Pharmacokinetic based study on "lagged stimulation" of Curcumae Longae Rhizoma - Piper nigrum couplet in their main active components' metabolism using UPLC-MS-MS.
BACKGROUND
Curcumae Longae Rhizoma is one of the commonly used traditional Chinese medicines, which has multiple biological activities such as relieving stagnation and stasis, pain alleviation, curing amenorrhea and wounds. However, its main active component-curcumin has poor absorption and very fast metabolism in body. To solve this problem, Piper nigrum was introduced for its ability to strengthen bioavailability of other compounds.


OBJECTIVE
In most cases of TCM couplets, all ingredients were prepared and taken simultaneously, which in our opinion did not take full advantage of their interactions. Therefore, order of administration should be adjusted according to pharmacokinetic parameters of the ingredients, which the ones act as supplement can first be taken, and main therapeutic components followed when the former reached its peak.


METHODS
the extract of Piper nigrum (containing at least 95% piperine) was taken by rats 6h before taking Curcumae Longae Rhizoma extract (containing at least 95% curcumin). Then, a UPLC-MS-MS method was developed to determine their content in plasma simultaneously. Determination was carried out by on a C18 column within 5min by isocratic elution using 0.2% formic acid and acetonitrile (50:50, v/v). Tandem mass detection was conducted by selective reaction monitoring (SRM) via electrospray ionization (ESI) source in positive mode. Samples were pre-treated by liquid-liquid extraction (LLE), and verapamil was used as internal standard (IS).


RESULTS
For both curcumin and piperine, the proposed method had good linearity (r2=0.999) within the concentration range of 1-1000ng/ml, with good recovery, precision and stability. The lower limit of quantification (LLOQ) was 1ng/ml. As pharmacokinetic data indicated, Maximum concentration (Cmax) of curcumin increased significantly to 394.06; the time reach maximum concentration (Tmax) and elimination half-life (T1/2) were 0.5 and 0.67h, respectively; CONCLUSION: The results provide a good strategy for the investigation of TCM formula especially the couplets, as well as a fast, selective and sensitive UPLC-MS-MS method determining active components in-vivo. Furthermore, the finding of "lagged stimulation" suggested that the use of complex formula should take pharmacokinetics into much more careful consideration.
Guangdong Province Engineering Technology Research Institute of T.C.M., 60 Hengfu rd., Guangzhou, 510095, China; Affiliated Guangdong second TCM hospital, Guangzhou University of Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China; Guangdong Provincial Key Laboratory of Research and Development in Traditional Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China.
Zhao
Z
Chen
Guangdong Province Engineering Technology Research Institute of T.C.M., 60 Hengfu rd., Guangzhou, 510095, China; Affiliated Guangdong second TCM hospital, Guangzhou University of Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China; Guangdong Provincial Key Laboratory of Research and Development in Traditional Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China. Electronic address: jbddjsw@163.com.
Dongmei
D
Sun
Guangdong Province Engineering Technology Research Institute of T.C.M., 60 Hengfu rd., Guangzhou, 510095, China; Affiliated Guangdong second TCM hospital, Guangzhou University of Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China; Guangdong Provincial Key Laboratory of Research and Development in Traditional Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China.
Xiaoli
X
Bi
Guangdong Province Engineering Technology Research Institute of T.C.M., 60 Hengfu rd., Guangzhou, 510095, China; Affiliated Guangdong second TCM hospital, Guangzhou University of Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China; Guangdong Provincial Key Laboratory of Research and Development in Traditional Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China.
Xiaohui
X
Zeng
Guangdong Province Engineering Technology Research Institute of T.C.M., 60 Hengfu rd., Guangzhou, 510095, China; Affiliated Guangdong second TCM hospital, Guangzhou University of Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China; Guangdong Provincial Key Laboratory of Research and Development in Traditional Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China.
Wenhui
W
Luo
Guangdong Province Engineering Technology Research Institute of T.C.M., 60 Hengfu rd., Guangzhou, 510095, China; Affiliated Guangdong second TCM hospital, Guangzhou University of Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China; Guangdong Provincial Key Laboratory of Research and Development in Traditional Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China.
Dake
D
Cai
Guangdong Province Engineering Technology Research Institute of T.C.M., 60 Hengfu rd., Guangzhou, 510095, China; Affiliated Guangdong second TCM hospital, Guangzhou University of Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China; Guangdong Provincial Key Laboratory of Research and Development in Traditional Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China.
Qiaohuang
Q
Zeng
Guangdong Province Engineering Technology Research Institute of T.C.M., 60 Hengfu rd., Guangzhou, 510095, China; Affiliated Guangdong second TCM hospital, Guangzhou University of Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China; Guangdong Provincial Key Laboratory of Research and Development in Traditional Chinese Medicine, 60 Hengfu rd., Guangzhou, 510095, China.
Aili
A
Xu
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D030024:Curcuma / Q000737:chemistry / Q000378:metabolism*; D003474:Curcumin / Q000493:pharmacokinetics*; D065607:Cytochrome P-450 Enzyme Inhibitors / Q000493:pharmacokinetics*; D004365:Drugs, Chinese Herbal / Q000493:pharmacokinetics; D008297:Male; D029222:Piper nigrum / Q000737:chemistry / Q000378:metabolism*; D010880:Piperidines / Q000493:pharmacokinetics*; D010936:Plant Extracts / Q000493:pharmacokinetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D027343:Rhizome / Q000737:chemistry; D053719:Tandem Mass Spectrometry
D065607:Cytochrome P-450 Enzyme Inhibitors; D004365:Drugs, Chinese Herbal; D010880:Piperidines; D010936:Plant Extracts; D003474:Curcumin
Curcumae Longae Rhizoma; Lagged stimulation; Pharmacokinetics; Piper nigrum; UHPLC-MS-MS