitrile is HPLC grades；

The water is HPLC grades；

The chromatographic column be 250 × 4.6mm of Chiral INA, 5 μm.

It is furthermore preferred that measuring method of the present invention, comprising the following steps:

(1) solution is prepared, prepares blank solution, system suitability solution, test solution and contrast solution respectively；The blank is molten Liquid is dilution, and the dilution is water；

System suitability solution: taking bupivacaine HCl reference substance 10mg, accurately weighed, sets in 100ml measuring bottle, adds dilution molten Scale is solved and be diluted to, is shaken up；Precision measures above-mentioned solution 2ml, sets in 20ml measuring bottle, adds diluted to scale, shake up； (concentration: 0.01mg/ml)

Test solution: precision measures Levobupivacaine HCL injection 2ml and sets in 100ml measuring bottle, adds diluted to scale, It shakes up；(concentration: 0.1mg/ml)

Contrast solution: precision measures test solution 1ml and sets in 200ml measuring bottle, adds diluted to scale, shakes up；(concentration: 0.5 μ g/ml)

(2) measuring method: after the system stabilizes, respectively by blank solution, system suitability solution, contrast solution, test solution Inject liquid chromatograph, record chromatogram, chromatographic condition is as follows: chromatographic column: amylose bonded silica gel is filler, flow velocity 1.0mL/min, column temperature: 25 DEG C, sample volume: 20 μ l, Detection wavelength: 215nm, mobile phase are that volumetric concentration is 0.1%DEA- acetonitrile (40:60)；

The chromatographic column be 250 × 4.6mm of Chiral INA, 5 μm；.

The method of inspection of the Levobupivacaine HCL injection optical isomer further includes method before testing Verifying, this method are verified as according to the chromatographic condition formally detected, measurement result are as follows:

Beneficial effect

From the above technical scheme, detection method of the present invention is to Levobupivacaine HCL injection optical isomer Chromatographic peak separating degree height, system suitability with higher, comply with standard in terms of repeatability.The present invention is using conveniently High performance liquid chromatography detects Levobupivacaine HCL injection optical isomer, can be used for monitoring the left cloth ratio of hydrochloric acid The quality of cacaine injection.The detection method of Levobupivacaine HCL injection optical isomer proposed by the present invention has special Attribute is strong, system suitability is good, precision is good, accuracy is good, linear relationship is good, good tolerance, is suitable for salt Sour chirocaine injection optical isomer detection, effectively to control the quality of product, while measuring method behaviour of the invention To make simply, the time used is shorter, and it is at low cost, it is time-consuming short.

Detailed description of the invention

Fig. 1 is the detection in " Chinese Pharmacopoeia " first enlarged edition of version in 2015 about Levobupivacaine HCL optical isomer The liquid chromatogram of method blank solution；

Fig. 2 is the detection method in " Chinese Pharmacopoeia " first enlarged edition of version in 2015 about Levobupivacaine HCL optical isomer The liquid chromatogram of system suitability solution；

Fig. 3 is the detection method in " Chinese Pharmacopoeia " first enlarged edition of version in 2015 about Levobupivacaine HCL optical isomer Test the liquid chromatogram of solution；

Fig. 4 is the liquid chromatogram of the detection method blank solution of Levobupivacaine HCL optical isomer；

Fig. 5 is the liquid chromatogram of the detection method system suitability solution of Levobupivacaine HCL optical isomer；

Fig. 6 is that the detection method of Levobupivacaine HCL optical isomer tests the liquid chromatogram of solution；

Fig. 7 is Levobupivacaine HCL linear relationship chart.

Specific embodiment

The present invention is explained further and illustrated with specific embodiment below, but do not limit the invention in any way Range.

Comparative test 1

(1) in " Chinese Pharmacopoeia " first enlarged edition of version in 2015 about the detection method of Levobupivacaine HCL optical isomer with The comparison of the application method

(2) injection procedure:

After the system stabilizes, sample introduction blank solution, system suitability solution, contrast solution, test each 1 needle of solution, record chromatography Figure, it is desirable that: the separating degree in chromatogram obtained by system suitability solution between right Bupivacaine and chirocaine chromatographic peak is answered ≥1.5。

It calculates: I%=×0.5

Wherein I% is optical isomer content

As is right Bupivacaine peak area in test solution

Ar is chirocaine peak area in contrast solution

As a result judge: should must not cross 0.5% containing right Bupivacaine in sample.

(3) testing result: test map as shown in figures 1 to 6, as a result as shown in the table

(4) conclusion

In the system suitability solution of method 1 and method 2, the separating degree of right Bupivacaine and chirocaine is not much different, Greater than 1.5.Method 1 is tested in solution, and right Bupivacaine and chirocaine fail to reach baseline separation, and method 2 tests solution In, right Bupivacaine and chirocaine separating degree are greater than 1.5, and separating degree is good.

Since method 1 is tested in solution, right Bupivacaine and chirocaine fail to reach baseline separation, even if being based on wind The maximum integral parameter in danger also fails to the right Bupivacaine content of actual response.

To sum up, the optical isomer of selecting method 2, i.e. the application method for Levobupivacaine HCL injection measures, The quality of product can more effectively be controlled.