[Material basis of kidney Yang deficiency rats before and after salt processing of Dipsacus asper based on spectrum-effect relationship].

This paper aims to study the spectrum-effect relationship between the fingerprints before and after salt processing of Dipsacus asper and the efficacy of warming and tonifying kidney Yang and find the main active components against kidney Yang deficiency before and after salt processing of D. asper, so as to provide the basis for clarifying the effect of salt processing on kidney Yang deficiency. The HPLC fingerprint before and after salt processing of D. asper was established by the HPLC-DAD. 15 common peaks were obtained, and 11 components were identified. The content changes of various components in rat serum were detected, and the difference in efficacy before and after salt processing was compared. The results of pharmacological experiments showed that salt processing of D. asper could enhance the kidney index. At the same dose, there was a significant difference between the raw D. asper and D. asper after salt processing groups. Compared with the model group, the contents of ACTH, cAMP, CORT, E_2, GH, Na~+-K~+-ATPase, T, and T4 in the serum of rats in the administration group increased to a certain extent, and the contents of cGMP and TNF-α decreased to a certain extent. Among them, there were significant differences in the above indexes in the serum of rats in the high-dose group of raw D. asper, middle-dose group of D. asper after salt processing, high-dose group of D. asper after salt processing, and the positive drug group. The overall results showed that D. asper after salt processing was more effective than raw D. asper in preventing kidney yang deficiency. The efficacy of D. asper was evaluated by grey correlation analysis, entropy method, and Pearson correlation analysis, and the components of D. asper after salt processing against kidney yang deficiency were screened out. According to the results of correlation degree ranking, the components with increased ranking before and after salt processing of D. asper were loganin, chlorogenic acid, dipsacoside A, asperosaponin Ⅵ, caffeic acid, and isochlorogenic acid B. It was preliminarily speculated that these compounds may be the potential pharmacodynamic components for the treatment of kidney yang deficiency before and after salt processing of D. asper. The changing components before and after the salt processing of D. asper were determined, which proved that D. asper after salt processing was superior to D. asper in the treatment of kidney yang deficiency. The spectrum-effect relationship between the efficacy of D. asper before and after salt processing and the treatment of kidney yang deficiency was established, which laid a foundation for the subsequent study on the pharmacodynamic components and molecular mechanism of salt processing of D. asper.

Salt processing: A unique and classic technology for Chinese medicine processing.

Traditional Chinese medicine (TCM) processing is the summary of long-term clinical experience, processing can change the property, taste and meridian of TCM, achieve the purpose of reducing toxicity and increasing efficiency, to ensure the safety of clinical medication. This paper summarizes the research advance of salt processing in recent years from the aspects of the types of excipients, salt processing methods, salt processing purpose and the influence of salt process on the chemical composition, pharmacodynamic effect and in vivo behavior of TCM, and analyzes and discusses the shortcomings of the current research, so as to provide ideas for the further research and development of salt processing of TCM. The literatures were classified and summarized by consulting the scientific database (such as SciFinder Scholar, CNKI, Google Scholar, Baidu Scholar, etc.), Chinese herbal classics and Chinese Pharmacopoeia and so on. The results show that salt processing is helpful to introduce drugs into the kidney channel and enhance the effect of nourishing Yin and falling fire. The pharmacological effect, chemical composition and in vivo behavior of TCM will change after salt processing. In the future, we should strengthen the research on standardizing the dosage of excipients, the quality standard after processing, the relationship between the change of chemical composition after salt processing and the enhancement of pharmacological efficacy, so as to deeply explain the principle of salt processing, and further improve and optimize the salt making process. By combing the influence of the of salt processing of TCM and analyzed the current problems, we hope to provide guidance for the in-depth study of the salt processing mechanism of TCM and the inheritance and innovation of TCM processing technology.

Study on HPLC Fingerprint of Alpiniae Oxyphyllae Fructus Before and After Salt-Processing Based on Chemical Pattern Recognition / 中药新药与临床药理

Objective A HPLC fingerprint method of Alpiniae Oxyphyllae Fructus(AOF)before and after salt-processing was established,to compare the differences of chemical components between raw and processed AOF combined with chemical pattern recognition.Methods HPLC method was used to establish the fingerprint of raw and salt-processed AOF.Principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)were applied to explore the different components of raw and salt-processed AOF in different batches.Results Totally 30 and 32 common peaks in the HPLC fingerprint from the raw and salt-processed AOF were detected,respectively.And 8 of them were identified by comparison with the standards.They were peak X2(5-hydroxymethylfurfural),peak 1(protocatechuic acid),peak 2(protocatechualdehyde),peak 4(epicatechin),peak 21(chrysin),peak 22(kaempferide),peak 25(tectochrysin)and peak 26(nootkatone).The results of PCA and OPLS-DA showed that raw and salt-processed AOF can be grouped into two categories.A total of 12 components,which were considered as differential markers of raw and salt-processed AOF,were screened by method of variable importance in projection(VIP).The 12 components were peak X1,peak 26(nootkatone),peak 16,peak 3,peak X2(5-hydroxymethylfurfural),peak 25(tectochrysin),peak 15,peak 12,peak 8,peak 10,peak 17 and peak 20.Conclusion The combination of HPLC fingerprint and chemical pattern recognition can be used to analyze the quality differences of AOF before and after salt-processing.

Pharmacokinetic investigation of principal active constituents in renal fibrotic rats after oral administration of crude and salt-processed eucommiae cortex extracts / 药学学报

A quantitative analysis method for six principal active constituents (acubin, geniposidic acid, chlorogenic acid, pinoresinol di-O-glucopyranoside, geniposide, and pinoresinol 4-O-glucopyranoside) of crude Eucommiae Cortex (EC) and its salt-processed product extracts was developed to investigate and compare their pharmacokinetic behaviors in adenine-induced renal fibrotic rats in vivo. UHPLC-QqQ-MS/MS technology was employed. Scan was conducted in negative ion mode and quantitative determination was carried out by MRM paired ion. The established method was fully validated by specificity, linearity, precision, accuracy, stability, recovery, and matrix effect, and the results of methodological investigation met the requirements of biological sample analysis. Then, a quick, sensitive, and accurate method was successfully established, which could simultaneously measure the contents of six active constituents of crude and salt-processed EC extracts in rat plasma. After a single administration to renal fibrotic rats of crude EC and its salt-processed product extracts, the plasma concentration of each constituent at different time points was measured, the pharmacokinetic parameters were calculated and the concentration time curves were structured. The experiment was approved by the experimental animal ethics committee from Nanjing University of Chinese Medicine (No. 202103A008). The results showed that compared to the crude Eucommiae Cortex group, the tmax of aucubin, pinoresinol di-O-glucopyranoside, geniposide, and pinoresinol 4-O-glucopyranoside in the salt-processed Eucommiae Cortex group rat plasma were significantly lower than those in the crude group (P < 0.05, P < 0.01); the Cmax and AUC0-48 h of chlorogenic acid, the Cmax, AUC0-48 h and AUC0-∞ of pinoresinol di-O-glucopyranoside, and the Cmax of geniposide and pinoresinol 4-O-glucopyranoside were significantly higher than those in the crude group (P < 0.05, P < 0.01). Our investigation found that compared to crude Eucommiae Cortex, a variety of active ingredients could play a role of quick effect with higher peak blood concentration and bioavailability after oral administration of salt-processed Eucommiae Cortex, which were consistent with the traditional Chinese medicine theory of "salt-processing enhancing drug into kidney meridian", providing an experimental basis for the selection of quality control indexes and the in-depth study of processing mechanisms and metabolic rules in vivo of Eucommiae Cortex and its salt-processed product.

Study on the processing chemistry of Fructus Psoraleae by a combination of untargeted and targeted metabolomics.

Fructus Psoralea is widely used to treat osteoporosis and skin inflammatory diseases. Because of the side effects on the liver, renal and cardiovascular systems, it is processed to salt-processed Fructus Psoraleae to meet the requirements of clinical use. However, the mechanisms involved in the transformation of the chemical components are unclear. In this study, ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry was used to analyze the chemical profiles of this herbal medicine and the chemical transformation mechanism involved during the salt processing was studied. A total of 83 compounds were identified. Principal component analysis and orthogonal partial least squares discriminate analysis were used to observe the distribution trend of all samples and visualize the difference. Raw and processed Fructus Psoraleae were clearly clustered into two groups. Furthermore, 17 marker compounds were identified as primary contributors to their differences based on t-test analysis (p < 0.01) and orthogonal partial least squares discriminate analysis (variable importance for the projection > 1). Finally, ultra-high performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry was used to evaluate the quality of Fructus Psoraleae by simultaneous analysis of 13 components highly related to efficacy. There were variations in the contents of 13 chemicals of Fructus Psoraleae and salt-processed products. The results of untargeted and targeted metabolomics revealed that salt processing affected the chemical composition of Fructus Psoraleae.

An emerging strategy for tracing the anti-erectile dysfunction components of raw and salt-processed semens of Astragali Complanati by combinatory liquid chromatography-mass spectrometry-based quantitative analysis, efficacy assessment on impotent rats, and partial least squares regression.

Semens of Astragali Complanati own anti-erectile dysfunction effect; however, the components which contribute to the anti-erectile dysfunction effect remain unclear. This work raised a strategy that integrates liquid chromatography coupled mass spectrometry-based quantitative analysis, anti-erectile dysfunction assessment on impotent rats, and their relationship analysis for pinpointing anti-erectile dysfunction components from semens of Astragali Complanati. For simultaneous quantification of seven major components in raw and salt-processed semens of Astragali Complanati, an accurate and reliable liquid chromatography-mass spectrometry method was developed under multiple reaction monitoring mode. Of note, chloramphenicol was employed as the internal standard. The method showed good linearity and repeatability, where the recovery rates of each component ranged from 98.1 to 104.7%, and the precisions of intra- and interday were all within 3.4%. The method has been used for quantification of the seven major components in 10 batches of raw and salt-processed semens of Astragali Complanati. Then, the anti-erectile dysfunction effects of raw and salt-processed semens of Astragali Complanati were evaluated on impotent rats. Gray relationship analysis and partial least squares regression were combined for elucidating the relationship. As a result, complanatuside, astragalin, complanatoside B, and kaempferol were found to be responsible for anti-erectile dysfunction effect of Astragali Complanati.

Study on the salt-processing technology of Rosa laevigata and its HPLC fingerprints and chromaticity values before and after salt-processing / 中国药房

OBJECTIV E To optimize the s alt-processing technology of Rosa laevigata ，and to study high performance liquid chromatography（HPLC）fingerprints and chromaticity values of R. laevigata before and after salt-processing. METHODS The comprehensive scoring method was adopted to optimize the salt-processing technology of R. laevigata using appearance character ， moisture and polysaccharide content as index. Fingerprints were established by HPLC method before and after salt-processing ，and chromaticity values （L*，a*，b*）of the powder before and after salt-processing were determined. The multivariate statistical analysis was carried out for raw product and salt-processing product of R. laevigata by using common peak areas and chromaticity values as index. RESULTS The optimal salt-processing technology of R. laevigata was to mix it with appropriate amount of salt water ，place them in the preheated frying wok at 140 ℃，fry them for 25 min，and rotate frying wok 20 times/min. Ten common peaks were calibrated by HPLC fingerprints before and after salt-processing ，and 3 components were identified ，such as gallic acid ，catechin and ellagic acid. The chromaticity values L*，b* and E* changed significantly after salt-processing. The multivariate statistical analysis method could distinguish raw products and salt-processing products into two categories ，in which peaks 1，5，6 and 10 and chromaticity values b* and E* were important characteristic factors. CONCLUSIONS The optimized salt-processing technology is stable and reliable ，and the established fingerprint has good repeatability and stability. Fingerprint and chromaticity values combined with multivariate statistical analysis can provide reference for the identification and quality analysis of R. laevigata before and after salt-processing.

Correlation analysis between extracts and endogenous metabolites to characterise the influence of salt-processing on compatibility mechanism of 'Psoraleae Fructus & Foeniculi Fructus'.

ETHNOPHARMACOLOGICAL RELEVANCE: 'Salt-processed Psoraleae Fructus & salt-processed Foeniculi Fructus' (sPF&sFF) is a common Chinese medicinal combination for treating diarrhoea. However, it is not clear how sPF and sFF work together, and why salt-processing is necessary. AIM OF THE STUDY: To investigate the compatibility mechanism of sPF&sFF and the influence of salt-processing on it. MATERIALS AND METHODS: Firstly, the metabolomics approach was appliedto screen the differential components between four (s)PF&(s)FF extracts, i.e., sPF&sFF, sPF&FF, PF&sFF, and PF&FF extracts. Then, an in vivo metabolomics study was carried out to filter critical metabolites reflecting the curative effects of (s)PF&(s)FF, and construct a metabolic network. Finally, a correlation analysis between chemical components in extracts and critical metabolites in vivo was performed to find out the synergistic and/or antagonistic effects between herbs as well as the influence of salt-processing. RESULTS: Salt-processing had a direct influence on the contents of chemical components in sPF and sFF extracts, and there existed positive/negative correlations between the content change of chemical components and the effects of critical metabolites. Therefore, salt-processing indirectly affected on these correlations and was (i) conducive to the positive effects of sPF and sFF on bile acids, making sFF play a synergistic role, thereby, sPF&sFF could perform better than sPF and other three combinations and effectively relieve the symptoms of fatty diarrhoea, osmotic diuresis, malnutrition, and weight loss; (ii) conducive to the positive effects of sPF on triacylglycerol, 12(S)-hydroxyeicosatetraenoic acid, cholesterol, and arachidonic acid, and adverse to that of sFF, making sFF play an antagonistic role, thereby, sPF&sFF could prevent a series of side effects caused by over-regulation and suitably relieve the symptoms of osmotic diuresis, polyuria, malnutrition, and weight loss; and (iii) adverse to the positive effects of sPF and sFF on thromboxane A2, sphinganine and sphingosine, making sFF play a synergistic role, thereby, sPF&sFF could prevent a series of side effects and moderately relieve the symptoms of metabolic diarrhoea and polyuria. CONCLUSIONS: Salt-processing indirectly affected on the correlations between chemical components in extracts and critical metabolites in vivo, and exhibited both conducive and adverse effects on the efficacy, making sPF and sFF cooperate with each other to moderately repair the metabolic disorders. Thereby, sPF&sFF could suitably relieve the diarrhoea and polyuria symptoms in the model and exert the most appropriate efficacy. Moreover, this novel strategy provided a feasible approach for further studying the compatibility mechanism of herbs.

Comparison of pharmacokinetics of phytoecdysones and triterpenoid saponins of monomer, crude and processed Radix Achyranthis Bidentatae by UHPLC-MS/MS.

1. The aim of this study was to develop a selective, rapid, accurate and sensitive ultrahigh performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method for pharmacokinetic (PK) studies of phytoecdysones and triterpenoid saponins after oral administration of five monomers, crude, wine-processed and salt-processed Radix Achyranthis bidentatae (RAB).2. A Thermo Hypersil GOLD C18 column (100 mm × 2.1 mm, 1.9 µm) coupled with a mobile phase of (A) acetonitrile and (B) water (both containing 0.3% acetic acid) was used for sample separation. The mass analysis was performed in a triple quadruple mass spectrometer using selected reaction monitoring (SRM) with negative scan mode.3. The results showed that this method exhibited desirable sensitivity, precision, stability and repeatability. The extraction recoveries of the compounds ranged from 94.2 to 99.8% and the matrix effects ranged from 93.3 to 100.5%. Comparing the Cmax and AUC of five analytes in those groups showed this tendency: salt-processed RAB > wine-processed RAB > crude RAB > monomer group. The results confirmed the feasibility of TCM theory to enhance the efficacy of processed RAB.

Modern research on theory of “salt-processing enhancing drug into kidney meridian” proposed by healer Jia-mo Chen from Xinan / 中草药

Jia-mo Chen who was a Xinan medical scientist put forward the theory of “Salt-processing enhancing drug into kidney meridian” recorded in “Enlightening Primer of Materia Medica”, which has great significance for the clinical application of Chinese medicine processing excipients. In recent years, scholars have done a lot of research work on the chemical composition and pharmacological effect changes of Chinese medicines and related prescriptions before and after salt-processing, and preliminarily explained the scientific connotation of the processing theory of “salt-processing enhancing drug into kidney meridian” of Chinese materia medica (CMM). According to the literatures of recent years studies, the research progress on processing theory of “Salt-processing enhancing drug into kidney meridian” of CMM was summarized and concluded from the perspective of chemical composition and pharmacological effects changes, and the problems of the present study were analyzed, which put forward new research idea for the theory of “salt-processing enhancing drug into kidney meridian” combined with modern research methods.

Research on rationality of salt-processed composition of Buguzhi Pills through efficacy evaluation / 中草药

Objective: To investigate the effects of Buguzhi Pills and three kinds of simulative Buguzhi Pills on diarrhea rats with spleen-kidney yang deficiency, and to confirm the rationality of the salt-processed composition of Buguzhi Pills. Methods: SD rats were randomly divided into nine groups: normal group, model group, positive drug group, salt-processed Psoraleae ructus group, raw Psoraleae Fructus group, Buguzhi Pills (salt-processed Psoraleae Fructus + salt-processed Foeniculi Fructus) group and three kinds of simulative Buguzhi Pills groups (salt-processed Psoraleae Fructus + raw Foeniculi Fructus; raw Psoraleae Fructus + salt-processed Foeniculi Fructus; raw Psoraleae Fructus + raw Foeniculi Fructus). A complex method of hydrocortisone and sennae folium was adopted to establish the diarrhea model due to spleen-kidney yang deficiency. After administration, the organ indexes of thymus, spleen, kidney, adrenal gland, and testis were measured; The pathological changes of corresponding organs were observed; The serum levels of motilin and testosterone were detected by ELISA. Results: Compared with the normal group, the food intake and body weight reduced; The organ indexes decreased in varying degrees; The above organs showed obvious pathological changes under the microscope; the level of motilin showed an upward trend, and the level of testosterone decreased significantly in model group. The above symptoms and indicators were improved in varying degrees after administration, and the effect of Buguzhi Pills was better than that of Psoraleae Fructus alone and three kinds of simulative Buguzhi Pills on the whole. Conclusion :The efficacy of Buguzhi Pills on warming kidney-spleen yang and relieving diarrhea was superior to that of Psoraleae Fructus alone, and the antidiarrheal effect can be enhanced by salt-processing.

A novel ultra high-performance liquid chromatography-tandem mass spectrometry method for the simultaneous determination of xanthones and steroidal saponins in crude and salt-processed Anemarrhenae Rhizoma aqueous extracts.

We established a rapid and sensitive ultra high-performance liquid chromatography tandem mass spectrometry method for the simultaneous quantification of xanthones and steroidal saponins in rat plasma. Chromatographic separation was achieved on a C18 column with a mobile phase comprising acetonitrile and 0.1% formic acid. The detection was performed by negative electrospray ionization in multiple reaction monitoring mode. The validated method showed good linearity within the tested range (r > 0.9945). The intra- and interday precision at high, medium, and low concentrations was less than 7.96%. The bias of accuracies ranged from -1.92 to 9.62%. The extraction recoveries of the compounds ranged from 84.78 to 88.69%, and the matrix effects ranged from 96.76 to 108.59%. This method was successfully applied to a pharmacokinetic comparison of crude and salt-processed Anemarrhenae Rhizoma aqueous extracts after oral administration in rats. The maximum plasma concentration and area under concentration-time curve of timosaponin BIII and timosaponin AIII increased significantly (P < 0.05 or 0.01) and those of timosaponin BII decreased significantly (P < 0.05) after processing. These results could contribute to the clinical application of crude and salt-processed Anemarrhenae Rhizoma and reveal the processing mechanism.

Analysis of Chemical Variations between Crude and Salt-Processed Anemarrhenae rhizoma Using Ultra-High-Performance Liquid Chromatography-Mass Spectrometry Methods.

The present study was designed to systematically investigate the chemical profile differences between crude Anemarrhenae rhizoma (CAR) and salt-processed Anemarrhenae rhizoma (SAR). Ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS), coupled with multivariate statistical analysis was used for the discrimination of chemical profiles and the identification of the differentiation of the chemical constitutions of CAR and SAR. In addition, seven main constituents of CAR and SAR were simultaneously determined by ultra-high-performance liquid chromatography-quadrupole mass spectrometry (UHPLC-MS) for analyzing the content variations. A total of 24 components were found to be the main contributors to the significant difference between CAR and SAR. The structures of the marker compounds were identified based on their chromatographic behaviors, intact precursor ions, and characteristic MS fragmentation patterns. The potential structural transformation mechanism of furostanol saponins during salt processing was explored. The results may provide a scientific foundation for deeply elucidating the processing mechanism of Anemarrhenae rhizoma.

Modern study on theory of "salt-processing enhancing drug into kidney meridian" / 中草药

"Salt-processing enhancing drug into kidney meridian" is one of the basic theories to guide the processing of Chinese materia medica (CMM). In recent years, scholars have done a lot of research work on salt-processing, which has effectively promoted the scientific explanation of the theory. In this paper, recent researches of salt-processed CMM from the perspective of "salt-processing enhancing drug into kidney meridian" were reviewed. Based on the analysis of the existing problems, this review provides a new idea for the mechanism study of salt-processing.

Evaluation of the influence of salt processing on pharmacokinetics of psoralen and isopsoralen in Psoralea corylifolia L.

A sensitive, specific and rapid ultra-high-pressure liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method has been developed to investigate pharmacokinetic properties of psoralen and isopsoralen, two compounds isolated from raw/salt-processed fruit of Psoralea corylifolia L. UHPLC-MS/MS was used with positive ion electrospray. The mobile phase was composed of acetonitrile and 0.1% formic acid aqueous solution and a gradient elution program at flow rate of 0.3 mL/min was applied. Multiple reaction monitoring mode was used for the quantification of psoralen, isopsoralen ([M + H](+) m/z 187.0 → m/z 131.0) and scoparone (m/z 207.0 → m/z 151.1). Scoparone served as an internal standard. The method was fully validated for its sensitivity, selectivity, stability, matrix effect and extraction recovery. The obtained results showed that salt-processed Buguzhi significantly promoted the absorption of psoralen and isopsoralen, and increased the bioavailability of these compounds.

Optimization of salt-processing technology for Phyllanthus emblica by comprehensive score method / 中草药

Objective: To choose the optimal salt-processing technology for Phyllanthus emblica. Methods: Taking the contents of gallic acid, querectin, ethanol soluble extraction, and water-soluble extract as the indexes, the factors of brine concentration, watering multiple, soaking time, and drying temperature were investigated. L9(34) orthogonal design and comprehensive score were used to optimize the salt-processing technology for P. emblica. Results: The optimal salt-processing technology was: 3% salt, adding 1.2-fold water, soaking for 10 d, and drying under 90°C. Conclusion: The optimal salt-processing technology is stable, reasonable, and feasible, and could be used to guide the standardization production.