To compare the in vitro bioactivity between a generic lactobacillin granules drug and three commercial lactobacillin granules drugs, this study investigated inhibition on pathogenic bacteria, the growth promoting effect on probiotics, and establish a method to evaluate the in vitro bioactivity consistency of drugs that regulate gut microbiota.

Two culture systems were set up to investigate the inhibitory effect on pathogenic bacteria and the growth promoting effect on probiotics by lactobacillin granules. The in vitro bioactivity consistency of four products was evaluated by microbial growth curve and analyzed by two-way analysis of variance with Dunnett-t test.

No significant difference (P>0.05) was observed on inhibition of Staphylococcus aureus and growth promotion of Lactobacillus rhamnosus between the generic lactobacillin granules and the commercial lactobacillin granules.

This method could be used to evaluate the in vitro bioactivity of drugs that regulate gut microbiota, and provided guidance on relevant drug development and quality evaluation.

To establish an inductively coupled plasma mass spectrometry (ICP-MS) method for the determination of 25 potentially risky elemental impurities in alfentanil hydrochloride injection.

The Agilent 7800 ICP-MS inductively coupled plasma mass spectrometer was used, the conventional tuning mode was used, the RF power was 1 550 W, the plasma gas flow rate was 15 L·min^-1, the matrix effect was eliminated by external standard, and the sample was diluted for direct injection.

The method can simultaneously determine the content of 25 kinds of elemental impurities, and its linear relationship was good (r >0.99). The RSD of the repeatability test was ≤10% (n=6), and the recovery rate was 80.0%-120.0% (n=9), all of which met the requirements of methodological validation.

The content of elemental impurities in alfentanil hydrochloride injection is lower than 30% of the limit specified in ICH·Q3D, which will not bring safety risks to the drug, providing a reference for the quality control and risk assessment of elemental impurities in other similar drugs.

To establish an HPLC method for the determination of potentially genotoxic impurity E, impurity I, and 2-chloromethyl-4-methoxy-3，5-dimethlpyridine in esomeprazole sodium.

The chromatographic conditions were as follows: impurity E, YMC-Triart C₁₈ column (250 mm×4.6 mm, 5 μm), mobile phase A 0.05 mol·L^-1 monopotassium phosphate buffer, mobile phase B was acetonitrile, the gradient elution program was used at the flow rate of 1.0 mL·min^-1, the detection wavelength was 302 nm and the column temperature was 30 ℃. Impurities I, Agilent Microspher C₁₈ column (100 mm×4.6 mm, 3 μm), mobile phase A was water-phosphate buffer (pH 7.6)-acetonitrile(80∶10∶10), mobile phase B was acetonitrile-phosphate buffer (pH 7.6)-water (80∶1∶19), the flow rate was 1.0 mL·min^-1, the detection wavelength was 302 nm and the column temperature was 30 ℃. 2-chloromethyl-4-methoxy-3，5-dimethlpyridine, GL Inertsil ODS-3 column (250 mm×4.6 mm, 5 μm), mobile phase A was 0.01 mol·L^-1 disodium phosphate solution (pH 6.5), mobile phase B was acetonitrile, the flow rate was 1.0 mL·min^-1, the detection wavelength was 265 nm and the column temperature was 30 ℃.

The linear ranges of impurity E, impurity I and 2-chloromethyl-4-methoxy-3，5-dimethlpyridine were 0.025 1-0.200 7, 0.020 2-0.302 7, 0.126 6-2.110 0 μg·mL^-1. The LOQ of impurity E, impurity I, 2-chloromethyl-4-methoxy-3，5-dimethlpyridine were 0.50, 0.40, 2.53 ng, and the LOD were 0.15, 0.12, 0.84 ng. The average recovery rate ranged from 96% to 104%, and the RSD was less than 2%. No potential toxic impurities were detected in the samples.

The method has the advantages of good repeatability, high precision, high accuracy and good linearity, and the analysis method are simple and efficient.

To establish and verify the helium mass spectrometry detection method to test the sealing integrity of the container closure system (CCS) of pressurized metered dose aerosols (pMDIs).

Determined the maximum allowable leakage limit (MALL) of pMDIs of CCS in the whole life cycle. The special test chambers and helium filling devices were designed and manufactured, and helium filling devices to test the negative and positive control samples under high pressure (absolute pressure 672 kPa) and normal pressure (absolute pressure 100 kPa) respectively. According to the requirements of methodology validation, the validation of detection limit, system suitability, precision, specificity and detection range indicators were completed.The accelerated sample sealing integrity testing for 3, 6 months and 3, 12, 24 months in the long-term stability inspection period was achieved.

Under both helium-filled pressure conditions, the methodological validation indicators were in line with the acceptable standards, and the results of the stability samples were less than the detection limits. This method can detect 100% that the equivalent pore diameter of pMDIs product CCS was 0.095 μm and above.

The helium mass spectrometry detection method can quickly and quantitatively investigate the leakage rate of pMDIs products, prove the seal integrity of CCS with high sensitivity, and meet the industry’s requirements for pMDIs product quality control.

To establish a high-performance liquid chromatography (HPLC) method for investigating the extraction of common antioxidants and extractable sulfur in medical rubber stoppers and the migration of antioxidants and extractable sulfur to propofol medium/long chain fat emulsion injection.

Waters Symmetry RP₁₈(250 mm×4.6 mm，5 μm) was used as chromatographic column with methanol-acetonitrile -1% acetic acid solution as the mobile phase. Detection wavelength, velocity of flow and column temperature were set to 277 nm, 1 mL·min^-1 and 35 ℃.

Good resolution and linear relationship (r≥0.999 6) in the range of 0.1-20 μg·mL^-1 were achieved. This method possesses superior precision, stability, repeatability, and all RSD were less than 5%. The rate of recovery was 93.3%-108.7%, and the RSD was 1.8%-12.5%. The migration of antioxidant BHT was detected in three batches of solution and the content was higher than its corresponding permitted daily exposure (PDE) value, which means there is a large safetyrisk.

The method represents high sensitivity and is easy to operate, which can effectively detect the migration of antioxidants in propofol medium/long chain fat emulsion injection.

To develop an inductively coupled plasma mass spectrometry (ICP-MS) matrix matching method for determination of migration of 11 elements in ketorolac tromethamine injection, including Al, As, B, Ca, Cd, Fe, Mn, Pb, Sb, Si, Ti.

Samples were diluted and capacitated with 1% nitric acid after precipitation with nitric acid. 2% ethanol was added into the standard solution as the matrix. The matrix effect was eliminated by the matrix matching method, and the polyatomic ion interferences were eliminated by the hydrogen collision reaction mode and the helium collision mode.

The linearity of 11 elements was good (r≥0.997 0). The limits of detection were 0. 049-133 ng·mL^-1. The average recoveries of all 11 elements were in the range of 83.8%-107.1%, and the RSD of repeatability was 2.6%-11.0%. Three batches of ketorolac tromethamine injection under acceleration were tested. The overall safety risk was low.

The established matrix matching ICP-MS method is simple, rapid and accurate. It can effectively eliminate the matrix effect and be used for the determination of eleven elemental impurities in ketorolac tromethamine injection, providing technical reference for risk assessment of drug packaging material compatibility.

To determine the polysaccharide hydrolysates from Polygonati Rhizoma under glycoside enzymatic hydrolysis’ HPLC fingerprint. Examine the variations among the polysaccharides produced by the various varieties of Polygonati Rhizoma and provide references for the assessment of the polysaccharide quality.

After the polysaccharide from Polygonati Rhizoma was hydrolyzed by fructose enzymes, its fingerprint was established by HPLC-ELSD. The fingerprint was then analyzed using similarity analysis (SA), hierarchical cluster analysis (HCA), and principal component analysis (PCA) to determine the differences between the polysaccharides from various Polygonati Rhizoma varieties.

Polysaccharides from various strains of Polygonati Rhizoma had different HPLC-ELSD fingerprints, and a total of 17 distinct oligosaccharide fragments were discovered, all of which contained fructose, glucose, and sucrose. Analysis revealed that there are significant intra species differences, minor differences, and high levels of similarity between the three types of Polygonati Rhizoma. The real Polygonati Rhizoma differs significantly from the imitation in several important ways.

Polygonati Rhizoma can be successfully classified according to varieties, using the fingerprint of polysaccharide hydrolysates of the various varieties of Polygonati Rhizoma and its adulterants. The developed HPLC method can be used for the differential analysis of polysaccharides in Polygonati Rhizoma and is straightforward, precise, and repeatable.

To study the preparation and quality control of 3,5-dihydroxy-7,4’-dimethoxyflavone candidate chemical reference substance from Zhuang medicine Amomum paratsaoko S. Q. Tong & Y. M. Xia.

3,5-dihydroxy-7,4’-dimethoxyflavone was separated and purified consecutively by silica gel, recrystallization as well as preparative HPLC. The structure of 3,5-dihydroxy-7,4’-dimethoxyflavone was identified by IR, UV, NMR and other comprehensive spectrum analytical methods with MS spectrum. Its purity was determined by HPLC and TLC, the ash content was determined by incandescent residue method, and the content of 3,5-dihydroxy-7,4’-dimethoxyflavone was calculated by mass balance method. The method for quality analysis of 3,5-dihydroxy-7,4’-dimethoxyflavone candidate reference materials was investigated by HPLC.

The mean value determined by HPLC of 3,5-dihydroxy-7,4’-dimethoxyflavone was 98.85%, the ash value was 0.02%, and the content of 3,5-dihydroxy-7,4’-dimethoxyflavone calculated by mass balance method was 99.83%. The established analytical method was specific and precise.

3,5-Dihydroxy-7,4’-dimethoxyflavone which was prepared in this study met the quality standard requirements of chemical reference substance, and could be used as a reference substance for quality control of medicinal materials and their preparations such as Amomum paratsaoko S. Q. Tong & Y. M. Xia and others. The analysis method was accurate and reliable, which provided a scientific basis for the research and formulation of quality standard of this reference substance.

To evaluate the quality of honeyed Eriobotryae Folium from different habitats and to select the best habitat of honeyed Eriobotryae Folium preferentially based on high performance liquid chromatography fingerprinting and chemical pattern recognition methods.

The detection was performed on an Acclaim^TM 120A C₁₈ (250 mm×4.6 mm, 5 μm) column with the mobile phase of 0.2% aqueous phosphoric acid (A)-acetonitrile (B) in gradient elution (0-5 min, 5%B; 5-6 min, 5%B→10%B; 6-20 min, 10%B; 20-50 min, 10%B→25%B; 50-60 min, 25%B). The volume flow rate was 1.0 mL·min^-1, the detection wavelength was 327 nm, the column temperature was 30 ℃, and the injection volume was 10 μL. The fingerprint profiles of 30 batches of honeyed Eriobotryae Folium from different habitats were established, and the fingerprint profiles combined with chemical pattern recognition were used to conduct comprehensive analysis of honeyed Eriobotryae Folium from different habitats. And cluster analysis(CA), principal component analysis (PCA) and comprehensive scoring were performed on honeyed Eriobotryae Folium from different habitats. Orthogonal partial least squares-discriminant analysis(OPLS-DA) was used to screen out the differential markers of honeyed Eriobotryae Folium from different habitats, and the habitats of honeyed Eriobotryae Folium were selected based on the comprehensive scoring.

The fingerprint profiles of 30 batches of honeyed Eriobotryae Folium were established. Twelve common peaks were identified, and 4 peaks were identified as neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid and auriculoside according to the control finger. CA divided the 30 batches of Honeyed Eriobotryae Folium samples into 6 categories. By PCA, 3 principal components were extracted, with a cumulative variance contribution of 84.315%. Six differential markers were obtained according to OPLS-DA, two of which were identified as chrysoside and chlorogenic acid. The better habitats of honeyed Eriobotryae Folium were screened as Sichuan, Guangxi, Guangdong and Shaanxi according to the comprehensive score.

Good precision, repeatability and stability results are obtained for fingerprinting and content determination. The combination of fingerprinting and chemical pattern recognition can comprehensively evaluate the quality of honeyed Eriobotryae Folium, and this method is stable and reliable, which can provide an effective reference basis for the habitat study of honeyed Eriobotryae Folium.

To improve the liquid chromatographic determination method of cefixime granules related substance.

High performance liquid chromatography was used, YMC-Triart C₁₈ column (250 mm×4.6 mm, 5 μm) was selected, 0.05 mol·L^-1 ammonium formate solution (pH 4.7)-methanol was used as mobile phase, flow rate was 1 mL·min^-1, and gradient washing was carried out.The injection volume was 10 μL. The detection wavelength was 254 nm.

This chromatographic condition was applied to the detection of cefixime granules. The differences between this method, the pharmacopeial method and the method of USP PF 2018 were compared, and the systematic methodological verification of specificity, linearity, accuracy, precision and durability were completed. Using pharmacopeial methods, baseline separation of degradation impurities A1~A4 or impurities B1~B4 cannot be reached, and current methods cannot be used to determine polymer B and polymer D. The method proposed in this article can make the resolution between cefixime and each specific impurities meet the requirements (R ≥1.5), and can detect and quantify polymer B and polymer D at the same time, and the resolution was better than the current method.

This method improves the separation between cefixime and impurities, more impurities is detected and can accurate quantify specific impurities. This method has high sensitivity and good repeatability, and is suitable for the quality control of cefixime.