技术领域technical field
本发明属于药品成分检测技术领域，尤其涉及一种滴眼液中苯扎氯铵的快速检测方法。The invention belongs to the technical field of drug component detection, in particular to a rapid detection method for benzalkonium chloride in eye drops.
背景技术Background technique
为防止眼用制剂在贮存及使用过程中受微生物污染，大部分滴眼液尤其多剂量滴眼液常加入抑菌剂。苯扎氯铵(BAC)因其低浓度时具有良好的杀菌效果、毒性小等特点，被作为滴眼液中常用的抑菌剂。目前苯扎氯铵的检测方法主要为高效液相色谱法，高效液相色谱法能有效测定苯扎氯铵含量，但其色谱峰易拖尾，柱效难以满足需求，分析样本时间长，一个样本的分析时间达20分钟，甚至更久，对于大批量样本的分析操作，流动相等试剂损耗大，分析成本高，操作强度大。另外，苯扎氯铵由3种正烷烃基取代基二甲基苄基氯化铵的同系物组成，会因对照品与滴眼液产品中同系物组成不一致对分析检测造成困扰。此外质谱分析法也用于苯扎氯铵含量测定，但其所用仪器昂贵，普及度小，且会破坏分析样品。In order to prevent ophthalmic preparations from being contaminated by microorganisms during storage and use, most eye drops, especially multi-dose eye drops, are often added with antibacterial agents. Benzalkonium chloride (BAC) is commonly used as a bacteriostatic agent in eye drops because of its good bactericidal effect and low toxicity at low concentrations. At present, the detection method of benzalkonium chloride is mainly high performance liquid chromatography. High performance liquid chromatography can effectively measure the content of benzalkonium chloride, but its chromatographic peak is easy to tail, and the column efficiency is difficult to meet the demand. It takes a long time to analyze the sample. The analysis time of the sample is 20 minutes or even longer. For the analysis operation of a large number of samples, the loss of flow equal reagents is large, the analysis cost is high, and the operation intensity is high. In addition, benzalkonium chloride is composed of three homologues of n-alkane substituent dimethyl benzyl ammonium chloride, which will cause problems for analysis and detection due to the inconsistency in the composition of the homologues in the reference substance and eye drop products. In addition, mass spectrometry is also used to determine the content of benzalkonium chloride, but the instruments used are expensive, the popularity is small, and the analysis samples will be destroyed.
近年来，化学计量学方法的快速发展给复杂样品中感兴趣物质分析带来新的生机，但用于分析滴眼液中抑菌组分的化学计量学方法非常缺乏。其中空间夹角判据作为一种新的化学计量学方法，因其具有所用样品数量少、预处理简单、方法简便、快速等优点得到较多应用。In recent years, the rapid development of chemometric methods has brought new vitality to the analysis of substances of interest in complex samples, but there is a lack of chemometric methods for the analysis of antibacterial components in eye drops. Among them, the spatial angle criterion, as a new chemometric method, has been widely used because of its advantages such as small number of samples used, simple pretreatment, simple and fast method, etc.
发明内容Contents of the invention
本发明的目的在于提供一种滴眼液中苯扎氯铵的快速检测方法，旨在解决传统检测技术存在的分析效率低、操作强度大以及分析成本高等不足，以及待测组分纯光谱信号难以获取的问题。The purpose of the present invention is to provide a rapid detection method for benzalkonium chloride in eye drops, aiming to solve the problems of low analysis efficiency, high operation intensity and high analysis cost of traditional detection technology, and the pure spectral signal of the components to be measured. Difficult to obtain problem.
本发明是这样实现的，一种滴眼液中苯扎氯铵的快速检测方法，所述快速检测方法通过斜投影的多变量体系中纯信号提取方法将待测组分十二烷基二甲基苄基氯化铵从苯扎氯铵混合物中提取；将提取的十二烷基二甲基苄基氯化铵纯信号与对应浓度值进行最小二乘拟合建立标准光谱数据库；然后采用空间夹角判据求得滴眼液样本中待测组分的含量。本发明中苯扎氯铵含量按十二烷基二甲基苄基氯化铵记。The present invention is achieved in this way, a rapid detection method of benzalkonium chloride in eye drops, said rapid detection method extracts the component to be measured, dodecyl dimethyl The benzyl benzyl ammonium chloride was extracted from the benzalkonium chloride mixture; the extracted dodecyl dimethyl benzyl ammonium chloride pure signal and the corresponding concentration value were subjected to least squares fitting to establish a standard spectral database; and then the spatial The included angle criterion was used to obtain the content of the component to be tested in the eye drop sample. In the present invention, the benzalkonium chloride content is recorded by dodecyl dimethyl benzyl ammonium chloride.
具体包括以下步骤：Specifically include the following steps:
第一步，苯扎氯铵溶液配制：The first step, preparation of benzalkonium chloride solution:
移取12.02mg/mL的苯扎氯铵，用二次蒸馏水定容，配制系列浓度为1～20μg/mL的苯扎氯铵标准溶液；Pipette 12.02 mg/mL of benzalkonium chloride, dilute to volume with twice distilled water, and prepare a series of standard solutions of benzalkonium chloride with a concentration of 1-20 μg/mL;
第二步，苯扎氯铵本底光谱数据库的建立：Second step, the establishment of benzalkonium chloride background spectrum database:
将配制的苯扎氯铵标准溶液过滤后经高效液相色谱分析，经紫外光谱仪采集系列苯扎氯铵标准溶液的多波长三维数据；采集所得的苯扎氯铵的多波长三维数据的光强度数据值转化成吸光度值；根据苯扎氯铵混合物中十二烷基二甲基苄基氯化铵、十四烷基二甲基苄基氯化铵的物质特性确定待测组分十二烷基二甲基苄基氯化铵的保留时间，根据苯扎氯铵色谱图，从苯扎氯铵标准溶液三维数据中扣除待测组分的光谱数据，将余光谱数据加和作为苯扎氯铵本底光谱数据库H；将所得的本底光谱数据库H进行奇异值分解，根据体系主成分数目将本底光谱数据库H进行重构，得到维数适当的本底光谱数据库H′；After the prepared benzalkonium chloride standard solution is filtered, it is analyzed by high performance liquid chromatography, and the multi-wavelength three-dimensional data of a series of benzalkonium chloride standard solutions are collected by an ultraviolet spectrometer; the light intensity of the multi-wavelength three-dimensional data of the collected benzalkonium chloride The data value is converted into an absorbance value; according to the material properties of dodecyl dimethyl benzyl ammonium chloride and tetradecyl dimethyl benzyl ammonium chloride in the benzalkonium chloride mixture, determine the component dodecyl According to the retention time of dimethyl benzyl ammonium chloride, according to the benzalkonium chloride chromatogram, the spectral data of the component to be measured is deducted from the three-dimensional data of the benzalkonium chloride standard solution, and the remaining spectral data are added as benzalkonium chloride Ammonium background spectrum database H; the obtained background spectrum database H is subjected to singular value decomposition, and the background spectrum database H is reconstructed according to the number of principal components of the system to obtain a background spectrum database H' with appropriate dimensions;
第三步，根据苯扎氯铵的色谱图找到待测组分的最高响应值对应的时间点，扣出待测组分光谱信号作为待测组分纯信号S；In the third step, according to the chromatogram of benzalkonium chloride, the time point corresponding to the highest response value of the component to be tested is found, and the spectral signal of the component to be tested is deducted as the pure signal S of the component to be tested;
第四步，采用光纤光谱仪对第一步制得的苯扎氯铵标准溶液进行光谱扫描，得到苯扎氯铵标准溶液的紫外光谱数据矩阵y；The 4th step, adopt fiber optic spectrometer to carry out spectral scanning to the benzalkonium chloride standard solution that the first step makes, obtain the ultraviolet spectral data matrix y of benzalkonium chloride standard solution;
第五步，将所得的苯扎氯铵本底光谱数据库H′、待测组分纯分析信号S和苯扎氯铵标准溶液紫外光谱数据矩阵y导入计算平台，应用基于斜投影的多变量体系中纯信号提取方法将待测组分十二烷基二甲基苄基氯化铵从苯扎氯铵混合物中提取，得到1～20μg/mL的系列浓度的稳定的待测组分纯光谱信号矩阵S′；The fifth step is to import the obtained benzalkonium chloride background spectrum database H', the pure analysis signal S of the component to be measured and the ultraviolet spectrum data matrix y of the standard solution of benzalkonium chloride into the computing platform, and apply the multivariable system based on oblique projection The medium-pure signal extraction method extracts the component to be tested, dodecyldimethylbenzyl ammonium chloride, from the benzalkonium chloride mixture, and obtains a stable pure spectral signal of the component to be tested with a series concentration of 1 to 20 μg/mL matrix S';
第六步，将得到的待测组分纯光谱信号与其对应的浓度值进行最小二乘拟合，得到单位浓度的十二烷基二甲基苄基氯化铵光谱数据，存入标准光谱数据库V；所得到的标准光谱库V可用于下一步计算。The sixth step is to perform least squares fitting on the pure spectral signal of the component to be measured and its corresponding concentration value to obtain the spectral data of dodecyl dimethyl benzyl ammonium chloride per unit concentration, and store it in the standard spectral database V; the obtained standard spectral library V can be used for the next calculation.
进一步，所述基于斜投影的多变量体系中纯信号提取方法具体包括：Further, the pure signal extraction method in the oblique projection-based multivariate system specifically includes:
步骤一，将求得的H′、S导入计算平台，求取纯信号分离模型，即斜投影算子，表达式为：Step 1. Import the obtained H′ and S into the computing platform to obtain the pure signal separation model, that is, the oblique projection operator. The expression is:
ESH＝S(STPH⊥S)-1STPH⊥，其中PH⊥＝I-PH＝I-H(HTH)-1HT；ESH = S(ST PH⊥ S)-1 ST PH⊥ , where PH⊥ = IPH = IH(HT H)-1 HT ;
在上式中，上标T表示矩阵的转置，I为与PH维数相同的单位矩阵；In the above formula, the superscript T represents the transposition of the matrix, and I is the identity matrix with the same dimension asPH ;
步骤二，将中得到y与斜投影算子ES|H相乘，即得到1～20μg/mL的系列浓度的十二烷基二甲基苄基氯化铵纯光谱信号S′。Step 2: Multiply the obtained y by the oblique projection operator ES|H to obtain the pure spectral signal S' of dodecyldimethylbenzylammonium chloride with a series concentration of 1-20 μg/mL.
进一步，所述建立标准光谱数据库之后需要通过液相-紫外联用构建样本本底光谱数据库；采用奇异值分解结合二阶差分法选取体系主成分数对本底数据库降维，得到数据量较小的本底光谱数据库；获取滴眼液样本紫外光谱数据。Further, after the establishment of the standard spectral database, it is necessary to construct the sample background spectral database through liquid phase-ultraviolet coupling; use the singular value decomposition combined with the second-order difference method to select the principal components of the system to reduce the dimension of the background database, and obtain a small amount of data. Background spectrum database; obtain the UV spectrum data of eye drop samples.
进一步，进一步包括以下步骤：Further, further comprising the following steps:
步骤一，移取滴眼液样本至10mL的容量瓶内，用二次蒸馏水稀释适当的倍数，制得滴眼液稀释液；Step 1, pipette the eye drop sample into a 10mL volumetric flask, and dilute it by an appropriate multiple with double distilled water to prepare the eye drop dilution;
步骤二，滴眼液样本本底光谱数据库的建立：Step 2, the establishment of the background spectrum database of eye drops samples:
步骤三，采用光纤光谱仪对制得的滴眼液稀释液进行光谱扫描，得到滴眼液稀释液的紫外光谱数据矩阵a。In step 3, the optical fiber spectrometer is used to perform spectral scanning on the prepared eye drop dilution to obtain the ultraviolet spectrum data matrix a of the eye drop dilution.
进一步，所述滴眼液样本本底光谱数据库的建立具体包括：Further, the establishment of the background spectrum database of the eye drop samples specifically includes:
第一步，获取滴眼液样本的液相-紫外联用数据，将不经稀释的滴眼液样本过滤后经高效液相色谱分析，经紫外光谱仪采集滴眼液样本的多波长三维数据。The first step is to obtain the liquid-ultraviolet data of the eye drop sample, filter the undiluted eye drop sample, analyze it by high-performance liquid chromatography, and collect the multi-wavelength three-dimensional data of the eye drop sample by ultraviolet spectrometer.
第二步，将采集所得的滴眼液样本多波长三维数据的光强度数据值转化成吸光度值，转化后的数据的每一行代表一个波长下的色谱数据，每一列代表一个时刻下洗脱出来物质的光谱数据；In the second step, the light intensity data value of the multi-wavelength three-dimensional data of the collected eye drop sample is converted into an absorbance value. Each row of the converted data represents the chromatographic data at one wavelength, and each column represents the elution at one time. Spectral data of the substance;
第三步，获取不含待测组分的本底光谱数据，根据确定的待测组分十二烷基二甲基苄基氯化铵的保留时间，从滴眼液样本三维数据中扣除待测组分的光谱数据，将其余光谱数据加和作为滴眼液本底光谱数据库H1；The third step is to obtain the background spectral data not containing the component to be measured, and to deduct the value to be tested from the three-dimensional data of the eye drop sample according to the retention time of the determined component to be tested, dodecyldimethylbenzyl ammonium chloride. Measure the spectral data of the components, and add the rest of the spectral data as the eye drops background spectral database H1;
第四步，本底光谱数据库的降维，将所得的本底光谱数据库H1进行奇异值分解，根据体系主成分数目将本底光谱数据库H1进行重构，得到维数适当的本底光谱数据库H1′。The fourth step is to reduce the dimensionality of the background spectrum database.The obtained background spectrum database H1 is subjected to singular value decomposition, and the background spectrum database H1 is reconstructed according to the number of principal components of the system to obtaina background spectrum with an appropriate dimension Database H1 '.
进一步，所述本地光谱数据库的降维中，体系主成分数目的确定方法为：将矩阵H、H1进行方差标准化后，添加白噪声掩蔽不均匀噪声和非线性因素的干扰，以二阶差分值序列转折点判断其独立变量数目，得到体系主成分数目N、N1；Further, in the dimensionality reduction of the local spectral database, the method for determining the number of principal components of the system is as follows: After standardizing the varianceof the matrices H and H1, adding white noise to mask the interference of uneven noise and nonlinear factors, and using second-order difference Judging the number of independent variables at the turning point of the value sequence, and obtaining the number of principal components of the system N, N1 ;
将本底光谱数据库H、H1进行重构的方法如下：The methodof reconstructing the background spectrum database H, H1 is as follows:
应用奇异值分解程序[US1V1]＝svd(H)对苯扎氯铵本底光谱数据库H进行奇异值分解降维，分解后得到m阶行正交矩阵U，n阶列正交矩阵V1和矩阵H的特征值矩阵S1，取U的前N列即为降维后的苯扎氯铵本底光谱数据库H′。Apply the singular value decomposition program [US1 V1 ]=svd(H) to carry out singular value decomposition and dimensionality reduction on the background spectrum database H of benzalkonium chloride, and obtain m-order row orthogonal matrix U and n-order column orthogonal matrix after decomposition V1 and the eigenvalue matrix S1 of the matrix H, taking the first N columns of U is the reduced-dimensional benzalkonium chloride background spectrum database H′.
同法得到H1′。H1 ′ can be obtained in the same way.
进一步，所述获取滴眼液样本紫外光谱数据之后需要对滴眼液样本中十二烷基二甲基苄基氯化铵含量测定，分别将所得的待测组分标准光谱数据库V、滴眼液本底光谱数据库H1′和滴眼液样本紫外光谱数据矩阵a导入计算平台，应用空间夹角判据从滴眼液样本稀释液光谱中扣减待测组分十二烷基二甲基苄基氯化铵的光谱，得出滴眼液稀释液中待测组分十二烷基二甲基苄基氯化铵的含量。Further, after the acquisition of the ultraviolet spectrum data of the eye drop sample, it is necessary to measure the content of dodecyldimethylbenzyl ammonium chloride in the eye drop sample, and the obtained component standard spectrum database V, eye drop The liquid background spectrum database H1 ′ and the eye drop sample ultraviolet spectrum data matrix a are imported into the computing platform, and the component to be measured, dodecyldimethyl The spectrum of benzyl ammonium chloride can be used to obtain the content of the component dodecyl dimethyl benzyl ammonium chloride to be measured in the eye drop dilution.
进一步，进一步包括以下步骤：Further, further comprising the following steps:
步骤一，依据定量精度设定扣减步长Δ；Step 1, set the deduction step size Δ according to the quantitative accuracy;
步骤二，在算式yi＝aix+bi中带入较大的x1值，得到v1；Step 2, enter a larger value of x1 into the formula yi =ai x+bi to obtain v1 ;
yi表示在i波长下十二烷基二甲基苄基氯化铵的吸光度值，ai、bi是常数，x表示十二烷基二甲基苄基氯化铵的浓度，v1表示在浓度为x1时十二烷基二甲基苄基氯化铵的多波长吸光度值，v1为所有的yi值组成的矩阵；yi represents the absorbance value of dodecyl dimethyl benzyl ammonium chloride at i wavelength, ai andbi are constants, x represents the concentration of dodecyl dimethyl benzyl ammonium chloride, v1 Represent the multi-wavelength absorbance value of dodecyl dimethyl benzyl ammonium chloride when the concentration is x1 , v1 is a matrix composed of all yi values;
步骤三，从滴眼液稀释液样本光谱数据a中扣除v1/Δ，扣除后的变量记为da；把本底光谱数据库H1′和变量da合并后记为对比空间M，计算对比空间M与v1夹角；Step 3: Deduct v1 /Δ from the spectral data a of the eye drop dilution sample, and record the deducted variable as da; combine the background spectral database H1 ′ with the variable da and record it as the contrast space M, and calculate the contrast space M Angle with v1 ;
步骤四，从滴眼液稀释液样本光谱数据a中逐步扣除v1后，重复步骤三；Step 4, after gradually deducting v1 from the spectral data a of the eye drop dilution sample, repeat step 3;
步骤五，当滴眼液稀释液样本中的十二烷基二甲基苄基氯化铵完全被扣除后，比对空间M和十二烷基二甲基苄基氯化铵的光谱向量v1的空间夹角值会出现最大值θmax，记录空间夹角最大值θmax相对应的扣减步数λ，通过十二烷基二甲基苄基氯化铵的浓度x1和扣减步数λ估算滴眼液稀释液样本中十二烷基二甲基苄基氯化铵的含量Y1，计算式为Y1＝x1/Δ×λ，得到的Y1即为滴眼液稀释液样本中十二烷基二甲基苄基氯化铵的含量值；若Y1值与x1值相差较大，则带入一个与Y1相接近且大于Y1的x2重新计算；Step five, when the dodecyldimethylbenzyl ammonium chloride in the eye drop dilution sample is completely deducted, compare the spectral vector v of the space M and dodecyldimethylbenzyl ammonium chloride The space angle value of1 will have the maximum value θmax , and record the deduction step number λ corresponding to the maximum value of the space angle θmax , through the concentration of dodecyl dimethyl benzyl ammonium chloride x1 and deduction The number of steps λ is to estimate the content Y1 of dodecyldimethylbenzyl ammonium chloride in the eye drop dilution sample, the calculation formula is Y1 = x1 /Δ×λ, and the obtained Y1 is the eye drop The content value of dodecyl dimethyl benzyl ammonium chloride in the diluent sample; if the value of Y1 is quite different from the value of x1 , then bring in a value of x2 that is close to Y1 and greater than Y1 to recalculate ;
步骤六，根据稀释倍数即求得实际滴眼液样本中十二烷基二甲基苄基氯化铵的含量。Step 6, obtain the content of dodecyl dimethyl benzyl ammonium chloride in the actual eye drop sample according to the dilution factor.
本发明提供的滴眼液中苯扎氯铵的快速检测方法，与现有技术相比，具有以下优势：Compared with the prior art, the rapid detection method of benzalkonium chloride in eye drops provided by the invention has the following advantages:
1.分析效率高、操作强度小：1. High analysis efficiency and low operation intensity:
由于本发明采用多波长紫外光谱直接定量苯扎氯铵，不需要分离，且样本测量及定量计算时间短，其分析效率较高，操作强度较小，非常适用于同类大批量样本的快速分析测定。Since the present invention uses multi-wavelength ultraviolet spectroscopy to directly quantify benzalkonium chloride, no separation is required, and the sample measurement and quantitative calculation time is short, the analysis efficiency is high, and the operation intensity is small, which is very suitable for rapid analysis and determination of similar large-scale samples .
2.分析成本低：2. Low analysis cost:
由于本发明对仪器并未提出特殊要求，只需简单设置便可沿用常见的一阶光谱仪器输出数据，没有对数据维数提出特别要求，数据采集后，通过本发明提出的数据处理步骤，即可通过数学分离方法实现十二烷基二甲基苄基氯化铵光谱信号提取，以及定量分析。因而不会提高仪器的制造难度和生产成本。而且试剂消耗量显著降低，降低了分析成本。Since the present invention has no special requirements for the instrument, the output data of the common first-order spectroscopic instrument can be used simply by simple setting, and there is no special requirement for the data dimension. After data collection, the data processing steps proposed by the present invention, namely Dodecyldimethylbenzylammonium chloride spectral signal extraction and quantitative analysis can be realized by mathematical separation method. Therefore, the manufacturing difficulty and production cost of the instrument will not be increased. Moreover, the consumption of reagents is significantly reduced, reducing the cost of analysis.
3.无需积累不含被测组分的本底数据库：3. There is no need to accumulate a background database that does not contain the measured components:
由于滴眼液种类繁多，原料复杂，建立完整的本底数据库工作量大，影响分析进程、分析精度及方法的适用性。而本发明借助高效液相色谱-紫外光谱联用的方法，获取滴眼液样本各个时刻、各个波长的光强三维数据，并获得不含被测组分的本底数据库。此方法不仅减小了工作强度，而且提高了分析精度，扩大了方法的适用性。Due to the variety of eye drops and complex raw materials, the workload of establishing a complete background database is heavy, which affects the analysis process, analysis accuracy and applicability of the method. However, the present invention obtains the three-dimensional data of light intensity at each moment and each wavelength of the eye drop sample by means of the method of high-performance liquid chromatography-ultraviolet spectroscopy, and obtains a background database without components to be measured. This method not only reduces the work intensity, but also improves the analysis precision and expands the applicability of the method.
4.可降低仪器和试剂损耗：4. It can reduce the loss of instruments and reagents:
对于同类样品的测定，本发明在进行一次色谱分离后，无需再利用联用仪器累积本底，仅通过光谱测量即可实现定量，从而降低了仪器及试剂损耗。For the determination of similar samples, after a chromatographic separation, the present invention does not need to use a combined instrument to accumulate the background, and can realize quantification only through spectral measurement, thereby reducing the loss of instruments and reagents.
5.稳健性好：5. Good robustness:
本发明与现有的单波长、双波长及若干个波长方法相比，抗干扰能力强，可辨识程度高，稳健性好。Compared with the existing single-wavelength, double-wavelength and several-wavelength methods, the present invention has strong anti-interference ability, high recognizability and good robustness.
6.本发明分析方法比较简单，无复杂步骤，易于推广应用。6. The analytical method of the present invention is relatively simple, without complicated steps, and easy to be popularized and applied.
附图说明Description of drawings
图1是本发明实施例提供的滴眼液中苯扎氯铵的快速检测方法流程图；Fig. 1 is the rapid detection method flowchart of benzalkonium chloride in the eye drop that the embodiment of the present invention provides;
图2是本发明实施例提供的斜投影提取的十二烷基二甲基苄基氯化铵紫外光谱图；Fig. 2 is the dodecyl dimethyl benzyl ammonium chloride ultraviolet spectrogram that the oblique projection provided by the embodiment of the present invention extracts;
图3是本发明实施例提供的空间夹角值与扣减步数的曲线图。Fig. 3 is a graph of the included space angle value and the number of deduction steps provided by the embodiment of the present invention.
具体实施方式Detailed ways
为了使本发明的目的、技术方案及优点更加清楚明白，以下结合实施例，对本发明进行进一步详细说明。应当理解，此处所描述的具体实施例仅仅用以解释本发明，并不用于限定本发明。In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.
下面结合附图对本发明的应用原理作详细的描述。The application principle of the present invention will be described in detail below in conjunction with the accompanying drawings.
本发明涉及一种滴眼液中成分组分的检测方法，采用滴眼液中混合物苯扎氯铵中的十二烷基二甲基苄基氯化铵完成含量检测。The invention relates to a method for detecting components in eye drops, which uses dodecyl dimethyl benzyl ammonium chloride in a mixture of benzalkonium chloride in the eye drops to complete content detection.
如图1所示，本发明实施例的滴眼液中苯扎氯铵的快速检测方法包括以下步骤：As shown in Figure 1, the rapid detection method of benzalkonium chloride in the eye drops of the embodiment of the present invention comprises the following steps:
S101：采用斜投影的多变量体系中纯信号提取方法将待测组分十二烷基二甲基苄基氯化铵从苯扎氯铵混合物中提取，建立标准光谱数据库；S101: Using the pure signal extraction method in the oblique projection multivariate system to extract the component to be measured, dodecyl dimethyl benzyl ammonium chloride, from the benzalkonium chloride mixture, and establish a standard spectral database;
S102：通过液相-紫外联用构建样本本底光谱数据库；获取滴眼液样本紫外光谱数据；S102: Construct a sample background spectrum database through liquid phase-ultraviolet combination; obtain the ultraviolet spectrum data of the eye drop sample;
S103：最后根据空间夹角判据求得滴眼液样本中待测组分十二烷基二甲基苄基氯化铵的含量，完成测定。S103: Finally, the content of the component to be tested in the eye drop sample, dodecyldimethylbenzyl ammonium chloride, is obtained according to the spatial angle criterion, and the determination is completed.
本发明实施例的具体步骤如下：The concrete steps of the embodiment of the present invention are as follows:
本发明实施例的一种滴眼液中苯扎氯铵的快速检测方法，包括以下步骤：A kind of rapid detection method of benzalkonium chloride in the eye drop of the embodiment of the present invention, comprises the following steps:
I、提取十二烷基二甲基苄基氯化铵纯信号，建立标准光谱数据库。I, extract the pure signal of dodecyl dimethyl benzyl ammonium chloride, and establish a standard spectral database.
A.苯扎氯铵溶液配制：A. Preparation of benzalkonium chloride solution:
精确移取适量12.02mg/mL的苯扎氯铵，用二次蒸馏水定容，配制系列浓度为1～20μg/mL的苯扎氯铵标准溶液。Accurately pipette an appropriate amount of 12.02 mg/mL benzalkonium chloride, dilute to volume with double distilled water, and prepare a series of benzalkonium chloride standard solutions with a concentration of 1-20 μg/mL.
所述的苯扎氯铵1～20μg/mL系列浓度其中苯扎氯铵按十二烷基二甲基苄基氯化铵记。The concentration of benzalkonium chloride is 1-20 μg/mL series, wherein benzalkonium chloride is recorded as dodecyl dimethyl benzyl ammonium chloride.
B.苯扎氯铵本底光谱数据库的建立：B. The establishment of the background spectrum database of benzalkonium chloride:
SB1.获取苯扎氯铵标准溶液的液相-紫外联用数据：SB1. Obtain the liquid phase-UV data of benzalkonium chloride standard solution:
将步骤A配制的苯扎氯铵标准溶液过滤后经高效液相色谱分析，经紫外光谱仪采集系列苯扎氯铵标准溶液的多波长三维数据。The benzalkonium chloride standard solution prepared in step A is filtered and analyzed by high-performance liquid chromatography, and the multi-wavelength three-dimensional data of a series of benzalkonium chloride standard solutions are collected by an ultraviolet spectrometer.
步骤SB1中高效液相色谱条件为：The high performance liquid chromatography condition in step SB1 is:
色谱柱：ODS-SP92689C18色谱柱(4.6mm*250mm)Chromatographic column: ODS-SP92689C18 chromatographic column (4.6mm*250mm)
等度洗脱，流动相：乙腈：0.02mol/L乙酸铵水溶液(每L中含三乙胺10mL，用冰醋酸调节pH至5.0±0.5)＝90∶10(V/V)Isocratic elution, mobile phase: acetonitrile: 0.02mol/L ammonium acetate aqueous solution (each L contains 10 mL of triethylamine, adjust the pH to 5.0±0.5 with glacial acetic acid) = 90:10 (V/V)
流速：1mL/minFlow rate: 1mL/min
检测波长：262nmDetection wavelength: 262nm
进样量：20uLInjection volume: 20uL
柱温：室温。Column temperature: room temperature.
步骤SB1中紫外光纤光谱仪条件为：The conditions of the ultraviolet fiber optic spectrometer in step SB1 are:
检测波长范围：196.93～400.36nm；光谱间隔：0.46nm；石英比色皿光程：1.0cm；以二次蒸馏水为参比样品；积分时间：8ms。Detection wavelength range: 196.93 ~ 400.36nm; spectral interval: 0.46nm; quartz cuvette optical path: 1.0cm; twice distilled water as a reference sample; integration time: 8ms.
SB2.转化数据格式：SB2. Conversion data format:
将步骤SB1中采集所得的苯扎氯铵的多波长三维数据的光强度数据值转化成吸光度值，转化后的数据的每一行代表一个波长下的色谱数据，每一列代表一个时刻下洗脱出来物质的光谱数据；The light intensity data value of the multi-wavelength three-dimensional data of benzalkonium chloride collected in step SB1 is converted into an absorbance value, each row of the converted data represents the chromatographic data at one wavelength, and each column represents the elution at one moment Spectral data of the substance;
SB3.获取不含待测组分的本底光谱数据：SB3. Obtain background spectral data without components to be measured:
根据苯扎氯铵混合物中十二烷基二甲基苄基氯化铵、十四烷基二甲基苄基氯化铵的物质特性、文献资料及实验操作确定待测组分十二烷基二甲基苄基氯化铵的保留时间，其中十二烷基二甲基苄基氯化铵的保留时间为8.31～9.02min，十四烷基二甲基苄基氯化铵的保留时间为11.35～11.79min。根据苯扎氯铵色谱图，从苯扎氯铵标准溶液三维数据中扣除待测组分十二烷基二甲基苄基氯化铵的光谱数据，将其余光谱数据加和作为苯扎氯铵本底光谱数据库H；According to the material properties, literature and experimental operation of dodecyl dimethyl benzyl ammonium chloride and tetradecyl dimethyl benzyl ammonium chloride in the benzalkonium chloride mixture, the component dodecyl to be measured is determined. The retention time of dimethyl benzyl ammonium chloride, wherein the retention time of dodecyl dimethyl benzyl ammonium chloride is 8.31 ~ 9.02min, the retention time of tetradecyl dimethyl benzyl ammonium chloride is 11.35～11.79min. According to the benzalkonium chloride chromatogram, the spectral data of the component dodecyl dimethyl benzyl ammonium chloride to be measured is deducted from the three-dimensional data of the benzalkonium chloride standard solution, and the remaining spectral data are summed as benzalkonium chloride Background spectrum database H;
SB4.本底数据库的降维：SB4. Dimensionality reduction of the background database:
由于步骤SB3得到的数据库H数据量大，将会导致计算量很大。因此将步骤SB3中所得的本底光谱数据库H进行奇异值分解，根据体系主成分数目将本底光谱数据库H进行重构，得到维数适当的本底光谱数据库H′。Since the database H obtained in step SB3 has a large amount of data, it will lead to a large amount of calculation. Therefore, the background spectrum database H obtained in step SB3 is subjected to singular value decomposition, and the background spectrum database H is reconstructed according to the number of principal components of the system to obtain a background spectrum database H' with an appropriate dimension.
C.获取待测组分的纯信号：C. Obtain the pure signal of the component to be measured:
根据苯扎氯铵的色谱图找到待测组分的最高响应值对应的时间点8.49min，从苯扎氯铵标准溶液三维数据中扣出待测组分光谱信号作为待测组分纯信号S。According to the chromatogram of benzalkonium chloride, the time point 8.49min corresponding to the highest response value of the component to be tested is found, and the spectral signal of the component to be tested is deducted from the three-dimensional data of the standard solution of benzalkonium chloride as the pure signal S of the component to be tested .
D.苯扎氯铵标准溶液光谱量测：D. Benzalkonium chloride standard solution spectrum measurement:
采用光纤光谱仪对步骤A制得1～20μg/mL的苯扎氯铵标准溶液进行光谱扫描，得到苯扎氯铵标准溶液的多波长紫外光谱数据矩阵y。Using a fiber optic spectrometer to perform spectral scanning on the 1-20 μg/mL benzalkonium chloride standard solution prepared in step A, to obtain the multi-wavelength ultraviolet spectrum data matrix y of the benzalkonium chloride standard solution.
E.提取待测组分纯信号：E. Extract the pure signal of the component to be tested:
将步骤SB4、C和D所得的苯扎氯铵本底光谱数据库H′、待测组分纯分析信号S和苯扎氯铵标准溶液紫外光谱数据矩阵y导入计算平台，应用基于斜投影的多变量体系中纯信号提取方法将待测组分十二烷基二甲基苄基氯化铵从苯扎氯铵混合物中提取，得到1～20μg/mL的系列浓度的稳定的待测组分纯光谱信号矩阵S′。Import the benzalkonium chloride background spectrum database H', the pure analysis signal S of the component to be measured and the ultraviolet spectrum data matrix y of the benzalkonium chloride standard solution obtained in steps SB4, C and D into the computing platform, and apply the multi- The pure signal extraction method in the variable system extracts the component to be tested, dodecyl dimethyl benzyl ammonium chloride, from the benzalkonium chloride mixture, and obtains the stable pure signal of the component to be tested with a series concentration of 1-20 μg/mL. Spectral signal matrix S'.
所述的基于斜投影的多变量体系中纯信号提取方法为：The pure signal extraction method in the multivariate system based on oblique projection is:
(1)将步骤SB4、C求得的H′、S导入计算平台，求取纯信号分离模型，即斜投影算子，该算子的表达式为：(1) Import the H' and S obtained in steps SB4 and C into the computing platform to obtain a pure signal separation model, that is, the oblique projection operator. The expression of this operator is:
ES|H＝S(STPH⊥S)-1STPH⊥，其中PH⊥＝I-PH＝I-H(HTH)-1HT；ES|H ＝S(ST PH⊥ S)-1 ST PH⊥ , where PH⊥ ＝IPH ＝IH(HT H)-1 HT ;
在上式中，上标T表示矩阵的转置，I为与PH维数相同的单位矩阵；In the above formula, the superscript T represents the transposition of the matrix, and I is the identity matrix with the same dimension asPH ;
(2)将步骤D中得到y与斜投影算子ES|H相乘，即得到1～20μg/mL的系列浓度的十二烷基二甲基苄基氯化铵纯光谱信号S′(参见图2)；(2) Multiplying y obtained in step D with the oblique projection operator ES|H , promptly obtains the dodecyldimethylbenzyl ammonium chloride pure spectral signal S'( See Figure 2);
F.待测组分标准光谱库的建立：F. Establishment of the standard spectral library of the components to be measured:
将步骤E中得到的待测组分纯光谱信号与其对应的浓度值进行最小二乘拟合，得到单位浓度的十二烷基二甲基苄基氯化铵光谱数据，存入标准光谱数据库V；Carry out least squares fitting with the pure spectral signal of the component to be measured obtained in step E and its corresponding concentration value, obtain the dodecyl dimethyl benzyl ammonium chloride spectral data of unit concentration, store in standard spectral database V ;
所得到的标准光谱库V可用于下一步计算。The obtained standard spectral library V can be used for the next calculation.
II、滴眼液样本中待测组分的含量测定。II. Determination of the content of the component to be tested in the eye drop sample.
G.滴眼液样本处理：G. Eye drop sample processing:
精确移取适量滴眼液样本至10mL的容量瓶内，用二次蒸馏水稀释适当的倍数，制得滴眼液稀释液。Accurately pipette an appropriate amount of eye drop sample into a 10mL volumetric flask, and dilute an appropriate multiple with double distilled water to prepare the eye drop dilution.
H.滴眼液样本本底光谱数据库的建立：H. Establishment of background spectrum database of eye drops samples:
SH1.获取滴眼液样本的液相-紫外联用数据：SH1. Obtain liquid phase-ultraviolet data of eye drop samples:
将不经稀释的滴眼液样本过滤后经高效液相色谱分析，经紫外光谱仪采集滴眼液样本的多波长三维数据。The undiluted eye drop sample was filtered and analyzed by high performance liquid chromatography, and the multi-wavelength three-dimensional data of the eye drop sample was collected by ultraviolet spectrometer.
步骤SH1中用到的高效液相色谱条件与步骤SB1中用到的高效液相色谱条件一致。The high performance liquid chromatography conditions used in step SH1 are consistent with the high performance liquid chromatography conditions used in step SB1.
步骤SH1中用到的紫外光纤光谱仪条件与步骤SB1中用到的紫外光纤光谱仪条件一致。The conditions of the ultraviolet fiber spectrometer used in step SH1 are consistent with the conditions of the ultraviolet fiber spectrometer used in step SB1.
SH2.转化数据格式：SH2. Conversion data format:
将步骤SH1中采集所得的滴眼液样本多波长三维数据的光强度数据值转化成吸光度值，转化后的数据的每一行代表一个波长下的色谱数据，每一列代表一个时刻下洗脱出来物质的光谱数据；The light intensity data value of the multi-wavelength three-dimensional data of the eye drop sample collected in step SH1 is converted into an absorbance value, each row of the converted data represents the chromatographic data at one wavelength, and each column represents the substance eluted at a time Spectral data;
SH3.获取不含待测组分的本底光谱数据：SH3. Obtain background spectral data without components to be measured:
根据步骤SB3确定的待测组分十二烷基二甲基苄基氯化铵的保留时间，从滴眼液样本三维数据中扣除待测组分的光谱数据，将其余光谱数据加和作为滴眼液本底光谱数据库H1；According to the retention time of the component to be measured in dodecyl dimethyl benzyl ammonium chloride determined in step SB3, the spectral data of the component to be measured is deducted from the three-dimensional data of the eye drop sample, and the remaining spectral data are summed as the drop Eye fluid background spectrum database H1 ;
SH4.本底光谱数据库的降维：SH4. Dimensionality reduction of background spectral database:
由于步骤SH3得到的数据库H1数据量大，将会导致计算量很大。因此将步骤SH3中所得的本底光谱数据库H1进行奇异值分解，根据体系主成分数目将本底光谱数据库H1进行重构，得到维数适当的本底光谱数据库H1′。Since the database H1 obtained in step SH3 hasa large amount of data, it will lead to a large amount of calculation. Therefore, the background spectrum database H1 obtained in step SH3 is subjected to singular value decomposition, and the background spectrum database H1 is reconstructed according to the numberof principal components of the system to obtaina background spectrum database H1' with an appropriate dimension.
I.滴眼液样本光谱量测：I. Spectral measurement of eye drops samples:
采用光纤光谱仪对步骤G中制得的滴眼液稀释液进行光谱扫描，得到滴眼液稀释液的紫外光谱数据矩阵a；Using a fiber optic spectrometer to perform spectral scanning on the eye drop dilution prepared in step G, to obtain the ultraviolet spectrum data matrix a of the eye drop dilution;
J.滴眼液样本中十二烷基二甲基苄基氯化铵含量测定：Determination of content of dodecyldimethylbenzyl ammonium chloride in J. eye drops sample:
分别将步骤F、SH4和I所得的待测组分标准光谱数据库V、滴眼液本底光谱数据库H1′和滴眼液样本紫外光谱数据矩阵a导入计算平台，应用空间夹角判据从滴眼液样本稀释液光谱中扣减待测组分十二烷基二甲基苄基氯化铵的光谱，得出滴眼液稀释液中待测组分十二烷基二甲基苄基氯化铵的含量；Import the standard spectral database V of the component to be measured obtained in steps F, SH4 and I, the background spectral database H1 ' of eye drops, and the UV spectral data matrix a of the eye drop sample into the computing platform, and apply the spatial angle criterion from Subtract the spectrum of the component to be measured, dodecyldimethylbenzyl ammonium chloride, from the spectrum of the eye drop sample diluent to obtain the concentration of the component to be measured in the eye drop dilution, dodecyldimethylbenzyl ammonium chloride content;
所述的滴眼液样本中十二烷基二甲基苄基氯化铵含量测定的具体步骤如下：The specific steps of dodecyl dimethyl benzyl ammonium chloride content determination in the described eye drop sample are as follows:
(1)依据定量精度设定扣减步长Δ(本实施例为1000)；(1) Set the deduction step size Δ (1000 in this embodiment) according to the quantitative accuracy;
(2)在算式yi＝aix+bi中带入较大的x1值，得到v1；(2) Bringing a larger value of x1 into the formula yi =ai x+bi to obtain v1 ;
所述的yi表示在i波长下十二烷基二甲基苄基氯化铵的吸光度值，ai、bi是常数，x表示十二烷基二甲基苄基氯化铵的浓度，v1表示在浓度为x1时十二烷基二甲基苄基氯化铵的多波长吸光度值，v1为所有的yi值组成的矩阵；Describedyi represents the absorbance value of dodecyl dimethyl benzyl ammonium chloride at i wavelength, ai ,bi are constants, and x represents the concentration of dodecyl dimethyl benzyl ammonium chloride , v1 represents the multi-wavelength absorbance value of dodecyldimethylbenzyl ammonium chloride when the concentration is x1 , and v1 is a matrix composed of all yi values;
(3)从滴眼液稀释液样本光谱数据a中扣除v1/Δ(本实施例为扣除v1/1000)，扣除后的变量记为da；把本底光谱数据库H1′和变量da合并后记为对比空间M，计算对比空间M与v1夹角；(3) Deduct v1 /Δ (deduct v 1 /1000) from the eye drop dilution sample spectral data a (in this embodiment, deduce v1 /1000), and denote the variable after the deduction as da; the background spectrum database H1 ' and the variable da After the combination, it is recorded as the comparison space M, and the angle between the comparison space M and v1 is calculated;
(4)从滴眼液稀释液样本光谱数据a中逐步扣除v1后，重复步骤(3)；(4) After gradually deducting v1 from the eye drop dilution sample spectral dataa , repeat step (3);
(5)当滴眼液稀释液样本中的十二烷基二甲基苄基氯化铵完全被扣除后，比对空间M和十二烷基二甲基苄基氯化铵的光谱向量v1的空间夹角值会出现最大值θmax，记录空间夹角最大值θmax相对应的扣减步数λ(参见图3，空间夹角最大值对应的扣减步数为843)，通过十二烷基二甲基苄基氯化铵的浓度x1和扣减步数λ估算滴眼液稀释液样本中十二烷基二甲基苄基氯化铵的含量Y1，计算式为Y1＝x1/Δ×λ，得到的Y1即为滴眼液稀释液样本中十二烷基二甲基苄基氯化铵的含量值。(5) After the dodecyldimethylbenzyl ammonium chloride in the eye drop dilution sample is completely deducted, compare the spectral vector v of the space M and dodecyldimethylbenzyl ammonium chloride The space angle value of1 will have the maximum value θmax , and record the deduction steps λ corresponding to the maximum value of the space angle θmax (see Figure 3, the number of deduction steps corresponding to the maximum value of the space angle is 843), through The concentration of dodecyl dimethyl benzyl ammonium chloride x1 and the number of deduction steps λ are used to estimate the content Y1 of dodecyl dimethyl benzyl ammonium chloride in the eye drop dilution sample, and the calculation formula is Y1 =x1 /Δ×λ, and the obtained Y1 is the content value of dodecyldimethylbenzyl ammonium chloride in the eye drop dilution sample.
若Y1值与x1值相差较大，则带入一个与Y1相接近且大于Y1的x2重新计算。If the value of Y1 is quite different from the value of x1 , bring in a value of x2 that is close to Y1 and greater than Y1 to recalculate.
(6)根据步骤G中的稀释倍数即可求得实际滴眼液样本中十二烷基二甲基苄基氯化铵的含量。(6) According to the dilution factor in step G, the content of dodecyldimethylbenzyl ammonium chloride in the actual eye drop sample can be obtained.
步骤SB4、SH4.本地光谱数据库的降维中，所述的体系主成分数目的确定方法为：Steps SB4, SH4. In the dimensionality reduction of the local spectral database, the method for determining the number of principal components of the system is:
将矩阵H、H1进行方差标准化后，添加一定强度的白噪声掩蔽不均匀噪声和非线性因素的干扰，以二阶差分值序列转折点判断其独立变量数目，得到体系主成分数目N、N1。After standardizing the variance of the matrices H and H1 , adding a certain intensity of white noise to mask the interference of uneven noise and nonlinear factors, judging the number of independent variables by the turning point of the second-order difference value sequence, and obtaining the number of principal components of the system N, N1 .
步骤SB4、SH4.本地光谱数据库的降维中，所述的将本底光谱数据库H、H1进行重构的方法如下：Steps SB4, SH4. In the dimensionality reductionof the local spectral database, the method for reconstructing the background spectral database H, H1 is as follows:
应用奇异值分解程序[US1V1]＝svd(H)对苯扎氯铵本底光谱数据库H进行奇异值分解降维，分解后得到m阶行正交矩阵U，n阶列正交矩阵V1和矩阵H的特征值矩阵S1，取U的前N列即为降维后的苯扎氯铵本底光谱数据库H′；Apply the singular value decomposition program [US1 V1 ]=svd(H) to carry out singular value decomposition and dimensionality reduction on the background spectrum database H of benzalkonium chloride, and obtain m-order row orthogonal matrix U and n-order column orthogonal matrix after decomposition V1 and the eigenvalue matrix S1 of the matrix H, taking the first N columns of U is the benzalkonium chloride background spectrum database H′ after dimensionality reduction;
同法得到滴眼液样本本底光谱数据库H1′Obtain the background spectral database H1 ′ of eye drops samples by the same method
为了验证本发明测定结果的准确性，选择了7种市售滴眼液样本，通过高效液相色谱-紫外光纤光谱仪联用建立本底数据库后，用本发明方法测定滴眼液样本中的含量，并与高效液相色谱法结果比较(参见表1)。In order to verify the accuracy of the measurement results of the present invention, 7 kinds of commercially available eye drop samples were selected, and after the background database was established through the combination of high performance liquid chromatography-ultraviolet fiber optic spectrometer, the method of the present invention was used to measure the content of the eye drop samples. , and compared with the results of high performance liquid chromatography (see Table 1).
表1滴眼液中含量的两种方法测定结果Two kinds of method determination results of content in table 1 eye drops
本发明还进行了回收率实验，结果如表2所示。The present invention has also carried out the recovery rate experiment, and the results are shown in Table 2.
表2回收率实验Table 2 Recovery experiment
对1#、3#样本进行精密度实验，RSD分别为2.78％、0.54％。The precision experiment is carried out on samples 1# and 3# , and the RSDs are 2.78% and 0.54% respectively.
以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present invention should be included in the protection of the present invention. within range.
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510539781.6ACN105181866A (en) | 2015-08-28 | 2015-08-28 | Method for rapid detection of benzalkonium chloride in eye drops |
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| CN201510539781.6ACN105181866A (en) | 2015-08-28 | 2015-08-28 | Method for rapid detection of benzalkonium chloride in eye drops |
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| CN201510539781.6APendingCN105181866A (en) | 2015-08-28 | 2015-08-28 | Method for rapid detection of benzalkonium chloride in eye drops |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105866268A (en)* | 2016-03-28 | 2016-08-17 | 上海市食品药品检验所 | Detection method for simultaneously determining various antibacterial agents in eye drops |
| CN105954393A (en)* | 2016-04-25 | 2016-09-21 | 广西壮族自治区梧州食品药品检验所 | Method for measuring content of disinfectant in wet tissue by using high performance liquid chromatography-evaporation light scattering |
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| CN109406698A (en)* | 2018-11-30 | 2019-03-01 | 广州中科检测技术服务有限公司 | A kind of method that HPLC measures single-stranded substituted dimethyl benzyl ammonium chloride content |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105866268A (en)* | 2016-03-28 | 2016-08-17 | 上海市食品药品检验所 | Detection method for simultaneously determining various antibacterial agents in eye drops |
| CN105954393A (en)* | 2016-04-25 | 2016-09-21 | 广西壮族自治区梧州食品药品检验所 | Method for measuring content of disinfectant in wet tissue by using high performance liquid chromatography-evaporation light scattering |
| CN106323891A (en)* | 2016-08-25 | 2017-01-11 | 广西科技大学 | Method of quick analysis of azo dyes synthesis process |
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| CN108680688A (en)* | 2018-05-08 | 2018-10-19 | 江苏安舜技术服务有限公司 | The assay method of BAC and DDAC in a kind of soil and plant-derived product |
| CN109406698A (en)* | 2018-11-30 | 2019-03-01 | 广州中科检测技术服务有限公司 | A kind of method that HPLC measures single-stranded substituted dimethyl benzyl ammonium chloride content |
| CN120253717A (en)* | 2025-03-27 | 2025-07-04 | 中检易兴元科技(北京)有限公司 | A quantitative analysis method, storage medium and device for alcohol substances in a mixture |
| Publication | Publication Date | Title |
|---|---|---|
| CN105181866A (en) | Method for rapid detection of benzalkonium chloride in eye drops | |
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