[關鍵詞]
[摘要]
目的 建立一測多評法(QAMS)測定藏茴香中6種咖啡酰奎寧酸類成分含量,并驗證該方法在藏茴香質(zhì)量評價中應用的可行性與適用性。方法 取藏茴香粉末(過三號篩)0.5 g,精密加入70%甲醇 20 mL ,超聲處理(250 W、頻率 53 kHz)30 min,制備供試品溶液;采用Phenomenex GeminiR C18(250 mm×4.6 mm,5 μm)色譜柱,以乙腈-0.1%甲酸水溶液為流動相,梯度洗脫,檢測波長為 330 nm,體積流量 1.0 mL·min-1,柱溫 30 ℃,進行專屬性、供試品提取條件、檢測波長選擇、色譜條件、線性關系、精密度、重復性、穩(wěn)定性、加樣回收率方法學考察,建立異綠原酸A、新綠原酸、綠原酸、隱綠原酸、異綠原酸 B、異綠原酸 C成分含量檢測的 HPLC法;以異綠原酸 A為內(nèi)參成分,分別計算新綠原酸、綠原酸、隱綠原酸、異綠原酸B、異綠原酸C 5種成分的相對校正因子,分別采用3種不同色譜儀和3種色譜柱進行相對校正因子、相對保留時間耐用性考察,對藏茴香樣品同時采用外標法與 QAMS 測定 6 種成分的質(zhì)量分數(shù),比較 2 種測定方法結果的差異。結果 建立的6種成分的HPLC檢測方法的專屬性、供試品提取條件、檢測波長選擇、色譜條件、線性關系、精密度、重復性、穩(wěn)定性、加樣回收率均符合要求;新綠原酸、綠原酸、隱綠原酸、異綠原酸B、異綠原酸C的相對校正因子平均值分別是 1.362、1.257、1.335、1.470、1.134,3種不同色譜儀和 3種色譜柱對相對校正因子、相對保留時間均無明顯影響;QAMS與外標法2種方法測定3批藏茴香樣品中6種成分得到的結果之間無顯著差異。結論 建立的QAMS簡便、準確、可靠,可用于藏茴香中6種咖啡??鼘幩犷惓煞帧惥G原酸A、新綠原酸、綠原酸、隱綠原酸、異綠原酸B、異綠原酸C的定量分析。
[Key word]
[Abstract]
Objective To establish a method for the content determination of six caffeoylquinic acids components in Carum carvi L. through quantitative analysis of multi-components with single marker (QAMS). To verify the feasibility and applicability of the method in its quality control. Methods Take 0.5 g of C. carvi powder (through the third sieve), accurately add 20 mL of 70% methanol, and conduct ultrasonic treatment (250 W, frequency 53 kHz) for 30 min to prepare the test solution. Phenomenex GeminiR C 18 column (250 mm × 4.6 mm, 5 μm) was used, acetonitrile-0.1% formic acid aqueous solution was used as mobile phase, gradient elution, flow rate was 1.0 mL·min-1, column temperature was 30 ℃, and detection wavelength was 330 nm. Conduct methodological studies on specificity, test article extraction conditions, detection wavelength selection, chromatographic conditions, linear relationship, precision, repeatability, stability, and sample addition recovery. With isochlorogenic acid A as the internal reference component, relative correction factors of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B and isochlorogenic acid C was established. Three different chromatographs and three chromatographic columns were used to investigate the relative correction factor and relative retention time durability. The external standard method and QAMS were used to simultaneously determine the mass fractions of six components in C. carvi samples, and the differences between the two determination results were compared. Results The specificity, extraction conditions, detection wavelength selection, chromatographic conditions, linear relationship, precision, repeatability, stability, and sample recovery of the established HPLC detection methods for the six components meet the requirements. The relative positive factors of isochlorogenic acid A and neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B and isochlorogenic acid C were 1.362, 1.257, 1.335, 1.470, and 1.134, respectively. Three different chromatographs and three chromatographic columns had no significant impact on the relative correction factor and relative retention time. There was no significant difference between the calculated values of six components in three batches of samples by the QAMS and the measured values of the external standard method. Conclusion The method established in the study is simple and stable, and can be used for the synchronous quality control of six components (isochlorogenic acid A, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B and isochlorogenic acid C) in C. carvi.
[中圖分類號]
R291.4;927.2
[基金項目]
新疆維吾爾自治區(qū)自然科學基金資助項目(2022D01E94);新疆維吾爾自治區(qū)天山英才培養(yǎng)計劃資助項目(2022TSYCCX0023)