CN103235072A

CN103235072A - Method for determining content of artemether in artemether injection with high performance liquid chromatography

Info

Publication number: CN103235072A
Application number: CN2013101759734A
Authority: CN
Inventors: 张梅; 彭学东; 赵金召; 祁亚楠
Original assignee: ZHANGJIAGANG WEISHENG BIOLOGICAL PHARMACEUTICAL CO Ltd
Current assignee: ZHANGJIAGANG WEISHENG BIOLOGICAL PHARMACEUTICAL CO Ltd
Priority date: 2013-05-14
Filing date: 2013-05-14
Publication date: 2013-08-07

Abstract

The invention discloses a method for determining the content of artemether in an artemether injection with high performance liquid chromatography. The method is characterized in that (a) a chromatographic column adopts octadecyl silane as a filler, a mobile phase is acetonitrile-water (75:25), the flow rate is 1.0ml/min, the column temperature is 40 DEG C, and the detection wavelength is 216nm; (b) during solution preparation, 2ml of artemether injection is measured precisely and added into a 100ml of volumetric flask, an appropriate amount of methanol and trichloromethane (9:1) mixed solvent is added and subjected to ultrasonic treatment till a solution is clear, and the methanol and trichloromethane (9:1) mixed solvent is added till the solution reaches the scale, and an artemether standard product is prepared by the same method; and (c) during determination: the content of artemether in the artemether injection is calculated with a peak area according to an external standard method. The invention provides a stable and reliable method for determining the content of artemether in the artemether injection with the high performance liquid chromatography. The method is easy and simple to operate, precise and reliable in determination result, high in specificity, and short in detection time, and the precise and reliable in content detection method is provided for quality control of the artemether injection.

Description

The method of Artemether content in a kind of high effective liquid chromatography for measuring Artemether parenteral solution

Technical field

The invention belongs to the pharmaceutical preparation analysis field, be specifically related to the method for Artemether content in a kind of high effective liquid chromatography for measuring Artemether parenteral solution.

Background technology

Artemether is a kind of efficient, quick-acting for anti-chloroquine malignant malaria and pernicious malaria medicine.Molecular formula is C ₁₆H ₂₆O ₅, molecular weight: 298.37; Artemether is white crystals or crystalline powder, odorless, mildly bitter flavor.Very easily dissolving is easily held in ethanol or ethyl acetate in acetone or methenyl choloride, and is almost insoluble in water.Its structural formula is seen accompanying drawing 1.

Research (Wang Yuhe according to existing Artemether injection manufacture craft, Artemether injection and preparation process thereof), the Artemether injection be use the hybrid injection formed by oil refining in the exquisiteness, tea oil and middle refining triglyceride with oil add Artemether after filtration, flow processs such as embedding, logical nitrogen, sterilization make.Owing to contain compositions such as tea oil in the injection composition, conventional Artemether compound method is inapplicable in the Artemether assay in the Artemether injection, Chinese Pharmacopoeia 2010 editions is not set up the coherent detection standard to the Artemether parenteral solution as yet, domestic research for this respect also seldom, Yao waits the (content of HPLC method mensuration Artemether parenteral solution quietly, Yao Jing, Cheng Qilei, Zhang Qiming etc., the Pharmaceutical Analysis magazine, 2008,28 (1): 91～92) once openly reported Artemether assay in a kind of HPLC method mensuration Artemether parenteral solution, and adopted C18 post (150mm * 4.6mm, 5um), flow and select acetonitrile-water (70: 30) mutually for use, flow velocity 1.0ml/min, sample dissolution is selected the absolute ethyl alcohol ultrasonic dissolution for use; This method selects for use absolute ethyl alcohol to carry out sample preparation, needs the ultrasonic processing of long period, and the sample preparation is more time-consuming, and under the normal temperature condition in the parenteral solution tea oil etc. separate out easily, testing result is unstable relatively; Establishing a kind of high performance liquid chromatography content more accurate, that measure Artemether in the Artemether parenteral solution reliably has great importance to the quality control of Artemether parenteral solution.

Summary of the invention

The objective of the invention is to set up the method for Artemether content in a kind of high effective liquid chromatography for measuring Artemether parenteral solution, specifically implement by following technical method:

(1) chromatographic condition:

It is filling material that chromatographic column is selected for use with the octadecyl silane, the present invention selects for use Thermo C18 post (4.6 * 250mm5 μ m), Thermo C18 post (4.6 * 150mm5 μ m) and Thermo C18 post (4.6 * 100mm5 μ m) to test respectively, preferred Thermo C18 post (4.6 * 250mm5 μ m) has separating effect preferably.

Flowing is acetonitrile-water mutually, and the acetonitrile ratio is 60%～80%, preferred 75%.

Flow velocity 1.0ml/min, 30～40 ℃ of column temperatures, preferred 40 ℃.

The present invention uses the Agilent-1260 high performance liquid chromatograph, and UV-detector detects wavelength 216nm.

(2) formulations prepared from solutions: precision is measured 2ml Artemether parenteral solution to the 100ml volumetric flask, and it is ultrasonic to the solution clarification to add an amount of methyl alcohol and methenyl choloride (9: 1) mixed solvent, uses methyl alcohol and methenyl choloride (9: 1) mixed solvent to be settled to scale mark again.Formulations prepared from solutions of the present invention also selects for use absolute ethyl alcohol to test, and testing sample solution was separated out easily when parenteral solution concentration was big slightly, and the stability of solution of preparation is poor, so the present invention selects methyl alcohol and methenyl choloride (9: 1) mixed solvent sample dissolution for use.The standard solution compound concentration is consistent with need testing solution, all uses methyl alcohol and methenyl choloride (9: 1) mixed solvent sample dissolution.

(3) determination method: measure Artemether standard items and need testing solution 20 μ l respectively, auto injection is measured, and presses external standard method with Artemether content in the calculated by peak area Artemether parenteral solution.

By above chromatographic condition, precision is measured 20 μ l Artemether standard solution and solution to be measured, inject liquid chromatograph respectively, the peak-to-peak degree of separation of Artemether peak and impurity is 7.25, and the main peak symmetrical factor is 1.03, and number of theoretical plate is calculated as 15000 by the Artemether peak, Artemether peak symmetrical factor is 1.05 in the injection liquid samples solution, theoretical cam curve is greater than 10000, and the minimum separation degree is 2.7 between any two adjacent peaks, meets the HPLC method and measures requirement.

Description of drawings

Fig. 1 is the Artemether structural formula

Fig. 2 is the efficient liquid phase collection of illustrative plates of Artemether standard items

Fig. 3 is the efficient liquid phase collection of illustrative plates of Artemether parenteral solution

Fig. 4 is Artemether standard items sampling volume and peak area linear relationship

Embodiment

Embodiment 1

Precision is measured Artemether parenteral solution (from the Pacific Ocean, Tianjin medicine company limited) 2ml and is placed the 100ml volumetric flask, it is ultrasonic (ultrasonic at twice to add an amount of methyl alcohol and methenyl choloride (9: 1) mixed solvent, about 15min altogether, 50Hz,) dissolve fully to grease, left standstill 5 minutes, and added methyl alcohol and methenyl choloride (9: 1) mixed solvent again and be settled to scale.Solution filters through 0.45 μ m aperture filter and namely gets need testing solution; 5 samples of need testing solution balance preparation.Precision weighing 40mg Artemether standard items place the 25ml volumetric flask, add the dissolving of methyl alcohol and methenyl choloride (9: 1) mixed solvent and are settled to scale with methyl alcohol and methenyl choloride (9: 1) mixed solvent, filter through 0.45 μ m aperture filter and namely get standard solution; 5 samples of standard solution preparation.

Chromatographic condition: select for use Sai Mo to fly the generation that C18 of company post (4.6 * 250mm5 μ m); Flowing is acetonitrile-water (75: 25) mutually, flow velocity 1.0ml/min; 40 ℃ of column temperatures; Detect wavelength 216m; Sampling volume 20 μ l.

By above-mentioned chromatographic condition, precision is measured each 5 of 20 μ l standard items and test samples respectively, sample introduction respectively, and the gained peak area value is as shown in table 1:

Table 1 standard items and test sample peak area

5 balance samples of standard solution sample introduction gained peak area RSD value respectively are 0.08%; Calculate Artemether content in the Artemether parenteral solution by external standard method, it is that to measure Artemether content RSD value 95.44%, 5 time be 0.17% that the need testing solution duplicate measurements records in the Artemether parenteral solution Artemether average content for 5 times.

Embodiment 2: methodological study

Artemether standard specimen continuous sample introduction 5 times, peak area RSD value is 0.17%, precision is good.The Artemether standard items are mixed with 1.0mg/ml solution by the inventive method, measured respectively at 0 o'clock, 2 o'clock, 4 o'clock, 8 o'clock, 12 o'clock, 18 o'clock, 24 o'clock, peak area RSD value is 0.58%, illustrates that the Artemether standard specimen was stable at least 24 hours, can be used for assay.By the need testing solution that same concentrations is made, respectively at 0 o'clock, 2 o'clock, 4 o'clock, 8 o'clock, 12 o'clock, 18 o'clock, 24 o'clock mensuration, peak area RSD value was 1.02%, illustrates that the Artemether parenteral solution was stable at least 24 hours, can be used for assay.

The accurate Artemether standard items that claim are made 1.0mg/ml solution by the inventive method, respectively sample introduction 1 μ l, 2 μ, 5 μ l, 10 μ l, 20 μ l, 30 μ l, 50 μ l, 70 μ l, 90 μ l, 100 μ l, result show that sample size has good linear relationship with peak area in 1 μ g～100 μ g scopes, and linear equation is y=34.928x+9.1303 (R ²=0.9999), sample size and peak area typical curve are seen Fig. 4.

Claims

1. the method for Artemether content in the high effective liquid chromatography for measuring Artemether parenteral solution is characterized in that:

(1) chromatographic condition: it is filling material that chromatographic column is selected for use with the octadecyl silane, and flowing is acetonitrile-water (75: 25) mutually, flow velocity 1.0ml/min, and 40 ℃ of column temperatures detect wavelength 216nm;

(2) formulations prepared from solutions: precision is measured 2ml Artemether parenteral solution to the 100ml volumetric flask, and it is ultrasonic to the solution clarification to add an amount of methyl alcohol and methenyl choloride (9: 1) mixed solvent, uses methyl alcohol and methenyl choloride (9: 1) mixed solvent to be settled to scale mark again;

2. according to Artemether content assaying method in the Artemether parenteral solution described in the claim 1, it is characterized in that chromatographic column selects C18 post (4.6 * 250mm5 μ m) for use, separating effect is better.

3. according to Artemether content assaying method in the Artemether parenteral solution described in the claim 1, ultrasonic 10min earlier when it is characterized in that preparing the need testing solution ultrasonic dissolution, leave standstill 5min more ultrasonic to the parenteral solution tea oil dissolve the solution clarification fully.

4. according to Artemether content assaying method in the Artemether parenteral solution described in the claim 1, it is characterized in that by external standard method with peak area when Artemether carries out assay in the Artemether parenteral solution, the need testing solution duplicate measurements records that Artemether content RSD value is 0.17% in the Artemether parenteral solution for 5 times.