Simultaneous determination of five alkaloids in extracted and non-extracted poppy APIs by HPLC.

Background: Poppy Active Pharmaceutical Ingredients (APIs) are mainly used in the production of antitussives, analgesics, and other preparations. The alkaloid components play a major role, but the use of some alkaloids can cause dependence and toxic side effects in patients.

Objective: The main aim of this study was to develop an effective and simple high-performance liquid chromatography (HPLC) for the simultaneous determination of five alkaloids in three APIs.

Method: Waters XBridge C18 column (4.6 mm × 150 mm, 3.5 μm) was used as the chromatographic column. Mobile phase A was composed of 5.5 mmol/L sodium heptane sulfonate solution-acetonitrile-methanol-phosphoric acid (83:7:10:0.22, v/v/v/v), and Mobile phase B was composed of 5.5 mmol/L sodium heptane sulfonate solution-acetonitrile-methanol-phosphoric acid (60:15:25:0.45, v/v/v/v). The flow rate was 1.2 mL/min; The detection wavelength was 230 nm; The column temperature was 40 °C.

Results: The HPLC method was suitable for the simultaneous determination of the contents of five alkaloids in three APIs. The verification experiment showed that the linear relationship of the five alkaloids in their respective ranges was good (R2 ≥ 0.998); The average recoveries ranged from 87.62 to 103.18%; The RSDs of repeatabilities were 0.3∼2.7%.

Conclusions: In this study, a new HPLC method was developed and successfully validated. Compared with the determination method of opium alkaloids specified in Pharmacopoeia, this HPLC method does not need a complex sample pretreatment process, improves detection efficiency, avoids the use of highly corrosive reagents, and reduces the experimental risk. It can simultaneously meet the determination of five alkaloids in three APIs.

Highlights: This method can simultaneously quantify five alkaloids in three poppy APIs without a complex sample pretreatment process and the application of strong corrosive solvents. It is applied to these APIs for the first time, expanding the scope of alkaloid analysis and providing new data for comparative studies of different API sources.