Quality detection method of traditional Chinese medicine water-honeyed pills

Abstract

The invention discloses a quality detection method of a traditional Chinese medicine preparation Xuefuzhuyu water-honeyed pill. Microscopic identification of angelica sinensis, peach kernels, rehmannia and platycodon grandiflorum, thin-layer identification of achyranthes bidentata, radix paeoniae rubra, platycodon grandiflorum, radix bupleuri and safflower carthamus, and peach kernel and safflower carthamus content determination methods are mainly added on the basis of an original detection method. The quality detection method is simple, convenient and rapid, results are accurate and reliable, and safety and effectiveness of medicine quality are guaranteed.

Description

TECHNICAL FIELD

[0001] The application relates to a detection method of traditional Chinese medicine, in particular to a quality detection method of Xuefuzhuyuwan Daimiwan. BACKGROUND

[0002] Xuefuzhuyuwan Daimiwan is composed of 11 medicinal materials, i.e., Angelica sinensis, Radix Paeoniae Rubra, Semen Persicae, Flos Carthami, Ligusticum chuanxiong, Rehmannia glutinosa, Achyranthes bidentata, Fructus Aurantii Immaturus (Fried with Wheat Bran), Platycodon grandiflorum, Bupleurum falcatum and Glycyrrhiza uralensis, and has the effects of activating blood and removing blood stasis, promoting qi and relieving pain. The quality standard of the preparation is recorded in the 2020 edition of the Chinese Pharmacopoeia. At present, microscopic and thin-layer qualitative identification and content determination of Radix Paeoniae Rubra and Fructus Aurantii Immaturus (Fried with Wheat Bran) are carried out in the quality control of Xuefuzhuyuwan Daimiwan. There is no content determination of main pharmacodynamic components in Semen Persicae and Flos Carthami and no microscopic identification method of Angelica sinensis, Rehmannia glutinosa, Semen Persicae and Platycodon grandiflorum in the original standard. In order to make up for the above-mentioned deficiencies and make the quality control technology more perfect and scientific, the microscopic identification method of Angelica sinensis, Rehmannia glutinosa, Semen Persicae and Platycodon grandiflorum is added to the quality standard detection method of Xuefuzhuyuwan Daimiwan, the proportion of the developing agent in the thin-layer identification method of Achyranthes bidentata and Radix Paeoniae Rubra is optimized, the thin-layer identification method of Platycodon grandiflorum is optimized, the thin-layer identification method of Bupleurum falcatum and Flos Carthami is added, and the content determination method of main pharmacodynamic components in the two monarch drugs Semen Persicae and Flos Carthami in the prescription is added. The optimized and added detection contents of the application are different from the quality standard of Xuefuzhuyuwan Daimiwan recorded in the 2020 edition of the Chinese Pharmacopoeia and have certain innovativeness. SUMMARY

[0003] In order to overcome the deficiencies of the prior art, the purpose of the application is to provide a quality detection method of Xuefuzhuyuwan Daimiwan, to increase the determination method of the product quality of Xuefuzhuyuwan Daimiwan through experimental method research, and to enhance the effectiveness and quality controllability of the product.

[0004] The purpose of the application is realized by the following technical scheme:

[0005] A quality detection method of Xuefuzhuyuwan Daimiwan, which comprises the following steps:

[0006] (1) Microscopic identification of Xuefuzhuyuwan Daimiwan:

[0007] Angelica sinensis: The parietal wall of the spindle-shaped parenchyma cell is slightly thick, and there are slightly inclined and interlaced textures.

[0008] Rehmannia glutinosa: The parenchyma tissue is grayish brown to black brown, and the cells are mostly shriveled and contain brown or black nuclear substances.

[0009] Semen Persicae: The stone cell is orange yellow, light yellow or orange red, and is shell-shaped or helmet-shaped in side view.

[0010] Platycodon grandiflorum: The connecting lactiferous duct contains small yellow oil droplets and fine granular substances.

[0011] (2) Thin-layer chromatography identification:

[0012] a. Optimization of the developing solvent for the thin-layer identification method of Achyranthes bidentata and Paeonia lactiflora: The original developing solvent ratio of chloroform-ethyl acetate-methanol-concentrated ammonia solution (2:4:8:1) was optimized to chloroform-ethyl acetate-methanol-concentrated ammonia solution (50:10:20:2.5);

[0013] b. Identification of Platycodon grandiflorus by thin-layer chromatography: Platycodon grandiflorus in the prescription of Xuefu Zhuyu Wan was identified by thin-layer chromatography with Platycodon grandiflorus as a reference standard.

[0014] c. Identification of Bupleurum by Thin-Layer Chromatography: Bupleurum in the prescription of Xuefu Zhuyu Pill was identified by thin-layer chromatography using Bupleurum as a reference standard.

[0015] d. Thin-layer chromatography for identification of safflower: Using thin-layer chromatography with safflower as a reference standard, the safflower in the prescription of Xuefu Zhuyu Pill was identified;

[0016] (3) Content determination:

[0017] a. Determination of peach kernel by high performance liquid chromatography: The peach kernel in Xuefu Zhuyu Pill was determined by high performance liquid chromatography with amygdalin as a reference standard.

[0018] b. Determination of safflower by high performance liquid chromatography: Safflower in Xuefu Zhuyu Pill was determined by high performance liquid chromatography with hydroxysafflower yellow A as a reference standard.

[0019] This invention provides a method for quality determination of Xuefu Zhuyu Pills (water-honey pills). This invention is a practical and repeatable method derived from extensive experimentation. The above-mentioned determination method can be incorporated into the quality control standards for Xuefu Zhuyu Pills (water-honey pills). This method can objectively reflect changes in the product's internal components and can comprehensively control the product's quality.

[0020] The present invention has the following advantages:

[0021] (1) The quality control method of the present invention is based on the Xuefu Zhuyu Pill of the 2020 edition of the Pharmacopoeia of the People's Republic of China. It adds microscopic identification methods for Angelica sinensis, Rehmannia glutinosa, Prunus persica and Platycodon grandiflorus, optimizes the developing agent ratio of the thin-layer identification method for Achyranthes bidentata and Paeonia lactiflora, optimizes the thin-layer identification method for Platycodon grandiflorus, adds the thin-layer identification method for Bupleurum chinense and Carthamus tinctorius, and adds the content determination of Prunus persica and Carthamus tinctorius. It controls the product quality of Xuefu Zhuyu Water-Honey Pill preparation from an overall and macroscopic perspective.

[0022] (2) The method of the present invention is simple, stable, highly precise, reproducible and easy to master. Detailed Implementation

[0023] The embodiments of the present invention are described in detail below. It should be noted that these embodiments are descriptive and not limiting, and should not be used to limit the scope of protection of the present invention.

[0024] Comparative Example 1

[0025] Thin-layer chromatography for the identification of Achyranthes bidentata and Paeonia lactiflora:

[0026] The developing solvent ratio for thin-layer chromatography identification of Achyranthes bidentata and Paeonia lactiflora was optimized. The original developing solvent ratio of chloroform-ethyl acetate-methanol-concentrated ammonia solution (2:4:8:1) was adjusted to chloroform-ethyl acetate-methanol-concentrated ammonia solution (50:10:20:2.5).

[0027] ① Take 5g of this product, grind it into a fine powder, add 50ml of methanol, heat under reflux for 60 minutes, cool, centrifuge for 5 minutes, take the supernatant, evaporate to dryness, add 20ml of water to dissolve the residue, add 10ml of 50% ethanol and 1ml of dilute hydrochloric acid, shake well, extract twice with ether (25ml each time), discard the ether extract, extract twice with ethyl acetate (25ml each time), discard the ethyl acetate extract, extract twice with water-saturated n-butanol (20ml each time), combine the n-butanol extracts, take half of the n-butanol solution, wash twice with ammonia solution (20ml each time), discard the ammonia washing solution, evaporate the n-butanol solution to dryness, add 1ml of methanol to dissolve the residue, and use it as the test solution.

[0028] ② Take 1g of Achyranthes bidentata reference material, add 25ml of 50% methanol, heat under reflux for 1 hour, cool, filter, add 20ml of water to the filtrate, and extract twice with water-saturated n-butanol, 25ml each time. Combine the n-butanol extracts, wash twice with water-saturated n-butanol, 25ml each time, discard the water washing liquid, evaporate the n-butanol solution to dryness, add 1ml of methanol to dissolve the residue, and use it as the reference material solution.

[0029] ③ Take β-ecdysterone reference standard and paeoniflorin reference standard, and add methanol to prepare solutions containing 1 mg per 1 ml, as reference solutions.

[0030] ④ Perform the thin-layer chromatography test (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0502). Take 4-10 μl of the test solution, 6 μl of the reference medicinal material solution, and 2 μl of the reference solution, and spot them separately on the same silica gel GF254 thin-layer plate. Use chloroform-ethyl acetate-methanol-concentrated ammonia solution (50:10:20:2.5) as the developing solvent, develop, remove, air dry, and examine under ultraviolet light (254 nm).

[0031] Comparative experiment: Take 4-10 μl of the test solution, 6 μl of the control herb solution and 2 μl of the control solution, and spot them separately on the same silica gel GF254 thin layer plate. Use chloroform-ethyl acetate-methanol-concentrated ammonia solution (2:4:8:1) as the developing solvent, develop, remove, air dry, and examine under ultraviolet light (254 nm).

[0032] ⑤ Results Analysis: Under ultraviolet light (254 nm), the chromatogram of the test sample showed spots of the same color at the corresponding positions as those of the reference medicinal material and β-ecdysterone reference standard. The chromatograms of paeoniflorin reference standard and negative sample showed no spots at the corresponding positions, thus not interfering with the identification of Achyranthes bidentata. Spray with 5% vanillin-sulfuric acid solution, heat until the spots are clearly visible, and examine under sunlight. The chromatogram of the test sample showed spots of the same color at the corresponding positions as those of the paeoniflorin reference standard. The chromatograms of β-ecdysterone reference standard, Achyranthes bidentata reference medicinal material, and negative sample showed no spots at the corresponding positions, thus not interfering with the identification of paeoniflorin.

[0033] Comparative Experiment: Under ultraviolet light (254 nm), the chromatogram of the test sample showed spots of the same color at the corresponding positions as those of the reference medicinal material and β-ecdysterone reference standard. The chromatograms of paeoniflorin reference standard and negative sample showed no spots at the corresponding positions, thus not interfering with the identification of Achyranthes bidentata. Spraying with 5% vanillin-sulfuric acid solution and heating until the spots were clearly visible, the chromatogram was examined under sunlight. The chromatogram of the test sample showed spots of the same color at the corresponding positions as those of the paeoniflorin reference standard. The chromatograms of β-ecdysterone reference standard, Achyranthes bidentata reference medicinal material, and negative sample showed no spots at the corresponding positions, thus not interfering with the identification of paeoniflorin. However, the spots were close to the solvent front, resulting in excessively large specific migration values. Therefore, the developing solvent ratio of the original method was optimized.

[0034] Comparative Example 2

[0035] Thin-layer chromatography for identifying Platycodon grandiflorus:

[0036] The thin-layer identification method has been optimized.

[0037] ① Take 5g of this product, grind it into a fine powder, add 20ml of dilute hydrochloric acid, heat under reflux for 45 minutes, cool, filter, and extract the filtrate twice with 20ml of chloroform each time. Combine the chloroform extracts, evaporate to dryness, and dissolve the residue in 1ml of methanol to prepare the test solution. Separately, take 1g of Platycodon grandiflorus reference material, add 20ml of dilute hydrochloric acid, and prepare a reference material solution using the same method.

[0038] Comparative experiment: Take 5g of this product, grind it into a fine powder, add 50ml of methanol, heat under reflux for 60 minutes, cool, centrifuge for 5 minutes, take the supernatant, recover the solvent under reduced pressure and evaporate to dryness, add 20ml of a mixture of 7% sulfuric acid ethanol solution and water (1:3) to the residue, heat under reflux in a water bath for 3 hours, cool, and extract twice with 20ml of chloroform each time. Combine the chloroform extracts, wash twice with 20ml of water each time, discard the washing liquid, dehydrate the chloroform solution with an appropriate amount of anhydrous sodium sulfate, filter, evaporate the filtrate to dryness, dissolve the residue in 1ml of methanol, and use it as the test solution. Separately, take 1g of Platycodon grandiflorus reference material, add 20ml of a mixture of 7% sulfuric acid ethanol solution and water (1:3), and prepare the reference material solution in the same way.

[0039] ② Perform the thin-layer chromatography test (Chinese Pharmacopoeia 2015 Edition, Part IV, General Chapter 0502). Take 10 μl of the test solution and 5-10 μl of the reference herb solution and spot them separately on the same silica gel GF254 thin-layer plate. Use chloroform-ethyl ether (1:1) as the developing solvent, develop, remove, air dry, and examine under ultraviolet light (254 nm).

[0040] ③Results Analysis: In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material. After spraying with 10% sulfuric acid ethanol solution and heating at 105℃ until the spots are clearly visible, examine under sunlight. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material.

[0041] Comparative experiment: Under ultraviolet light (254nm) and sunlight, no spots appeared in the chromatogram of the control medicinal material, while the spots in the chromatogram of the test sample were not obvious. Therefore, the method was optimized.

[0042] Comparative Example 3

[0043] Identification of Bupleurum chinense by thin-layer chromatography:

[0044] ① Take 5g of this product, grind it into a fine powder, add 30ml of methanol, sonicate for 30 minutes, cool, filter, evaporate the filtrate to dryness, add 30ml of water to dissolve the residue, extract three times with 20ml of water-saturated n-butanol each time, combine the n-butanol solutions, wash with 50ml of ammonia test solution, then wash with 50ml of water saturated with n-butanol, discard the aqueous solution, evaporate the n-butanol solution to dryness, add 2ml of methanol to dissolve the residue, and use this as the test solution.

[0045] ② Take another 0.5g of Bupleurum reference material and prepare a reference material solution using the same method.

[0046] ③ Perform the thin-layer chromatography test (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0502). Take 5-10 μl of each of the above two solutions and spot them separately on the same silica gel G thin-layer plate. Use the lower layer solution of chloroform-methanol-water (13:5:2) placed below 10℃ as the developing solvent. Develop, remove, air dry, spray with 2% p-dimethylaminobenzaldehyde in 40% sulfuric acid solution, and heat until the spots are clearly visible.

[0047] ④ Results analysis: In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material; when examined under ultraviolet light (365nm), fluorescent spots of the same color appear in the chromatogram of the test sample at the corresponding positions as in the chromatogram of the reference medicinal material.

[0048] Comparative Example 4

[0049] Thin-layer chromatography for the identification of safflower:

[0050] ① Take 2.5g of this product, grind it into a fine powder, add 10ml of 80% acetone solution, seal tightly, shake for 15 minutes, let stand, and take the supernatant as the test solution.

[0051] ② Take another 0.5g of safflower reference material and prepare a reference material solution using the same method.

[0052] ③ Perform the thin-layer chromatography test (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0502). Take 5-10 μl of each of the above two solutions and spot them separately on the same silica gel G thin-layer plate. Use ethyl acetate-methanol-formic acid-water (7:0.4:2:3) as the developing solvent, develop, remove, and air dry.

[0053] ④ Results analysis: In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material.

[0054] Example 1

[0055] Determination of peach kernel by high performance liquid chromatography:

[0056] The peach kernels in Xuefu Zhuyu Wan (a traditional Chinese medicine formula) were determined by high performance liquid chromatography (HPLC), with amygdalin as a reference standard.

[0057] ① Preparation of standard solution: Take an appropriate amount of amygdalin reference standard and add methanol to prepare a solution containing about 0.4 mg of amygdalin per ml, which is used as the reference solution.

[0058] ② Preparation of the test solution: Take an appropriate amount of this product, grind it into a fine powder, mix it well, take about 2g, weigh it accurately, place it in a stoppered conical flask, add 50ml of methanol accurately, weigh it, heat it under reflux for 1 hour, cool it, weigh it again, make up the weight loss with methanol, shake it well, filter it, and take the filtrate to obtain the test solution.

[0059] ③ Preparation of negative sample solution: All medicinal materials except peach kernel were taken according to the prescription, and samples were prepared according to the preparation method. Then, a negative sample solution was prepared according to the method for preparing the test sample solution. Analysis was performed under the same chromatographic conditions. No chromatographic peak was observed at the corresponding retention time of amygdalin in the negative sample. The results indicate that the negative sample did not interfere with the detection.

[0060] ④ Setting of liquid chromatography conditions: The chromatographic column is octadecylsilane-bonded silica gel as the packing material, the mobile phase is 0.1% phosphoric acid solution (A)-methanol (B), gradient elution: 0-30 min 10-25% B%, 30-40 min 25-100% B%, 40-50 min 100-10% B%, detection wavelength is 210 nm, flow rate is 1.0 ml / min, and the theoretical plate number calculated based on amygdalin should not be less than 3000.

[0061] ⑤ Determination: Accurately pipette 10 μl each of the reference solution and the test solution into the liquid chromatograph and detect them to obtain the results.

[0062] ⑥ Results Analysis: The retention time confirmed the presence of amygdalin in the test sample, thus revealing the content of peach kernel in Xuefu Zhuyu Wan (a traditional Chinese medicine formula). The content limit was defined as amygdalin (C0.05) per 1g of peach kernel. 20 H 27 NO 11 The result was 1.66mg, which is greater than 1.50mg, so it is a qualified product.

[0063] Example 2

[0064] Determination of safflower by high performance liquid chromatography:

[0065] The safflower in Xuefu Zhuyu Wan (a traditional Chinese medicine formula) was determined by high performance liquid chromatography (HPLC), with hydroxysafflower yellow A as a reference standard.

[0066] ① Preparation of standard solution: Take an appropriate amount of hydroxysaffron yellow A reference standard and add 25% methanol to prepare a solution containing about 1.0 mg of amygdalin per 1 ml, which is used as the reference solution.

[0067] ② Preparation of the test solution: Take an appropriate amount of this product, grind it into a fine powder, mix it well, take about 4g, weigh it accurately, place it in a stoppered conical flask, accurately add 50ml of 25% methanol, weigh it, sonicate it (power 300W, frequency 50kHz) for 40 minutes, cool it, weigh it again, make up the lost weight with 25% methanol, shake it well, filter it, and take the filtrate to obtain the test solution.

[0068] ③ Preparation of negative sample solution: All medicinal materials except safflower were taken according to the prescription, and samples were prepared according to the preparation method. Then, a negative sample solution was prepared according to the method for preparing the test sample solution. Analysis was performed under the same chromatographic conditions. No chromatographic peak was observed at the corresponding retention time of hydroxysafflower yellow pigment A in the negative sample. The results indicate that the negative sample did not interfere with the detection.

[0069] ④ Setting of liquid chromatography conditions: The chromatographic column is octadecylsilane-bonded silica gel as the packing material, the mobile phase is 70% acetonitrile (A)-0.1% phosphoric acid solution (B), gradient elution: 0-8 min 76-71% B, 8-15 min 71-0% B, 15-25 min 0-76% B, detection wavelength is 403 nm, flow rate is 1.0 ml / min, and the theoretical plate number calculated based on hydroxysafflower yellow A should not be less than 3000.

[0070] ⑤ Determination: Accurately pipette 10 μl each of the reference solution and the test solution into the liquid chromatograph and detect them to obtain the results.

[0071] ⑥ Results Analysis: The retention time confirmed the presence of hydroxysaffron yellow A in the test sample, thus determining the safflower content in Xuefu Zhuyu Wan (a traditional Chinese medicine formula). The content limit was set at hydroxysaffron yellow A (C40) per gram of safflower. 27 H 32 O 16 The result was 0.80mg, which is greater than 0.75mg, so it is a qualified product.

[0072] Example 3

[0073] A quality control method for a traditional Chinese medicine preparation, Xuefu Zhuyu Water-Honey Pill, comprising the following steps:

[0074] (1) Microscopic identification of Xuefu Zhuyu Pill (water-honey pill):

[0075] Angelica sinensis: thin-walled cells are spindle-shaped with slightly thick walls and very fine oblique interlacing textures;

[0076] Rehmannia glutinosa: The parenchyma is grayish-brown to dark brown, the cells are mostly shrunken, and contain brown or black nuclei;

[0077] Peach kernel: Stone cells are orange-yellow, pale yellow or orange-red, shell-shaped or helmet-shaped when viewed from the side, with denser pits and grooves on the thinner side and more obvious stratification on the thicker side.

[0078] Platycodon grandiflorus: The connecting ducts contain tiny pale yellow oil droplets and fine granular substances.

[0079] (2) Thin-layer chromatography identification:

[0080] a. Optimization of the developing solvent for the thin-layer identification method of Achyranthes bidentata and Paeonia lactiflora: The original developing solvent ratio of chloroform-ethyl acetate-methanol-concentrated ammonia solution (2:4:8:1) was optimized to chloroform-ethyl acetate-methanol-concentrated ammonia solution (50:10:20:2.5);

[0081] b. Identification of Platycodon grandiflorus by thin-layer chromatography: Platycodon grandiflorus in the prescription of Xuefu Zhuyu Wan was identified by thin-layer chromatography with Platycodon grandiflorus as a reference standard.

[0082] c. Identification of Bupleurum by Thin-Layer Chromatography: Bupleurum in the prescription of Xuefu Zhuyu Pill was identified by thin-layer chromatography using Bupleurum as a reference standard.

[0083] d. Thin-layer chromatography for identification of safflower: Using thin-layer chromatography with safflower as a reference standard, the safflower in the prescription of Xuefu Zhuyu Pill was identified;

[0084] (3) Content determination:

[0085] a. Determination of peach kernel by high performance liquid chromatography: Peach kernel in Xuefu Zhuyu Pills was determined by high performance liquid chromatography using amygdalin as a reference standard. The content of peach kernel in Xuefu Zhuyu Pills (water-honey pills) was found to be 1g per gram of peach kernel containing amygdalin (C... 20 H 27 NO 11 The total dose is 1.66 mg.

[0086] b. Determination of safflower by high performance liquid chromatography: The safflower content in Xuefu Zhuyu Shuimi Pills was determined by high performance liquid chromatography using hydroxysafflower yellow A as a reference standard. The determination was based on the safflower content per 1g of pill. 27 H 32 O 16 The total dose is 0.88 mg.

Claims

1. A quality testing method for a traditional Chinese medicine preparation, Xuefu Zhuyu Water-Honey Pill, characterized by the following steps: (1) Microscopic identification of Xuefu Zhuyu Pill (water-honey pill): Angelica sinensis: thin-walled cells are spindle-shaped with slightly thick walls and very fine oblique interlacing textures; Rehmannia glutinosa: The parenchyma is grayish-brown to dark brown, the cells are mostly shrunken, and contain brown or black nuclei; Peach kernel: Stone cells are orange-yellow, pale yellow or orange-red, shell-shaped or helmet-shaped when viewed from the side, with denser pits and grooves on the thinner side and more obvious stratification on the thicker side. Platycodon grandiflorus: The connecting laticifers contain tiny pale yellow oil droplets and fine granular substances; (2) Thin-layer chromatography identification: a. Optimization of the developing solvent for the thin-layer identification method of Achyranthes bidentata and Paeonia lactiflora: The original developing solvent ratio of chloroform-ethyl acetate-methanol-concentrated ammonia solution (2:4:8:1) was optimized to chloroform-ethyl acetate-methanol-concentrated ammonia solution (50:10:20:2.5); b. Identification of Platycodon grandiflorus by thin-layer chromatography: Platycodon grandiflorus in the prescription of Xuefu Zhuyu Wan was identified by thin-layer chromatography with Platycodon grandiflorus as a reference standard. c. Identification of Bupleurum by Thin-Layer Chromatography: Bupleurum in the prescription of Xuefu Zhuyu Pill was identified by thin-layer chromatography using Bupleurum as a reference standard. d. Thin-layer chromatography for identification of safflower: Using thin-layer chromatography with safflower as a reference standard, the safflower in the prescription of Xuefu Zhuyu Pill was identified; (3) Content determination: a. Determination of peach kernel by high performance liquid chromatography: Peach kernel in Xuefu Zhuyu Pills was determined by high performance liquid chromatography using amygdalin as a reference standard. The limit for peach kernel content in Xuefu Zhuyu Pills (water-honey pills) is defined as follows: per 1g of peach kernel, amygdalin (C... 20 H 27 NO 11 The total amount should not be less than 1.50 mg. b. Determination of safflower by high performance liquid chromatography: The safflower content in Xuefu Zhuyu Pills was determined using high performance liquid chromatography with hydroxysafflower yellow A as a reference standard. The limit for safflower content in Xuefu Zhuyu Pills (water-honey pills) is 1g per gram of safflower as determined by hydroxysafflower yellow A (C... 27 H 32 O 16 The total amount should not be less than 0.75mg.