To establish an HPLC method of the content and related substances of compound amino acid injection(3AA).

RP-HPLC was adopted to determine compound amino acid injection(3AA), combining with the use of two-dimensional column switching-LC/MSⁿ method was applied to separate and identify the impurities. The determination was performed on Capcell PAK AQ C₁₈(250 mm×4.6 mm, 3 μm) column with 0.2 mol·L^-1 sodium dihydrogen phosphate solution (adjusted pH to 2.8 with phosphoric acid) -acetonitrile (98∶2) as mobile phase at the flow rate of 1.0 mL·min^-1. The column temperature was 40 ℃, and the detection wavelength was 210 nm. And the injection volume was 20 μL. The LC/MSⁿ method was performed on a Thermo Accucore AQ C₁₈ (100 mm×4.6 mm, 2.6 μm) column with 0.1% formic acid solution as mobile phase A, 0.1% formic acid solution-acetonitrile as mobile phase B, at a flow rate of 0.4 mL·min^-1, and at a column temperature of 40 ℃. The mass spectrometry conditions were performed using an ESI ionisation source in the positive-ion scanning mode with a scan range of m/z 100-1 000, and the secondary mass spectrum was carried out by data-dependent scanning.

The related substances were completely separated from the main constituents in RP-HPLC. The standard curve of valine was linear over the range of 1.263-5.050 mg·mL^-1, with the average recovery of 99.0% (n=9). The standard curve of isoleucine was linear over the range of 1.350-5.402 mg·mL^-1, with the average recovery of 99.4%(n=9). The standard curve of leucine was linear over the range of 1.647-6.588 mg·mL^-1, with the average recovery of 99.5%(n=9). The main impurities in the three batches of samples were all process impurities introduced from the raw materials, with methionine content of 4.344 μg·mL^-1, 3.751 μg·mL^-1, 4.503 μg·mL^-1, respectively, phenylalanine content of 4.636 μg·mL^-1, 4.889 μg·mL^-1, 4.753 μg·mL^-1, respectively. The maximum single impurity contents were 0.01%, 0.02% and 0.01%, respectively.

The method is proved by the methodology validation that it can be used for the quality control of compound amino acid injection(3AA).

To analyze and optimize stir-baked with vinegar technology of Genkwa Flos decoction pieces by multi-index weighted scoring method based on orthogonal experiment, and to provide technical index on standardization of stir-baked with vinegar technology of Genkwa Flos decoction pieces.

The orthogonal experiment was employed to analyze and evaluate the four key factors related to stir-baked with vinegar technology including the amount of vinegar, dampening time, temperature and stir-baked time by the six main indicators including the total contents of chlorogenic acid, tiliroside, luteolin, apigenin, hydroxygenkwanin and genkwanin, the content of alcohol extractive and appearance characters.

The amount of vinegar and stir-baked temperature had notable effects, while the dampening time and stir-baked time had no notable effects on the test results. Considering comprehensively the effects of four key factors, the optimized stir-baked with vinegar technology was as follows: add 0.3 kg of vinegar to 1 kg of Genkwa Flos, and fry at 160 ℃ for 10 min after dampening for 15 min.

This study offers guidance and reference for standardizing the stir-baked with vinegar technology of Genkwa Flos decoction pieces.

To establish a solid-phase extraction-high performance liquid chromatography(SPE-HPLC) method for the rapid detection of related substances of vitamin D₃, trans vitamin D₃ (impurity A), 7-dehydrocholesterol (impurity B), lumisterol 3 (impurity C), and isotachysterol 3 (impurity D), in vitamin D drops (soft capsules).

The content, which was approximately equivalent to vitamin D₃ 1 548 IU, was taken in about 1.2 g, weighed accurately, put into a test tube, 4 mL of isooctane was added to it, and it was swirled and mixed well. Purification was performed using solid phase extraction columns, with n-hexane-ethyl acetate(85∶15) as the elution agent and anhydrous ethanol as the desorption agent. A Luna Silica(2) 100 Å silica gel column(150 mm×4.6 mm, 3 μm) was used, with n-hexane-n-pentanol (996.5∶3.5) as the mobile phase. The content of vitamin D₃ impurities was calculated using the adding standard calibration factors principal component self-control method.

The separation degree between pre-vitamin D₃ and vegetable oil was greater than 1.5. The linear range of vitamin D₃ was 0.02-0.80 μg·mL^-1, with r=0.999 5. The linear range of impurity A was 0.02 -0.80 μg·mL^-1, with r=0.999 9. The linear range of impurity B was 0.02-0.80 μg·mL^-1, with r=0.999 9. The linear range of impurity C was 0.02-0.80 μg·mL^-1, with r=0.999 9. The linear range of impurity D was 0.02-0.80 μg·mL^-1, with r=0.999 9. The average recovery rate of impurities A-D (n=9) was 93.2%-102.9%. The detection results of 3 batches of vitamin D drops (soft capsules) showed that the content of known impurities and other maximum single impurities were less than 0.5%, and the total content of impurities was less than 1.0%.

The results indicate that the established method is suitable for the detection of vitamin D₃ related substances in vitamin D drops (soft capsules). It has the advantages of being easy to operate, providing rapid and accurate results, and providing technical assurance for the quality control of fat-soluble vitamin D₃ preparations.

To establish a rapid molecular identification method for DNA fingerprinting-immunocolloidal gold test strips of Chinese herbal medicine human placentophagy genetic markers and to develop an integrated quality control gene detection kit.

Self-developed method and reagents could be for template DNA extraction from human placentophagy. Species-specific primers were designed according to the human mtDNA cytb gene, and the 5’ ends of the primers were labeled with FAM and Biotin, respectively. The optimal annealing temperature and cycle number of PCR were screened to establish a DNA fingerprinting molecular identification method. To develop PCR gene detection reagent premixes, to prepare DNA clones of control herbs using molecular cloning and gene sequencing techniques and to utilize immunocolloidal gold test strips for result detection. Finally, a rapid gene detection kit integrating DNA extraction reagent, PCR gene detection reagent premixes, control herb DNA clone solution, blank control solution and test strips was developed, and evaluated for specificity, reproducibility, sensitivity and stability.

The template DNA extracted by self-developed DNA extraction reagents were all in the concentration of 150-280 ng·μL^-1, the purities were all in the range of 1.8-1.9, and there were no trail in the agarose gel electrophoresis. And the immunocolloidal gold test strips showed two red bands for human placentophagy control herb and authentic product, and one red band for easy-mix product and blank control at annealing temperature of 59 ℃ and 20 cycles for human placentophagy specific primers. The agarose gel electrophoresis was verified to be correct. The DNA sequencing results of the control herbs were 100% homologous with human mtDNA cytb gene. The self-developed integrated rapid gene detection kit was specific, reproducible, stable and sensitive at 0.1 ng·μL^-1.

The DNA fingerprinting-immunocolloidal gold test strip molecular identification method can specifically, accurately, rapidly and visually identify the authenticity of human placentophagy, and the integrated rapid gene detection kit provides a regulated and standardized testing method for quality control of human placentophagy. Therefore, it is more suitable for on-site field testing and universal promotion.

To find out identification characteristics of Baijiangcao from different origins by pharmacognostical study. To discuss the establishment of whole herbs Chinese medicine standard.

Principles of plant taxonomy, macroscopic identification and microscopic identification were used to compare the differences among Baijiangcao from different origins and commercial counterfeit.

The macroscopic characteristics, such as the surface of stem, the cross section of stem, the shape of leaves, hairy on inflorescence and odour. And the microscopic characteristics of transverse section of stem could be used to distinguish Baijiangcao from different origins and commercial counterfeit.

This research establishes the identification method of Baijiangcao from different origins and commercial counterfeit, and provides references for the improvement of quality standard of Baijiangcao from different origins.

To establish a fast, accurate and sensitive method by UPLC-DAD-Q-TOF-MS/MS for the detection of 20 kinds of illegally added chemicals such as azilsartan, nifedipin, lacidipine、nebivolo in anti-hypertensive health foods.

Waters HSS T3 C₁₈（100 mm×2.1 mm, 1.8 μm ） column was used with mobile phase of 0.01 mol·L^-1 ammonium acetate aqueous solution-methanol and gradient elution（0 min, 5% A; 0-8 min, 5% A→95% A; 8-11 min, 95% A; 11-11.2 min, 95% A→5% A; 11.2-12 min, 5%). The flow rate was 0.3 mL·min^-1. The injection volume was 1 μL. ESI⁺, ESI^- scan and MS/MS mode were applied. Qualitative and quantitative analyses were carried out by UPLC-DAD-Q-TOF-MS/MS.

The method was validated. A good linear relationship was showed in the concentration of 1-50 μg·mL^-1, （RSD<3%）and the recovery was obtained in range of 80.5%-98.7%. Illegally added chemical drugs were found in 15 batches of samples tested by this method. Two or more types of drugs were detected in 14 batches of samples. Torsemide, candesartan cilexetil, lacidipine were detected in 9 samples. Lacidipine and nebivolol were found in one of the sample.

This method is fast, accurate and sensitive with a high degree of separation. It can be used as a powerful tool for the fast detection of illegally added chemicals in anti-hypertensive health foods.

To establish and optimize a rapid direct analysis of real time tandem mass spectrometry (DART-MS/MS) method for the rapid detection of 12 benzodiazepines in blood and urine that can be used in forensic toxicology work.

A DART ion source was used in conjunction with an API4000 Q Trap mass spectrometer. A DART 12Dip-It^TM autosampling module with a module travel speed of 0.6 mm·s^-1, a sample volume of 5 μL, and a gate voltage of 200 V were applied. The mass spectrometry section scans in positive ion mode using multiple reaction monitoring (MRM) mode. After further optimization, the DART-MS/MS method was validated and applied to real case samples.

Ethyl acetate was selected as the extractant for liquid-liquid extraction and the temperature of the carrier gas heater was optimized. The method has good selectivity and does not interfere with delayed effects. The linearity was good, and the limits of detection (LODs) for the targets in blood and urine were in the ranges of 0.5-10 ng·mL^-1 and 0.2-2 ng·mL^-1, respectively. And the limits of quantification (LOQs) were in the ranges of 1-50 ng·mL^-1 and 0.5-5 ng·mL^-1, respectively. The recoveries ranged from 78.8% to 119%, and the matrix effects ranged from -17.5% to 18.5%. The intra- and inter-day precisions were not greater than 14.4% for the high and intermediate concentrations, and not greater than 18.1% at the limits of quantification. This method enables fast and accurate examination of case samples.

The method is fast and convenient, with good sensitivity, and can be applied to the research and work of rapid detection of toxicants to improve the efficiency of detection.

To compare the differences of the volatile oil components of Pimpinella thellungiana Wolff extracted with different methods by GC-MS combined with retention index.

Volatile oil of Pimpinella thellungiana Wolff were extracted using steam distillation, salting-out assisted steam distillation and enzymatic hydrolysis-assisted steam distillation. The chemical components of volatile oil extracted by different methods were analyzed by GC-MS combined with retention index, while the relative contents of volatile oil components were calculated by peak area normalization method.

Among the three extraction methods, the extraction rate of volatile oil was as follows, enzymatic hydrolysis-assisted steam distillation > salting-out assisted steam distillation ≈ steam distillation method. The most comprehensive types of volatile oil were extracted by salting-out assisted steam distillation, followed by enzymatic hydrolysis-assisted steam distillation, and finally by steam distillation. The main types of the volatile oil components of Pimpinella thellungiana Wolff extracted with different methods were basically unchanged, but the relative contents of various compounds were different. A total of 47 compounds were identified from the volatile oil extracted with the three methods, including 35 common compounds. The main chemical structural types of these compounds were terpenes, terpenes, terpenoids, aromatics and aliphatics, among which the higher content compounds were β-bisabolene（17.25%-19.42%）, caryophyllene oxide （12.90%-15.70%）, 1-(3-methyl-2-butenoxy)-4-(1-propenyl) benzene （5.02%-9.36%） and β-pinene （5.31%-6.62%）.

The chemical composition types and relative contents of the volatile oil of Pimpinella thellungiana Wolff extracted with different methods are different, but the differences are not significant. The results can provide reference for the selection of suitable extraction methods and further utilization of volatile oil from Pimpinella thellungiana Wolff.

To establish a chromatogram-effect correlated quality evaluation system for the antibacterial activity of Flos Lonicerae extract by data analysis of components.

The best liquid chromatographic conditions were optimized by investigating the conditions of mobile phase, wavelength and flow rate, and the quantitative fingerprint of 50 batches of Flos Lonicerae extract was established. The best antibacterial concentration of the extract was obtained by microdilution method, and the antibacterial rates of 50 batches of samples were determined. Similarity analysis, cluster analysis, principal component analysis, grey correlation analysis and mathematical model establishment by support vector machine were used to analyze the quantitative fingerprints and antibacterial rates of 50 batches of samples.

Eighteen common peaks were selected from the 50 quantitative fingerprints of Flos Lonicerae extract, and the similarities were between 0.608-1. Six chemical components were identified (peak 4: neochlorogenic acid, peak 8: chlorogenic acid, peak 9: cryptochlorogenic acid, peak 16: isochlorogenic acid B, peak 17: isochlorogenic acid A and peak 18: isochlorogenic acid C). The average antibacterial rates of 50 batches of extracts were between 3.93%-70.50%. The results of principal component analysis and cluster analysis were highly consistent, and the results of grey correlation analysis showed that all components were positively correlated with antibacterial effect. The relative deviations between the predicted data of the mathematical model and the experimental data were all below 2%.

The HPLC conditions of Flos Lonicerae extract are stable and reliable, and the quality evaluation system of antibacterial activity of Flos Lonicerae extract established can evaluate its quality based on antibacterial rate.