Study On Quality Standard Of Fructus Cannabis And Processed Products Of Traditional Chinese Medicine

Purpose:Hemp seed is the dry ripe fruit of Cannabis sativa L., which has a special curative effect on astriction by moisturizing pathogenic dryness to loosen the bowel. Hemp seed has both high medicinal value and high nutritional value. However, few researches were carried out on this traditional Chinese medicine, the herb's quality control standard has not yet been established, and the modern processing research of hemp seed is zero. In order to efficiently control the quality of hemp seed and its processed products, we studied the change of the components and pharmacology effect before and after processing by "fried Act" and "microwave" processing technology.Material and method:Material: Hemp seed was purchased from Anguo Chinese herbal medicines market and identified as the dry ripe fruit of Cannabis sativa L. by Prof. Zhai yan-jun in Liaoning University of TCM.Method:(1) The single factor and the response surface methodology were used to develop an optimum extraction procedure for hemp oil from hemp seed. The orthogonal design was used to select an optimum extraction procedure for cannabinoids of hemp seed.(2) HPLC method was developed to simultaneous determine cannabinoids in hemp seed collected from 13 different geographic locations.(3) HPLC method was developed to simultaneous determine fatty acids in hemp seed collected from 13 different geographic locations.(4) "Fried Act" and "microwave" processing technology were carried out to compare the change of the index components before and after processing.(5) Mice constipation model and mice hot-plate method was established to examine laxative pharmacology effect and analgesic pharmacology effect, respectively. (6) The quality standards of hemp seed and its processed products were established preliminarily.Results:(1) The optimum extraction procedure for hemp oil from hemp seed was as follows: petroleum ether 75 mL, ultrasonic time 30 min and ultrasonic temperature 20℃. An HPLC method for simultaneous determination of flax acid, linoleic acid, hexadecanoic acid, oleic acid and stearic acid in hemp seed was established. The method is of high sensitivity and good reproducibility and it can be used for quality control of hemp seed.(2) The optimum extraction procedure for cannabinoids was as follows: methanol 100 mL, ultrasonic time 30 min and extraction repetition 3 times.(3) A silica gel column chromatography-HPLC method for simultaneous determination of cannabidiol, cannabinol and tetrehydrocannabinol in hemp seed was established. The method is of high sensitivity and good reproducibility and it can be used for quality control of hemp seed.(4) Both the "Fried Act" and "microwave" processing showed the same result: the contents of cannabinoids material were reduced and the contents of fatty acid substances were increased.(5) Pharmacodynamics experiments showed that hemp seed's ethanol extraction had the significant analgesic effect, while the processed products'decreased. On the other hand, both the petroleum ether extract of the hemp seed and processed products had the laxative effect in mice, and the processed was slightly better than the raw.(6) The quality standards of hemp seed and its processed products were established preliminarily. Tentative: calculated on dry goods, including three cannabinoids (Δ⁹-tetrehydrocannabinol, cannabinol and cannabidiol) in hemp seed not less than 2.54μg·g^-1, including five fatty acids (linoleic acid,α-flax acid, hexadecanoic acid, oleic acid and stearic acid) not less than 20%; including three cannabinoids in processed products not less than 0.94μg·g^-1, including five fatty acids not less than 27 %. Conclusions:(1) An optimum extraction procedure for hemp oil from hemp seed was established by response surface methodology. An optimum extraction procedure for cannabinoids of hemp seed was established by orthogonal design.(2) Some accurate, reliable and sensitive methods for determination of cannabinoids and fatty acids in hemp seed were established by HPLC, which could be used as quality control of hemp seed.(3) "Fried Act" and "microwave" processing method was carried out to process hemp seed, which could provide a valuable reference to establish hemp seed's processing procedure.(4) The pharmacodynamics research could provide experimental basis for clinical drug safety and standardization of processing of hemp seed.(5) The quality standards of hemp seed and its processed products are stable and feasible.