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Extraction of active ingredients of saffron

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					Extraction of active ingredients of saffron

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Key words HPLC method
Abstract: Objective: To screen the active ingredient in the extraction
process of safflower. Methods: RPHPLC determination of safflower
extract HSYA A as quantitative indicators, and the uniform application
of single-factor experimental design, selection extraction safflower
active ingredients. Results: The extraction of water as solvent,
reflux 2 times, each time 20 min, each time the amount of water to 13
times the amount of medicine. Per gram of safflower extract herbs HSYA
A was the highest. Conclusion: The extraction process is stable and
feasible.
Keywords: Safflower; active ingredient; RPHPLC method; HSYA A
To studies of extraction methods of utility portion in Carthamus
tinctorius
YANG Xiaojun, WU Guirong, WANG Yan
(College of Pharmacy, Xinjiang Medical University, Urumqi, 830054
China)
Abstract: Objective: To screen the extracting method of utility
portion in Carthamus tinctorius. Methods: A RPHPLC method was
developed to determinate the content of hydroxysafflor yellow A in
extracting portin of safflor yellow. According to the mono factor
design and uniform design. We screened the extracting method of
utility portion in Carthamus tinctorius. Results: The extracting
solvent was water, repeat 2 times, every time add 13 times water and
the extracting lasts 20 min to the Chinese medicine drug. The content
of hydroxysafflor yellow A was highest in extraction for every gram
Carthamus tinctorius. Conclusion: The method is simple, stabilization
and feasible.
Key words: Carthamus tinctorius; utility portion; RPHPLC;
hydroxysafflor yellow A
Safflower (Carthmus tinctorins L.) is a dry tubular flowers of
Compositae [1], only contained in the "Kai Bao Materia
Medica," "Lord of postpartum blood supply Koujin, not the
bad blood intra-abdominal, cramps, abdominal fetal death in, and
Jiuzhu service, also in Gudu of blood. " With the expansion of
its coronary artery, blood pressure, hypoxia, blood circulation,
anti-inflammatory analgesic and other pharmacological effects [2].
Germany and Japan in 1906, high-level turtles first was isolated from
the red pigment in safflower (C12H22O11) content of 0.3% to 0.6%;
yellow pigment mixture containing a variety of chalcones ketones
(Safflor yellow, SY), the content of 20% to 30 %. Pharmacological
experiments confirmed that [3] SY is the main active ingredients of
effective, SY hydroxyl yellow A (hydroxysafflor yellow A, HSYA) were
higher, with a typical pharmacological effects [4].
In the land rich in natural resources, plenty of sunshine, the
production of all-natural, pollution-free good base of green herbs,
safflower production accounting for about 80%. In this study, water as
solvent extraction, the single factor and uniform experimental design
and the active ingredients of saffron Optimum extraction process,
designed to provide resources for the development of the scientific
basis of Xinjiang safflower.
An instrument and reagent
1.1 Instrument HP 1100 liquid chromatograph (USA), Shimadzu UV2201 UV
spectrophotometer (Japan), high-speed centrifuge (Shanghai Anting),
YKH§º liquid fast mixer (Jiangxi), SYZ550 quartz sub-boiling water
distiller (Jintan), the United States Thermo Zirchrom C18 column (5
¦Ìm, 250 mm ¡Á 4.6 mm) (Tianjin Chen Hang Technology Co., Ltd.), a
single disk optical analytical balance (Beijing Optical Instrument
Factory, d = 0.01 mg) , electric thermostat set (Shanghai Medical
Equipment Factory.)
1.2 HSYA reagent A (providing medical department of Peking University,
content 98.05%), methanol HPLC grade phosphoric acid were of
analytical grade, safflower (Tacheng), ultra-pure water (Shihezi
University Hospital Affiliated to self).
2 Methods and Results
2.1 HSYA A determination of chromatographic conditions: mobile phase:
methanol -0.5% phosphoric acid (42:58); detection wavelength 400 nm;
column temperature 25 ¡æ; flow rate of 1.0 ml / min; injection 5.0 ¦Ìl.
External standard method: the number of theoretical plates shall not
be less than 6500, resolution greater than 1.5; standard curve (Y =
0.000 375Ai +0.000 526, r = 0.999 8, n = 3), linear relations in the
10 ~ 160 ¦Ìg / ml concentration range can be detected, every 1.0 hours
in the day with a known concentration of high, medium and low HSYA
standard sample Sample (n = 9) measured the precision (RSD = 1.1%), in
each Determination of the test, the sample every 2 hours for Reference
Standard HSYA and stability of the test determination (RSD = 1.1%) and
reproducibility (RSD = 1.2%), application of high, medium and low
concentration of 3 the test for recovery rate experiment, recovery was
101.2%, indicating that the method stable and feasible. Record HSYA
chromatograms (Figure 1A).
2.2 Preparation and determination of the test weighed amount of
safflower medicine, according to the design extraction process were
precise amount of extract 0.5 ml, with a 0.45 ¦Ìm microporous membrane
filtration, the filtrate obtained sample determined. Record the test
of HSYA chromatograms (Figure 1B).
Figure 1HPLC chromatography (HSYA) slightly
2.3 Extraction Process
2.3.1 Single-factor experiment
2.3.1.1 Different indicators of water addition on the extraction times
fixed 1, extraction time 1 h, do not soak, each with herbs
6,8,10,15,20 times for reflux extraction of water, HSYA determination
results were 0.88%, 1.04%, 1.57%, 1.67%, 1.40%.
2.3.1.2 Indicators of different extraction time on extraction times of
1 fixed, extraction of water for the herbs to 15 times (w / v), do not
soak, respectively 20,40,60,80,100 min to reflux extraction, HSYA
determination results were 1.32%, 1.49%, 1.45%, 1.44%, 1.25%.
2.3.1.3 Different extraction times on the index of a fixed amount of
water is 15 times, the extraction time was 40 min, not soaking,
respectively, were extracted by 1,2,3,4 times. HSYA determination
results were 1.50%, 1.68%, 1.69%, 1.66%.
2.3.1.4 immersion time on the index of a fixed amount of water is 15
times, the extraction time was 40 min, extraction times of 1 second,
respectively, after soaking 0.5,1,2,4,6,8,11 h extraction. HSYA
determination results were 1.42%, 1.43%, 1.42%, 1.36%, 1.42%, 1.32%,
1.31%.
2.3.2 Extraction of uniform design
2.3.2.1 Extraction of uniform experimental design can be seen from the
single factor experiments solvent volume and extraction time have a
greater impact on the index, and extraction times with little effect
of immersion time, it will be fixed at 2 times the number of
extraction, not immersion. The solvent volume, extraction time is
investigated factors, using uniform experiment design selection
process. Factor levels in Table 1, experiments, and results in Table
2.
Table 1 Experimental design factors uniform level of form factors
[omitted]

Table 2U5 (52) uniform experiment design table NO. [Omitted]
2.3.2.2 the results of data processing using uniform experiment,
Shenyang Pharmaceutical University, uniform design procedures for
regression, have equation: Y =- 0.576 0 +0.365 9X1-0.014 2X21
inspection of the equation, residual standard deviation S = 0.009 5,
multiple correlation coefficient R2 = 0.994 3, the regression equation
test of significance of F = 87.239 6, equation significantly (P
<0.05).
Optimum conditions: X1 (plus water) = 12.9 times, X2 (extraction time)
= 20 min, the theoretical prediction with HSYA 1.78% safflower,
prediction interval: 1.62% ~ 1.96%.
After a uniform design of experiments and data analysis: determine the
process of this study were: the amount of added solvent 13 times the
amount of medicine, 100 ¡æ back to the water extraction for 2 times,
each time 20 min.
2.3.2.3 verification experiment to take medicine three copies,
respectively, determined by this study to extract and sample handling
process, using established RPHPLC method for the determination HSYA
content of the samples were 1.69% (RSD = 1.18%, n = 3), 1.67% (RSD =
1.25%, n = 3), 1.68% (RSD = 1.58%, n = 3). 3 samples HSYA average
content of 1.68% (RSD = 0.60%, n = 3).
3 Discussion
3.1 In the light of the test chemicals and chemical stability in
aqueous solution HSYA the visit, the refrigerator -3 ¡æ save 15 d by
the ultraviolet and the reproducibility of qualitative and
quantitative HPLC chromatogram of the determination of the RSD
<2.0%, but room temperature, light, then 48 h chromatographic
impurity peak appears, and its UV absorbance and chromatographic peak
area of significant variation, RSD> 8.0%. HSYA structure because
contains 4 hydroxyl groups, so that stability is affected by
temperature, light and other factors larger.
3.2 The research based on the strong yellow water-soluble, water
extraction by selecting the first solvent to the active ingredient in
HSYA safflower as index, on the extraction process was the
experimental study. First screened by single factor indicator of
greater impact factor - HSYA (%), and then the level of each factor
based on a uniform experimental design, and process optimization. The
results show that: Extraction of active ingredients of saffron in the
extraction of content can HSYA 1.6% to 1.7%, the method is simple,
high yield, time-saving, stable, viable, worthy of promotion, with the
development of resources in Xinjiang safflower important application
value.
References:
[1] National Pharmacopoeia Committee. The Chinese Pharmacopoeia [S].
2000 edition. A. Beijing: Chemical Industry Press, 2000. 119.
[2] Song Liren, Hong Xun, Ding Xu, et al. Dictionary of Modern
Medicine [M]. Beijing: People Health Press, 2001.924928.
[3] Li Guangsheng. On the comprehensive utilization of red flowers
[J]. Shanxi Pharmaceutical Industry, 1992, 11 (4): 40.
[4] Takahashi Y, Miyassaka N, Tasaka SH, et al. Constitution of two
coloring matters in the flower petals of carthamus tinctorius L [J].
Tetrahedron Lett, 1982,23 (49): 5.