To establish an HPLC method for the determination of related substances in nifedipine.

HPLC was adopted on a PFP column (250 mm×4.6 mm, 5 μm) with a gradient elution system of 20 mmol·L^-1 potassium dihydrogen phosphate solution and methanol, the flow rate was 1.0 mL·min^-1, and the column temperature was maintained at 30 ℃. The detection wavelength was set at 265 nm.

The resolutions were good between the peaks of nifedipine and ten known impurities, including impurity D, 2-nitrobenzaldehyde, monoamide, hydroxy dehydro lactone, impurity C, dehydro-N-oxide, impurity A, impurity B, m-nifedipine, p-nifedi-pine. The resolutions between the known impurity peaks were not less than 1.5, the resolutions between the main peak of nifedipine and it’s front and back impurity peaks were not less than 2.0. The calibration curves of mass concentration of above known impurities were linear respectively in their concentration range of 0.000 2-0.015 mg·mL^-1(r>0.999, n=7). The correlation coefficients of above known impurities were 1.000, 1.000, 1.000, 1.000, 0.999 9, 0.999 9, 0.999 9, 1.000, 0.999 9, 0.999 9, respectively. The average recovery rates of above known impurities were 93.1%(RSD=2.3%), 110.6%(RSD=1.9%), 109.2%(RSD=2.0%), 111.0%(RSD=2.1%), 108.1%(RSD=1.9%), 112.4%(RSD=1.8%), 110.8%(RSD=1.9%), 91.5%(RSD=3.1%), 98.9%(RSD=2.7%), 110.1%(RSD=2.6%), respectively. The detection limit of above known impurities was 0.000 06 mg·mL^-1, the quantification limit of above known impurities was 0.000 2 mg·mL^-1. The impurity determination results of the three batches of nifedipine samples showed that the content of the known impurities and the maximum single unknown impurity were less than 0.1%, the total impurities contents were less than 0.5%.

The method has good sensitivity and specificity, and it is suitable for the quality control of nifedipine.

To investigate different batches of Runbi Tongqiao drops by fingerprint and multi-index component content determination method, and to provide basis for its quality evaluation.

Acclaim TM-C₁₈ chromatographic column(250 mm×4.6 mm, 5 μm) was used with methanol(A)-acetonitrile(B) -0.2% formic acid aqueous solution(C) as the mobile phase with gradient elution. The flow rate was 0.8-1.0 mL·min^-1. The injection volume was 10 μL and the detection wavelength was 327 nm. The HPLC fingerprint of Runbi Tongqiao drops was established, and the quality consistency of Runbi Tongqiao drops was compared by stoichiometric method. The contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, luteoloside and isochlorogenic acid C were determined by HPLC.

A total of 14 common peaks were calibrated in the fingerprints of 10 batches of samples, and the similarities were ≥0.933. Seven common peaks of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, luteoloside and isochlorogenic acid C were identified by reference substances. The contents of seven components in Runbi Tongqiao drops were determined simultaneously, which were 0.131 5-0.133 8 mg·mL^-1, 0.095 5-0.098 9 mg·mL^-1, 0.087 4-0.090 1 mg·mL^-1, 0.012 8-0.015 8 mg·mL^-1, 0.010 3-0.013 7 mg·mL^-1, 0.022 7-0.024 9 mg·mL^-1 and 0.006 8-0.008 6 mg·mL^-1, respectively.

The established fingerprint of Runbi Tongqiao drops is stable and reliable, and the simultaneous determination method of multi-component content is simple and fast. It can be used for the quality control of Runbi Tongqiao drops, which lays a solid foundation for the follow-up study of Runbi Tongqiao drops.

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 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).

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 study the related substances in salbutamol sulfate inhalation aerosol during national drug sampling and testing, and to compare the impurity content and evaluate the quality between samples from various enterprises.

HPLC external standard method was utilized to simultaneously determine the content of related substances A, B, C, D, E, F, G, H, I, J, N, and other unknown impurities in salbutamol sulfate inhalation aerosol. Thermo Synchronis C₈ chromatography column (250 mm×4.6 mm, 5 μm) was used. Sodium heptane sulfonate solution-acetonitrile was used as the mobile phase. Linear gradient elution was performed and the flow rate was 1.0 mL·min^-1. Column temperature was 40 ℃ and detection wavelength was 220 nm. Injection volume was 20 μL. The sources of impurities through forced degradation experiments were explored. Toxicity prediction software for impurity toxicity assessment was applied.

After method validation, the specificity of the method was good, and the separation degree between each impurity peak was greater than 1.5. RSDs of precision test were 0.30%-1.7%(n=6)；mass concentrations of linear range were from 0.050 to 5.000 μg·mL^-1(r=0.999 9). Limit of quantitative was in the range of 0.025-0.200 μg·mL^-1, limit of detection was in the range of 0.008-0.070 μg·mL^-1. The repeatability RSD of raw materials was 0.80%-3.8%, and the recovery rate was 95.2%-104.8%. The repeatability RSD of inhaled aerosol was 1.2%-2.9%, and the recovery rate was 98.7%-102.8%. The forced degradation test showed that impurities D, F, I, J, and N were all degradation impurities. 110 batches of samples were checked and the results of the relevant substances met the regulations. In the samples of diverse enterprises, impurities C, D, F, and N were detected more frequently, while impurities E, G, and H were not detected. Impurity J was only detected in one batch. The predicted impurity D by QSAR software falls to ICH M7 (R2) level 2.

The established method is sensitive and accurate, and can accurately quantify the content of related substances in salbutamol sulfate aerosol, providing effective technical support for systematic supervision. Further toxicity studies should be conducted on impurity D and reasonable limits should be established.

To analyze the fragmentation rule and pathway of pelargonidin, cyanidin, delphinidin, peonidin, petunidin and malvinidol under UHPLC-QTOF-MS positive mode electric spray, identify the characteristic product ions of six anthocyanins, and provide a theoretical basis for the establishment of mass spectrometry database and detection methods.

The chromatographic conditions were as follows: chromatographic column, Fusion-RPC₁₈ (50 mm×2.0 mm, 4 μm), mobile phase 0.1% formic acid aqueous solution (A) and methanol (B), gradient elution (0-1 min, 95%A; 1-5 min, 95%A→10%A; 5-6 min, 1%A; 6-7 min, 95%A), flow rate 0.3 mL·min^-1, column temperature 40 ℃, and injection volume 10 μL. The mass spectrometry conditions were as follows: TOF MS-IDA MS/MS, curtain gas, 0.20 MPa, collision gas CAD 7 MPa, IS voltage, 5 500 V/-4500 V, ion source temperature, 500 ℃, nebulizer gas, GAS1, 0.38 MPa, auxiliary, GAS2, 0.48 MPa, DP voltage, ±60 V, fragmentation voltage, (35±15) V, and time 0.2 s. Under the condition of positive mode of electrospray, the mass spectrometry data of six anthocyanins were measured. According to the pyrylium ions formed by the 2-phenylchromogenic structure of anthocyanins, and combined with the auxiliary analysis of the mass spectrometer database, the possible product ions were deduced.

The results showed that the six anthocyanins mainly undergo cleavage reactions on the pyranium ring, ultimately generating intermediate ions of pyrogallol and benzyl alcohol. On the other way, it occured α cracking, σ cracking causes the loss of functional groups on the ring, ultimately resulting in the formation of Chain hydrocarbons without functional groups.

The research results can provide support for the mass spectrometry characteristic ion data of six anthocyanins, which can be used to establish ion library data for anthocyanin products and also provide reference for the development and research of detection methods.

To establish an HPLC method for determination of related substances in apixaban API.

The analytical column was an ACE Excel3 C₁₈-PFP (150 mm×4.6 mm, 3 μm）. The mobile phase A was buffer(30 mmol·L^-1 ammonium acetate in water)-acetonitrile（90∶10） and the mobile phase B was buffer(30 mmol·L^-1 ammonium acetate in water)-acetonitrile（5∶95）. The whole run carried out by gradient elution at a flow rate of 1.2 mL·min^-1. The detection wavelength was set at 280 nm, the column temperature was 40 ℃ and the injection volume was 10 μL.

Apixaban was separated completely from the impurities and degradation products(the resolution>2.0). The test solution was stable for at least 48 h. The LOQs of apixaban, methyl ester product, ethyl ester product, chlorine impurity, dehydrogenation impurity, bihydrolytic impurity, ringopen methyl ester product, cyclate, impurity D, hydrolytic impurity, ringopen acid impurity, ringopen amide impurity and 5-chlorhexyl chloride derived impurity, were all 0.05%. The linear correlation coefficients of apixaban, methyl ester product, ethyl ester product, hydrolytic impurity, ringopen acid impurity, ringopen amide impurity and 5-chlorhexyl chloride derived impurity were all more than 0.99. The range were from the LOQ for impurity content to 150% of the target concentration. The average recoveries（RSD）(n=9) of methyl ester product, ethyl ester product, hydrolytic impurity, ringopen acid impurity, ringopen amide impurity and 5-chlorhexyl chloride derived impurity were 102.0%（2.7%）, 106.4%（2.2%）, 111.2%（4.0%）, 104.4%（2.9%）, 102.9%（2.7%）, 101.8%（2.9%）. The repeatability and intermediate precision completely met the requirements. The impurities contents in three batches of apixaban API 6 months accelerate stability test completely met the requirements, respectively.

This method is simple, rapid, sensitive and specific to be used for the determination of related substances in apixaban API.

To establish an UPLC-MS method for the analysis of salidroside, methyloleoside, specnuezhenide, acteoside, oleuropein, and G13 in plasma, and the differences in pharmacokinetic profiles of five different concoctions of Ligustri Lucidi Fructus, wine Ligustri Lucidi Fructus, vinegar Ligustri Lucidi Fructus, salt Ligustri Lucidi Fructus and steamed Ligustri Lucidi Fructus in vivo of rats were investigated.

SPF-grade male SD rats were randomly divided into 6 groups, and were given 4.2 g·kg^-1 of aqueous extracts of different concoctions of Ligustri Lucidi Fructus by gavage (for the amount of raw drug), and the plasma samples were methanol-precipitated proteins with geniposide as the internal standard, and the plasma samples were used to determine salidroside, methyloleoside, specnuezhenide, acteoside, oleuropein, and G13 in the plasma of the rats in the different time points by using the negative-ion SIM mode of UPLC-MS/MS. Kinetica 5.1 software was used to calculate the pharmacokinetic parameters, and GraphPad Prism 8.4.0 software was used to analyze the data.

The mass concentrations of salidroside, methyloleoside, specnuezhenide, acteoside and oleuropein were in the range of 2.00-1 385 ng·mL^-1, and the mass concentration of G13 was in the range of 1.30-650 ng·mL^-1, with good linearity, the relative standard deviations of the precision were all less than 10%, the recoveries of the extracts were all in the range of 85%-105%, and the matrix effect and stability were in accordance with the requirements of biological samples. The results of pharmacokinetic study showed that the AUC_0-∞, MRT_0-t, t_1/2, and C_max of acteoside were significantly higher in wine Ligustri Lucidi Fructus (P<0.01) compared to raw Ligustri Lucidi Fructus, which was 751.36 ng·mL^-1·h, 5.87 h, 377.82 h, and 38.11 ng·mL^-1, respectively. C_max, AUC_0-∞, and t_1/2 were significantly higher (P<0.01) for specnuezhenide and G13 in salt Ligustri Lucidi Fructus compared to raw Ligustri Lucidi Fructus, with C_max of specnuezhenide and G13 in salt Ligustri Lucidi Fructus were 66.45 ng·mL^-1 and 204.27 ng·mL^-1, respectively. The AUC_0-∞ were 342.69 ng·mL^-1 and 423.44 ng·mL^-1·h, and t_1/2 were 101.64 h and 15.98 h, respectively. Compared to raw Ligustri Lucidi Fructus, the C_max of oleuropein was significantly higher (P<0.05) in vinegar Ligustri Lucidi Fructus, wine Ligustri Lucidi Fructus, and steamed Ligustri Lucidi Fructus with 66.81 ng·mL^-1, 68.00 ng·mL^-1, and 66.38 ng·mL^-1, respectively. The AUC_0-∞ of salidroside and methyloleoside was the highest in vinegar Ligustri Lucidi Fructus, which was 5 782.74 ng·mL^-1 and 545.26 ng·mL^-1·h, respectively.

This study reveals the changing law of the six active ingredients in different artillery products of Ligustri Lucidi Fructus in vivo and their different characteristics, which provides a basis for the clinical application of different artillery products of Ligustri Lucidi Fructus.