Determination of nucleosides and nucleobases in Isatidis Radix by HILIC-UPLC-MS/MS

Abstract

To establish an ultra-high performance liquid chromatography coupled with triple quadrupole mass (HILIC-UPLC-MS/MS) for determination of sixteen nucleosides and nucleobases in Isatidis Radix. Chromatographic separation was carried out on a Acquity UPLC BEH Amide column with gradient elution of acetonitrile (containing 0.1% acetic acid) and water (containing 0.8% acetic acid and 10 mmol L À1 ammonium acetate) at a flow rate of 0.3 mL min À1 , the column temperature was set at 35 C; Waters XevoÔ TQ worked in multiple reaction monitoring mode. Each component were separating in 11 min. All calibration curves were linear (r 2 > 0.997) over the tested ranges. The limits of detection of these compounds were 0.02-42.54 ng mL À1 . The limits of quantitation of these compounds were 0.05-98.18 ng mL À1 . The average recoveries were in the range of 93.81-105.77% with RSD value less than 7.5%. The result showed that almost all of these Isatidis Radix were rich in nucleosides and nucleobases, but their contents were obviously various. The method was simple and fast with high precision, sensitivity and repeatability, which can be used for determination of this type class of nucleosides and nucleobases in other medicinal herbs.

Recent applications of hydrophilic interaction liquid chromatography in pharmaceutical analysis

“…Nucleosides, nucleotides, and nucleobases, a group of highly active compounds, are involved in the regulation and modulation of various physiological processes in human body . As shown in Table , various natural product samples, including functional foods , medicinal plants , medicine and food dual purpose products , are containing nucleosides‐based compounds. It is clear that UAE is the common sample preparation method for compound extraction, and due to the high sensitivity and wide linear dynamic range for quantification, triple quadrupole MS (TQ‐MS) is frequently used for detection.…”

“…Taking advantage of the rapid and excellent separation performance of HILIC–ESI‐TQ‐MS/MS, nucleosides, and nucleobases in medicinal plants Cordyceps and Radix Isatidis were also analyzed. After optimization and validation, the developed method was applied for the analysis of real samples that were collected from different locations in China.…”

Simultaneous Quantification of Three Chemical Types Bioactive Compounds in Radix Isatidis and Its Relevant Pharmaceutical Dosage Forms by HPLC-DAD

“…In our previous research, it was found that the nucleosides were difficult to obtain good separation in high proportion of organic solvents because of their high and similar polarity. 5 So the proportion of organic phase slowly increased from 3 to 10% in 12 min. As shown in Figure 3, the nucleosides were completely separated with good peak symmetry.…”

“…16 Many analytical methods including high performance liquid chromatography (HPLC), 17,18 capillary zone electrophoresis (CZE) 19 and ultra-high performance liquid chromatography coupled with triple quadrupole mass have been developed for the determination of one of the three chemical types bioactive compounds (alkaloids, nucleosides and phenylpropanoids) in Radix Isatidis or its pharmaceutical dosage forms. 5,20 However, to the best of our knowledge, no method has been developed for simultaneous analysis of alkaloids, phenylpropanoids, nucleosides in Radix Isatidis or its relevant pharmaceutical dosage forms.…”

“…1 Uracil, cytidine, hypoxanthine, uridine, guanosine and adenosine are the main nucleosides in Radix Isatidis. 5 The latest studies showed that nucleosides were important bioactive compounds related to anti-inflammatory, immunoregulation, antitumor and antivirus activities. [6][7][8][9] Clemastanin B, indigoticoside A, isolariciresinol and syringin are the major phenylpropanoids isolated from Radix Isatidis.…”