Establishment of HPLC Fingerprint and Content Determination of 5 Components in Papaveris Pericarpium / 中国药房

OBJECTIVE：To establish the fingerprint of Papaveris Pericarpium， and to determine the contents of 5 components，such as morphine ，codeine，thebaine，papaverine and narcotine. METHODS ：HPLC method was adopted. The determination was performed on a Agilent ZORBAX Eclipse XDB-C18 column with mobile phase consisted methanol -sodium heptanesulfonate with gradient elution at the flow rate of 1.0 mL/min. The detection wavelength were set at 238 nm（papaverine） and 216 nm（morphine，codeine，narcotine，thebaine）. The column temperature was 20 ℃，and sample size was 10 µL. HPLC fingerprints of 15 batches of Papaveris Pericarpium were established by using the Similarity Evaluation System of TCM Chromatographic Fingerprints （2012 edition），and the common peaks were determined in combination with the spectra of mixed control. The contents of morphine ，codeine，thebaine，papaverine and narcotine were determined simultaneously by the same method. The cluster analysis was conducted by using SPSS 19.0 software. RESULTS ：There were 13 common peaks in 15 batches of Papaveris Pericarpium ，and the similarity was greater than 0.99. Five chromatographic peaks were identified ，which were morphine，codeine，thebaine，papaverine and narcotin. The results of cluster analysis showed that 15 batches of Papaveris Pericarpium could be clustered into two categories ，S1-S7 and S 8-S15，belonging to two cities. The linear ranges of five components were 10.21-102.10，10.43-104.30，1.54-30.70，2.36-47.28，2.32-57.90 μg/mL，respectively（all r＞0.998）. RSDs of precision，stability（24 h）and repeatability tests were all less than 2％. The average recoveries were 99.46％（RSD＝1.08％，n＝ 6），97.84％（RSD＝1.55％，n＝6），91.10％（RSD＝1.74％，n＝6），96.43％（RSD＝1.25％，n＝6）、94.82％（RSD＝1.20％，n＝6）， respectively. The contents of 5 components were 2.342 9-4.082 2，0.430 4-0.889 7，0.055 2-0.090 4，0.299 3-0.558 8，0.343 2- 0.656 2 mg/g. CONCLUSIONS ：The established HPLC fingerprint and content determination method is simple ，feasible，sensitive and accurate. It combined with the cluster analysis could reflect characteristics and int ernal quality of chemical components in Papaveris Pericarpium . Papaveris Pericarpium in different cities possess regionalization characteristics ，but its quality isbasically stable.

The Pharmacokinetics of Morphine and Codeine in Human Plasma and Urine after Oral Administration of Qiangli Pipa Syrup.

Papaveris pericarpium, a natural source of morphine and codeine, is the principal active component in many antitussive traditional Chinese medicines. We herein report the first PK study of papaveris pericarpium in human plasma and urine following oral administration of single (15, 30, 60 mL) and multiple dose (15 mL) of Qiangli Pipa Syrup (MOR 0.1 mg/mL, COD 0.028 mg/mL) by monitoring morphine and codeine using a HPLC-MS/MS method. Their Tmax and t1/2 values are independent of dosages, while the AUC0-t linearly increased with higher dosages, indicating linear PK characteristics. AUC0-t increased obviously after multiple doses, indicating possible risk of accumulative toxicity. Urine studies suggested risks of positive opiate drug tests with a cutoff of 300 ng/mL, which lasted 6-14 h at different doses. These results provide important information for clinical safety, efficacy and rational drug use of Qiangli Pipa Syrup and also guide the related judicial expertise of its administration.

Improvement and Optimization of Quality Standard for Qiangli Pipa Distillate Formula / 中国药师

Objective: To improve the quality standard for Qiangli Pipa distillate formula. Methods: The chemical reactions for i-dentification in the old standard were deleted. TLC methods were established to identify Papaveris pericarpium, Eriobotryae folium and Platycodonis radix. The contents of morphine and codeine phosphate from Papaveris pericarpium were determined by HPLC. The chro-matographic conditions for morphine content determination were as follows: an Inertsil ODS-3 chromatographic column (250 mm × 4. 6 mm, 5 μ m ) was used, the mobile phase was acetonitrile-the mixture of 0. 01 mol·L-1sodium heptane sulfonate and 0. 02 mol ·L-1potassium dihydrogen phosphate with equal amount (10% phosphoric acid was used to adjust pH to 2. 8) ( 13: 87 ), the detec-tion wavelength was 220 nm ,the column temperature was 30 ℃, and the flow rate was 1. 0 ml·min-1. The chromatographic condi-tions for codeine phosphate content determination were as follows : an Inertsil ODS-3 chromatographic column (250 mm×4. 6 mm, 5 μ m ) was used, the mobile phase was acetonitrile-the mixture of 0. 01 mol·L-1sodium heptane sulfonate and 0. 02 mol·L-1s potas-sium dihydrogen phosphate with equal amount (10% phosphoric acid was used to adjust pH to 2. 8) ( 15: 85 ), the detection wave-length was 220 nm, the column temperature was 30 ℃, and the flow rate was 1. 0 ml·min-1. Results: TLC had obvious characteris-tics with clear and well-separated spots. Morphine showed a good linear relationship within the range of 3. 14-62. 8 μg·ml-1( r=1. 000 0) with the average recovery of 96. 69% (RSD=2. 41% , n=9). Codeine phosphate showed a good linear relationship within the range of 3. 52-87. 94 μg·ml-1(r=1. 000 0) with the average recovery of 95. 68% (RSD=2. 64% , n=9). Conclusion: The quality standard for Qiangli Pipa distillate formula is improved. The methods are easy-operated and accurate with good specificity, which are suitable for the quality control of Qiangli Pipa distillate formula.

Identification of major bioactive components and their metabolites in rat plasma after oral administration of Zhikebao tablet by UHPLC-QTOF-MS / 药学学报

An ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) method was established to identify the metabolites in rat plasma after oral administration of Zhikebao tablets. The high-resolution mass spectrometer was operated in positive and negative ion mode, respectively. First, full-scan was applied, which was dependent on a multiple mass defect filter (MMDF) combined with dynamic background subtraction (DBS). These were utilized to trigger the information dependent acquisition (IDA) function in the experiment. For the IDA criteria, the eight most intense candidate ions of per cycle were selected to do a product ion scan. Then Metabolite Pilot 2.0 software was utilized to load data to seek possible metabolites. The analytical models employed by Metabolite pilot 2.0 were established for representative compounds of the Papaveris Pericarpium and licorice in Zhikebao tablet. Finally, metabolites were identified according to accurate mass measurement and retention time. 38 components from the rat plasma after oral administration of the drug have been found, including 5 prototype opium alkaloids, liquiritin, glycyrrhizic acid and 31 relative metabolites. The metabolic transformation of Zhikebao tablet in rats was mainly induced by glucuronidation, sulfation, methylation, amine to carboxylic acid, hydrolysis and so on. In this paper, the metabolites of the main active components of Zhikebao tablet were tentatively identified, and the metabolic pathway was compared with that of single chemical drugs. Moreover, it laid the fundamental elucidation of further metabolism study of Zhikebao tablet or other compound traditional Chinese medicine preparations which containing Papaveris Pericarpium or licorice.