Four-week intravenous repeated dose toxicity study of vitacamphorae injection in rats.

The study was undertaken to evaluate the safety of vitacamphorae (VCP) injection in Sprague-Dawley (SD) rats. Rats were intravenously administered with VCP at the doses of 0, 5, 15, and 50 mg/kg/day (equivalent to 0, 5, 15, and 50 times the clinical equivalent dose) for 4 weeks, respectively. In addition, we also tested oxidative stress-related parameters and cytokine levels in rat serum. In the current study, intravenous administration of VCP at a dose of 50 mg/kg/day caused significant pathophysiological responses in rats. Compared with the control group, different doses of VCP exposure had no significant effect on body weight, food consumption, and clinic pathology of rats after 4 weeks of VCP administration. Rats in high-dose group (50 mg/kg/day) showed general symptoms of convulsions after VCP administration. The toxicological significance of VCP exposure in the spleen of high-dose female rats was observed, which showed a significant increase in the relative spleen weights (P < 0.01) and mild lymphocyte proliferation in splenic pathology. Furthermore, the results of oxidative stress and cytokine detection showed that the levels of antioxidant enzymes SOD increased in each administration group, but the levels of a series of pro-inflammatory cytokines IL-1ß, IL-6, IL-8, IL-12, and IFN-γ also increased in these groups. Above changes caused by VCP exposure can be reversed after 4 weeks of recovery. Overall, the results showed that the no-observed-adverse-effect-level (NOAEL) of VCP injection for 4-week toxicity was 15 mg/kg/day.

Cocrystal and its Application in the Field of Active Pharmaceutical Ingredients and Food Ingredients.

BACKGROUND: The development of solid drug dosage form and food ingredients is constrained by their low solubility, low dissolution, low bioavailability and poor physicochemical properties. Formation of cocrystal is a novel and promising method to enhance and improve the properties of materials without breaking the covalent bonds. METHODS: The goal of this review is to summarize the cocrystals and their applications in the field of Active Pharmaceutical Ingredients (APIs) and food ingredients (AFIs), mainly on the effective improvements of APIs' and AFIs' pharmacokinetic, physicochemical and mechanical properties by the formation of cocrystals. RESULTS: After years of research and development on cocrystals in the area of pharmaceutical and food industries, significant progress has been made. Formation of cocrystal is an efficient method for improving the solubility, dissolution rate, permeability and in vivo bioavailability of APIs and AFIs, as well as for enhancing stability and mechanical properties. CONCLUSION: Cocrystals exhibit complex structures which can conspicuously affect the physical and chemical properties of original substance, with good clinical performance and outstanding stability during processing and storage.

[Identification of Sun-dried and Sulfur-fumigated Chuanxiong Rhizoma by FTIR].

OBJECTIVE: To evaluate the sulfur-fumigation effects on the chemical constituents of Chuanxiong Rhizoma. METHODS: Alcoholic and aqueous extracts of sun-dried and sulfur-fumigated Chuanxiong Rhizoma samples were analyzed by FIIR. FTIR spectra were acquired and disposed by software Omnic 8. 0, second derivative IR spectra were analyzed by software OPUS 6. 5 through smoothing and differentiation treatment to FTIR spectra, and the absorption frequencies of alcoholic and aqueous extracts of sun-dried and sulfur-fumigated Chuanxiong Rhizoma samples were assessed by using principal component analysis with software SPSS. RESULTS: There were significant differences between sun-dried and sulfur-fumigated Chuanxiong Rhizoma samples in the region ranging from 2 000 cm-1 to 750 cm-1. Some absorption peaks weakened or disappeared, and some newly emerged after the processing of sulfur-fumigation. CONCLUSION: The processing of sulfur-fumigation can influence the effective constituents of Chuanxiong Rhizoma. The established method of FTIR combined with second derivative and principal component analysis has been proved to be an effective, intuitional and rapid approach to distinguish sun-dried and sulfur-fumigated Chuanxiong Rhizoma.

Evaluation of the influence of sulfur fumigation on the pharmacokinetics of four active ingredients in Si Wu Tang.

Sulfur fumigation may induce the decrease or the chemical transformation of some active ingredients of traditional Chinese medicines in vitro. Whether sulfur fumigation can cause the pharmacokinetic changes of the active ingredients in vivo is related to the efficacy and the safety of Chinese medicines' application clinically. A sensitive, specific, and accurate method for the simultaneous determination of paeoniflorin, ferulic acid, senkyunolide A, and senkyunolide I in rat plasma by ultra high performance liquid chromatography coupled with triple quadrupole mass spectrometry was developed to evaluate the influence of sulfur fumigation to Si Wu Tang for the first time. Each compound was extracted from plasma samples by liquid-liquid extraction with ethyl acetate, and the chromatographic separation was accomplished on an Agilent Extend C18 column with a linear gradient elution. The mass spectrometric detection and analysis were performed by using an AB Sciex triple quadrupole 5500 mass spectrometer in multiple reaction monitoring mode. The validated method was successfully applied to a pharmacokinetic study of four compounds in rats after oral administration of sun-dried and sulfur-fumigated Si Wu Tang. The results provided a meaningful basis for evaluating the affection of sulfur fumigation to the clinical application and the efficacy of Si Wu Tang.

Analysis of the influence of sulfur-fumigation on the volatile components of Angelicae sinensis Radix by comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry.

BACKGROUND: Sulfur-fumigation of Angelicae sinensis Radix causes changes in the structure and composition of volatile components. These changes alter the curative effect and the quality of A. sinensis Radix. MATERIALS AND METHODS: In this study, comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (GC × GC-TOFMS) was employed to investigate the influence of sulfur-fumigation on the volatile components, and to characterize and quantify the chemical composition of the volatile oil of A. sinensis Radix. RESULTS: The present study has shown that sulfur-fumigated A. sinensis Radix samples had significant loss of the main active compounds and a more destructive fingerprint profile compared to non-fumigated samples. CONCLUSION: From this study, it can be concluded that the combination of GC × GC and TOFMS has potential as a quality monitoring tool in herbal medicine and food processing industries.

Novel characterization of Radix Angelicae Dahuricae before and after the sulfur-fumigation process by combining high performance liquid chromatographic fingerprint and multi-ingredients determination.

BACKGROUND: Harmful sulfur-fumigation processing method is abused during Radix Angelicae Dahuricae preparation. However, the analytical technique characterizing Radix Angelicae Dahuricae before and after the sulfur-fumigation process is absent. MATERIALS AND METHODS: The high performance liquid chromatography (HPLC) technique was adopted to develop methods combining finger-print analysis and multi-ingredients simultaneous determination for quality evaluation of Radix Angelicae Dahuricae before and after the sulfur-fumigation process. The chromatographic fingerprint method was established for qualitative analysis coupled with statistical cluster analysis basing on Euclidean distance. Additionally, a determination method was developed for quantitative analysis, which was able to assay the concentrations of the major coumarins including imperatorin, isoimperatorin, xanthotoxin, xanthotoxol, isoimpinellin, oxypeucedanin, and bergapten in Radix Angelicae Dahuricae simultaneously. The separations of the two methods were both achieved on a Hypersil octadecylsilyl C18 column (250 mm × 4.6 mm, 5 µm) at 35°C under different strategic gradient elution programs. The detection wavelength was set at 254 nm all the time. Method validation data indicated that the methods were both reliable and applicable. They were then used to assay different Radix Angelicae Dahuricae samples collected from good agricultural practice (GAP) bases and local herbal markets. RESULTS: The successful application demonstrated that the combination of HPLC fingerprint and simultaneous quantification of multi-ingredients offers an efficient approach for quality evaluation of Radix Angelicae Dahuricae before and after the sulfur-fumigation process. CONCLUSION: In order to discriminate Radix Angelicae Dahuricae before and after the sulfur-fumigation process, oxypeucedanin, and xanthotoxol were the most sensitive biomarkers and should be determined.

Multi-component analysis in sun-dried and sulfur-fumigated Angelicae Sinensis Radix by single marker quantitation and chemometric discrimination.

BACKGROUND: A new method has been developed for the simultaneous determination of ferulic acid, senkyunolide A, and Z-ligustilide in Angelicae Sinensis Radix before and after sulfur-fumigation using quantitative analysis of multi-components by a single marker (QAMS). MATERIALS AND METHODS: The feasibility and accuracy of QAMS were checked by the external standard method, and various high-performance liquid chromatographic instruments and chromatographic conditions were investigated to verify its applicability. Using ferulic acid as the internal reference substance, and the contents of senkyunolide A and Z-ligustilide were calculated according to relative correction factors by high-performance liquid chromatography. Meanwhile, the influence of sulfur-fumigation on these chemical components in Angelicae Sinensis Radix were evaluated and discriminated by chromatographic fingerprint and chemometrics. RESULTS: There was no significant difference observed between the QAMS method and the external standard method. Furthermore, sulfur-fumigation reduced the contents of ferulic acid, senkyunolide A, and Z-ligustilide in Angelicae Sinensis Radix by some degree, and the sun-drying and sulfur-fumigation processing could be easily discriminated by chromatographic fingerprint and chemometrics. CONCLUSION: QAMS is a convenient and accurate approach to analyzing multi-component when reference substances are unavailable, simultaneously, chemometrics is an effective way to discriminate sun-dried and sulfur-fumigated Angelicae Sinensis Radix.

Element analysis and characteristic identification of non-fumigated and sulfur-fumigated Fritillaria thunbergii Miq. using microwave digestion-inductively coupled plasma atomic emission spectrometry combined with Fourier transform infrared spectrometry.

BACKGROUND: Sulfur-fumigation may induce chemical transformation of traditional Chinese medicines leading to harmful effects following patient ingestion. For quality control, it is urgently needed to develop a reliable and efficient method for sulfur-fumigation identification. MATERIALS AND METHODS: The spectrochemical identification of non-fumigated and sulfur-fumigated Fritillaria thunbergii Miq. was carried out to evaluate inorganic elements and organic components. The concentrations of 12 elements, including Zn, Mn, Cu, Fe, Li, Mg, Sr, Pb, As, Cd, Hg, and S of samples were determined by microwave digestion - inductively coupled plasma atomic emission spectrometry (ICP-AES). Meanwhile, Fourier transform infrared spectrometry (FTIR) was used for the study of chemical group characteristic reactions after sulfur-fumigation. RESULTS: The concentrations of Fe, Mg, Hg, and S elements showed significant differences between non-fumigated and sulfur-fumigated Fritillaria thunbergii Miq. The characteristic stretching vibrations of some groups in FTIR spectra, such as -OH, -S = O and -S-O, provided the identification basis for the discrimination of non-fumigated and sulfur-fumigated Fritillaria thunbergii Miq. CONCLUSION: The application of microwave digestion - ICP-AES was successfully used in combination with FTIR to authenticate and evaluate the quality of medicinal Fritillaria thunbergii Miq. Further applications of this technique should be explored.

[Influence of sulfur-fumigated Paeoniae Radix Alba on contents of ten indicative components in Guizhi Decoction].

OBJECTIVE: To investigate the influence of sulfur-fumigated Paeoniae Radix Alba on the contents of main constituents in Guizhi Decoction. METHODS: HPLC was applied to the simultaneous determination and comparative analysis of ten main indicative components in Guizhi Decoction contained sun-dried and sulfur-fumigated Paeoniae Radix Alba. The analysis was performed on a Kromasil C18 column (250 mm x 4.6 mm, 5 µm) with the mobile phase consisting of acetonitrile-0.1% aqueous formic acid. The flow rate was 1.0 mL/min and the column temperature was 30 °C. The detection wavelengths were set at 254 nm and 280 nm, respectively. 20 µL was injected for analysis with gradient elution. RESULTS: There were obvious differences existed between the concentrations of ten main indicative components in Guizhi Decoction contained sun-dried and sulfur-fumigated Paeoniae Radix Alba. CONCLUSION: Sulfur-fumigated Paeoniae Radix Alba can influence the main effective constituents contained in Guizhi Decoction in quantity. The absence in the study of prescriptions affected by sulfur-fumigation is filled up through the investigation of the chemical basis of Guizhi Decoction influenced by sulfur-fumigated Paeoniae Radix Alba, and also strong evidences are offered to the further regulation and supervision of sulfur-fumigation in herbal medicines.

Characterization of Chemical Composition of Pericarpium Citri Reticulatae Volatile Oil by Comprehensive Two-Dimensional Gas Chromatography with High-Resolution Time-of-Flight Mass Spectrometry.

Pericarpium Citri Reticulatae (Chenpi in Chinese) has been widely used as an herbal medicine in Korea, China, and Japan. Chenpi extracts are used to treat indigestion and inflammatory syndromes of the respiratory tract such as bronchitis and asthma. This thesis will analyze chemical compositions of Chenpi volatile oil, which was performed by comprehensive two-dimensional gas chromatography with high-resolution time-of-flight mass spectrometry (GC × GC-HR-TOFMS). One hundred and sixty-seven components were tentatively identified, and terpene compounds are the main components of Chenpi volatile oil, a significant larger number than in previous studies. The majority of the eluted compounds, which were identified, were well separated as a result of high-resolution capability of the GC × GC method, which significantly reduces, the coelution. ß -Elemene is tentatively qualified by means of GC × GC in tandem with high-resolution TOFMS detection, which plays an important role in enhancing the effects of many anticancer drugs and in reducing the side effects of chemotherapy. This study suggests that GC × GC-HR-TOFMS is suitable for routine characterization of chemical composition of volatile oil in herbal medicines.

Profiling and characterization of volatile components from non-fumigated and sulfur-fumigated Flos Lonicerae Japonicae using comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry coupled with chemical group separation.

Flos Lonicerae Japonicae (FLJ) is a popular herb used for many centuries in Traditional Chinese Medicine as a treatment of fever and inflammation. Non-fumigated processing of FLJ has been the traditional approach used in post-harvest preparation of the commodity for commercial use. However, in recent years, natural drying processing of FLJ has been replaced by sulfur-fumigation for efficiency and pest control. Sulfur-fumigation can induce changes in the volatile compounds of the herb, altering its medicinal properties. A comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GC×GC-TOF/MS) method was established for the resolution and determination of volatile components in non-fumigated and sulfur-fumigated FLJ. In this paper, analysis of the volatile oils in non-fumigated and sulfur-fumigated (including lab-prepared sulfur-fumigated and industrial sulfur-fumigated) FLJ was performed using GC×GC-TOF/MS. Seventy-three representative volatile components were identified, including furans, alkalies, acids, aldehydes, ketones, alcohols, terpenes, esters, and others, as the main components of FLJ volatile oils. The proposed method was successfully applied for rapid and accurate quality evaluation of FLJ and its related medicinal materials and preparations.

[Sulfur-fumigation, maintenance method of Chinese herbal medicine-discard or inheritance].

Sulfur-fumigation processing technology is an ancient maintenance method, which plays a certain role in storage and preservation for Chinese herbal medicine. But in recent years, with the further explanation of sulfur-fumigation processing mechanism and more attention to the safety of drugs, such traditional maintenance method of Chinese herbal medicine is now being questioned by more and more people. The authors think we should have selective inheritance rather than abslute discard to this ancient processing technology after reviewing the literatures published in recent 20 years, and some suggestions are also put forward, which can supply some references for related drug supervision departments.

[Quick identification and discrimination of sun-dried and sulfur-fumigated cimicifugae rhizoma by Fourier transform infrared spectroscopy].

OBJECTIVE: To develop a method for quick identification of sun-dried and sulfur-fumigated Cimicifugae Rhizoma by Fourier transform infrared spectroscopy (FTIR). METHODS: The alcoholic and aqueous extracts of sun-dried and sulfur-fumigated Cimicifugae Rhizoma were analyzed and compared by FTIR combined with second derivative infrared spectroscopy. RESULTS: FTIR spectra showed that there were some differences in the positions of infrared absorption peaks and the relative intensities in the alcoholic and aqueous extracts of sun-dried and sulfur-fumigated Cimicifugae Rhizoma, and the second derivative IR spectra clearly enhanced the spectral resolution of their differences. FTIR spectra showed that the new absorption peaks of Cimicifugae Rhizoma appeared and a part of original absorption peaks disappeared after sulfur-fumigation in aqueous extracts, while a lot of new absorption peaks appeared and the intensities of almost all absorption peaks significantly decreased after sulfur-fumigation in alcoholic extracts. Second derivative IR spectra showed that both sun-dried and sulfur-fumigated Cimicifugae Rhizoma extracted by water differed significantly from each other ranging from about 3 950 to 3 940 cm(-1), 3 850 to 3 800 cm(-1), 1 800 to 1 750 cm(-1), as well as from 1 400 to 1 350 cm(-1); Differences also existed between sun-dried and sulfur-fumigated Cimicifugae Rhizoma extracted by ethanol ranging from about 3 980 to 3 960 cm(-1), 3 850 to 3 800 cm(-1), and 1 500 to 1 460 cm(-1). CONCLUSION: The FTIR method combined with the second derivative IR spectrum can be used to analyze and distinguish sun-dried and sulfur-fumigated Cimicifugae Rhizoma quickly and accurately. The developed method provides an efficient approach for the quality control of Chinese herbal medicines with its simplicity and strong specificity.

[Quick identification of sun-dried and sulfur-fumigated angelicae sinensis radix by Fourier transform infrared spectroscopy].

OBJECTIVE: To develop a quick identification method for the sun-dried and sulfur-fumigated Angelicae Sinensis Radix used by Fourier transform infrared spectroscopy (FTIR) combined with second derivative infrared spectroscopy. METHOD: The alcoholic and aqueous extracts of sun-dried and sulfur-fumigated Angelicae Sinensis Radix were analyzed by using FTIR, the further analysis was used by second derivative infrared spectroscopy. RESULT: There existed differences between their infrared spectra either extracted by ethanol or water, while the distinctions were more obvious after analyzing their alcoholic and aqueous extracts through high resolution of second derivative infrared spectroscopy. Infrared spectra showed that the absorption peaks of Angelicae Sinensis Radix were significantly reduced and a new absorption peak appeared after sulfur-fumigated process in alcoholic extracts, while both of them changed markedly in the "fingerprint region" ranging from 1 000 to 400 cm(-1) in aqueous extracts. Second derivative spectra showed that the absorption peaks of sulfur-fumigated Angelicae Sinensis Radix extracted by ethanol weakened and disappeared at about 3 578 cm(-1) and 3 541 cm(-1), while both of them differed significantly from each other ranging from 1 400 to 1 200 cm(-1) as well as 800 cm(-1) to 600 cm(-1), difference also existed between them extracted by water ranging from about 3 900 to 3 850 cm(-1) and 3 800 to 3 750 cm(-1). CONCLUSION: The FTIS method combined with second derivative can be utilized to distinguish sun-dried and sulfur-fumigated Angelicae Sinensis Radix efficiently, conveniently and accurately, and provide a basis for identification and quality control of Angelicae Sinensis Radix.

Global detection and analysis of volatile components from sun-dried and sulfur-fumigated herbal medicine by comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry.

The sulfur-fumigation process can induce changes in the contents of volatile compounds and the chemical transformation of herbal medicines. Although literature has reported many methods for analyzing volatile target compounds from herbal medicine, all of them are largely limited to target compounds and sun-dried samples. This study provides a comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry (GC×GC-TOF/MS) method based on a chemical profiling approach to identify non-target and target volatile compounds from sun-dried and sulfur-fumigated herbal medicine. Using Chrysanthemum morifolium as a model herbal medicine, the combined power of this approach is illustrated by the identification of 209 and 111 volatile compounds with match quality >80% from sun-dried and sulfur-fumigated Chrysanthemum morifolium, respectively. The study has also shown that sulfur-fumigated samples showed a significant loss of the main active compounds and a more destructive fingerprint profile compared to the sun-dried ones. 50 volatile compounds were lost in the sulfur-fumigated Chrysanthemum morifolium sample. The approach and methodology reported in this paper would be useful for identifying complicated target and non-target components from various complex mixtures such as herbal medicine and its preparations, biological and environmental samples. Furthermore, it can be applied for the intrinsic quality control of herbal medicine and its preparations.

Study on chemical fingerprinting of crude and processed Atractylodes macrocephala from different locations in Zhejiang province by reversed-phase high-performance liquid chromatography coupled with hierarchical cluster analysis.

BACKGROUND: In China, Atractylodes macrocephala is mainly distributed in Zhejiang province. It is, therefore, desirable to determine a reliable and accurate methodology to differentiate the samples collected from Zhejiang province. Although some studies on the fingerprints of Atractylodes macrocephala using HPLC have been published, none of them compared the processed product of Atractylodes macrocephala from different areas of Zhejiang province. MATERIALS AND METHODS: Reversed-phase high-performance liquid chromatography (RP-HPLC) coupled with hierarchical cluster analysis was employed in the fingerprint analysis of Atractylodes macrocephala from Zhejiang province, China. The LC assay was performed on a reversed-phase C18 column with linear gradient elution using water and acetonitrile. The LC data showed considerable variation of chemical constituents among Atractylodes macrocephala populations. RESULTS: 21 and 22 characteristic peaks in the 14 grants of Atractylodes macrocephala and its processed product were determined in samples from different habitats of Zhejiang province, respectively. Their chromatographic patterns were generally consistent although their contents of chemical compositions were greatly different. The results of hierarchical cluster analysis showed that the samples could be divided into four groups; it was able to select excellent resources from the groups. CONCLUSION: This was the first report of hierarchical cluster analysis of crude and processed Atractylodes macrocephala according to their chemical fingerprints and could be applied to the intrinsic quality control of crude and processed Atractylodes macrocephala. The processing technique of Atractylodes macrocephala through the pilot-scale experiment was first studied and is simple and suitable for prepared yin pian of Atractylodes macrocephala industrial manufacture.