Berberidis cortex, the dry bark of Berberis L., is used to treat diabetes and contains at least three bioactive components: berberine (BBR), berbamine (BBM) and magnoflorine (MGF). BBR in turn is metabolized into berberrubine (BRB). Although it is possible to quantify each of these components individually in serum, there are currently no methods for simultaneously quantifying all four. Here, we developed a specific and rapid method for simultaneously quantifying BBR, BBM, MGF and BRB in mouse serum using ultra high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Samples were pretreated by protein precipitation, separated using an ACQUITY UPLC® BEH C18 column and detected by a triple quadrupole mass spectrometer with electrospray ionization. The compound [9,10-(OC2H3)2]-BBR (d6-BBR) was used as internal standard for BBR and BRB, boldine (BOL) for MGF and tetrandrine (TET) for BBM. The m/z transitions for precursor/product ion pairs were 336.1/320.2 for BBR, 305.2/566.3 for BBM, 342.0/297.1 for MGF, 322.1/307.2 for BRB, 342.2/294.3 for d6-BBR, 312.2/580.3 for TET and 328.1/265.2 for BOL. We validated our method in terms of selectivity, linearity and lower limit of quantification, accuracy, precision, matrix effect and recovery, dilution integrity and stability. This method showed good linearity from 0.1 to 40 ng/mL for BBR, 8 to 3200 ng/mL for BBM, 5 to 2000 ng/mL for MGF and 0.2 to 80 ng/mL for BRB. The chromatographic run time was 3.9 min, and sample preparation took approximately 15 min per batch. Finally, we used our method to measure BBR, BBM, MGF and BRB in serum from diabetic mice after gavage administration of BBR hydrochloride, BBM hydrochloride, and MGF. This method is precise, accurate and suitable for high-throughput sample analysis.

中文翻译：

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一种使用UPLC-MS / MS同时定量小鼠血清中小ber碱，小rb碱，木兰花碱和小ber红素的快速方法。

小ber的干燥树皮小ber皮被用于治疗糖尿病，并包含至少三种生物活性成分：小ber碱（BBR），贝巴明（BBM）和木兰花素（MGF）。BBR依次代谢为小ber红素（BRB）。尽管可以对血清中的这些成分分别进行定量，但目前尚无同时定量所有四种成分的方法。在这里，我们开发了一种特定的快速方法，可使用超高效液相色谱-串联质谱（UPLC-MS / MS）同时定量小鼠血清中的BBR，BBM，MGF和BRB。样品通过蛋白质沉淀进行预处理，使用ACQUITYUPLC®BEH C18色谱柱分离，并通过带电喷雾电离的三重四极杆质谱仪进行检测。化合物[9，10-（OC2H3）2] -BBR（d6-BBR）用作BBR和BRB的内标，丁二烯（BOL）用作MGF，粉防己碱（TET）用作BBM。BBR的前体/产物离子对的m / z跃迁为336.1 / 320.2，BBM为305.2 / 566.3，MGF为342.0 / 297.1，BRB为322.1 / 307.2，d6-BBR为342.2 / 294.3，TET为312.2 / 580.3适用于BOL的328.1 / 265.2。我们在选择性，线性和定量下限，准确性，精密度，基质效应和回收率，稀释完整性和稳定性方面验证了我们的方法。该方法显示出良好的线性，BBR为0.1至40 ng / mL，BBM为8至3200 ng / mL，MGF为5至2000 ng / mL，BRB为0.2至80 ng / mL。色谱运行时间为3.9分钟，每批样品制备大约花费15分钟。最后，我们用我们的方法来测量BBR，BBM，糖尿病小鼠血浆中的BGF盐酸盐，BBM盐酸盐和MGF灌胃后的MGF和BRB。该方法精确，准确，适用于高通量样品分析。