目的 建立UPLC波長切換法同時(shí)測定雙黃連注射劑［雙黃連注射液、注射用雙黃連（凍干）、雙黃連粉針劑］中5種特征成分，并對87批樣品（注射液63批，凍干劑15批，粉針劑9批）進(jìn)行測定，對批次間、企業(yè)間及劑型間樣品結(jié)果差異性進(jìn)行分析。方法 樣品經(jīng)提取后，采用UPLC波長切換法，同時(shí)測定了綠原酸、咖啡酸、連翹酯苷A、黃芩苷、連翹苷5種特征成分，色譜柱 ：Waters HSS T3 C₁₈（100 mm×2.1 mm，1.7 μm），以乙腈-0.1%磷酸溶液為流動相進(jìn)行梯度洗脫，體積流量0.3 mL·min^-1，綠原酸、咖啡酸及連翹酯苷 A檢測波長為 324 nm，黃芩苷檢測波長為 276 nm，連翹苷檢測波長為228 nm；進(jìn)行專屬性、線性關(guān)系、精密度、重復(fù)性、準(zhǔn)確度、穩(wěn)定性方法學(xué)考察；取樣品3批，分別按建立的方法、現(xiàn)行質(zhì)量標(biāo)準(zhǔn)測定方法檢測，對比2種方法的檢測結(jié)果；采用建立的方法對87批樣品進(jìn)行測定。繪制各測定指標(biāo)結(jié)果的頻率分布直方圖或箱式圖，進(jìn)行注射液不同企業(yè)間、批次間各成分含量差異性分析；繪制5個(gè)成分及綠原酸和咖啡酸總量的平均含量分布情況雷達(dá)圖，進(jìn)行不同劑型之間結(jié)果差異性分析；采用 minitab軟件對 87批樣品測定結(jié)果進(jìn)行主成分分析。結(jié)果 建立 UPLC波長切換法經(jīng)方法學(xué)考察符合要求；與現(xiàn)行質(zhì)量標(biāo)準(zhǔn)測定方法比對，建立的方法檢測結(jié)果無差異。注射液 2個(gè)生產(chǎn)企業(yè)樣本數(shù)相當(dāng)，綠原酸和咖啡酸總量的均值及離散程度無差異，但不同企業(yè)2種成分比例存在差別；黃芩苷、連翹苷及連翹酯苷A含量均值一致，離散程度一致，不同生產(chǎn)企業(yè)、不同批次間無顯著性差異。雷達(dá)圖結(jié)果顯示，凍干劑與粉針劑工藝除干燥方式外完全一致，其綠原酸、咖啡酸、兩者總量及黃芩苷含量比例落點(diǎn)幾乎重合；連翹苷與連翹酯苷A在粉針劑中含量是凍干劑的近2倍，且批次間差異性大；縱向比較3個(gè)劑型，除連翹苷含量相近外，其余成分含量差別均較大。主成分分析結(jié)果顯示，3個(gè)不同劑型被明顯區(qū)分，現(xiàn)行質(zhì)量標(biāo)準(zhǔn)控制項(xiàng)目越多的劑型，樣品越集中。結(jié)論 建立的方法準(zhǔn)確可靠，在多樣本測定的基礎(chǔ)上為標(biāo)準(zhǔn)統(tǒng)一及藥品質(zhì)量標(biāo)準(zhǔn)制定的合理性提供參考。

Objective To establish a UPLC method for simultaneous determination of five components in Shuanghuanglian Injection (injection, lyophilized powder and powder-injection) by switching wavelength, and 87 batches of samples were determined. The differences of results among batches, enterprises and dosage forms were analyzed. Methods The sample were ultrasound-treated with appropriate solvents, UPLC was used to simultaneously determine five components of chlorogenic acid, caffeic acid, forsythiaside A, baicalin and forsythin. Waters HSS T3 C₁₈ (100 mm × 2.1 mm, 1.7 μm) column was used for gradient elution with acetonitrile-0.1% phosphoric acid as mobile phase at a flow rate of 0.3 mL·min^-1. The detection wavelength of chlorogenic acid, caffeic acid and forsythiaside A was 324 nm, baicalin was 276 nm, forsythin was 228 nm. Conduct methodological studies on specificity, linear relationship, precision, repeatability, accuracy, and stability was carry on. Three batches of samples were tested according to the established method and the current quality standard measurement method, and the test results of the two methods were compared. 87 batches of samples were determined. The established method was used to determine 87 batches of samples. Draw a frequency distribution histogram or box chart of the results of each measurement index, and analyze the content differences of various components among different enterprises and batches of injection. The average content distribution of five components and the total amount of chlorogenic acid and caffeic acid was plotted using a radar chart, and the difference in results between different dosage forms was analyzed. The results of 87 batches of samples were analyzed by principal component analysis using minitab software. Results The establishment of UPLC wavelength switching method meets the requirements through methodological investigation. Compared with the current quality standard measurement method, the established method has no difference in detection results. The sample numbers of the two injection production enterprises were similar, and there was no difference in the mean value and dispersion degree of the total amount of chlorogenic acid and caffeic acid, but there were differences in the proportion of components between different enterprises. The mean values of baicalin, forsythrin, and forsythrin A content were consistent, and the degree of dispersion was consistent. There was no significant difference between different manufacturers and different batches. The radar chart results show that the freeze-dried agent and powder injection process are completely consistent except for the drying method, with the total amount of chlorogenic acid, caffeic acid, and baicalin content ratio falling points almost coincident. The content of forsythrin and forsythrin A2 in powder injection was nearly twice as high as that in freeze-dried formulation, and there was significant difference between batches. Longitudinal comparison of the three dosage forms showed that except for the similar content of phillyrin, the content of other components varied significantly. The results of principal component analysis showed that three different dosage forms were clearly distinguished, and the more dosage forms controlled by current quality standards, the more concentrated the samples. Conclusion The method established in this study is accurate and reliable, and provides a basis for the unification of standards and the rationality of drug quality standards on the basis of multi-sample determination.

R284.1