鹿角胶

English text reference: Chinese Pharmacopoeia (2020 Edition)

Overview

Deer Horn Glue is the solid glue made by decocting and concentrating deer horn.

Processing

Cut deer horn into sections, wash and soak, decoct in water in several portions, filter, combine the filtrate (or add a small amount of alum powder), let it stand, filter the glue liquid, concentrate (may add appropriate amount of yellow rice wine, rock sugar, and soybean oil) to a thick paste, condense, cut into pieces, and dry to obtain the product.

Description

It is flat square or cuboid. Yellowish-brown or reddish-brown, semi-transparent, some with yellowish-white foam on the surface. Brittle, easily broken, with a shiny fracture. Slight odor, slightly sweet taste.

Identification

To 0.1 g of the powder add 50 ml of 1% ammonium bicarbonate solution, treat with ultrasound for 30 minutes, filter through a microporous membrane, take 100 μl of the filtrate, place it in a microsampling bottle, add 10 μl of trypsin solution (take trypsin for sequence analysis, prepare a solution containing 1 mg per ml with 1% ammonium bicarbonate solution before use), shake well, and incubate at 37°C for 12 hours as the test solution. Take 0.1 g of the reference drug deer horn glue and prepare the reference drug solution by the same method. Perform the test according to the high-performance liquid chromatography-mass spectrometry method (General Rule 0512 and General Rule 0431), using octadecylsilane-bonded silica gel as the filling material (column inner diameter 2.1 mm); use acetonitrile as mobile phase A and 0.1% formic acid solution as mobile phase B, and perform gradient elution according to the specified conditions in the table; the flow rate is 0.3 ml per minute. Use a mass spectrometry detector, electrospray ionization positive ion mode (ESI+), and perform multiple reaction monitoring (MRM), selecting the mass-to-charge ratio (m/z) 765.4 (double charge)→554.0 and m/z 765.4 (double charge)→733.0 as the detection ion pairs. Inject 5 μl of the deer horn glue reference drug solution, and the signal-to-noise ratio of the MRM chromatographic peak determined by the above detection ion pairs should be greater than 3:1.

Take 5 μl of the test solution, inject it into the high-performance liquid chromatography-mass spectrometry system, and determine it. In the ion flow chromatogram extracted by the mass-to-charge ratio (m/z) 765.4 (double charge)→554.0 and m/z 765.4 (double charge)→733.0 ion pairs, a chromatographic peak that is consistent with the retention time of the reference drug should be simultaneously present.

Examination

Water

Weigh accurately 1 g of the product, add 2 ml of water, dissolve by heating, place it on a water bath and evaporate until the thickness does not exceed 2 mm, and determine the water content according to the method for water determination (General Rule 0832, method 2), which should not exceed 15.0%.

Total ash

Take 1.0 g of the product and examine it according to the regulations (General Rule 2302), which should not exceed 3.0%.

Heavy metals

Take the residue under the total ash item and examine it according to the regulations (General Rule 0821, method 2), which should not exceed 30 mg/kg.

Arsenic salts

Take 1.0 g of the product, add 1 g of calcium hydroxide, mix, add a small amount of water, stir well, dry, first carbonize with low heat, then incinerate at 500-600°C to complete ashing, cool, add 5 ml of hydrochloric acid and 2 ml of water, and examine it according to the regulations (General Rule 0822), which should not exceed 2 mg/kg.

Insoluble matter in water

Weigh accurately 1.0 g of the product, add 10 ml of water, dissolve by heating, transfer the solution to a 10 ml centrifuge tube that has been weighed, centrifuge, remove the oil floating on the tube wall, pour off the supernatant, add warm water along the tube wall to the mark, centrifuge, wash 3 times as before, pour off the supernatant, heat the centrifuge tube at 105°C for 2 hours, take it out, cool it in a desiccator for 30 minutes, weigh accurately, and calculate to obtain the result.

The insoluble matter in water of the product should not exceed 2.0%.

Others

It should comply with the relevant regulations under the adhesive item (General Rule 0184).

Assay

Determine by high-performance liquid chromatography (General Rule 0512).

Chromatographic conditions and system suitability test

Use octadecylsilane-bonded silica gel as the filling material; use acetonitrile-0.1 mol/L sodium acetate solution (adjust the pH value to 6.5 with acetic acid) (7:93) as mobile phase A, and use acetonitrile-water (4:1) as mobile phase B. Perform gradient elution according to the specified conditions in the table; the detection wavelength is 254 nm. The column temperature is 43°C. The theoretical plate number calculated based on the peak of L-hydroxyproline should not be less than 4000.

Preparation of reference solution

Take an appropriate amount of L-hydroxyproline reference substance, glycine reference substance, alanine reference substance, and L-proline reference substance, weigh accurately, and add 0.1 mol/L hydrochloric acid solution to prepare a mixed solution containing 70 μg of L-hydroxyproline, 0.14 mg of glycine, 60 μg of alanine, and 70 μg of L-proline per 1ml.

Preparation of test solution

Take about 0.25 g of the crude powder of the product, weigh accurately, place it in a 25 ml volumetric flask, add 20 ml of 0.1 mol/L hydrochloric acid solution, treat with ultrasound (power 300 W, frequency 40 kHz) for 30 minutes, cool, add 0.1 mol/L hydrochloric acid solution to the mark, shake well. Accurately measure 2 ml, place it in a 5 ml ampoule, add 2ml of hydrochloric acid, hydrolyze at 150°C for 1 hour, cool, transfer to an evaporating dish, wash with water in portions of 10 ml, pour the washing liquid into the evaporating dish, evaporate to dryness, dissolve the residue in 0.1 mol/L hydrochloric acid solution, transfer to a 25 ml volumetric flask, add 0.1 mol/L hydrochloric acid solution to the mark, shake well.

Accurately measure 5 ml of the above reference solution and test solution, place them in separate 25 ml volumetric flasks, add 2.5 ml of acetonitrile solution of 0.1 mol/L phenyl isothiocyanate (PITC) and 2.5 ml of acetonitrile solution of 1 mol/L triethylamine, shake well, and let it stand at room temperature for 1 hour. Add 50% acetonitrile to the mark, shake well. Take 10 ml, add 10 ml of n-hexane, shake, let it stand for 10 minutes, take the lower layer solution, filter, and take the subsequent filtrate.

Assay method

Accurately suck 5 μl of the derivatized reference solution and test solution, inject them into the liquid chromatograph, and determine.

Calculated on the basis of the dried product, the content of L-hydroxyproline should not be less than 6.6%, glycine should not be less than 13.3%, alanine should not be less than 5.2%, and L-proline should not be less than 7.5%.

Property

Warm.

Flavor

Sweet and salty.

Meridian tropism

Kidney and liver meridians.

Actions

It warms and tonifies the liver and kidney, nourishes essence and blood.

Indications

It is used for lumbago and coldness of the waist and knees caused by liver and kidney deficiency, impotence and premature ejaculation, weakness and emaciation, excessive uterine bleeding, bloody stools, bloody urine, and swelling and pain of carbuncle.

Dosage

3-6 g.

Administration

Decocted and taken orally.

Specification

Each piece weighs 6g.

Storage

Store in a sealed container.