To establish an HPLC-CAD method for the determination of related substances in peramivir injection and to analyze the impurity profile.

Welch Ultimate Polar RP column (250 mm×4.6 mm, 3.5 μm) was used with 5 mmol·L^-1 ammonium formate solution (pH adjusted to 4.6 with formic acid) -acetonitrile (95∶5) as mobile phase A and mobile phase A-acetonitrile (50∶50) as mobile phase B under gradient elution at the flow rate of 0.8 mL·min^-1. Column temperature was 30 ℃. CAD atomizer temperature was 50 ℃, acquisition frequency was 5 Hz and filter was 3.6 s. The degradation pathway and impurity structure of peramivir were estimated by LC-MS/MS.

The separation between peramivir and the impurities was good. The mass concentration of peramivir and 7 impurities showed good linear relationships with the peak area in the ranges of 0.2-12 μg·mL^-1. The limits of quantitation were 4.32-12.8 ng, and the limits of detection were 2.16-6.44 ng. The average recovery (n=9) of impurity Ⅲ was 96.2%, RSD was 1.8%. The control solution and the test product solution were stable for 102 hours at room temperature. After fine-tuning the parameters of liquid chromatography, there was no influence on the detection results of related substances. Based on LC-MS/MS, the structure and possible degradation pathway of the degraded related substances were deduced.

This method is accurate, reliable and specific for the determination of related substances in peramivir injection, and provides a basis for the establishment of quality standards for this variety.

To determine the contents of 28 elements in Xianzhuli under processes of dry distillation and fire preparation by inductively coupled plasma mass spectrometry (ICP-MS), and evaluate the rationality of the current temperature used in the dry distillation process as well as whether the preparation process of commercial samples conforms to the traditional or modern one.

The samples of Xianzhuli were pre-treated with nitric acid before microwave digestion. The contents of 28 elements in 59 batches of Xianzhuli samples were determined by inductively coupled plasma mass spectrometry(ICP-MS), and the methodology was investigated.

The standard curve of 28 elements had a good linear relationship with r≥0.999 2. The detection limits were 0.007 1-1.249 5 ng·mL^-1, the RSDs for precision, repeatability and repeatability tests were 0.30%-3.5%, 0.69%-6.4% and 1.1%-3.3%, respectively and the recovery rates were 88.6%-105.5% with RSDs ranged from 1.0% to 3.2%. The contents of Na, Mg, K, Ca, Mn, Fe, Zn and Rb in 59 batches of Xianzhuli were high, and the contents of heavy metal elements did not exceed the limit requirements. The results of cluster analysis showed that the samples prepared by dry distillation and fire preparation were clustered into one class. By principal component analysis, 7 principal components were obtained, and the cumulative variance rate was 75.6%. Mg, K, Ca, Cr, Fe, Co, Cu, Rb, Cd, Ba, Tl and Pb were identified as the characteristic elements of Xianzhuli.

Based on the contents of beneficial and harmful elements, it is found that the contents of toxic and harmful elements in the sample processed with 160 ℃ dry distillation are lower than processed with fire preparation. And the contents of beneficial elements are similar in two kinds of samples. This temperature is consistent with the dry distillation process temperature approved by provincial drug regulatory authorities for currently certified enterprises. Most of the commercially available samples meet the requirements of traditional or modern processes, and a few may have quality differences due to different processes. The determination and analysis of heavy metals and other elements in Xianzhuli under different processes can provided scientific basis for safe production and rational use of Xianzhuli.

To establish the HPLC fingerprint and multi-component quantitative analysis of Hanchuang Zupa granules, and evaluate the quality of multiple batches of Hanchuang Zupa granules by chemical pattern recognition technology.

The sample pretreatment conditions and chromatographic analysis conditions of Hanchuang Zupa granules were optimized, and the optimal HPLC fingerprint and multi-component quantitative analysis method were established as follows: stationary phase was YMC-Pack ODS-A column (250 mm×4.6 mm, 5 μm, 12 nm) was adopted, and the mobile phase was acetonitrile-water (containing 0.1% phosphoric acid) with gradient elution, the detection wavelength was 220 nm, the column temperature was 30 ℃, the flow rate was 1.0 mL·min^-1. Hierarchical cluster analysis (HCA), principal components analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were applied to evaluate the quality of 17 batches of Hanchuang Zupa granules.

Methodological investigation of HPLC fingerprint and content determination were well verified and met the analysis requirements. A total of 25 common peaks were obtained by full peak matching, and eight of them were identified by comparing with the retention time of mixed reference substances. The similarities of 17 batches of samples were above 0.90, which showed good consistency and stability between the samples. Seventeen samples could be classified into three clusters. Three principal components from 21 common peaks were extracted by PCA. Six quality differential compounds were presented in the fingerprints by OPLS-DA, including rutoside, gallic acid, ammonium glycyrrhizinate, chlorogenic acid and so on. The resolution and linear relationship of eight components in quantitative analysis were good. The average recovery rates were 98.0%-99.1% with RSD≤2.0%.

In this study, the qualitative analysis of HPLC fingerprint and quantitative analysis of multiple index components is specific, simple and accurate, which can provide a reference for the quality control and quality evaluation of Hanchuang Zupa granules.

To identify the authenticity of the Longgu, and reveal the scientific illustration for superior quality of white Longgu.

In this study, the quality of 88 batches of Longgu was evaluated by cluster analysis, principal component analysis and correlation analysis based on character, ultraviolet fluorescence and results obtained with inductively coupled plasma mass spectrometry, color analyzer and nuclear radiation detector.

The Longgu spots on the surface of Longgu were the key identification points of genuine Longgu, and the Longgu spots were mainly composed of Mn elements. Bright blue fluorescence could be seen in the fresh section of keel under 365 nm. Among the 42 elements determined, only Mg and Zn were higher in the fake Longgu than in the genuine Longgu. Other elements were lower in the fake Longgu than in the genuine Longgu. Heavy metals and harmful elements in counterfeit samples were below limits in pharmacopoeia, and cluster analysis showed that all counterfeit samples were grouped into one group separately. The Longgu could be divided into four color types: white, cyan, yellow and brown. The larger a * was, the redder the color of Longgu was, the higher the contents of Fe and harmful elements (As, Cu, Pb and Cd) were. And the contents of element V were strongly correlated with those of As, Cu, Pb and U, which could be used as index element of heavy metals and harmful elements. The contents of radioactive element U and the radiation values were highest in brown Longgu, second highest in yellow Longgu was the, lower in cyan Longgu, and lowest in white Longgu. The average content of U in brown Longgu was 0.021% with the highest content of 0.09%, and the average radiation value was 0.33 μSv·h^-1 (Background value was 0.18 μSv·h^-1) with the highest value of 0.47 μSv·h^-1. The average content of U in yellow Longgu was 0.012% with the highest content of 0.021%, and the average radiation value was 0.32 μSv·h^-1 with the highest value of 0.39 μSv·h^-1. The average content of U in cyan Longgu was 0.008 9% with the highest content of 0.012%, and the average radiation value was 0.30 μSv·h^-1 with the highest value of 0.35 μSv·h^-1. The average content of U in white Longgu was 0.006 7% with the highest content of 0.009 1%, and the average radiation value was 0.28 μSv·h^-1 with the highest value of 0.32 μSv·h^-1.

Considering the harmful elements and radiation safety, the white Longgu has better quality, and the brown Longgu has worse quality.

To establish a quantitative analysis of multi-components by single marker (QAMS) method to simultaneously determine the contents of kalangin, diphenylheptane A, galangin, kaempferin and galangin-3-O-methyl ether.

Hypersil ODS2 C₁₈ (250 mm×4.6 mm, 5 μm) was used by high performance liquid chromatography (HPLC) and was eluted with 0.3% acetic acid in water (A) and acetonitrile (B) (0-2 min, 10%B→15%B；2-5 min, 15%B→20%B；5-20 min, 20%B→35%B；20-30 min, 35%B→40%B；30-55 min, 40%B；55-60 min, 40%B→100%B), the column temperature was 30 ℃, the volume flow rate was 0.8 mL·min^-1, and the detection wavelength was 254 nm. The relative correction factors of each chemical component were calculated with gingerin as the internal reference, and the content was determined, which was compared with the results of the external standard method, and the quality of galangal from different origins was evaluated by chemical pattern recognition.

Kaempferol, diphenylheptane A, galangin, kaempferin and galangin-3-O-methyl ether had a good linear relationship in the ranges of 0.005 5-0.110 0, 0.140 0-2.800 0, 0.149 6-2.992 0, 0.021 5-0.430 0 and 0.022 2-0.444 0 μg, respectively, and the average sample recoveries were 100.7%, 101.4%, 99.9%, 100.9% and 101.7%, respectively. The RSDs were 1.3%, 2.8%, 0.83%, 1.4% and 1.8%, respectively. Quantitative analysis of multicomponents by single marker method and external standard method was no significant difference in the results.

This method is rapid, accurate and specific, which can provide a reference for the quality control of galangal.

To establish a method for the simultaneous determination of six components (rutin, naringin, neohesperidin, quercetin, purpurin and mollugin) in Huazhi tablets and Huazhi capsules by HPLC.

The assay was performed on an Agilent ZORBAX SB-Aq (150 mm×4.6 mm, 5 μm) and the sample was eluted by mobile phase consisting of acetonitrile(A) -0.1% phosphoric acid(B) with a gradient at a flow rate of 1.0 mL·min^-1 and the column temperature was 30 ℃. The detection wavelengths were set at 250 nm for rutin during 0-20 min, 283 nm for naringin and neohesperidin during 20-37 min, and 250 nm for quercetin, purpurin and mollugin during 37-60 min.

Rutin, naringin, neohesperidin, quercetin, purpurin and mollugin exhibited good linearity(r>0.999 0) in the ranges of 37.26-1 863.20 μg·mL^-1, 11.27-563.70 μg·mL^-1, 9.58-479.04 μg·mL^-1, 1.92-95.90 μg·mL^-1, 0.52-25.83 μg·mL^-1 and 0.90-45.10 μg·mL^-1, respectively. The average recoveries of the above mentioned six components in Huazhi tablets (n=6) were 103.2%(RSD=1.4%), 99.5%(RSD=1.7%), 97.7%(RSD=1.2%), 95.2%(RSD=1.1%), 104.2% (RSD=1.2%) and 104.2%(RSD=0.80%), respectively, and the average recoveries of the above mentioned six components in Huazhi capsules (n=6) were 102.5% (RSD=1.3%), 96.9%(RSD=0.48%), 97.1%(RSD=1.1%), 96.9%(RSD=0.78%), 102.3%(RSD=1.4%) and 101.8%(RSD=1.2%), respectively. The content ranges (mean ± SD) of rutin, naringin, neohesperidin, quercetin, purpurin and mollugin in the 17 batches of Huazhi tablets and Huazhi capsules were 31.14-98.25(44.33±15.65) mg·g^-1, 5.12-20.85(12.96±5.85) mg·g^-1, 4.03-17.00(10.14±5.17) mg·g^-1, 0.63-7.17(1.97±1.49) mg·g^-1, 0.23-1.32(0.57±0.28) mg·g^-1 and 0.67-1.72(1.08±0.29) mg·g^-1, respectively.

The established method can simultaneously determine six components in Huazhi tablets and Huazhi capsules and can provide a reference for the quality control of Huazhi tablets and Huazhi capsules.

To establish a quality evaluation method of Erdi Xiaoke mixture based on HPLC fingerprint, multi-component determination and chemometric analysis.

The separation was performed on a 30 ℃ thermostatic Acutfex PW-C₁₈ (250 mm×4.6 mm, 5 μm) column, with the mobile phase comprising of acetonitrile -0.1% phosphoric acid flowing at 1.0 mL·min^-1 in a gradient elution manner, and the detection wavelength was set at 285 nm to establish HPLC fingerprint of Erdi Xiaoke mixture. Contents of 11 components [5-hydroxymethyl furfural(5-HMF), salvianic acid A, chlorogenic acid, caffeic acid, calycosin-7-glucoside, acteoside, isochlorogenic acid B, isochlorogenic acid C, rosmarinic acid, salvianolic acid B and formononetin] in 12 batches of Erdi Xiaoke mixture were determined, and similarity evaluation, cluster analysis, principal component analysis and orthogonal partial least squares discriminant analysis were used to comprehensively evaluate the quality of Erdi Xiaoke mixture.

The 11 constituents showed good linear relationships within their respective ranges(r≥0.999 6), with average recovery rates of 95.3%-101.5%, and the RSDs were in the range of 1.1%-2.3%. There were 18 common peaks in the fingerprints for 12 batches of samples with the similarities above 0.960. The range of the content of the above 11 components in the 12 batches of samples were as follows as 43.40-65.70, 219.43-274.70, 119.00-164.78, 11.87-18.17, 42.80-56.86, 10.01-20.71, 8.21-14.73, 40.15-57.39, 43.29-60.29, 755.22-1 208.50, 2.47-4.44 μg·mL^-1. And 12 batches of Erdi Xiaoke mixture could be clustered into three categories with some differences in sample quality among different batches. In addition, 5-HMF, isochlorogenic acid B and chlorogenic acid were the main potential markers affecting the quality of Erdi Xiaoke mixture.

This method is stable, reliable, accurate and sensitive, and can be used to control and comprehensively evaluate the quality of Erdi Xiaoke mixture.

To develop a flow-through cell method for the dissolution test of omega-3-acid ethyl ester 90 soft capsules and compare the dissolution behaviors from different manufacturers.

The medium (surfactant and its concentration, pH, dosage of pepsin), flow rate and system mode (closed versus open) were investigated. The samples were collected at the specified time and determined by HPLC. The similarity of the dissolution curves between generic drugs and reference listed drug was evaluated by similarity factor (f₂).

A closed-loop mode of flow-through cell apparatus was employed, with 0.01 mol·L^-1 hydrochloric acid solution containing 4.0% Triton X-100 as the dissolution medium, and the flow rate was 2.0 mL·min^-1. The dissolution curves of the samples that have passed consistency evaluation are similar to those of the reference and the samples produced by enterprise in the declaration stage are partly similar. The method has effective distinguish ability for product quality and different prescriptions.

The newly established method can be used for the quality control of omega-3-acid ethyl ester 90 soft capsules, and can provide references for further consistency evaluation and the dissolution method development of lipid-filled soft gelatin capsule (SGC).