Pharmacokinetics and metabolism of ligustilide, a major bioactive component in Rhizoma Chuanxiong, in the rat.

Ligustilide is the most abundant bioactive ingredient in Rhizoma Chuanxiong, a Chinese medicinal herb commonly used for the treatment of cardiovascular ailments. The present study reported, for the first time, the pharmacokinetics of ligustilide, administered in its pure form and in an herbal extract, in rats. After i.v. administration of pure ligustilide, it was distributed extensively (V(d), 3.76 +/- 1.23 l/kg) and eliminated rapidly (t(1/2), 0.31 +/- 0.12 h). The i.v. clearance (CL) of ligustilide after Chuanxiong extract administration was significantly higher than that dosed in its pure form [CL, 20.35 +/- 3.05 versus 9.14 +/- 1.27 l/h/kg, p < 0.01; area under the curve (AUC), 0.79 +/- 0.10 versus 1.81 +/- 0.24 mg x h/l, p < 0.01], suggesting significant interaction between ligustilide and components present in the extract. Dose-dependent pharmacokinetics was observed after i.p. administration, and a significantly higher dose-normalized AUC (1.77 +/- 0.23 mg x h/l) at 52 mg/kg was obtained than that at 26 mg/kg (0.93 +/- 0.07 mg x h/l, p < 0.05). Oral bioavailability of ligustilide was low (2.6%), which was partly because of extensive first-pass metabolism in the liver. Seven metabolites of ligustilide were identified, and three of them were unequivocally characterized as butylidenephthalide, senkyunolide I, and senkyunolide H. These three compounds also occurred naturally in the herb and were reported to be bioactive.

A method for the analysis of ginsenosides, malonyl ginsenosides, and hydrolyzed ginsenosides using high-performance liquid chromatography with ultraviolet and positive mode electrospray ionization mass spectrometric detection.

A high-performance liquid chromatographic separation coupled to diode array absorbance and positive mode electrospray mass spectrometric detection has been developed for the analysis of ginsenosides, malonyl ginsenosides, and hydrolyzed ginsenosides in extracts of Asian ginseng (Panax ginseng) and American ginseng (P. quinquefolius). The method is capable of separating, identifying, and quantifying the predominant ginsenosides found in heated alcoholic extracts of Asian and American ginseng roots routinely sold as nutraceuticals. It also separates and identifies the malonyl ginsenosides often found in cold alcoholic extracts of ginseng root and has the potential to quantify these compounds if pure standards are available. Furthermore, it can separate and identify ginsenoside hydrolysis products such as those readily produced in situations mimicking gastric situations, including those used for dissolution studies (i.e., 0.1 N HCl, 37 degrees C).

Simultaneous quantification of 12 bioactive components of Ligusticum chuanxiong Hort. by high-performance liquid chromatography.

A sensitive and specific HPLC-UV method has been developed, for the first time, to simultaneously quantify 12 bioactive ingredients in Ligusticum chuanxiong Hort. (Rhizoma Chuanxiong). This assay was fully validated in respect to precision, accuracy and sensitivity. This method was successfully applied to quantify twelve ingredients in six different Chuanxiong samples. The results demonstrated significant variations in the total content and quantity of each of the main bioactive compounds in different herbs, indicating that quality control of bioactive ingredients in Chuanxiong is critical to ensure its clinical benefits. This assay can be readily utilized as quality control method for Chuanxiong.

[Study on fingerprint of rhizoma chuanxiong by HPLC-DAD-MS].

AIM: To establish a high performance liquid chromatographic fingerprint for the quality control of rhizoma Chuanxiong, a traditional Chinese medicine derived from the root of Ligusticum chuanxiong Hort.. METHODS: An on-line optimized HPLC-DAD-MS technique was employed. The HPLC analysis was performed on a Waters Symmetry C18 column (150 mm x4. 6 mm ID, 5 microm) with a Waters Spherisorb S5 ODS2 (10 mm x 4.6 mm) guard column. The mobile phase consisted of A (methanol) and B (0.25% acetic acid). Components were separated using the following gradient profile: 32% B at 0-3 min, 32%-85% B at 3-33 min, 85%-100% B at 33-52 min; flow rate was 0.7 mL x min(-1). DAD was set from 190 to 400 nm, the fingerprint was monitored at 294 nm. All mass spectra were acquired in the positive ion mode with electrospray ionization; the full scan mass spectrum was recorded over the range of m/z 100-800. Nine samples from three companies were analyzed; the main characteristic peaks were identified based on the comparison of UV and MS spectra of each analyte with that of authentic compounds and literature data. RESULTS: The HPLC fingerprint was established based on the analysis of nine rhizoma Chuanxiong herbal samples supplied by three companies. Twenty-one characteristic peaks were found in all nine samples. These peaks were classified into four groups: group I at 0-12 min, three peaks were found, and the marker peak 3 was confirmed as ferulic acid; group II at 12-24 min, four peaks were found, and the marker peaks 4 and 5 were identified as senkyunolide I and senkyunolide H; group III at 24-32 min, there were seven peaks, and the marker peaks 9, 11, 13 and 14 were elucidated as senkyunolide A, coniferylferulate, ligustilide and 3-butylidenephthalide, respectively; group IV at 32-50 min, seven peaks were observed, and the marker peaks 15 and 17 were identified as riligustilide and levistolide A. The peak areas of 13 main peaks with normalized peak area (1% were determined. Using the most abundant peak 13 as the reference peak, the calculated relative retention times (tR of the characteristic peak/tR of the reference peak) among nine samples were consistent (RSD < or = 1%), while the calculated relative peak areas (peak area of the characteristic peak/peak area of the reference peak) among nine samples were significantly different (P < 0.001), indicating that all nine samples tested contain similar 13 main components with different quantities. CONCLUSION: The established HPLC fingerprint is very specific, and can be used to evaluate the quality consistency of different rhizoma Chuanxiong herbs.

Metabolic conversion from co-existing ingredient leading to significant systemic exposure of Z-butylidenephthalide, a minor ingredient in Chuanxiong Rhizoma in rats.

Pharmacokinetic (PK) study of medicinal herbs is a great challenge, because which component(s) is(are) the bioactive ingredients is largely unknown. Most of the reported PK studies of herbs focused on the major ingredients regardless of their in vivo bioactivities, while PK of components with low content in herbs is often ignored. The present study demonstrates how PK study can reveal potential importance of a low content ingredient to the herbal bioactivities using Z-butylidenephthalide (BuPh), a bioactive phthalide present in a significantly low quantity in medicinal herb Chuanxiong Rhizoma, as an example. PK of BuPh was investigated in rats using Chuanxiong extract, fraction containing BuPh and ligustilide, and pure BuPh, respectively. The results demonstrated that remarkable blood concentrations of BuPh were observed after administration of the herbal extract and its systemic exposure was significantly different between BuPh given in pure and mixed forms. More interestingly, AUC of BuPh via intake of fraction (9.3-fold) and extract (4.5-fold) was significantly greater than that obtained from pure BuPh, which was further evidenced to be mainly due to metabolic conversion from ligustilide, a major component in Chuanxiong. Our findings revealed that although it naturally occurred in low amount, BuPh reached significant systemic concentrations via metabolic conversion from ligustilide. Moreover, our results demonstrated that PK study is one of crucial and inevitable steps for revealing in vivo bioactive ingredients of herbal medicines, and such studies should be more appropriate to focus on in vivo profile of the ingredients co-existing in herbs rather than only studying them individually.

Post-harvest alteration of the main chemical ingredients in Ligusticum chuanxiong Hort. (Rhizoma Chuanxiong).

Rhizoma Chuanxiong (Ligusticum chuanxiong HORT.) is a commonly used traditional Chinese medicinal herb for the treatment of cardiovascular disorders. Significant variations of the main components in this herb were observed in commercial samples. The present study investigated effects of post-harvest drying and processing methods on nine main components in the herb. Results showed that drying at 60 degrees C or under the sun the contents of three major constituents, namely senkyunolide A (4), coniferylferulate (5) and Z-ligustilide (6), decreased significantly, while the contents of ferulic acid (1), riligustilide (8) and levistolide A (9) increased significantly. Senkyunolide I (2) and senkyunolide H (3), which were not detected in fresh herbs, appeared in dried samples. Similar chemical alterations, such as decrease in the contents of three major ingredients and increase in the contents of compounds 1, 2, 3, 8 and 9, were also observed in differently processed herbal samples. The possible converting mechanisms of these components were clarified by employing pure major components treated under the same conditions.

Low oral bioavailability and pharmacokinetics of senkyunolide a, a major bioactive component in Rhizoma Chuanxiong, in the rat.

The pharmacokinetics of senkyunolide A, one of the major bioactive ingredients in the traditional Chinese medicinal herb Rhizoma Chuanxiong, which is commonly used for the treatment of cardiovascular diseases, was studied in rats. After intravenous (IV) administration, senkyunolide A was extensively distributed (Vd/F: 6.74 +/- 0.73 L/kg) and rapidly eliminated from the plasma (CL/F: 7.20 +/- 0.48 L/h per kilogram and t1/2: 0.65 +/- 0.06 hr). Hepatic metabolism was suggested as the major route of senkyunolide A elimination as indicated by the results of in vitro S9 fraction study. After intraperitoneal (IP) administration, senkyunolide A exhibited dose-independent pharmacokinetics. The absorption after IP administration was rapid (Tmax: 0.04 +/- 0.01 hours), and the bioavailability was 75%. After oral administration, senkyunolide A was also absorbed rapidly (Tmax: 0.21 +/- 0.08 hours); however, its oral bioavailability was low (approximately 8%). The contributing factors were determined to be instability in the gastrointestinal tract (accounting for 67% of the loss) and hepatic first-pass metabolism (accounting for another 25%). Pharmacokinetics of senkyunolide A were unaltered when Chuanxiong extract was administered, which suggests that components in the extract have insignificant effects on senkyunolide A pharmacokinetics.

Time-course accumulation of main bioactive components in the rhizome of Ligusticum chuanxiong.

Fresh rhizomes of Ligusticum chuanxiong, a commonly used traditional Chinese medicinal herb, were collected monthly from a cultivating base in China practicing good agriculture practice (GAP). These samples were analyzed by HPLC-UV for their main chemical ingredients. Senkyunolide A (6), coniferyl ferulate (7) and Z-ligustilide (8) were identified as the major ingredients. The accumulation of the main ingredients with time in the herb was elucidated. Both individual and total contents of all main components gradually increased from the beginning of October to the middle of next April. The weight of a single rhizome reached a plateau at the end of May, whilst the content of the major ingredients peaked in the middle of April. Based on these results, it is recommended that Rhizoma Chuanxiong be harvested between the middle of April and the end of May.

Simultaneous analysis of seventeen chemical ingredients of Ligusticum chuanxiong by on-line high performance liquid chromatography-diode array detector-mass spectrometry.

An on-line high performance liquid chromatography (HPLC)-diode array detector (DAD)-mass spectrometry (MS) analytical method has been developed to simultaneously separate and identify seventeen main constituents of Chuanxiong ( Ligusticum chuanxiong Hort.), a traditional Chinese medicinal herb. In three Chuanxiong samples, nine compounds were unequivocally determined as vanillin (1), ferulic acid (2), senkyunolide I (4), senkyunolide H (5), senkyunolide A (6), coniferyl ferulate (7), Z-ligustilide (8), neocnidilide (9) and 3-butylidenephthalide (10) by comparing their t R, UV, and MS data with those obtained for the authentic compounds. Furthermore, additional eight compounds were tentatively identified as senkyunolide J (11), senkyunolide F (12), 3-butylphthalide (13), cnidilide (14), riligustilide (15) or Z,Z'-6,8',7,3'-diligustilide (16), tokinolide B (17), levistolide A (18) and senkyunolide P (19), based on their MS data and the comparison of their UV spectra with the published references. This is the first report to describe the development of an on-line HPLC-DAD-MS method to simultaneously analyse seventeen main constituents present in Chuanxiong and to construct the chemical profiles of various Chuanxiong samples.