HPLC-MS/MS测定麦冬和参麦注射液中龙脑苷和龙脑次苷的含量

Determination of Bornenol-7-O-[β-D-apiofuranosyl-(1®6)]-β-D-glucopyranoside and Borneol-7-O-β-D- glucopyranoside in Ophiopogonis Radix and Shenmai injection by HPLC-MS/MS

关键词:  麦冬  参麦注射液  龙脑苷  龙脑次苷  高效液相色谱-串联质谱法

Key words:  Ophiopogonis Radix  Shenmai injection  bornenol-7-O-[β-D-apiofuranosyl-(1→6)]-β-D-glucopyranoside  borneol-7-O-β-D-glucopyranoside  HPLC-MS/MS

摘要:

目的 采用HPLC-MS/MS测定麦冬和参麦注射液中龙脑苷和龙脑次苷含量。方法 以马钱苷为内标，色谱柱为Agilent Poroshell 120 EC-C₁₈(4.6 mm×100 mm，2.7 μm)，以乙腈(A相)-0.1%甲酸水溶液(B相)为流动相，梯度洗脱，流速为0.5 mL·min^–1，柱温为30 ℃。选用电喷雾离子源，在正离子模式下进行检测，定量分析选用多反应监测模式。龙脑苷、龙脑次苷和内标的离子对分别为m/z 449.5→137.1，317.1→137.1，391.2→179.1。结果 龙脑苷和龙脑次苷检测质量浓度的线性范围分别为0.20~10.32 μg·mL^–1(r=0.995 3，n=5)，0.02~1.51 μg·mL^–1(r=0.998 0，n=6)，定量限分别为8.1，3.5 ng·mL^–1。日内、日间精密度，重复性试验结果良好(RSD<10%)，龙脑苷和龙脑次苷24 h内稳定(RSD<10%)，平均回收率(n=3)分别为99.9%(RSD=1.93%)和99.6%(RSD=3.09%)。结论 该方法准确、简便、重复性好，可用于测定麦冬和参麦注射液中龙脑苷和龙脑次苷的含量。

Abstract:

OBJECTIVE To establish a liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) method to assay bornenol-7-O-[β-D-apiofuranosyl(1®6)]-β-D-glucopyranoside(BAG) and borneol-7-O-β-D-glucopyranoside(BG) in Ophiopogonis Radix and Shenmai injection. METHODS Loganin was used as internal standard. The separation was performed on Agilent Poroshell 120 EC-C₁₈ column(4.6 mm×100 mm, 2.7 μm) with gradient elution of acetonitrile(A)-0.1% formic acid solution(B) at a flow rate of 0.5 mL·min^-1. The column temperature was 30℃. All the samples were determined using positive electrospray ionization with multiple reaction minitoring mode. The ion pairs of BAG, BG and internal standard were m/z 449.5→137.1, 317.1→137.1, 391.2→179.1, respectively. RESULTS The linear ranges of BAG and BG were 0.20-10.32 μg·mL^-1(r=0.995 3, n=5), 0.02-1.51 μg·mL^-1(r=0.998 0, n=6), and the quantification limits were 8.1, 3.5 μg·mL^-1, respectively. Intra-day and inter-day precision and repeatability tests were good(RSD<10%), and BAG and BG were stable within 24 h(RSD<10%). The average recoveries(n=3) were 99.9%(RSD=1.93%) and 99.6%(RSD=3.09%), respectively. CONCLUSION The established method is accurate, simple and reproducible, which can be used in separation and determination of BAG and BG in Ophiopogonis Radix and Shenmai injection.