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To establish an high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the simultaneous determination of 45 additives in oral liquid preparations of traditional Chinese medicine.
The samples were ultrasonically extracted by acetonitrile-methanol (9:1) (containing 0.1% formic acid). The separation was carried out on an Agilent Eclipse Plus C18 chromatographic column (150 mm×3.0 mm，1.8μm) using gradient elution of methanol -5 mmol·L-1 ammonium acetate. The compounds were scanned and detected simultaneously by electrospray ionization（ESI）ion source in both positive ion and negative ion mode under dynamic multiple reaction monitoring. The retention time and ion ratio were used for qualitative analysis and the external standard method was adopted for quantification.
The good linear relationship of peak area was observed for the 45 additives with correlation coefficients ≥0.992 in the concentration range of 5-2 000 ng·mL-1，and the limits of quantitation were between 0.2 mg·kg-1 and 4.0 mg·kg-1 under the above chromatographic and mass spectrometric conditions. The average recoveries of the blank samples at different added levels ranged from 75.4% to 118.4% with RSDs of 0.70%-9.8%. The method was used to detect 20 batches of oral liquid preparations of traditional Chinese medicines purchased from pharmacies. Benzoic acid was detected in 6 batches with the contents of benzoic acid from 0.13% to 0.27%，and sorbic acid，trans-cinnamic acid，molasses，ethyl 4-hydroxybenzoate，acesulfame potassium，dehydroacetic acid and saccharin sodium were detected in 7 batches of samples respectively.
The established high-throughput detection method is sensitive，simple and fast in pre-treatment，with high accuracy，stable recovery and reduced detection cost effectively. It can be used for the simultaneous rapid screening of multiple additives in oral liquid preparations of traditional Chinese medicine.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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