safety and quality control of pharmaceutical excipients

Safety and Quality Control of Pharmaceutical Excipients for Injection

National Institutes for Food and Drug ControlSong Xiaosong

2017-4-25

http://www.nicpbp.org.cn

01

03

02

Introduction

Current status of excipients for injection in China

Key points of quality control for injectable excipients

04

05

Examples of safety and qualitycontrol of the excipients forinjection

Summary


API Pharmaceutical

preparation

process

Pharmaceutical

container

Pharmaceutical

excipients

Pharmaceutical

products Quality control

1. Introduction


• Functions of excipients

• 63 classifications of pharmaceutical excipients based on fuctions:pH adjusting agent, chelating agent, coating agent, coating agent, protective agent, humectant, disintegrating agent, surface

active agent, precipitating agent, film forming material, flavoring agent, carbon dioxide adsorbent, foaming agent,preservative, excipient, desiccant, curing agent, buffer, slow release material, flavoring agent, antioxidant, antioxidant, anti-sticking agent, air displacer, condensing agent, gel materials, polishes, solvents, softeners, emulsifiers, ointment bases, softcapsule materials, lubricants, wetting agents, penetration enhancers, osmotic pressure regulators, suppository bases, sweeteners,fillers, pellets additives, stabilizers, adsorbents, absorbents, diluents, defoamers, flocculants, plaster bases, inks, thickeners,solubilizers, plasticizers, binders, filter aid, cosolvent, cosolvent, suspending agent, coloring agent.

• an excipient may have multiple functions Lactose：Solid preparations- dispersant, flavoring agent ; Injection-cryoprotectant；Inhalation-carrier； Glycerin：Liquid preparations- solvent, solubilizer, sweetener; Emulsion- moisturizer 、Transdermal preparations - penetration enhancers 。

1. Introduction


1. Introduction

Injection：delivering drug liquid into body directlywithout first-pass. it is a high-risky dosage form.

The adverse effect of excipients is more serious via injection route than via oral administration.


• Benzyl alcohol：Usually used as

solvent or preservative.

• Adverse effects：may induce

intraventricular hemorrhage,

hemolysis, hypotension, local

stimulation and gluteal muscle

contracture in infants

1、Injections containing benzyl

alcohol must be labeled “Prohibited

for children”;

2、 lyophilized powder for injection

which need benzyl alcohol as solvent

must be labeled “Prohibited for

children”Benzyl alcohol

Adverse effects of pharmaceutical excipients for injection

1. Introduction


Safety issue of alcoholic excipientsUse and dosage Adverse effect

Ethanol widely used with large dosage

Drug interaction, alcoholism (25mg / dl), cardiotoxicity

Benzyl alcohol

preservative（0.9-2%）、Solubilizer（＞5%）

Nerve effect

Propylene glycol

Solvent for intraperitoneal injectionpreservative forinhalation

cardiovascular effects, neurological effects, pyruvate, lactate, ototoxicity, thrombophlebitis

PEG SolventNasal suction enhancer

Local stimulation

1. Introduction


Other safety issue excipients

Intolerance Gluten

Lactose

Sensitization Propyl galate

Thimerosal

Parabens

Contact dermatitis/allergy Propylene glycol

Cetyl alcohol

Chloroacetamide

Polyethylene glycol

Benzalkonium Cl

Lanolin

Phenolated Vaseline

BHT

Photosensitivity Cinnamon oil

1. Introduction

Pharmaceutical excipients have activities in vivo.The quality of excipients may significant effect on the

safety of drug products, especially of injections.


2. Current status of excipients for injection in China

drug manufacturer

5%

food additive manufacturer

6%sugar

manufacturer21%

others1%

chemistry manufacturer

67%

Pharmaceutical excipient manufacturers in China

Only about 110 professional pharmaceutical excipient manufacturers in China


Before Ch.P 2015 ： over 140 excipients were used in injectionmanufacturing，but only 2 of them has injection level standards,others are common level or chemical level.

Safety requirements of excipients：should be based on their different applications

Excipients used in injection manufacturing in Chinapropylene glycol

sodium hydroxide

glycerinpolysorbate

80EDTA‐2Na ethanol

anhydrousethanol

PEG400sodium benzoate

sodium chloride

potassium chloride

Anhydrous sodium sulfite

ammonia sulfuric acidsodium

pyrosulfitecitric acid

calcium chloride

meglumine

sodium bicarbonate

chloroform mannitoldisodium hydrogen phosphate

cholesterolanhydrous sodium acetate

glycinesodium sulfate

sodium carboxymethylcellulose

polysorbate

20

sodium hydrogen sulfite

magnesium sulfate

sodium citrate

glutamic aciddipotassiumphosphate

lactic acid

phenollauryl sodium sulfate

methioninebenzyl alcohol

ethylenediamine soya bean

lecithin

lactosesodium

thiosulfatemalic acid

potassium hydroxide

diluted hydrochlori

c acid

Egg yolk lecithin

Merthiolate sucrosesodium

dihydrogenphosphate

sorbitol chloretonephosphoric acid

dextran soybean oil

2. Current status of excipients for injection in China

How to control the quality of excipients？

Improving the monograph of excipients in national pharmacopeia


02

2

23

201520102005200019951990

glycerin（for injection）

glycerin（for injection）

sodium chloride （for injection）PLGA（5050）（for injection）PLGA（7525）（for injection）PLGA（8515）（for injection）PEG300，PEG400（for injection）Polysorbate 80（for injection）Soya bean lecithin （for injection）egg yolk lecithin （for injection）activated charcoal （for injection）propylene glycol （for injection）

Trehalosebenzyl alcoholsodium oleatesodium citratexylitol

00

Tromethaminepoloxamer 188dipotassium phosphatedipotassium hydrogen phosphate trihydratePolyoxyl(35) Castor Oil

2. Current status of excipients for injection in ChinaCh.P 2015 collected more monographs of the excipients for injection


What’s new in Ch.P 2015

Monographesof excipients for injection

More class and monographs

Guidance of FRCs study

1.

New methods

2.

3. 4.

Ch.P 2015

PLGA（for injection）PEG（for injection）Soya bean lecithin（for injection）Egg yolk lecithin（for injection）activated charcoal （for injection）propylene glycol （for injection）sodium chloride （for injection）

…21 new（140using）

Molecular weight

NMR

Ch.P 2010：31 Classes

Ch.P 2015：66 Classes


Management of the excipients for injection by CFDA

excipients Use

1 2,6‐ditertbutyl‐4 methylphenol For injection、ophthalmic use

2 propylene glycol For Injection3 sodium hyaluronate For Injection、ophthalmic use4 poloxamer188 For Injection5 camellia oil（for injection） For Injection6 Soya Lecithin （for injection） For Injection7 Soybean Oil（for injection） For Injection8 Cholesterol For Injection9 egg yolk lecithin（for injection） For Injection

10 glycerin(for injection) For Injection11 refined corn oil For Injection12 Polysorbate 80 For Injection13 PEG 400 For Injection14 PEG 600 For Injection

Excipients under License Management


15 PEG 200 For Injection16 PEG 300 For Injection17 Egg yolk lecithin（for injection） For Injection18 maltose For Injection19 gelatin20 Vacant Gelatin Capsules

21 Enteric Vacant Gelatin Capsules22 Hydroxypropyl Betadex（for injection） For Injection23 lactose For Injection, inhalation

24 Anhydrous lactose For Injection, inhalation

25 Anhydrous ethanol（for injection）For Injection26 PLGA(25:75)27 oleic acid For Injection28 sodium oleate For Injection

Management of the excipients for injection by CFDA

Excipients under License Management


3. Key points of quality control for injectable excipients

Name

Chemical structure

Molecular fomulation

CAS No.

Source and manufacture

Appearance and solubility

Identification

Specific test

Assay

Classification

Packaging and storage

specific items for injectable excipients Items of quality specification

Source and manufacture

Endotoxin and Sterility

allergic impurities

Substances cause Haemolysis oragglutination

undue toxicity

Substances cause hypertension orhypotension

stimulating substance …


ingredients %

PC 73.0%PE 15.0%SPM 2.5%

PI 0.9%

LPC 5.8%

LPE 2.1%

OTHERS 0.9%

ingredients %

PC 57.8%PE 13.3%PI 2.2%

LPC 3.2%

LPE 0.6%

OTHERS 22.9%

Lecithin

The difference in composition and content would effect on thefunction and safety of excipients, and effect on the efficiency andsafety of injection.

The source of excipients for injection needs to be noticed.


Egg yolk lecithin for injection

Endotoxin

Sterility

For the process of preparations without terminal sterilization

May cause fever or other side effect


Allergic impurities in excipients for injection

Proteins in lactoseSource: whey fermented from milk

Allergen: residual protein in whey

residual protein in lactose %

α-Lactalbumin 19.7%

β-Lactoglbulin 43.6%

BSA 4.7%

免疫球蛋白（Ig） 3%

Other proteins 29%

The residual proteins may induce immune response and allergy, causing serious safety issue.


Phosphatidylcholine(PC)Phosphatidylethanolamine (PE)

Lysophosphatidylcholine (LPC) Lysophosphatidylethanolamine (LPE)

Lose a fatty acid chain(R2) Lose a fatty acid chain(R2)

Substances cause Haemolysis or agglutinationlysophosphatide in lecithin


Ketan Hippalgaonkar et. al. , AAPS PharmSciTech, (2010)

•LPC/LPE: hydrolysis from PC/PE•During producing process；•During sterilization process of emulsion

Substances cause Haemolysis or agglutinationlysophosphatide in lecithin


PC LPC

棘皮红细胞显微镜图

棘皮细胞

Compared with phospholipids, the hydrophilic and hydrophobic property of lysophospholipids are more equivalent, which accelerate the exchange between cell membrane and cytoplasm. Red blood cells become to echinoderms cells. When the concentration of lysophospholipids reaches 2×10-4M, haemolysis occurs.

Substances cause Haemolysis or agglutination——lysophosphatide in

lecithin


270 pharmaceutical excipients in Ch.P 2015

23 for injection:Sodium chloride

PLGA（5050）PLGA（7525）PLGA（8515）PEG300，PEG400Activated charcoal

Soya/egg yolk LecithinPolysorbate 80Propylene glycol

Tromethamine

Poloxamer188dipotassium phosphateDipotassium Hydrogen Phosphate Trihydrate

Polyoxyl(35)caster oil

TrehaloseBenzyl alcoholSodium oleateSodium citrateXylitol

《中国药典》2010年版品种（含修订后）

132个

《中国药典》2015年版新增品种

138个1醋酸钠 1丙氨酸2富马酸 2甘氨酸3果糖 3谷氨酸钠4三乙醇胺 4精氨酸5纤维醋法酯 5酪氨酸6乙基纤维素 6亮氨酸7巴西棕榈蜡 7牛磺酸8玉米淀粉 8色氨酸9果胶 9门冬氨酸10黄凡士林 10门冬酰胺11黄原胶 11缬氨酸12磷酸二氢钾 12组氨酸13磷酸氢二钾 13氨丁三醇14磷酸氢二钾三水合物 14琥珀酸15磷酸氢二钠 15马来酸16无水磷酸氢二钠 16磷酸氢二铵17琼脂 17乙基纤维素水分散体

18乙酸乙酯 18乙基纤维素水分散体（B型）

19异丙醇 19可溶性淀粉20预胶化淀粉 20阿拉伯半乳聚糖21硫酸 21微晶蜡22三油酸山梨坦（司盘85） 22西黄蓍胶23硬脂山梨坦（司盘 60） 23烟酸24依地酸二钠 24烟酰胺25单糖浆 25正丁醇26羊毛脂 26淀粉水解寡糖27油酸乙酯 27焦糖28阿司帕坦 28可压性蔗糖29苯甲酸钠 29磷酸淀粉钠30环拉酸钠 30磷酸钙31羧甲淀粉钠 31马铃薯淀粉32聚甲丙烯酸铵酯Ⅰ 32木薯淀粉33聚甲丙烯酸铵酯Ⅱ 33无水磷酸氢钙34十二烷基硫酸钠 34小麦淀粉35稀盐酸 35磷酸36盐酸 36滑石粉37白凡士林 37麝香草酚38麦芽糖 38苯甲醇39二甲基亚砜 39丁香酚40焦亚硫酸钠 40丁香油41麦芽糊精 41丁香茎叶油42氢氧化钠 42硫酸羟喹啉43轻质氧化镁 43氯甲酚44白陶土 44三氯蔗糖45石蜡 45油酸钠46玉米朊 46纯化水47二氧化钛 47十六醇

48轻质液状石蜡 48丙烯酸乙酯‐甲基丙烯酸甲酯共聚物水分散体

49山嵛酸甘油酯 49麦芽酚50阿拉伯胶 50甘油磷酸钙51油酸山梨坦（司盘80） 51甘油三乙酯52月桂山梨坦（司盘20） 52海藻糖53棕榈山梨坦（司盘 40） 53环甲基硅酮54硅酸镁铝 54碱石灰55羟苯丙酯 55D‐木糖56羟苯丙酯钠 56木糖醇57羟苯丁酯 57尿素

61醋酸纤维素 61氧化锌62羟丙甲纤维素 62腺嘌呤63羟丙纤维素 63硼砂64羧甲基纤维素钠 64硼酸65微晶纤维素 65油酸聚烃氧乙烯酯66大豆油 66液状石蜡67二氧化硅 67枸橼酸三乙酯68精制玉米油 68油酰聚氧乙烯甘油酯

69甜菊素 69月桂酰聚氧乙烯（6）甘油酯

70蔗糖硬脂酸酯 70月桂酰聚氧乙烯（8）甘油酯

71三氯叔丁醇 71月桂酰聚氧乙烯（12）甘油酯

72胆固醇 72月桂酰聚氧乙烯（32）甘油酯

73山梨酸 73羟苯苄酯74月桂氮卓酮 74可可酯75二甲硅油 75胶态二氧化硅76聚乙烯醇 76粉状纤维素77硬脂酸钙 77没食子酸78硬脂酸 78蔗糖八醋酸酯79硬脂酸镁 79去氧胆酸钠80海藻酸钠 80苯扎氯铵81黑氧化铁 81苯扎溴铵

82红氧化铁 82维生素E琥珀酸聚乙二醇酯

83黄氧化铁 83碳酸丙烯酯84紫氧化铁 84辛酸85棕氧化铁 85辛酸钠86枸橼酸一水合物 86三硅酸镁87无水枸橼酸 87稀磷酸88硫酸钙 88氯化钙89交联聚维酮 89氯化钾90聚维酮K30 90氯化镁91DL‐酒石酸 91稀醋酸92DL‐苹果酸 92无水碳酸钠93L‐苹果酸 93十八醇94醋酸 94硬脂酸锌95浓氨溶液 95硫酸铵96无水亚硫酸钠 96硫酸铝97亚硫酸氢钠 97海藻酸98胶囊用明胶 98枸橼酸钠99肠溶明胶空心胶囊 99氯化钠（供注射用）

100交联羧甲基纤维素钠 100碳酸氢钾

101硬脂酸聚烃氧（40）酯 101碳酸氢钠102泊洛沙姆188 102蔗糖丸芯103聚乙二醇1000 103冰醋酸104聚乙二醇1500 104薄荷脑105聚乙二醇4000 105二氧化碳106聚乙二醇600 106酒石酸钠

107聚乙二醇6000 1072,6‐二叔丁基‐4‐甲基苯酚（BHT）

108聚乙二醇400 108低取代羟丙基纤维素

109卡波姆 109丙交酯乙交酯共聚物（5050）（供注射用）

110甲基纤维素 110丙交酯乙交酯共聚物（7525）（供注射用）

111聚山梨酯 20 111丙交酯乙交酯共聚物（8515）（供注射用）

112聚山梨酯 40 112泊洛沙姆407

113聚山梨酯 60 113醋酸羟丙甲纤维素琥珀酸酯

114聚山梨酯 80 114硅化微晶纤维素115丙二醇 115聚氧乙烯116甘油（供注射用） 116卡波姆共聚物117聚丙烯酸树脂Ⅱ 117壳聚糖

118聚丙烯酸树脂Ⅲ 118羟丙甲纤维素邻苯二甲酸酯

119聚丙烯酸树脂Ⅳ 119羟乙纤维素120白蜂蜡 120羧甲纤维素钙121橄榄油 121聚乙二醇300（供注射用）122大豆磷脂 122聚乙二醇400（供注射用）123蛋黄卵磷脂 123聚山梨酯80（供注射用）124混合脂肪酸甘油酯（硬脂） 124甘油125氢化蓖麻油 125乙醇126氢化大豆油 126大豆磷脂（供注射用）127乳糖 127蛋黄卵磷脂（供注射用）128蔗糖 128活性炭（供注射用）129倍他环糊精 129枸橼酸三正丁酯


4. Examples of safety and quality control of the excipients for injection

§1、Activated charcoal（for injection）

§2、Egg yolk lecithin（for injection）

§3、Polysorbate 80（for injection）


4. Examples of safety and quality control of the excipients for injection

pretreatment

grinding

carbonization

activation

grinding

screening

source Charcoal


drug

excipient

diarrhea, flatulence, uremia, hyperuricemia

decolorization, pyrogen removement

materials preparation adsorbtion decarburization filtration sterilization filling

Van der Waals' force Transfer or sharing of electrons

Activated charcoal


Activated charcoalChP2015

Not for injectionChP2015

For injectionUSP39‐NF34 EP8.0

Appearance and solubility Fine black powder, odorless Fine black powder, odorless Fine black powder, odorless Fine black powder, odorless

Identification + + —— +

test

Acidity and alkalinity neutral neutral neutral NMT 0.75ml 0.02M HCL

Chloride ≤0.1% ≤0.1% ≤0.2% ——

Sulfate ≤0.05% ≤0.05% ≤0.2% ——

Uncarbonizedconstituent

+ + + +

sulfide —— + + +cyanide —— + + ——

Solubles in ethanol —— + —— ≤0.5%

Fluorescent substance —— + —— +

Solubles in acid ≤1.0% ≤0.8% ≤3.5% ≤3%

Alkali‐soluble colouredsubstances

—— —— —— +

Loss on drying ≤10.0% ≤10.0% ≤15% ≤15%

Sulphated ash≤3.0% ≤3.0% ≤4.0% ≤5.0%

Ferric ≤0.05% ≤0.02% —— ——

Zinc ≤0.02% ≤0.005% —— ≤25ppm

Copper —— —— —— ≤25ppmLead —— —— —— ≤10ppm

Heavy metal ≤30ppm ≤30ppm —— ——

Adsorptive power + + + +

Bacterial endotoxin —— + —— ——Microbial

enumeration tests—— + + +

Sterility —— + —— ——


Activated charcoal

Ch.P 2010Not for injection

Ch.P 2015For injection

GB/T13803.4‐1999 USP39‐NF34 EP8.0

Adsorptivepower

（1）No turbidity is produced.(100ml 0.12％quinine sulfate)




NLT 40g antipyrinumadsorbed by100g activated charcoal

(2) NLT 1.2ml iodine(0.05mol/L)




Adsorptive power is themost important functionalparameter of activatedcharcoal for injection,should be higher than ofnon-injection use product.


Activated charcoal

Ch.P 2010 Ch.P 2015 GB/T13803.4‐1999 USP39‐NF34 EP8.0

Bacterial endotoxin

——

(1) Background value: ≤2EU/g

—— ————

(2)Adsorptive power for bacterial endotoxin: ≥99%

Activated charcoal isused to removeendotoxin, if itselfhas endotoxin, maycontaminate drug

adsorptivepowder


Lecithin

Mixture of PC, PE, LPC, LPE, PI and SM, extracted from soybean or egg yolk

Used as emulsifier or preparing liposomes


Egg yolk lecithinChP2015 ChP2015

USP39‐NF34Not for injection For injection

Appearance and solubility + + +

Acid value ≤20.0 ≤20.0 ‐‐

Iodine value 60‐73 60‐73 ‐‐

Saponification value 195‐212 195‐212 ‐‐

Peroxide ≤3.0 ≤3.0 ‐‐

Identification + + ‐‐

test

LPC ‐‐ ≤3.5% ≤3.0%LPE ‐‐ ≤1.0% ‐‐

SPM ‐‐ ≤3.0% ‐‐triglyceride ≤3% ≤3% ‐‐

cholesterol ≤2% ≤2% ‐‐

palmitic acid ≤0.2% ≤0.2% ‐‐

free fatty acid ≤1% ≤1% ‐‐

residual solvent + + ‐‐

water determination ≤3% ≤3% ≤6.0%

protein ‐‐ + ‐‐

Heavy metal ≤5ppm ≤5ppm ‐‐arsenic ≤2ppm ≤2ppm ‐‐

Bacterial endotoxin ‐‐ ≤2.0EU/g ≤6 USP units/g

Microbial enumeration tests

+ + +

Sterility ‐‐ + ‐‐nitrogen 1.75%‐1.95% 1.75%‐1.95% ‐‐

phosphorus 3.5%‐4.1% 3.5%‐4.1% ‐‐PC ≥68% ≥68% ‐‐PE ≤20% ≤20% ‐‐


The ADI of Cholesterol is NMT 300mg.

Assume daily clinical use of emulsion injection is 500ml containing 6g egg

yolk lecithin, the limit of 2% cholesterol is acceptable.

Egg yolk lecithin

ChP2015Not for injection

ChP2015 For injection

USP39‐NF34

Cholesterol ≤2 ≤2 ‐‐


Because of haemolysis, the content of lysophospholipids in lecithin for

injection needs to be limited. However, the gastro‐intestic digestive

production of phospholipids are lysophospholipids, so oral take of

lysophospholipids is much more safer.

Egg yolk lecithin



USP39‐NF34

LPC——

≤3.5% ——

LPE—— ≤1.0%

——

LPC+LPE ≤4.0% ————


Egg yolk lecithin is extracted from egg yolk, so the protein may

residual in lecithin. Protein may cause allergy, so it needs to

be limited for injections. However, oral take of protein is safe.



USP39‐NF34

Protein ‐‐ + ‐‐

Egg yolk lecithin


ChP2015 ChP2015

USP39‐NF34Not for injection For injection

Appearance and solubility + + +

Acid value ≤2.0 ≤1.0 ≤2.0

Iodine value 18‐24 18‐24

Saponification value 45‐55 45‐55 45‐55

Peroxide value ≤10 ≤3 ≤10

hydroxyl value 65‐80 65‐80 65‐80

Specific gravity 1.06‐1.09 1.06‐1.09 1.06‐1.09

Viscosity 300‐450mm2/s 300‐450mm2/s 300‐500mm2/s

Identification + + ‐‐

test

pH 5.0‐7.5 5.0‐7.5 ‐‐

UV abs ‐‐ + ‐‐

ethylene glycol ≤0.01% ≤0.01% ‐‐

diethylene glycol ≤0.01% ≤0.01% ‐‐

triethylene glycol ‐‐ ≤0.01% ‐‐

ethylene oxide ≤1ppm ≤1ppm ≤1ppm

dioxane ≤10ppm ≤10ppm ≤10ppm

freezing test + + ‐‐

water determination ≤3.0% ≤0.5% ≤3.0%

residue on ignition ≤0.2% ≤0.1% ≤0.25%

Heavy metal ≤10ppm ≤10ppm ≤10ppm

arsenic ≤2ppm ≤2ppm ‐‐

oleic acid ≥58.0% ≥98.0% ≥58.0%

myristic acid ≤5.0% ≤0.5% ≤5.0%

palmitic acid ≤16.0% ≤0.5% ≤16.0%

palmitoleic acid ≤8.0% ≤0.5% ≤8.0%

stearic acid ≤6.0% ≤0.5% ≤6.0%

linoleic acid ≤18.0% ≤0.5% ≤18.0%

linolenic acid ≤4.0% ≤0.5% ≤4.0%

Bacterial endotoxin ‐‐ 0.012EU/mg ‐‐

Sterility ‐‐ + ‐‐

Polysorbate 80


Polysorbate 80 produced via old

manufacturing process has

adsorption at 234nm, indicating

impurities.

Polysorbate 80 produced via new

manufacturing process has no

adsorption at 234nm.

234nm

New process

Old process

Polysorbate 80

UV absorption: manufacturing process and impurities


Polysorbate 80

ChP2015 ChP2015 USP39‐NF34

Not for injection For injectionoleic acid ≥58.0% ≥98.0% ≥58.0%

myristic acid ≤5.0% ≤0.5% ≤5.0%palmitic acid ≤16.0% ≤0.5% ≤16.0%palmitoleic acid ≤8.0% ≤0.5% ≤8.0%stearic acid ≤6.0% ≤0.5% ≤6.0%linoleic acid ≤18.0% ≤0.5% ≤18.0%linolenic acid ≤4.0% ≤0.5% ≤4.0%

Fatty acids composition is an important function and safety parameter of polysorbate 80 for injection.More oleic acid in polysorbate 80, the more colorless and transparent.


impurities

qualitypurity

Illegal additive hydrogen peroxide

Polysorbate 80 is a mixture

colorless and transparent

component separation structure confirmation


Hydrogen peroxide test

hydrogen peroxide

ceriometry

iodometry

permanganate titration

Color of titration end-point is not

obvious

Iodine

淀粉


A

50 mL water+sample37

5minAdd 30 mL of sulfuric acid (3 mol·L-1)

Add 200 uL of phenanthroline indicator

Cerium sulfate titrant（0.001 mol·L-1）

hydrogen peroxide and other reducing

substances

B50 mL water+sample+

0.1% catalase

37

5minAdd 30 mL of sulfuric acid (3 mol·L-1)

Add 200 uL of phenanthroline indicator

Cerium sulfate titrant（0.001 mol·L-1）

other reducing substances




Result based on 10 batches of polysorbate 80


Component separation and structure confirmation

Theoretical structure of polysorbate 80

Complex ingredient ：different in

①parent nucleus（sorbitan、isosorbide）

②fatty acids（oleic acid, linoleic acid, palmitic

acid , Stearic acid, etc.）

③esterification of oleic acid (monoeaster,

diester, triester, tetraester)

④polymerization of ethylene oxide

X+Y+Z=20


optimization

Polysorbate 80

Chromatographic systemColumn：Agilent SB-C8 (250x4.6mm,5um)Mobile phase A：2% formic acid in waterMobile phase B：2% formic acid in isopropanol

A C

B

D

E

F

G

Column：Aiglent Eclipse XDB-C18（4.6x150mm, 5μm) ； temperature ： 30 ； Detector ： AlltechELSD； Mobile phase：A= methanol B=THF；Gradient elute:0~4.9min:100%B过渡到90%B;5.0~18.9min:90%B过渡到20%B;19~19.1:20%B过渡到100%B。



Polyoxyethylene

sorbitan

monooleate

PSM



peaks No. compounds

A

1 polyoxyethylene sorbitan（PS）2 Polyoxyethylene sorbitan monooleate （PSM）

3 Polyoxyethylene sorbitan dioleate （PSD）

B 4 Polyoxyethylene sorbitan trioleate （PSTri）

C 5 Polyoxyethylene sorbitan tetraoleate （PSTetra）

D 6 Polyoxyethylene isosorbitan（PI）

E 7 Polyoxyethylene isosorbitan monooleate （PIM）

F 8 Polyoxyethylene sorbitan dioleate （PID）

G 9 polyethylene glycol（PEG）

A C

B

D

EF

G

杂质峰

polar components

oleate



classification moleculepolymerization of ethylene oxide/ n

compounds

polyoxyethylene sorbitan（PS）

C6H12O5(OC2H4)n 19～30 12

Polyoxyethylene sorbitanmonooleate（PSM）

C24H44O6(OC2H4)n 15～3319

Polyoxyethylene sorbitandioleate（PSD）

C42H76O7(OC2H4)n 11～3525

Polyoxyethylene sorbitantrioleate（PSTri）

C60H108O8(OC2H4)n 17～3519

Polyoxyethylene sorbitantetraoleate（PSTetra）

C78H140O9(OC2H4)n 28～358

Polyoxyethylene isosorbitan（PI）

C6H10O4(OC2H4)n 11～25 15

Polyoxyethylene isosorbitanmonooleate（PIM）

C24H42O5(OC2H4)n 5～2723

Polyoxyethylene sorbitandioleate（PID）

C42H74O6(OC2H4)n 6～2520

polyethylene glycol（PEG） H (OCH2CH2)nOH 5～18 14

9 classes including 155 compounds


Composition and structure of Polysorbate 80

（PSM）（PSD）

（PSTri）（PSTetra）（PS）

（PI）

（PIM）（PID）

（PEG）

H (OCH2CH2)nOH

Difference in hydrophilic-lipophilic nature

X+Y+Z+W=20


Function and safety study of each composition is ongoing


Toxicity test on zebra fishes

manufacturers use batcheslethality

%

1 WEFor

injection20121023 26.7

2 WEFor


3 NOFFor

injection807367E 30.0

4 NOFFor


5 SEPPICNot for

injectionL08013 56.7

6 SEPPICNot for

injectionL10313 36.7

7 EKNot for


8 QMNot for


9 SYNot for

injection100414M 46.6

10 WENot for



Sensitization test on Beagles

Batches Manufacturing process

sensitizationtest on Beagles

Histamine（nM）

A 20100112 new low 3.588±11.09

B 20091019 new low 0.734±1.76

C 100414M old high71.14±15.28

D 20100404 Old high 65.11425±20.14

E 20100501 old high39.87375±31.04

分子量范围段：1、MW=500-1000；2、MW=1000-1500；3、MW=1500-2000；4、MW=2000-2500；5、MW=2500-3000；6、MW=3000-3500；7、MW=3500-4000；8、MW=4000-5000；9、MW=5000-6000；10、MW=6000-8000；11、MW=8000-10000；

Polysorbate 80 with MW distribution of

1000-3000 showed low sensitization on

beagles， indicating sensitization

might be predicted via MW

determination?


Summary

1. Injection is one of the most high‐risky dosage forms due to its clinical

application, so the quality of pharmaceutical excipients used for injection

may effect on the safety of injections directly.

2. Injection manufacturers should use the excipients for injection rather

than common excipients, in order to ensure the quality and safety.

3. The requirement of the excipients for injection is different from the

excipients for common use, especially in safety parameters, such like allergic

protein, impurities, bacterial endotoxin, microorganism, and sterility, etc.

4. In China, The revise of Ch.P 2020 is ongoing, more monographs of

excipients for injections will be collected.

5. Use excipients for injection in manufacturing injection can guarantee the

quality and safety of injection products。


Thanks

[email protected]

safety and quality control of pharmaceutical excipients

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